399 results on '"Automation, Laboratory instrumentation"'
Search Results
2. Analytical and Clinical Validation of a Non-Ristocetin Based VWF Assay on 2 Automated Analyzers in a Large Reference Laboratory.
- Author
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Kumar V, Goode D, Worfolk LA, Rhea-McManus J, Mitsios JV, and Wong ECC
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- Humans, Blood Coagulation Tests instrumentation, Blood Coagulation Tests methods, Blood Coagulation Tests standards, Reproducibility of Results, von Willebrand Diseases blood, von Willebrand Diseases diagnosis, Automation, Laboratory instrumentation, Limit of Detection, von Willebrand Factor analysis, Ristocetin pharmacology
- Abstract
Background: Historically, von Willebrand factor (VWF) activity assays utilized ristocetin despite limitations including poor limits of detection and high imprecision. Newer VWF activity assays such as the INNOVANCE® VWF Ac assay, however, do not rely on ristocetin to measure platelet-dependent VWF function. The purpose of this study was to evaluate the analytical and clinical performance of the Siemens Healthineers INNOVANCE VWF Ac Assay on the Siemens BCS® XP and the Sysmex® CS-2500 systems in a large reference laboratory setting., Methods: Performance indicators for the INNOVANCE VWF Ac assay were the limit of quantitation (LoQ), precision, and method comparison. Method comparison studies were performed using remnant plasma patient samples from routine coagulation tests and analyzed using both the INNOVANCE VWF Ac assay and the Siemens Healthineers ristocetin-dependent BC von Willebrand Reagent., Results: Evaluation of the INNOVANCE VWF Ac assay on the BCS® XP and CS-2500 systems demonstrated good precision and a lower LoQ compared to the BC von Willebrand Reagent. Method comparisons support the use of the INNOVANCE VWF Ac assay on the BCS® XP and CS-2500 systems to measure platelet-dependent VWF function. The INNOVANCE VWF Ac assay was able to further assist in von Willebrand disease classification in 6/7 (86%) samples when the result was below the LoQ for the BC von Willebrand Reagent (ristocetin cofactor activity)., Conclusions: These data are consistent with the 2021 American Society of Hematology/International Society on Thrombosis and Haemostasis/National Hemophilia Foundation/World Federation of Hemophilia von Willebrand disease guidelines that suggest using newer assays such as the INNOVANCE VWF Ac assay in place of ristocetin cofactor activity assays., (© Association for Diagnostics & Laboratory Medicine 2024. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)
- Published
- 2024
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3. Development and transfer of automated methods in neuroscience: The DADTA.
- Author
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Ullis D
- Subjects
- History, 20th Century, Automation methods, Automation instrumentation, Automation, Laboratory instrumentation, Automation, Laboratory methods, Automation, Laboratory history, North America, Neurosciences methods, Neurosciences history, Neurosciences instrumentation
- Abstract
In the second half of the 20th century, neuroscientists across North America developed automated systems for use in their research laboratories. Their decisions to do so were complex and contingent, partly a result of global reasons, such as the need to increase efficiency and flexibility, and partly a result of local reasons, such as the need to amend perceived biases of earlier research methodologies. Automated methods were advancements but raised several challenges. Transferring a system from one location to another required that certain components of the system be standardized, such as the hardware, software, and programming language. This proved difficult as commercial manufacturers lacked incentives to create standardized products for the few neuroscientists working towards automation. Additionally, investing in automated systems required massive amounts of time, labor, funding, and computer expertise. Moreover, neuroscientists did not agree on the value of automation. My brief history investigates Karl Pribram's decisions to expand his newly created automated system by standardizing equipment, programming, and protocols. Although he was an eminent Stanford neuroscientist with strong institutional support and computer know-how, the development and transfer of his automated behavioral testing system was riddled with challenges. For Pribram and neuroscience more generally, automation was not so automatic., (Copyright © 2024 The Author. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2024
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4. Piston-driven automated liquid handlers.
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Schuster J, Kamuju V, Zhou J, and Mathaes R
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- Humans, Workflow, Automation, Laboratory methods, Automation, Laboratory instrumentation, Specimen Handling methods
- Abstract
Laboratory capacities are often limited by time-consuming manual repetitive procedures rather than analysis time itself. While modern instruments are typically equipped with an autosampler, sample preparation often follows manual procedures including many labor-intensive, monotonous tasks. Particularly, for a high number of samples, well plates, and low microliter pipetting, manual preparation is error-prone often requiring repeated experiments. Sampling and sample preparation can account for greater analytical variability than instrument analysis. Repetitive tasks such as liquid handling benefit strongly from technological advances and led to the increasing applications of various automated liquid handlers (ALHs). In this review, we discuss the considerations for ALHs in the microliter range and highlight advantages and challenges when transforming from manual to automated workflows. We strongly focused on differences in liquid handling and outlined advantages due to sensor-controlled pipetting. ALHs can substantially improve costs-effectiveness and laboratory capacity. This is a consequence of increased efficiency, and throughput of laboratories while simultaneously raising data quality. Additionally, ALHs can improve safety, documentation of data, and sustainability. While automation requires careful consideration and resource demanding implementation, we believe it offers numerous advantages and can help to transform modern laboratories., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)
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- 2024
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5. AI Driven Lab-on-Chip Cartridge for Automated Urinalysis.
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Sahu A, Kandaswamy S, Singh DV, Thyagarajan E, Parthasarathy AK, Naganna S, and Dastidar TR
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- Humans, Artificial Intelligence, Urinalysis instrumentation, Urinalysis methods, Automation, Laboratory instrumentation, Automation, Laboratory methods, Lab-On-A-Chip Devices
- Abstract
After haematology, urinalysis is the most common biological test performed in clinical settings. Hence, simplified workflow and automated analysis of urine elements are of absolute necessities. In the present work, a novel lab-on-chip cartridge (Gravity Sedimentation Cartridge) for the auto analysis of urine elements is developed. The GSC consists of a capillary chamber that uptakes a raw urine sample by capillary force and performs particles and cells enrichment within 5 min through a gravity sedimentation process for the microscopic examination. Centrifugation, which is necessary for enrichment in the conventional method, was circumvented in this approach. The AI100 device (Image based autoanalyzer) captures microscopic images from the cartridge at 40x magnification and uploads them into the cloud. Further, these images were auto-analyzed using an AI-based object detection model, which delivers the reports. These reports were available for expert review on a web-based platform that enables evidence-based tele reporting. A comparative analysis was carried out for various analytical parameters of the data generated through GSC (manual microscopy, tele reporting, and AI model) with the gold standard method. The presented approach makes it a viable product for automated urinalysis in point-of-care and large-scale settings., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)
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- 2024
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6. High-Throughput Selection and Characterisation of Aptamers on Optical Next-Generation Sequencers.
- Author
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Drees A and Fischer M
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- Automation, Laboratory instrumentation, Automation, Laboratory methods, High-Throughput Nucleotide Sequencing instrumentation, Mutagenesis, Optical Devices, Sequence Analysis, DNA instrumentation, Aptamers, Nucleotide chemistry, High-Throughput Nucleotide Sequencing methods, Sequence Analysis, DNA methods
- Abstract
Aptamers feature a number of advantages, compared to antibodies. However, their application has been limited so far, mainly because of the complex selection process. 'High-throughput sequencing fluorescent ligand interaction profiling' (HiTS-FLIP) significantly increases the selection efficiency and is consequently a very powerful and versatile technology for the selection of high-performance aptamers. It is the first experiment to allow the direct and quantitative measurement of the affinity and specificity of millions of aptamers simultaneously by harnessing the potential of optical next-generation sequencing platforms to perform fluorescence-based binding assays on the clusters displayed on the flow cells and determining their sequence and position in regular high-throughput sequencing. Many variants of the experiment have been developed that allow automation and in situ conversion of DNA clusters into base-modified DNA, RNA, peptides, and even proteins. In addition, the information from mutational assays, performed with HiTS-FLIP, provides deep insights into the relationship between the sequence, structure, and function of aptamers. This enables a detailed understanding of the sequence-specific rules that determine affinity, and thus, supports the evolution of aptamers. Current variants of the HiTS-FLIP experiment and its application in the field of aptamer selection, characterisation, and optimisation are presented in this review.
- Published
- 2021
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7. Cytomegalovirus Viral Load in Transplanted Patients Using the NeuMoDx™ (Qiagen) Automated System: A 1-Month Experience Feedback.
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Luciani L, Mongin D, Ninove L, Nougairède A, Bardy K, Gazin C, Charrel RN, and Zandotti C
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- Automation, Laboratory instrumentation, Automation, Laboratory methods, Cytomegalovirus isolation & purification, Cytomegalovirus Infections blood, Cytomegalovirus Infections diagnosis, Feedback, France, Humans, Laboratories, Clinical, Latent Infection virology, Prospective Studies, Viral Load methods, Viral Load statistics & numerical data, Automation, Laboratory standards, Cytomegalovirus genetics, Cytomegalovirus Infections virology, DNA, Viral genetics, Transplant Recipients statistics & numerical data, Viral Load instrumentation
- Abstract
Cytomegalovirus (CMV) reactivations represent a significant morbidity and mortality problem in transplant patients. Reliable and rapid measurement of CMV viral load is a key issue for optimal patient management. We report here the evaluation of NeuMoDx™ (Qiagen) in a routine hospital setting (University Hospitals of Marseille, France) in comparison with our classical reference technique R-GENE. During one month, 719 CMV viral loads from 507 patients were measured in parallel in both techniques. Using the ROC (receiver operating characteristic) curve and our biological experience we suggest that values <52 IU/mL (geometric mean) correspond to negative samples, values >140 IU/mL (Fowlkes-Mallows index) correspond to quantifiable positive results and values ranging from 52 to 140 IU/mL represent non-quantifiable positive results. Follow-up of 15 transplant patients who developed CMV reactivation during the study showed that NeuMoDx™ provided higher viral load measurement during the first two weeks of follow-up for three patients. These important intra-individual variations resulted in a significant median increase considering the whole data set (6.7 points of difference expressed as a percentage of the initial viral load). However, no difference between the two techniques was noticeable after two weeks of treatment. Subsequent to this first study we conclude that NeuMoDx™, used with optimized logistics and an adapted threshold, allows a rapid CMV viral load measurement and that its use does not lead to any difference in patient management compared to the reference technique R-GENE
® .- Published
- 2021
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8. GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering.
- Author
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Tait A, Proctor T, Hamilton NJI, Birchall MA, and Lowdell MW
- Subjects
- 3T3 Cells, Animals, Automation, Laboratory instrumentation, Automation, Laboratory methods, Cells, Cultured, Cryopreservation methods, Cryopreservation standards, Culture Media chemistry, Epithelial Cells metabolism, Feeder Cells cytology, Feeder Cells metabolism, Fibroblasts metabolism, Humans, Mice, Practice Guidelines as Topic, Tissue Engineering standards, Epithelial Cells cytology, Fibroblasts cytology, Mouth Mucosa cytology, Tissue Engineering methods
- Abstract
Engineered epithelial cell sheets for clinical replacement of non-functional upper aerodigestive tract mucosa are regulated as medicinal products and should be manufactured to the standards of good manufacturing practice (GMP). The current gold standard for growth of epithelial cells for research utilises growth arrested murine 3T3 J2 feeder layers, which are not available for use as a GMP compliant raw material. Using porcine mucosal tissue, we demonstrate a new method for obtaining and growing non-keratinised squamous epithelial cells and fibroblast cells from a single biopsy, replacing the 3T3 J2 with a growth arrested primary fibroblast feeder layer and using pooled Human Platelet lysate (HPL) as the media serum supplement to replace foetal bovine serum (FBS). The initial isolation of the cells was semi-automated using an Octodissociator and the resultant cell suspension cryopreservation for future use. When compared to the gold standard of 3T3 J2 and FBS containing medium there was no reduction in growth, viability, stem cell population or ability to differentiate to mature epithelial cells. Furthermore, this method was replicated with Human buccal tissue, providing cells of sufficient quality and number to create a tissue engineered sheet.
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- 2021
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9. The 3D reconstructed skin micronucleus assay using imaging flow cytometry and deep learning: A proof-of-principle investigation.
- Author
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Allemang A, Thacker R, DeMarco RA, Rodrigues MA, and Pfuhler S
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- Artificial Intelligence, Automation, Laboratory instrumentation, Automation, Laboratory methods, False Positive Reactions, Feasibility Studies, Flow Cytometry instrumentation, Humans, Image Processing, Computer-Assisted instrumentation, Micronucleus Tests instrumentation, Micronucleus Tests methods, Mitomycin toxicity, Models, Biological, Mutagenicity Tests instrumentation, Mutagenicity Tests methods, Predictive Value of Tests, Proof of Concept Study, Skin diagnostic imaging, Skin, Artificial, Software, Tissue Scaffolds, Deep Learning, Flow Cytometry methods, Image Processing, Computer-Assisted methods, Skin pathology
- Abstract
The reconstructed skin micronucleus (RSMN) assay was developed in 2006, as an in vitro alternative for genotoxicity evaluation of dermally applied chemicals or products. In the years since, significant progress has been made in the optimization of the assay, including publication of a standard protocol and extensive validation. However, the diverse morphology of skin cells makes cell preparation and scoring of micronuclei (MN) tedious and subjective, thus requiring a high level of technical expertise for evaluation. This ultimately has a negative impact on throughput and the assay would benefit by the development of an automated method which could reduce scoring subjectivity while also improving the robustness of the assay by increasing the number of cells that can be scored. Imaging flow cytometry (IFC) with the ImageStream
®X Mk II can capture high-resolution transmission and fluorescent imagery of cells in suspension. This proof-of-principle study describes protocol modifications that enable such automated measurement in 3D skin cells following exposure to mitomycin C and colchicine. IFC was then used for automated image capture and the Amnis® Artificial Intelligence (AAI) software permitted identification of binucleated (BN) cells with 91% precision. On average, three times as many BN cells from control samples were evaluated using IFC compared to the standard manual analysis. When IFC MNBN cells were visually scored from within the BN cell images, their frequency compared well with manual slide scoring, showing that IFC technology can be applied to the RSMN assay. This method enables faster time to result than microscope-based scoring and the initial studies presented here demonstrate its capability for the detection of statistically significant increases in MNBN frequencies. This work therefore demonstrates the feasibility of combining IFC and AAI to automate scoring for the RSMN assay and to improve its throughput and statistical robustness., (Copyright © 2021 Elsevier B.V. All rights reserved.)- Published
- 2021
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10. Rapid Throughput Concentration-Response Analysis of Human Taste Discrimination.
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Palmer RK, Stewart MM, and Talley J
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- Adult, Automation, Laboratory instrumentation, Automation, Laboratory methods, Dose-Response Relationship, Drug, Female, High-Throughput Screening Assays instrumentation, Humans, Male, Middle Aged, Receptors, G-Protein-Coupled metabolism, Sucrose pharmacology, Taste Perception, Tongue drug effects, Tongue metabolism, Tongue physiology, Discrimination, Psychological, High-Throughput Screening Assays methods, Taste Threshold
- Abstract
Human taste threshold measurements often are used to infer tastant receptor functionality. However, taste thresholds can be influenced by receptor-independent variables. Examination of the full range of taste-active concentrations by taste discrimination has been hampered by logistics of testing multiple concentrations in replicate with human subjects. We developed an automated rapid throughput operant methodology for taste discrimination and applied it to concentration-response analysis of human taste. Tastant solutions (200 µl) drawn from a 96-well plate and self-administered to the tongue served as discriminative stimuli for money-reinforced responses on a touch-sensitive display. Robust concentration-response functions for "basic taste" stimuli were established, with particular focus on agonists of the taste 1 receptor member 2-taste 1 receptor member 3 heterodimer receptor (TAS1R2/R3). With a training cue of 100 mM sucrose, EC50 values of 56, 79, and 310 µM and 40 mM were obtained for rebaudioside A, sucralose, acesulfame potassium, and sucrose, respectively. Changing the sucrose training cue to 300 mM had no impact, but changing to 30 mM resulted in slight leftward shifts in potencies. A signal detection method also was used to determine values of d' , a probabilistic value for discriminability, which indicated that 5 mM was near the limits of detection for sucrose. With repeated testing, both EC50 values and 5 mM sucrose d' values were established for each individual subject. The results showed little correspondence between threshold sensitivities and EC50 values for sucrose. We conclude that concentration-response analysis of taste discrimination provides a more reliable means of inferring receptor function than measurement of discriminability at the lowest detectable tastant concentrations. SIGNIFICANCE STATEMENT: Many inferences about human tastant receptor functionality have been made from taste threshold measurements, which can be influenced by variables unrelated to receptors. We herein report a new methodology that enables rigorous concentration-response analysis of human taste discrimination and its use toward quantitative characterization of tastant agonist activity. Our data suggest that taste discrimination concentration-response functions are a more reliable reflection of underlying receptor activity than threshold measures obtained at the lowest detectable tastant concentrations., (Copyright © 2021 The Author(s).)
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- 2021
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11. The design and application of an automated microscope developed based on deep learning for fungal detection in dermatology.
- Author
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Gao W, Li M, Wu R, Du W, Zhang S, Yin S, Chen Z, and Huang H
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- Hair microbiology, Humans, Image Processing, Computer-Assisted methods, Microscopy, Fluorescence, Nails microbiology, Sensitivity and Specificity, Automation, Laboratory instrumentation, Automation, Laboratory methods, Deep Learning, Fungi cytology, Microscopy methods, Skin microbiology
- Abstract
Background: Light microscopy to study the infection of fungi in skin specimens is time-consuming and requires automation., Objective: We aimed to design and explore the application of an automated microscope for fungal detection in skin specimens., Methods: An automated microscope was designed, and a deep learning model was selected. Skin, nail and hair samples were collected. The sensitivity and the specificity of the automated microscope for fungal detection were calculated by taking the results of human inspectors as the gold standard., Results: An automated microscope was built, and an image processing model based on the ResNet-50 was trained. A total of 292 samples were collected including 236 skin samples, 50 nail samples and six hair samples. The sensitivities of the automated microscope for fungal detection in skin, nails and hair were 99.5%, 95.2% and 60%, respectively, and the specificities were 91.4%, 100% and 100%, respectively., Conclusion: The automated microscope we developed is as skilful as human inspectors for fungal detection in skin and nail samples; however, its performance in hair samples needs to be improved., (© 2020 Wiley-VCH GmbH.)
- Published
- 2021
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12. Leveraging Automation toward Development of a High-Throughput Gene Expression Profiling Platform.
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Chen J, Futran A, Crithary A, Li S, Wolicki A, Fogarty K, Baldick J, Chase P, Arey BJ, and Lippy J
- Subjects
- Antigens, Neoplasm genetics, Antigens, Neoplasm immunology, Automation, Laboratory instrumentation, Baculoviral IAP Repeat-Containing 3 Protein genetics, Baculoviral IAP Repeat-Containing 3 Protein immunology, Cell Line, Tumor, Chemokine CCL3 genetics, Chemokine CCL3 immunology, Drug Discovery methods, Gene Expression Profiling instrumentation, Gene Expression Profiling methods, Gene Expression Regulation, HCT116 Cells, High-Throughput Screening Assays instrumentation, Humans, Membrane Proteins genetics, Membrane Proteins immunology, Neoplasm Proteins genetics, Neoplasm Proteins immunology, Osteoblasts cytology, Osteoblasts metabolism, Primary Cell Culture, Real-Time Polymerase Chain Reaction standards, Ribosomal Proteins genetics, Ribosomal Proteins immunology, T-Lymphocytes cytology, T-Lymphocytes metabolism, Automation, Laboratory standards, Gene Expression Profiling standards, High-Throughput Screening Assays methods, Real-Time Polymerase Chain Reaction methods
- Abstract
We previously developed a panel of one-step real-time quantitative reverse transcription PCR (one-step qRT-PCR; hereafter referred to as qRT-PCR) assays to assess compound efficacy. However, these high-cost, conventional qRT-PCR manual assays are not amenable to high-throughput screen (HTS) analysis in a time-sensitive and complex drug discovery process. Here, we report the establishment of an automated gene expression platform using in-house lysis conditions that allows the study of various cell lines, including primary T cells. This process innovation provides the opportunity to perform genotypic profiling in both immunology and oncology therapeutic areas with quantitative studies as part of routine drug discovery program support. This newly instituted platform also enables a panel screening strategy to efficiently connect HTS, lead identification, and lead optimization in parallel.
- Published
- 2021
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13. High diversity droplet microfluidic libraries generated with a commercial liquid spotter.
- Author
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Zhang JQ, Siltanen CA, Dolatmoradi A, Sun C, Chang KC, Cole RH, Gartner ZJ, and Abate AR
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- Automation, Laboratory instrumentation, Emulsions chemistry, High-Throughput Screening Assays instrumentation, High-Throughput Screening Assays methods, Lipids chemistry, Microfluidics instrumentation, Small Molecule Libraries chemistry, Automation, Laboratory methods, Microfluidics methods
- Abstract
Droplet libraries consisting of many reagents encapsulated in separate droplets are necessary for applications of microfluidics, including combinatorial chemical synthesis, DNA-encoded libraries, and massively multiplexed PCR. However, existing approaches for generating them are laborious and impractical. Here, we describe an automated approach using a commercial array spotter. The approach can controllably emulsify hundreds of different reagents in a fraction of the time of manual operation of a microfluidic device, and without any user intervention. We demonstrate that the droplets produced by the spotter are similarly uniform to those produced by microfluidics and automate the generation of a ~ 2 mL emulsion containing 192 different reagents in ~ 4 h. The ease with which it can generate high diversity droplet libraries should make combinatorial applications more feasible in droplet microfluidics. Moreover, the instrument serves as an automated droplet generator, allowing execution of droplet reactions without microfluidic expertise.
- Published
- 2021
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14. Reduced activity of B lymphocytes, recognised by Sysmex XN-2000™ haematology analyser, predicts mortality in patients with coronavirus disease 2019.
- Author
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Rolla R, Vidali M, Puricelli C, Scotta AM, Pedrinelli A, Pergolini P, Pirisi M, Dianzani U, and Rigamonti C
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- Aged, Aged, 80 and over, Automation, Laboratory methods, B-Lymphocytes cytology, Blood Cell Count, COVID-19 epidemiology, COVID-19 virology, Creatinine blood, Epidemics, Female, Hematology methods, Hospital Mortality, Humans, Male, Middle Aged, Patient Admission statistics & numerical data, Prognosis, SARS-CoV-2 physiology, Automation, Laboratory instrumentation, B-Lymphocytes metabolism, COVID-19 diagnosis, Hematology instrumentation, SARS-CoV-2 isolation & purification
- Published
- 2021
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15. Analytical Performance Evaluation of Automated Coagulation Analyzer CP3000 for Routine and Special Coagulation Assays.
- Author
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Kim B, Seo MS, and Park R
- Subjects
- Anticoagulants analysis, Blood Coagulation physiology, Blood Coagulation Tests methods, Clinical Laboratory Services, Fibrin Fibrinogen Degradation Products analysis, Fibrinogen analysis, Humans, Partial Thromboplastin Time instrumentation, Partial Thromboplastin Time methods, Prothrombin analysis, Reference Values, Reproducibility of Results, Thrombin Time instrumentation, Thrombin Time methods, Automation, Laboratory instrumentation, Blood Coagulation Tests instrumentation
- Abstract
CP3000 coagulation analyzer is a high-throughput, fully automated coagulation analyzer. The objective of this study was to evaluate the analytical performance of CP3000 coagulation system for general and special coagulation analyses. Quality control materials and patient samples were used to evaluate the analytical performance of CP3000 coagulation system. Precision, carryover, linearity, comparability with ACL-TOP 700 coagulation system, and verification of reference range were evaluated or performed according to Clinical and Laboratory Standards Institute guidelines. Within-run and between-run precisions were below 5% for both normal and abnormal ranges. There was no detectable carryover. The linearity of antithrombin and fibrinogen were excellent. The comparability between CP3000 and ACL-TOP 700 coagulation systems was acceptable except for activated partial thromboplastin time and thrombin time due to differences in reagent composition. Reference ranges proposed by the manufacturer were verified to be acceptable. CP3000 coagulation system is a reliable system that can be used to perform routine and special coagulation tests rapidly and accurately. Because of its small footprint as an additional advantage, the implementation of CP3000 coagulation system can be efficient in hospital laboratories of various sizes., (Copyright © 2021 by the Association of Clinical Scientists, Inc.)
- Published
- 2021
16. Automated behavioral analysis.
- Author
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Vogt N
- Subjects
- Animals, Automation, Laboratory methods, Pattern Recognition, Automated, Algorithms, Automation, Laboratory instrumentation, Behavior, Animal, Disease Models, Animal, Neurosciences
- Published
- 2021
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17. Comparison of a Point-of-Care Assay and a High-Complexity Assay for Detection of SARS-CoV-2 RNA.
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Stevens B, Hogan CA, Sahoo MK, Huang C, Garamani N, Zehnder J, Kurzer J, and Pinsky BA
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- Automation, Laboratory instrumentation, Betacoronavirus genetics, COVID-19, COVID-19 Testing, Clinical Laboratory Techniques statistics & numerical data, Coronavirus Infections epidemiology, Coronavirus Infections virology, Humans, Molecular Diagnostic Techniques statistics & numerical data, Multiplex Polymerase Chain Reaction instrumentation, Pandemics, Pneumonia, Viral epidemiology, Pneumonia, Viral virology, Point-of-Care Systems statistics & numerical data, Reagent Kits, Diagnostic statistics & numerical data, Real-Time Polymerase Chain Reaction instrumentation, Reproducibility of Results, SARS-CoV-2, Time Factors, United States epidemiology, Betacoronavirus isolation & purification, Clinical Laboratory Techniques instrumentation, Coronavirus Infections diagnosis, Molecular Diagnostic Techniques instrumentation, Pneumonia, Viral diagnosis, RNA, Viral isolation & purification
- Abstract
Background: Numerous nucleic acid amplification assays utilizing different target genes of the SARS-CoV-2 genome have received emergency use authorization (EUA) by the United States Food and Drug Administration (FDA). Limited data are available comparing the test performance characteristics of these assays., Methods: A diagnostic comparison study was performed to evaluate the performance of the Cepheid Xpert Xpress SARS-CoV-2 assay compared to the Hologic Panther Fusion SARS-CoV-2 assay using clinical nasopharyngeal specimens. Agreement between the two assays was assessed by overall, positive, and negative percent agreement and Cohen's kappa coefficient., Results: A total of 104 (54 positive and 50 negative) clinical nasopharyngeal samples were tested by both assays. Using the Panther Fusion as a reference standard, the Xpert demonstrated an overall agreement of 99.0% [95% confidence interval (CI): 94.8-100], positive percent agreement of 98.1% (95% CI: 90.1-100), and a negative percent agreement of 100% (95% CI: 94.2-100). The kappa coefficient was 0.98 (95% CI: 0.94-1.0). One sample positive by the Panther Fusion with a cycle threshold (Ct) of 38.6 was found to be reproducibly negative by the Xpert assay., Conclusions: The Cepheid Xpert Xpress SARS-CoV-2 assay provides test performance comparable to the Hologic Panther Fusion SARS-CoV-2 assay while offering laboratories rapid, on-demand testing capacity., (© American Association for Clinical Chemistry 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2020
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18. A solution-free crystal-mounting platform for native SAD.
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Yu J, Shinoda A, Kato K, Tanaka I, and Yao M
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- Protein Conformation, Automation, Laboratory instrumentation, Automation, Laboratory methods, Crystallography, X-Ray instrumentation, Crystallography, X-Ray methods, Models, Molecular, Proteins chemistry
- Abstract
The native SAD phasing method uses the anomalous scattering signals from the S atoms contained in most proteins, the P atoms in nucleic acids or other light atoms derived from the solution used for crystallization. These signals are very weak and careful data collection is required, which makes this method very difficult. One way to enhance the anomalous signal is to use long-wavelength X-rays; however, these wavelengths are more strongly absorbed by the materials in the pathway. Therefore, a crystal-mounting platform for native SAD data collection that removes solution around the crystals has been developed. This platform includes a novel solution-free mounting tool and an automatic robot, which extracts the surrounding solution, flash-cools the crystal and inserts the loop into a UniPuck cassette for use in the synchrotron. Eight protein structures (including two new structures) have been successfully solved by the native SAD method from crystals prepared using this platform.
- Published
- 2020
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19. Image analysis and artificial intelligence in infectious disease diagnostics.
- Author
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Smith KP and Kirby JE
- Subjects
- Algorithms, Artificial Intelligence, Automation, Laboratory instrumentation, Humans, Automation, Laboratory methods, Communicable Diseases diagnosis, Image Processing, Computer-Assisted methods, Machine Learning
- Abstract
Background: Microbiologists are valued for their time-honed skills in image analysis, including identification of pathogens and inflammatory context in Gram stains, ova and parasite preparations, blood smears and histopathologic slides. They also must classify colony growth on a variety of agar plates for triage and assessment. Recent advances in image analysis, in particular application of artificial intelligence (AI), have the potential to automate these processes and support more timely and accurate diagnoses., Objectives: To review current AI-based image analysis as applied to clinical microbiology; and to discuss future trends in the field., Sources: Material sourced for this review included peer-reviewed literature annotated in the PubMed or Google Scholar databases and preprint articles from bioRxiv. Articles describing use of AI for analysis of images used in infectious disease diagnostics were reviewed., Content: We describe application of machine learning towards analysis of different types of microbiologic image data. Specifically, we outline progress in smear and plate interpretation as well as the potential for AI diagnostic applications in the clinical microbiology laboratory., Implications: Combined with automation, we predict that AI algorithms will be used in the future to prescreen and preclassify image data, thereby increasing productivity and enabling more accurate diagnoses through collaboration between the AI and the microbiologist. Once developed, image-based AI analysis is inexpensive and amenable to local and remote diagnostic use., (Copyright © 2020 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2020
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20. [Use of « Reference change value » in medical biology: about two examples: total PSA and hemoglobin].
- Author
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Bouverot C, Blachier S, Roquigny N, and Guerber F
- Subjects
- Automation, Laboratory instrumentation, Automation, Laboratory standards, Data Interpretation, Statistical, Diagnostic Tests, Routine instrumentation, Diagnostic Tests, Routine methods, Female, Humans, Laboratory Proficiency Testing, Male, Meta-Analysis as Topic, Observer Variation, Prostatic Neoplasms blood, Prostatic Neoplasms diagnosis, Reference Standards, Reference Values, Reproducibility of Results, Sensitivity and Specificity, Biological Variation, Individual, Clinical Laboratory Services standards, Diagnostic Tests, Routine standards, Hemoglobins analysis, Prostate-Specific Antigen analysis
- Abstract
The interpretation of the variation between the results of two dosages performed on the same patient is generally quite empirical. It is usually based on the experience of the biologist or physician. Through two examples, total PSA and hemoglobin, we hoped to set up an indicator of the significance variation between results: The Reference change value or RCV to provide assistance to the validator biologist and prescriber based on measured statistical arguments. This article describes the methodology used for the RCV calculation, the formatting on analysis reports and the limitations of the system.
- Published
- 2020
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21. [Digital morphology analyzers in hematology: a French survey].
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Grimon N, Ceccomarini T, Maurel-Ribes A, Rieu JB, Mémier V, and Dubois-Galopin F
- Subjects
- Attitude of Health Personnel, Computers, Diagnostic Tests, Routine instrumentation, Diagnostic Tests, Routine methods, Diagnostic Tests, Routine statistics & numerical data, Diagnostic Tests, Routine trends, France epidemiology, Hematologic Tests methods, Hematologic Tests statistics & numerical data, Hematologic Tests trends, Hematology methods, Hematology statistics & numerical data, Hematology trends, Humans, Job Satisfaction, Microscopy methods, Microscopy statistics & numerical data, Microscopy trends, Professional Practice statistics & numerical data, Professional Practice trends, Quality Control, Surveys and Questionnaires, Automation, Laboratory instrumentation, Automation, Laboratory methods, Automation, Laboratory statistics & numerical data, Hematologic Tests instrumentation, Hematology instrumentation, Image Processing, Computer-Assisted instrumentation, Image Processing, Computer-Assisted methods, Image Processing, Computer-Assisted statistics & numerical data, Image Processing, Computer-Assisted trends, Microscopy instrumentation
- Abstract
Digital morphology hematology analyzers are becoming more prevalent in laboratories Aims: investigate practices and assess the benefits and limits of digital automated microscopy in hematology., Methods: questionnaire sent by e-mail in 2018 to French public and private laboratories., Results: out of 118 responses (56 private, 62 public), 117 participants had a CellaVision® microscope, 1 had a West Medica®. Practices were sometimes different, especially in the choice of smears to be digitized or for quality controls (16.1% had internal quality controls, 48.3% external quality controls); 62.1% never used the red blood cell (RBC) characterization tool; the number of cells counted varied from 100 to 400. The study reported a high rate of agreement for these benefits: traceability (95.7%), staff training (94.1%), eye strain (91.4%), risk of error (87.2%), time saving (83.6%). Among the disadvantages, apart from the inadequate search for platelets clumps (93.2%), the agreement rates were often lower: adaptation to digital images (61.2%), difficult assessment of atypical morphologies (49.6%) or RBC morphology (49.6%)., Conclusion: despite well-established benefits, standardization of practices and technical improvement are still needed.
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- 2020
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22. Hemoglobin point-of-care testing in rural Gambia: Comparing accuracy of HemoCue and Aptus with an automated hematology analyzer.
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Nass SA, Hossain I, Sanyang C, Baldeh B, and Pereira DIA
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- Adolescent, Adult, Anemia diagnosis, Automation, Laboratory instrumentation, Automation, Laboratory standards, Female, Gambia, Hemoglobinometry instrumentation, Hemoglobinometry standards, Humans, Male, Rural Population, Sensitivity and Specificity, Anemia blood, Point-of-Care Testing standards
- Abstract
Background: Anemia is one of the most impactful nutrient deficiencies in the world and disproportionately affects children in low-resource settings. Point-of-care devices (PoCDs) measuring blood hemoglobin (Hb) are widely used in such settings to screen for anemia due to their low cost, speed, and convenience. Here we present the first iteration of Aptus, a new PoCD which measures Hb and hematocrit (HCT)., Aim: To evaluate the accuracy of Aptus and HemoCue® Hb 301 against an automated hematology analyzer (Medonic®) in Gambian children aged 6-35 months and the Aptus' usage in the field., Methods: Aptus, HemoCue® and Medonic® were compared using venous blood (n = 180), and Aptus and HemoCue® additionally using capillary blood (n = 506). Agreement was estimated using Bland-Altman analysis and Lin's concordance. Usage was assessed by error occurrence and user experience., Results: Mean Hb values in venous blood did not significantly differ between Aptus and HemoCue® (10.44±1.05 vs 10.56±0.93g/dl, p>0.05), but both measured higher Hb concentrations than Medonic® (9.75±0.99g/dl, p<0.0001). Lin's coefficient between Aptus and Medonic® was rc = 0.548, between HemoCue® and Medonic® rc = 0.636. Mean bias between the PoCDs venous measurements was -0.11g/dl with limits of agreement (LoA) -1.63 and 1.40g/dl. The bias was larger for the comparisons between the Medonic® and both Aptus (0.69g/dl, LoA 0.92 and 2.31g/dl) and HemoCue® (0.81g/dl, LoA 0.17 and 1.78g/dl). ROC curves showed an AUC of 0.933 in HemoCue® and 0.799 in Aptus. Capillary Hb was higher with Aptus than HemoCue® (10.33±1.11g/dl vs 10.01±1.07g/dl, p<0.0001). Mean bias was 0.32g/dl with LoA of -1.91 and 2.54g/dl. Aptus' usage proved intuitive, yet time-to-results and cuvettes could be improved., Conclusion: Both PoCDs showed a relatively limited bias but large LoA. Aptus and HemoCue® showed similar accuracy, while both overestimated Hb levels. Aptus showed promise, with its operation unimpaired by field conditions as well as being able to show HCT values., Competing Interests: The authors have declared that no competing interests exist. D.I.A.P. is one of the inventors of the IHAT iron supplementation technology, for which she could receive future awards to inventors through the MRC Awards to Inventor scheme. D.I.A.P. has served as a consultant for Vifor Pharma UK, Entia Ltd, Danone Nutritia, UN Food and Agriculture Organization (FAO) and Nemysis Ltd. D.I.A.P. has since moved to full employment with Vifor Pharma UK. S.A.N received a travel award from Entia Ltd. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
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- 2020
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23. Flagging performance of Sysmex XN-10 haematology analyser for malaria detection.
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Dumas C, Tirard-Collet P, Mestrallet F, Girard S, Jallades L, Picot S, and Bienvenu AL
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- Automation, Laboratory instrumentation, Blood Cell Count methods, Erythrocytes parasitology, Flow Cytometry methods, Hematology methods, Humans, Malaria blood, Sensitivity and Specificity, Blood Cell Count instrumentation, Flow Cytometry instrumentation, Hematology instrumentation, Malaria diagnosis
- Abstract
Aim: The aim was to assess the flagging performance of Sysmex XN-10 haematology analyser for malaria detection through the parasitic red blood cell ('pRBC') alarm., Methods: We retrospectively studied 584 blood samples performed on the Sysmex XN-10 analyser that were tested for malaria. Sensitivity, specificity, positive and negative predictive values, and prevalence were established for the pRBC alarm., Results: Sensitivity, specificity, and positive and negative predictive values for the pRBC flag were 7.8%, 100%, 100% and 87.7%, respectively. The prevalence of pRBC flag of 0.026% in the overall population was significantly different from the prevalence of 1.027% in the population tested for malaria., Conclusions: Considering the excellent specificity and the low prevalence of the flag in the overall population, we suggest, in case of the presence of pRBC flag, the implementation of a rapid review of the blood smear looking for Plasmodium , mostly if the patient had fever and had not been tested for malaria., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)
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- 2020
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24. Evaluation of the Risk of Laboratory Microbial Contamination during Routine Testing in Automated Clinical Chemistry and Microbiology Laboratories.
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Farnsworth CW, Wallace MA, Liu A, Gronowski AM, Burnham CD, and Yarbrough ML
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- Clinical Laboratory Techniques instrumentation, Clinical Laboratory Techniques methods, Fluorescence, Fluorescent Dyes chemistry, Hand Hygiene, Humans, Laboratories, Levivirus, Microbiological Techniques instrumentation, Microbiological Techniques methods, Risk Assessment, Specimen Handling, Automation, Laboratory instrumentation, Equipment Contamination, Fomites virology
- Abstract
Background: Every clinical specimen is potentially infectious, but data regarding risk for contamination of the laboratory environment during routine testing are scarce. We assessed contamination during routine sample analysis in automated clinical chemistry and microbiology laboratories., Methods: A fluorescent marker was applied to specimen container exteriors to assess the impact of gross contamination. Nonpathogenic MS2 virus was added to remnant blood, urine, and ESwab matrices as a biomarker of cross-contamination. Samples were processed and analyzed using Roche Cobas 8100 and ISE, c502, e602, and c702 modules (blood) and BD Kiestra total laboratory automation (blood, urine, ESwabs) over 3 experiments. Fluorescence transfer to laboratory surfaces and personnel was visualized using ultraviolet light. Surfaces were swabbed and assessed for MS2 cross-contamination by RT-PCR. Adherence to standard precautions by laboratory staff was assessed by observation., Results: Fluorescence was observed on 49 of 165 (30%) laboratory surfaces and personnel and 21 of 93 (23%) total laboratory automation instruments. Fluorescence transferred most frequently to gloves (31/40), computer accessories (9/18), and specimen loading racks (12/12). None of 123 areas swabbed were positive for MS2. Improper personal protective equipment use occurred at a rate of 0.36 and 0.15 events per staff per hour in the chemistry and microbiology laboratories, respectively. Hand-washing compliance was observed for 61 of 132 (46%) staff members evaluated., Conclusions: Analysis of grossly contaminated specimens on automated chemistry and microbiology equipment elicits a low likelihood of instrument contamination. However, handling contaminated specimen containers can result in contamination of environmental laboratory surfaces, representing a source of risk that is heightened by low adherence to appropriate personal protective equipment., (©American Association for Clinical Chemistry 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
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- 2020
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25. A machine-vision approach for automated pain measurement at millisecond timescales.
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Jones JM, Foster W, Twomey CR, Burdge J, Ahmed OM, Pereira TD, Wojick JA, Corder G, Plotkin JB, and Abdus-Saboor I
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- Animals, Female, Male, Mice, Time Factors, Automation, Laboratory instrumentation, Pain Measurement methods
- Abstract
Objective and automatic measurement of pain in mice remains a barrier for discovery in neuroscience. Here, we capture paw kinematics during pain behavior in mice with high-speed videography and automated paw tracking with machine and deep learning approaches. Our statistical software platform, PAWS (Pain Assessment at Withdrawal Speeds), uses a univariate projection of paw position over time to automatically quantify seven behavioral features that are combined into a single, univariate pain score. Automated paw tracking combined with PAWS reveals a behaviorally divergent mouse strain that displays hypersensitivity to mechanical stimuli. To demonstrate the efficacy of PAWS for detecting spinally versus centrally mediated behavioral responses, we chemogenetically activated nociceptive neurons in the amygdala, which further separated the pain-related behavioral features and the resulting pain score. Taken together, this automated pain quantification approach will increase objectivity in collecting rigorous behavioral data, and it is compatible with other neural circuit dissection tools for determining the mouse pain state., Competing Interests: JJ, WF, CT, JB, OA, TP, JW, GC, JP, IA No competing interests declared, (© 2020, Jones et al.)
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- 2020
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26. Hematological performance of a last generation automated blood cell counter: The Mindray BC-6800 Plus.
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Zini G, Cantelli F, Scavone F, Barbagallo O, and Ciminello A
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- Adult, Female, Humans, Male, Automation, Laboratory instrumentation, Blood Cell Count instrumentation, Hematologic Diseases blood
- Abstract
Introduction: Hematology analyzers produce reliable, reproducible, precise, accurate results, as well as a premicroscopic characterization of abnormal samples. We have evaluated the clinical performance of a new blood cell counter, which has been temporarily made available to our hematology laboratory., Methods: Over four months, we analyzed with the Mindray BC-6800 Plus more than 1000 samples with a high incidence of hematological abnormalities, using recommended ICSH and CLSI protocols. We have also assessed flagging efficiency for abnormal cells and scattergram cell distribution., Results: From a quantitative point of view, our assessment has identified state-of-the-art level reproducibility, excellent linearity, stability over 48°C at 4°C for the conventional parameters, lack of carry-over (<0.2%), and comparability with the routine instruments. These features would make the instrument suitable for immediate and smooth introduction in the hematology laboratory. Flags for abnormal cells are efficient; flag for blast cells has high sensitivity and predictive value of negative results. Additional benefits are provided by a competent interpretation of cell distribution scattergrams in samples from patients with specific hematological disorders., Conclusion: We have demonstrated good analytical and useful diagnostic performance of this new instrument, including effective selection of abnormal samples for informed microscope morphological analysis., (© 2020 John Wiley & Sons Ltd.)
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- 2020
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27. The application of fully automated dried blood spot analysis for liquid chromatography-tandem mass spectrometry using the CAMAG DBS-MS 500 autosampler.
- Author
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Luginbühl M and Gaugler S
- Subjects
- Chromatography, Liquid methods, Diagnostic Tests, Routine instrumentation, Diagnostic Tests, Routine methods, Drug Monitoring, Humans, Infant, Newborn, Neonatal Screening, Substance Abuse Detection, Automation, Laboratory instrumentation, Automation, Laboratory methods, Dried Blood Spot Testing instrumentation, Dried Blood Spot Testing methods, Tandem Mass Spectrometry methods
- Abstract
In the past decade, dried blood spot (DBS) sampling has been used increasingly for microsampling in various fields. This is predominantly driven by the significant advantages DBS offers regarding simple sample retrieval and shipment, combined with increased analyte stability. However, the manual handling of DBS samples is laborsome and prevents the use of a high-capacity bioanalytical workflow. The recent introduction of robotic DBS extraction systems in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has enabled the full automation of the analytical process. This results in overall higher sample throughput, minimal user interaction, and a significant reduction in consumables. Different instrumental setups are currently available which differ with respect to the extraction process, extract processing strategy, and internal standard application. This review article provides an overview of fully automated DBS analysis for one of these instruments, the DBS-MS 500 autosampler from CAMAG. The automated processes are described in detail and various applications are presented. Emphasis is placed on the advantages that the use of DBS, in combination with automation, brings - such as speed, reliability, and user-friendliness. Discussing DBS solutions for newborn screening, workplace drug testing, forensic screening, direct alcohol marker analysis, antiretroviral drugs, anti-epileptic drugs, and mass drug administration, the versatility and applicability of DBS are demonstrated in detail. In conclusion, this article shows how and why fully automated DBS analysis has penetrated the routine laboratory environment., (Copyright © 2020 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)
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- 2020
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28. High-throughput in situ experimental phasing.
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Lawrence JM, Orlans J, Evans G, Orville AM, Foadi J, and Aller P
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- Crystallography, X-Ray, Synchrotrons instrumentation, Automation, Laboratory instrumentation, Automation, Laboratory methods, Endopeptidase K chemistry, Macromolecular Substances chemistry, Muramidase chemistry
- Abstract
In this article, a new approach to experimental phasing for macromolecular crystallography (MX) at synchrotrons is introduced and described for the first time. It makes use of automated robotics applied to a multi-crystal framework in which human intervention is reduced to a minimum. Hundreds of samples are automatically soaked in heavy-atom solutions, using a Labcyte Inc. Echo 550 Liquid Handler, in a highly controlled and optimized fashion in order to generate derivatized and isomorphous crystals. Partial data sets obtained on MX beamlines using an in situ setup for data collection are processed with the aim of producing good-quality anomalous signal leading to successful experimental phasing., (open access.)
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- 2020
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29. Hepatitis Testing Performance of an Analyzer Integrated into Another Manufacturer's Automation System.
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Baste CL, Loeffler DM, Brana-Mulero C, Shah A, and Jin M
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- Hepatitis Antibodies immunology, Hepatitis, Viral, Human blood, Hepatitis, Viral, Human immunology, Hepatitis, Viral, Human virology, Humans, Serologic Tests methods, Software, Automation, Laboratory instrumentation, Hepatitis Antibodies blood, Hepatitis Viruses immunology, Hepatitis, Viral, Human diagnosis, Serologic Tests instrumentation
- Abstract
Background: The capacity to integrate platforms across vendors and disciplines has become an essential feature in the design of total laboratory automation (TLA) due space and test menu constraints. However, data on its performance are lacking. We aim to evaluate an integrated third-party immunoassay platform to the TLA system for the performance of hepatitis testing using turnaround time (TAT)., Methods: We use the Beckman Power Express (PE) system with linked 2 Beckman AU5800, 2 Beckman DxI 800, 2 Abbott Architect i2000, and other accessory components. The PE system is managed and interfaced to the laboratory information system (LIS) through Beckman Remisol (middleware) and Cennexus (track software). The hepatitis tests are performed on the Abbott Architect i2000 using Abbott Instrument Manager (middleware) for test results and this is interfaced with LIS and Cennexus. Using Viewics and Microsoft Excel, the test volumes and TAT of hepatitis results were analyzed before (February 2017 to January 2018) and after (February 2018 to January 2019) integration., Results: The TAT for each hepatitis test has decreased significantly, ranging from 13 to 81-minute reductions (P value <0.0001 for all tests) after instrument integration. The standard deviations of the TAT also decreased for each test. In addition, savings in labor expenditure of around 2 hours per day were observed. There were no laboratory space savings identified. Instead, 47.6 square foot more of space was utilized by the track connection lines., Conclusions: Our findings show significant improvement of TAT of hepatitis testing with the integration of the third-party Abbott Architect i2000 to Beckman PE system. In addition, the synchronization of multiple middleware for specimen management and result reporting allow the laboratory to achieve new efficiencies handling reflex tests and managing human resources., (© American Association for Clinical Chemistry 2020. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
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- 2020
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30. Experimental setup and image processing method for automatic enumeration of bacterial colonies on agar plates.
- Author
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Hogekamp L, Hogekamp SH, and Stahl MR
- Subjects
- Agar, Automation, Laboratory instrumentation, Colony Count, Microbial instrumentation, Escherichia coli, Image Processing, Computer-Assisted instrumentation, Pattern Recognition, Automated methods, Software, Automation, Laboratory methods, Colony Count, Microbial methods, Image Processing, Computer-Assisted methods
- Abstract
Automated colony counting methods have long been known in Microbiology. Numerous methods for automated image analysis have been described and a wide range of commercial products exists. Known advantages are saving cost by reducing enumeration time, automatic documentation, reproducibility, and operator independence. Still, even today the realization of all advantages of automated image analysis makes it necessary to either invest in an expensive, high performance commercial system, or to acquire expert knowledge in image processing. This is a considerable obstacle for many laboratories, and the reason why manual colony counting is still done frequently. This article describes an easy to apply automatic colony counting system-including suggestions for sample preparation-that can be put into operation with basic knowledge of image processing and low budget., Competing Interests: The authors have read the journal's policy and have the following competing interests: author SH is employed by the private, commercial company PALAS GmbH. There are no patents, products in development, or marketed products to declare. This does not alter the authors’ adherence to all PLOS ONE policies on sharing data and materials.
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- 2020
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31. SwiftReg cluster registration automatically reduces flow cytometry data variability including batch effects.
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Rebhahn JA, Quataert SA, Sharma G, and Mosmann TR
- Subjects
- Automation, Laboratory instrumentation, Computational Biology methods, Algorithms, Data Accuracy, Electronic Data Processing methods, Flow Cytometry statistics & numerical data, Immunologic Techniques methods
- Abstract
Biological differences of interest in large, high-dimensional flow cytometry datasets are often obscured by undesired variations caused by differences in cytometers, reagents, or operators. Each variation type requires a different correction strategy, and their unknown contributions to overall variability hinder automated correction. We now describe swiftReg, an automated method that reduces undesired sources of variability between samples and particularly between batches. A high-resolution cluster map representing the multidimensional data is generated using the SWIFT algorithm, and shifts in cluster positions between samples are measured. Subpopulations are aligned between samples by displacing cell parameter values according to registration vectors derived from independent or locally-averaged cluster shifts. Batch variation is addressed by registering batch control or consensus samples, and applying the resulting shifts to individual samples. swiftReg selectively reduces batch variation, enhancing detection of biological differences. swiftReg outputs registered datasets as standard .FCS files to facilitate further analysis by other tools.
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- 2020
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32. Repeatability imprecision from analysis of duplicates of patient samples and control materials.
- Author
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Kallner A and Theodorsson E
- Subjects
- Analysis of Variance, Automation, Laboratory instrumentation, Automation, Laboratory methods, Bias, Case-Control Studies, Chemistry, Clinical methods, Chemistry, Clinical statistics & numerical data, Humans, Quality Control, Reference Values, Automation, Laboratory standards, Chemistry, Clinical standards, Observer Variation, Reproducibility of Results, Uncertainty
- Abstract
Measurement imprecision is usually calculated from measurement results of the same stabilized control material(s) obtained over time, and is therefore, principally, only valid at the concentration(s) of the selected control material(s). The resulting uncertainty has been obtained under reproducibility conditions and corresponds to the conventional analytical goals. Furthermore, the commutability of the control materials used determines whether the imprecision calculated from the control materials reflects the imprecision of measuring patient samples. Imprecision estimated by measurements of patient samples uses fully commutable samples, freely available in the laboratories. It is commonly performed by calculating the results of routine patient samples measured twice each. Since the duplicates are usually analysed throughout the entire concentration interval of the patient samples processed in the laboratory, the result will be a weighted average of the repeatability imprecision measured in the chosen measurement intervals or throughout the entire interval of concentrations encountered in patient care. In contrast, the uncertainty derived from many measurements of control materials over periods of weeks is usually made under reproducibility conditions. Consequently, the repeatability and reproducibility imprecision play different roles in the inference of results in clinical medicine. The purpose of the present review is to detail the properties of the imprecision calculated by duplicates of natural samples, to explain how it differs from imprecision calculated from single concentrations of control materials, and to elucidate what precautions need to be taken in case of bias, e.g. due to carry-over effects.
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- 2020
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33. An exploration of the advantages of automated titration testing: low inter-instrument variability and equivalent accuracy for ABO and non-ABO antibody titres relative to tube testing.
- Author
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Adkins BD, Arnold Egloff SA, Fahey-Ahrndt K, Kjell AL, Cohn CS, and Young PP
- Subjects
- Automation, Laboratory instrumentation, Automation, Laboratory standards, Humans, Reproducibility of Results, Serologic Tests instrumentation, Serologic Tests standards, ABO Blood-Group System immunology, Automation, Laboratory methods, Serologic Tests methods
- Abstract
Background and Objectives: Obtaining IgM and IgG titres is important in numerous clinical situations, including solid-organ transplant, obstetrics, and for testing of out-of-group plasma-containing components. Tube method is the most prevalent testing modality, though it is both labour-intensive and known for intra- and inter-laboratory variability. The utility of automated gel testing as a method to improve both inter- and intra-laboratory reproducibility is unknown., Materials and Methods: Two academic centres participated in a study evaluating automated gel titreing. Group O plasma samples were used to measure titres of antibodies against ABO (IgM) with buffered gel cards and 4 minor and minor red-blood-cell antigens (IgG) anti-IgG gel cards. Multiple ORTHO VISION automated analyzers were used to assess inter-instrument variation. A subset of ABO (IgM) samples were compared between laboratories to evaluate inter-laboratory variability. Multiple samples were titred by tube and by automated gel technology to determine similarity of results., Results: Testing demonstrated no significant difference between analysers or between sites when performing automated titrations (P ≥ 0·99). Non-ABO IgG titres were evaluated and demonstrated little inter-instrument variability. The IgM anti-A and -B titres obtained by automated gel testing were neither consistently higher nor lower than tube titres. Greater than 90% of titre values were within one dilution., Conclusion: Based on this study, our data suggest that titreing by automated gel testing is both highly reproducible (IgM and IgG) and does not differ significantly from manual tube testing results of direct agglutination (IgM)., (© 2020 International Society of Blood Transfusion.)
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- 2020
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34. Measurement repeatability profiles of eight frequently requested measurands in clinical chemistry determined by duplicate measurements of patient samples.
- Author
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Kallner A, Petersmann A, Nauck M, and Theodorsson E
- Subjects
- Alanine Transaminase blood, Analysis of Variance, Aspartate Aminotransferases blood, Automation, Laboratory instrumentation, Automation, Laboratory methods, Biomarkers blood, C-Reactive Protein metabolism, Calcium blood, Chemistry, Clinical methods, Chemistry, Clinical statistics & numerical data, Cholesterol blood, Cohort Studies, Creatinine blood, Humans, Quality Control, Reference Values, Thyrotropin blood, Triglycerides blood, Automation, Laboratory standards, Chemistry, Clinical standards, Observer Variation, Reproducibility of Results, Uncertainty
- Abstract
Measurement uncertainties in clinical chemistry are commonly regarded as heteroscedastic - having a constant relative standard deviation irrespective of the concentration of the measurand. The uncertainty is usually determined at two concentrations using stabilized control materials and assumed to represent the analytical goal. The purpose of the present study was to use duplicates of unselected patient samples to calculate the absolute and relative repeatability component of the intra-laboratory measurement uncertainty from duplicates, using the Dahlberg formula and analysis of variance components. Estimates were made at five different concentration intervals of ALT, AST, Calcium, Cholesterol, Creatinine, CRP, Triglycerides and TSH covering the entire concentration interval of the patient cohort. This partioning allows detailing their repeatability profiles. The calculations of the profiles were based on randomly selected results from sets of duplicates ranging from 12,000 to 65,000 pairs. The repeatability of the measurands showed substantial variability within the measuring interval. Therefore, characterizing imprecision profiles as purely homo- or heteroscedastic or by a single number may not be optimal for the intended use. The present data make a case for nuancing the evaluation of analytical goals and minimal differences of measurement results by establishing uncertainty profiles under repeatability conditions, using natural patient samples.
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- 2020
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35. Disc diffusion AST automation: one of the last pieces missing for full microbiology laboratory automation.
- Author
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Dauwalder O and Vandenesch F
- Subjects
- Anti-Bacterial Agents pharmacology, Anti-Bacterial Agents therapeutic use, Diagnostic Tests, Routine instrumentation, Diagnostic Tests, Routine trends, Disk Diffusion Antimicrobial Tests trends, Humans, Laboratories organization & administration, Microbial Sensitivity Tests instrumentation, Microbial Sensitivity Tests trends, Workflow, Automation, Laboratory instrumentation, Disk Diffusion Antimicrobial Tests instrumentation
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- 2020
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36. Analytical validation of a quantitative method for therapeutic drug monitoring on the Alinity ® c Abbott.
- Author
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Dubois N, Sqalli G, Gilson M, and Charlier C
- Subjects
- Amikacin analysis, Amikacin blood, Automation, Laboratory instrumentation, Automation, Laboratory methods, Colorimetry instrumentation, Colorimetry methods, Gentamicins analysis, Gentamicins blood, Humans, Immunoassay instrumentation, Immunoassay methods, Phenobarbital analysis, Phenobarbital blood, Phenytoin analysis, Phenytoin blood, Reproducibility of Results, Theophylline analysis, Theophylline blood, Valproic Acid analysis, Valproic Acid blood, Vancomycin analysis, Vancomycin blood, Drug Monitoring instrumentation, Drug Monitoring methods, Nephelometry and Turbidimetry instrumentation, Nephelometry and Turbidimetry methods
- Abstract
Objective: The aim of this study was to evaluate the analytical performance of the Alinity
® c Abbott compared to the Architect® immunoassay system for the determination of drugs having a narrow therapeutic index., Methods: Valproic acid, amikacin, gentamicin, phenobarbital and vancomycin were analyzed using Particle-Enhanced Turbidimetric Inhibitor Immunoassay (Petinia), phenytoin and theophylline were analyzed using an immunoenzymatic method and a colorimetric method was performed to quantify lithium. The methods were validated according to the total error approach. Seven validation standards were analyzed in quintuplet during four days to establish the limits of the methods. Dilution integrity and interferences (hemolysis and high concentrations of bilirubin and lipids) were also tested. Depending on the analyte, the results obtained for twenty to forty patients on the Alinity® were compared to those obtained on the Architect® ., Results: The bias and the coefficients of variation for repeatability and for intermediate precision were lower than 15% for all drugs. Accuracy profiles were acceptable (acceptance limits fixed at 30%) in the validated ranges. The lower limits of quantification (LLOQ) were similar to those determined by Abbott except for gentamicin for which we determined a LLOQ at 1.22 mg/L while Abbott determined it at 0.5 mg/L. All assays diluted linear and analyte concentrations were not affected by interferences. Concentrations obtained for real samples on the Alinity® c are comparable to those obtained on the Architect® ci., Conclusions: The analytical validation of a method suitable for therapeutic drug monitoring of drugs on the Alinity® c meets the requirements of European Medicines Agency.- Published
- 2020
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37. Multi-site reproducibility of a human immunophenotyping assay in whole blood and peripheral blood mononuclear cells preparations using CyTOF technology coupled with Maxpar Pathsetter, an automated data analysis system.
- Author
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Bagwell CB, Hunsberger B, Hill B, Herbert D, Bray C, Selvanantham T, Li S, Villasboas JC, Pavelko K, Strausbauch M, Rahman A, Kelly G, Asgharzadeh S, Gomez-Cabrero A, Behbehani G, Chang H, Lyberger J, Montgomery R, Zhao Y, Inokuma M, Goldberger O, and Stelzer G
- Subjects
- Automation, Laboratory instrumentation, Automation, Laboratory methods, Automation, Laboratory standards, Canada, Data Analysis, Humans, Laboratory Proficiency Testing, Leukocytes, Mononuclear cytology, Pattern Recognition, Automated methods, Pattern Recognition, Automated standards, Reference Standards, Reproducibility of Results, United States, Blood Cells cytology, Flow Cytometry instrumentation, Flow Cytometry methods, Flow Cytometry standards, Immunophenotyping instrumentation, Immunophenotyping methods, Immunophenotyping standards
- Abstract
High-dimensional mass cytometry data potentially enable a comprehensive characterization of immune cells. In order to positively affect clinical trials and translational clinical research, this advanced technology needs to demonstrate a high reproducibility of results across multiple sites for both peripheral blood mononuclear cells (PBMC) and whole blood preparations. A dry 30-marker broad immunophenotyping panel and customized automated analysis software were recently engineered and are commercially available as the Fluidigm® Maxpar® Direct™ Immune Profiling Assay™. In this study, seven sites received whole blood and six sites received PBMC samples from single donors over a 2-week interval. Each site labeled replicate samples and acquired data on Helios™ instruments using an assay-specific acquisition template. All acquired sample files were then automatically analyzed by Maxpar Pathsetter™ software. A cleanup step eliminated debris, dead cells, aggregates, and normalization beads. The second step automatically enumerated 37 immune cell populations and performed label intensity assessments on all 30 markers. The inter-site reproducibility of the 37 quantified cell populations had consistent population frequencies, with an average %CV of 14.4% for whole blood and 17.7% for PBMC. The dry reagent coupled with automated data analysis is not only convenient but also provides a high degree of reproducibility within and among multiple test sites resulting in a comprehensive yet practical solution for deep immune phenotyping., (© 2019 International Clinical Cytometry Society.)
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- 2020
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38. Robotic automation of a UHPLC/MS-MS method profiling one-carbon metabolites, amino acids, and precursors in plasma.
- Author
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Andraos S, Goy M, Albert BB, Kussmann M, Thorstensen EB, and O'Sullivan JM
- Subjects
- Humans, Amino Acids blood, Automation, Laboratory instrumentation, Chromatography, High Pressure Liquid instrumentation, Robotics, Tandem Mass Spectrometry instrumentation
- Abstract
Amino acids (AAs) and one-carbon (1-C) metabolism compounds are involved in a range of key metabolic pathways, and mediate numerous health and disease processes in the human body. Previous assays have quantified a limited selection of these compounds and typically require extensive manual handling. Here, we describe the robotic automation of an analytical method for the simultaneous quantification of 37 1-C metabolites, amino acids, and precursors using reversed-phase ultra-high-pressure liquid chromatography coupled with tandem mass spectrometry (UHPLC/MS-MS). Compound extraction from human plasma was tested manually before being robotically automated. The final automated analytical panel was validated on human plasma samples. Our automated and multiplexed method holds promise for application to large cohort studies., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)
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- 2020
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39. Mindray SF-Cube technology: An effective way for correcting platelet count in individuals with EDTA dependent pseudo thrombocytopenia.
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Deng J, Chen Y, Zhang S, Li L, Shi Q, Liu M, and Yu X
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- Adult, Blood Platelets, Female, Humans, Platelet Aggregation, Platelet Count, Automation, Laboratory instrumentation, Edetic Acid chemistry, Hematologic Tests instrumentation, Thrombocytopenia blood
- Abstract
Several strategies are applied to determine the precise platelet count in individuals with ethylenediaminetetraacetic acid dependent pseudo thrombocytopenia (EDTA-PTCP) caused by in vitro aggregation of platelets in daily laboratory practice. None of them proves optimal for routine purposes. Thus, Mindray has developed the SF-Cube technology coupled with the CDR mode in the Mindray hematology analyzer to overcome the problem of EDTA-PTCP. With Mindray SF-Cube technology, platelet aggregates dissociate effectively and platelets are correctly counted in the CDR mode without pre-analytical management. In our studies, the EDTA-PTCP blood samples when analyzed with the CDR mode of Mindray BC-6800 plus analyzer, yield a markedly higher platelet count compared to those obtained with PLT-I on Sysmex XN-9000 hematology analyzer. We conclude that in patients with known or suspected EDTA-PTCP Mindray SF-Cube technology is a straightforward and effective way of determining the platelet count in EDTA-anticoagulated blood., Competing Interests: Declaration of Competing Interest The authors declare that no competing interests exist., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2020
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40. [Mastering the analytical process of the Complete Blood Count: how and why?]
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Bouhsane D, Bernous N, Mokrane R, Chaguetmi H, Ouchetati I, Houar I, Chouia F, Bouzenda K, Zouiten R, and Reggam K
- Subjects
- Automation, Laboratory instrumentation, Automation, Laboratory methods, Automation, Laboratory standards, Hematology methods, Hospitals, University standards, Humans, Leukocyte Count instrumentation, Leukocyte Count methods, Leukocyte Count standards, Pre-Analytical Phase methods, Pre-Analytical Phase standards, Prospective Studies, Quality Control, Reproducibility of Results, Transfusion Medicine methods, Transfusion Medicine standards, Blood Cell Count instrumentation, Blood Cell Count methods, Blood Cell Count standards, Hematology standards, Laboratories, Hospital standards
- Abstract
This is a prospective study realized at the level of the hematology department and blood transfusion center of the University Hospital Center (CHU) of Dr Ben Badis of Constantine and spread out over a period of one year (from January 1st to December 31st). The work focused on the analytical processes mastery of the NFS needs a compulsory step concerning technical and organizational laboratory skills respecting the ISO 15189 laws going through a mastery of support processes (humain resourses, informatics, materials, documents, management) indispensable for the good function of analytic proceedings, a performance evaluation of the hematology analyzer Advia (2120 I and II and 560) and quality control management (intern, extern). The analytic performance evaluation of Advia gives reliable results reproductible and stable for use of the routine automatisation good inter-machine correlation and laboratory performance in terms of the quality extern evaluation with military hospital laboratory.
- Published
- 2020
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41. Evaluation of Bio-Rad® discs for antimicrobial susceptibility testing by disc diffusion and the ADAGIO™ system for the automatic reading and interpretation of results.
- Author
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Strauss M, Zoabi K, Sagas D, Reznik-Gitlitz B, and Colodner R
- Subjects
- Bacteria drug effects, Humans, Reproducibility of Results, Sensitivity and Specificity, Anti-Bacterial Agents pharmacology, Automation, Laboratory instrumentation, Automation, Laboratory methods, Disk Diffusion Antimicrobial Tests methods, Disk Diffusion Antimicrobial Tests standards
- Abstract
The disc diffusion test is used for antimicrobial susceptibility testing worldwide. In this study, the performance of both Bio-Rad® antibiotic discs (as compared with Oxoid® discs) and the ADAGIO™ automated system for the reading of disc diffusion test results was evaluated with American Type Culture Collection (ATCC) quality control (QC) and wild strains of bacteria. Inhibition zones of both disc brands were read manually and through use of the ADAGIO™ system. Categorized interpretation of the results for each strain and antibiotic combination was summarized according to the Clinical Laboratory Standards Institute MS-100 (2017 update) manual and ADAGIO™ readings. Eight ATCC QC strains and 120 different wild strains were evaluated, to give a total of 1226 antibiotic/bacteria combinations and 2486 manual readings. One major error and four minor errors (0.08% and 0.34%, respectively) were detected via manual readings of the Bio-Rad® discs as compared with the Oxoid® discs. For the same number of antibiotic/bacteria combinations, five minor errors and one major error (0.42% and 0.08%, respectively) were detected with the Bio-Rad® discs read by the ADAGIO™ system. In addition, the number of times the automatic reading needed manual edition with Bio-Rad® discs was statistically lower than it did with Oxoid® discs (3.7% vs. 5.7%, p < 0.05). These findings support the hypothesis that Bio-Rad discs are not inferior to Oxoid® discs, and the performance of the ADAGIO™ system is comparable to that of manual readings with both disc brands.
- Published
- 2020
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42. Automated buffer preparation using quaternary valve in fast performance liquid chromatography for protein purification from a cell membrane.
- Author
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Lee JY, Dang K, Liu A, and Alba BM
- Subjects
- Buffers, Chromatography, Liquid methods, Equipment Design, Humans, Automation, Laboratory instrumentation, Cell Membrane chemistry, Chromatography, Liquid instrumentation, Membrane Proteins isolation & purification
- Abstract
There is a great need for high-throughput protein purification to produce protein molecules for research and therapeutics. Although there have been significant advancements made in automated multi-step chromatography and preparative in-process design-of-experiment (DOE) capabilities in commercial fast performance liquid chromatography (FPLC) instruments, almost all commercial FPLCs rely on a binary buffer mixing system, which hinders automated buffer preparation. Nevertheless, current-generation FPLCs are equipped with a quaternary mixer designed for limited in-line buffer preparation and preparative pH scouting DOE experiments. We decided to leverage the quaternary mixing capability by extending and re-programming AkTA Avant's quaternary valve into an automated in-process buffer preparation system to simplify automated purification requiring complex washing steps. We accomplished this by using two extra inlet valves, a sample valve, and versatile valve to split inputs of the quaternary valve into software-selectable stock solutions of pH buffers, salts, eluents, and additives. We also devised a new flow scheme to perform automated two-step chromatography using only one versatile valve. This was accomplished by using only stock parts and software to facilitate reproduction. To demonstrate the versatility and capability of the system, we purified a transmembrane protein that requires a detergent to stay soluble and needs an in-column, high-salt washing step to achieve high purity., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2020
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43. End-to-End Platform for Human Pluripotent Stem Cell Manufacturing.
- Author
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Pandey PR, Tomney A, Woon MT, Uth N, Shafighi F, Ngabo I, Vallabhaneni H, Levinson Y, Abraham E, and Friedrich Ben-Nun I
- Subjects
- Automation, Laboratory instrumentation, Bioreactors, Cell Differentiation, Cell Proliferation, Cells, Cultured, Humans, Induced Pluripotent Stem Cells metabolism, Induced Pluripotent Stem Cells physiology, Myocytes, Cardiac cytology, Myocytes, Cardiac metabolism, Myocytes, Cardiac physiology, Primary Cell Culture instrumentation, Automation, Laboratory methods, Cellular Reprogramming Techniques methods, Induced Pluripotent Stem Cells cytology, Primary Cell Culture methods
- Abstract
Industrialization of stem-cell based therapies requires innovative solutions to close the gap between research and commercialization. Scalable cell production platforms are needed to reliably deliver the cell quantities needed during the various stages of development and commercial supply. Human pluripotent stem cells (hPSCs) are a key source material for generating therapeutic cell types. We have developed a closed, automated and scalable stirred tank bioreactor platform, capable of sustaining high fold expansion of hPSCs. Such a platform could facilitate the in-process monitoring and integration of online monitoring systems, leading to significantly reduced labor requirements and contamination risk. hPSCs are expanded in a controlled bioreactor using perfused xeno-free media. Cell harvest and concentration are performed in closed steps. The hPSCs can be cryopreserved to generate a bank of cells, or further processed as needed. Cryopreserved cells can be thawed into a two-dimensional (2D) tissue culture platform or a three-dimensional (3D) bioreactor to initiate a new expansion phase, or be differentiated to the clinically relevant cell type. The expanded hPSCs express hPSC-specific markers, have a normal karyotype and the ability to differentiate to the cells of the three germ layers. This end-to-end platform allows a large scale expansion of high quality hPSCs that can support the required cell demand for various clinical indications.
- Published
- 2019
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44. Evaluation of a new Rapid Antimicrobial Susceptibility system for Gram-negative and Gram-positive bloodstream infections: speed and accuracy of Alfred 60AST.
- Author
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Anton-Vazquez V, Adjepong S, Suarez C, and Planche T
- Subjects
- Automation, Laboratory instrumentation, Automation, Laboratory methods, Bacteremia diagnosis, Blood Culture methods, Clinical Laboratory Techniques methods, Gram-Negative Bacterial Infections diagnosis, Gram-Negative Bacterial Infections microbiology, Gram-Positive Bacterial Infections diagnosis, Gram-Positive Bacterial Infections microbiology, Humans, Microbial Sensitivity Tests, Prospective Studies, Reproducibility of Results, Time Factors, Anti-Bacterial Agents pharmacology, Bacteremia microbiology, Clinical Laboratory Techniques instrumentation, Gram-Negative Bacteria drug effects, Gram-Positive Bacteria drug effects
- Abstract
Background: Blood stream infections (BSIs) are a major cause of morbidity and mortality. The time from taking blood cultures to obtain results of antibiotic sensitivity can be up to five days which impacts patient care. The Alfred 60 AST™ can reduce laboratory time from positive culture bottle to susceptibility results from 16 to 25 h to 5-6 h, transforming patient care. To evaluate the diagnostic accuracy of a rapid antimicrobial susceptibility system, the Alfred 60 AST™, in clinical isolates from patients with BSIs and confirm time to results. 301 Gram-negative and 86 Gram-positive isolates were analysed directly from positive blood culture bottles following Gram staining. Antimicrobial susceptibility results and time-to-results obtained by rapid Alfred 60 AST system and BD Phoenix were compared ., Results: A total of 2196 antimicrobial susceptibility test results (AST) were performed: 1863 Gram-negative and 333 Gram-positive. AST categorical agreement (CA) for Alfred 60 AST™ was 95% (1772/1863) for Gram-negative and 89% (295/333) for Gram-positive isolates. Gram-negative CA: ampicillin 96% (290/301); ciprofloxacin 95% (283/297); ceftriaxone 96% (75/78); meropenem 97% (288/297); piperacillin-tazobactam 95% (280/295); gentamicin 94% (279/297) and amikacin 93% (277/298). The median time to susceptibility results from blood culture flagging positive was 6.3 h vs 20 h (p < 0.01) for Alfred system vs BD Phoenix™., Conclusion: Alfred 60 AST system greatly reduced time to antimicrobial susceptibility results in Gram-negative and Gram-positive BSIs with good performance and cost, particularly for Gram-negative bacteraemia.
- Published
- 2019
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45. Multicenter performance evaluation of the Abbott Alinity hq hematology analyzer.
- Author
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Slim CL, Wevers BA, Demmers MWHJ, Lakos G, Hoffmann JJML, Adriaansen HJ, Kooren JA, and Storm H
- Subjects
- Automation, Laboratory instrumentation, Blood Cell Count methods, Clinical Laboratory Services, Hematology methods, Humans, Laboratories, Leukocyte Count, Leukocytes, Netherlands, Reproducibility of Results, Blood Cell Count instrumentation, Hematology instrumentation
- Abstract
Background Alinity hq (Abbott) is a new high-throughput hematology analyzer that exclusively employs optical principles for detecting and enumerating blood cells. It reports 29 parameters, including a six-part white blood cell (WBC) differential. The aim of this multicenter study was to evaluate the analytical and clinical performance of the Alinity hq. Methods Complete blood count (CBC) results and morphological flagging were compared to that of CELL-DYN Sapphire (Abbott) and 2 × 200-cell manual differential results, on 1473 whole-blood samples from a well-defined patient population from three different clinical laboratories in the Netherlands. In addition, within-run and within-laboratory precision, linearity, limit of quantitation, carryover and sample stability were assessed. External quality assessment samples were also evaluated. Results Data analysis demonstrated strong concordance of Alinity hq results with those of CELL-DYN Sapphire for all CBC parameters, except for basophil granulocytes. Alinity hq WBC differential showed high level of agreement with manual differential results and exhibited a better agreement with manual basophil results than CELL-DYN Sapphire. The sensitivity of the Alinity hq Blast flag was 57.6%, equal to the 57.6% sensitivity of the CELL-DYN Sapphire's Blast Alert. When considering samples with ≥5% blasts, the sensitivity of the Alinity hq Blast flag was 70.0%. Analytical performance of Alinity hq was shown to be consistent with state-of-the-art (SOTA) performance characteristics. Conclusions Alinity hq CBC measurands demonstrated good overall agreement with results obtained with CELL-DYN Sapphire, as well as manual WBC differential. The analytical and clinical performance characteristics of Alinity hq make it well suited for clinical laboratories.
- Published
- 2019
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46. Protocol development for discovery of angiogenesis inhibitors via automated methods using zebrafish.
- Author
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Mauro A, Ng R, Li JY, Guan R, Wang Y, Singh KK, and Wen XY
- Subjects
- Algorithms, Animals, Animals, Genetically Modified, Automation, Laboratory instrumentation, Dose-Response Relationship, Drug, Drug Discovery instrumentation, Drug Evaluation, Preclinical methods, Embryo, Nonmammalian, Endothelial Cells drug effects, Equipment Design, High-Throughput Screening Assays instrumentation, Indoles pharmacology, Neovascularization, Physiologic drug effects, Oximes pharmacology, Angiogenesis Inhibitors pharmacology, Automation, Laboratory methods, Drug Discovery methods, High-Throughput Screening Assays methods, Zebrafish
- Abstract
Their optical clarity as larvae and embryos, small size, and high fecundity make zebrafish ideal for whole animal high throughput screening. A high-throughput drug discovery platform (HTP) has been built to perform fully automated screens of compound libraries with zebrafish embryos. A Tg(kdrl:EGFP) line, marking endothelial cell cytoplasm, was used in this work to help develop protocols and functional algorithms for the system, with the intent of screening for angiogenesis inhibitors. Indirubin 3' Monoxime (I3M), a known angiogenesis inhibitor, was used at various concentrations to validate the protocols. Consistent with previous studies, a dose dependant inhibitory effect of I3M on angiogenesis was confirmed. The methods and protocols developed here could significantly increase the throughput of drug screens, while limiting human errors. These methods are expected to facilitate the discovery of novel anti-angiogenesis compounds and can be adapted for many other applications in which samples have a good fluorescent signal., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2019
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47. Performance of Five Commercial Identification Platforms for Identification of Staphylococcus delphini.
- Author
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Canver MC, Tekle T, Compton ST, Callan K, Burd EM, Zimmer BL, Bemis DA, Carroll KC, and Westblade LF
- Subjects
- Animals, Automation, Laboratory methods, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Staphylococcal Infections diagnosis, Staphylococcal Infections microbiology, Staphylococcus aureus isolation & purification, Staphylococcus hyicus isolation & purification, Staphylococcus intermedius isolation & purification, Automation, Laboratory instrumentation, Automation, Laboratory standards, Staphylococcal Infections veterinary, Staphylococcus chemistry, Staphylococcus isolation & purification
- Abstract
The Staphylococcus intermedius group (SIG) is a collection of coagulase-positive staphylococci consisting of four distinct species, namely, Staphylococcus cornubiensis , Staphylococcus delphini , Staphylococcus intermedius , and Staphylococcus pseudintermedius SIG members are animal pathogens and rare causes of human infection. Accurate identification of S. pseudintermedius has important implications for interpretation of antimicrobial susceptibility testing data and may be important for other members of the group. Therefore, we sought to evaluate the performance of five commercially available identification platforms with 21 S. delphini isolates obtained from a variety of animal and geographic sources. Here, we show that automated biochemical platforms were unable to identify S. delphini to the species level, a function of its omission from their databases, but could identify isolates to the SIG level with various degrees of success. However, all automated systems misidentified at least one isolate as Staphylococcus aureus One matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) system was able to identify S. delphini to the species level, suggesting that MALDI-TOF MS is the best option for distinguishing members of the SIG. With the exception of S. pseudintermedius , it is unclear if other SIG members should be routinely identified to the species level; however, as our understanding of their role in animal and human diseases increases, it may be necessary and important to do so., (Copyright © 2019 American Society for Microbiology.)
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- 2019
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48. Performance evaluation of Revohem ™ FVIII chromogenic and Revohem ™ FIX chromogenic in the CS-5100 autoanalyser.
- Author
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Suzuki A, Suzuki N, Kanematsu T, Shinohara S, Arai N, Kikuchi R, and Matsushita T
- Subjects
- Automation, Laboratory methods, Blood Coagulation Tests methods, Hemophilia A blood, Hemophilia A diagnosis, Hemophilia A metabolism, Hemophilia B blood, Hemophilia B diagnosis, Hemophilia B metabolism, Humans, Lupus Coagulation Inhibitor blood, Lupus Coagulation Inhibitor metabolism, Reproducibility of Results, Sensitivity and Specificity, von Willebrand Diseases blood, von Willebrand Diseases diagnosis, von Willebrand Diseases metabolism, Automation, Laboratory instrumentation, Blood Coagulation Tests instrumentation, Chromogenic Compounds metabolism, Factor IX metabolism, Factor VIII metabolism
- Abstract
Introduction: Chromogenic substrate assay (CSA) reagents Revohem
™ FVIII and Revohem™ FIX are now available as in vitro diagnostic reagents for autoanalysers in Japan. In this study, we evaluated the performance of these reagents in the CS-5100 automated coagulation analyser., Methods: We assessed within-run and between-day imprecision, on-board stability and frozen-storage stability of Revohem FVIII and FIX. Sensitivity to lupus anticoagulant (LA) was examined using LA-positive patient plasma. Correlations were analysed using plasma samples from normal individuals and patients with haemophilia A (HA) or B (HB) or von Willebrand disease (VWD)., Results: Imprecision was <2% for Revohem FVIII and <6.5% for Revohem FIX. On-board storage of Revohem FVIII resulted in a <10% decrease in FVIII levels from baseline at 24 hours, whereas Revohem FIX showed a >10% decrease at 8 hours. Revohem FVIII showed good stability while frozen for 22 days. Although Revohem FIX showed degradation due to freeze-thawing, a new calibration improved stability up to 22 days. Interference from LA was not observed with Revohem FVIII or FIX. The FVIII CSA-CSA correlation was excellent in normal (r = 0.9924), HA (r = 0.9945) and VWD (r = 0.9914). The FVIII CSA-OSA correlation was good in normal (r = 0.8468) and excellent in HA (r = 0.975) and VWD (r = 0.9936). The FIX CSA-OSA correlation was fair in normal (r = 0.4791) and excellent in HB (r = 0.9501)., Conclusion: Revohem FVIII and FIX both showed excellent performance in the CS-5100 analyser. These reagents could be useful in routine laboratory testing for diagnosing and treating haemophilia., (© 2019 John Wiley & Sons Ltd.)- Published
- 2019
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49. Automated differential white blood cell count and cytological analysis can detect near-tetraploid cells in chronic lymphoproliferative disorders.
- Author
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Settegrana C, Chapiro E, Frere C, Davi F, Choquet S, Leblond V, Cosson A, Nguyen-Khac F, and Maloum K
- Subjects
- Aged, Aged, 80 and over, Automation, Laboratory instrumentation, Chronic Disease, Cytogenetic Analysis instrumentation, Female, Humans, Karyotype, Leukemia, Lymphocytic, Chronic, B-Cell blood, Leukemia, Lymphocytic, Chronic, B-Cell diagnosis, Leukemia, Lymphocytic, Chronic, B-Cell genetics, Leukocyte Count instrumentation, Lymphoproliferative Disorders blood, Male, Middle Aged, Reproducibility of Results, Sensitivity and Specificity, Automation, Laboratory methods, Cytogenetic Analysis methods, Leukocyte Count methods, Lymphoproliferative Disorders diagnosis, Lymphoproliferative Disorders genetics, Tetraploidy
- Published
- 2019
- Full Text
- View/download PDF
50. Blast flagging of the Sysmex XN-10 hematology analyzer with supervised cell image analysis: Impact on quality parameters.
- Author
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Petrone J, Jackups R Jr, Eby CS, Shimer G, Anderson J, and Frater JL
- Subjects
- Automation, Laboratory methods, Blood Cell Count methods, Hematology methods, Hematopoietic Stem Cells metabolism, Humans, Leukocyte Count, Leukocytes cytology, Leukocytes metabolism, Reproducibility of Results, Automation, Laboratory instrumentation, Blood Cell Count instrumentation, Hematology instrumentation, Hematopoietic Stem Cells cytology
- Abstract
Introduction: The Sysmex XN-10 automated hematology analyzer (Sysmex Corporation) is routinely used in hematology laboratories to perform complete blood cell count with differential (CBC w/ diff). The sensitivity of this system for blast detection is unclear, since many prior studies evaluating the blast flagging capabilities of Sysmex XN series used the white precursor cell (WPC) channel, which is not cleared for use in the United States., Methods: We assessed the blast flagging capabilities of the Sysmex XN-10 compared with CellaVision (a cell image analyzer)-assisted visual hematology results. We evaluated the following flags: "blasts?/abnormal lymph?" and "immature granulocytes present" and compared differences in turnaround time between methods., Results: We collected data on 2239 CBC w/ diff Sysmex automated analyzer differential and CellaVision-assisted visual differential from the inpatient hematology-oncology population of a tertiary care medical center. Solely analyzing the first CBC/diff from each unique patient, both flags had a combined sensitivity of 100%, specificity of 50.2%, PPV of 21.7%, and NPV of 100%. The mean turnaround time for the automated differential was 19.5 minutes (SD 35.9 minutes) compared with 66.4 minutes for the CellaVision-assisted visual differential (SD 68.5 minutes; P < 0.001; Figure 1)., Conclusion: The Sysmex XN-10 abnormal lymphocyte/blast and immature granulocytes flags had excellent sensitivity and acceptable specificity in detecting circulating blasts with shorter turnaround time than the CellaVision-assisted visual differential. Our study suggests that automated differentials performed on Sysmex XN-10 can replace visual differentials as a first-line screening method for blast detection with improved turnaround time in hematology-oncology populations., (© 2019 John Wiley & Sons Ltd.)
- Published
- 2019
- Full Text
- View/download PDF
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