19 results on '"Youchun Xu"'
Search Results
2. SMART: A Swing-Assisted Multiplexed Analyzer for Point-of-Care Respiratory Tract Infection Testing
- Author
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Li Zhang, Xu Wang, Dongchen Liu, Yu Wu, Li Feng, Chunyan Han, Jiajia Liu, Ying Lu, Dmitriy V. Sotnikov, Youchun Xu, and Jing Cheng
- Subjects
LAMP ,point-of-care ,SARS-CoV-2 ,Clinical Biochemistry ,Biomedical Engineering ,microfluidics ,multiplexed detection ,General Medicine ,Instrumentation ,Engineering (miscellaneous) ,Analytical Chemistry ,Biotechnology - Abstract
Respiratory tract infections such as the ongoing coronavirus disease 2019 (COVID-19) has seriously threatened public health in the last decades. The experience of fighting against the epidemic highlights the importance of user-friendly and accessible point-of-care systems for nucleic acid (NA) detection. To realize low-cost and multiplexed point-of-care NA detection, a swing-assisted multiplexed analyzer for point-of-care respiratory tract infection testing (SMART) was proposed to detect multiple respiratory tract pathogens using visible loop-mediated isothermal amplification. By performing hand-swing movements to generate acceleration force to distribute samples into reaction chambers, the design of the SMART system was greatly simplified. By using different format of chips and integrating into a suitcase, this system can be applied to on-site multitarget and multi-sample testing. Three targets including the N and Orf genes of SARS-CoV-2 and the internal control were simultaneously analyzed (limit of detection: 2000 copies/mL for raw sample; 200 copies/mL for extracted sample). Twenty-three clinical samples with eight types of respiratory bacteria and twelve COVID-19 clinical samples were successfully detected. These results indicate that the SMART system has the potential to be further developed as a versatile tool in the diagnosis of respiratory tract infection.
- Published
- 2023
- Full Text
- View/download PDF
3. A fully integrated SNP genotyping system for hereditary hearing-loss detection
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Nan Li, Yuanyue Zhang, Minjie Shen, and Youchun Xu
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Genotype ,DNA Mutational Analysis ,Infant, Newborn ,Biomedical Engineering ,Bioengineering ,General Chemistry ,Deafness ,Biochemistry ,Connexins ,Sulfate Transporters ,Mutation ,otorhinolaryngologic diseases ,Humans ,Hearing Loss - Abstract
Hereditary hearing loss is one of the most common human neurosensory disorders, and there is a great need for early intervention methods such as genetically screening newborns. Single nucleotide polymorphisms (SNPs) are the major genetic targets for hearing-loss screening. In this study, a fully integrated SNP genotyping system was constructed to identify hereditary hearing loss-related genetic markers from human whole blood. The entire detection process, including blood cell lysis, nucleic acid extraction, the reaction mixture distribution, the chambers sealing and the two-colour multiplex competitive allele-specific polymerase chain reaction (KASP), can be automatically conducted in a self-contained cassette within 3 hours. To critically evaluate the performance of the system, its specificity, sensitivity and stability were assessed. Then, 13 clinical samples were genotyped with this fluidic cassette system to detect seven hotspot deafness-associated mutations in three genes (
- Published
- 2022
4. Silent Antibodies Start Talking: Enhanced Lateral Flow Serodiagnosis with Two-Stage Incorporation of Labels into Immune Complexes
- Author
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Dmitriy V. Sotnikov, Nadezhda A. Byzova, Anatoly V. Zherdev, Youchun Xu, and Boris B. Dzantiev
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Immunoassay ,SARS-CoV-2 ,Clinical Biochemistry ,Biomedical Engineering ,COVID-19 ,Metal Nanoparticles ,General Medicine ,Antigen-Antibody Complex ,Analytical Chemistry ,Limit of Detection ,immunochromatography ,test strips ,RBD protein ,coronavirus ,Humans ,Serologic Tests ,Gold ,Instrumentation ,Engineering (miscellaneous) ,Biotechnology ,biotechnology - Abstract
Nowadays, the presence of pathogen-specific antibodies in the blood is widely controlled by a serodiagnostic technique based on the lateral flow immunoassay (LFIA). However, its common one-stage format with an antigen immobilized in the binding zone of a test strip and a nanodis-persed label conjugated with immunoglobulin-binding proteins is associated with risks of very low analytical signals. It is caused by the presence of non-specific immunoglobulins in very large excess to the target antibodies in the tested samples thus decreasing their binding with the detected labels. In this study, the first stage of the immunochromatographic serodiagnosis was carried out in its traditional format using a conjugate of gold nanoparticles with staphylococcal immunoglobulin-binding protein A and an antigen immobilized on a working membrane. At the second stage, a labeled immunoglobulin-binding protein was added, which enhanced the coloration of the bound immune complexes. The use of two separated steps, binding of specific antibodies, and further coloration of the formed complexes allowed a significant reducing the influence of non-specific immunoglobulins on the assay results. The proposed approach was ap-plied for the serodiagnosis using a recombinant RBD protein of SARS-CoV-2. As a result, an in-crease in the intensity of test zone coloration by more than two orders of magnitude was demonstrated, which enabled to significantly reduce false-negative results. When testing a panel of 16 positive and 8 negative serum samples, the diagnostic sensitivity of the LFIA was 62.5% for the common format and 100% for the enhanced format; the diagnostic specificity of both variants was 100%.
- Published
- 2022
- Full Text
- View/download PDF
5. An enhanced centrifugation-assisted lateral flow immunoassay for the point-of-care detection of protein biomarkers
- Author
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Minjie Shen, Nan Li, Jing Cheng, Youchun Xu, and Ying Lu
- Subjects
Male ,Protein biomarkers ,Point-of-Care Systems ,Biomedical Engineering ,Centrifugation ,Enzyme-Linked Immunosorbent Assay ,Bioengineering ,02 engineering and technology ,01 natural sciences ,Biochemistry ,law.invention ,law ,medicine ,Humans ,Sample preparation ,Chemiluminescence ,Point of care ,Immunoassay ,Detection limit ,Chromatography ,medicine.diagnostic_test ,Chemistry ,010401 analytical chemistry ,General Chemistry ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,0210 nano-technology ,Biomarkers ,Lateral flow immunoassay - Abstract
Protein biomarkers are widely used for disease diagnosis, but the current detection methods utilized in centralized laboratories are mainly based on enzyme-linked immunosorbent assay (ELISA)-derived sandwich-type immunoassays such as chemiluminescent or electrochemiluminescent immunoassays, which suffer from long detection times and cumbersome instruments. For the point-of-care (POC) detection of protein biomarkers, various test strips for lateral flow immunoassay (LFIA) have been manufactured, but their detection sensitivities and capabilities for raw samples are limited. In this study, an enhanced centrifugation-assisted lateral flow immunoassay (ECLFIA) was established to rapidly detect protein biomarkers in whole blood with a higher sensitivity than LFIA. By inserting a nitrocellulose membrane into a centrifugal disc, fully automated operations, including sample preparation, active lateral flow actuation, washing, and signal amplification, which could hardly be performed in conventional LFIA, were enabled on the centrifugal platform for ECLFIA. The entire process for detecting human prostate specific antigen (PSA) in a drop of blood (20 μL) could be completed in 15 min. The limit of detection for our ECLFIA system was 0.028 ng mL-1, showing a 21.4-fold improvement compared to that of LFIA. Moreover, this system was utilized to detect PSA in 34 clinical samples. The results were compared to those measured using a commercial instrument used in the hospital, and a good correlation coefficient of 0.986 was obtained, demonstrating the practicality of this ECLFIA system. In summary, the ECLFIA system established in this study can be an efficient tool for the POC detection of protein biomarkers with comprehensive advantages in sensitivity, simplicity and speed.
- Published
- 2020
6. A novel electromagnet-triggered pillar valve and its application in immunoassay on a centrifugal platform
- Author
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Yunzeng Zhu, Youchun Xu, Minjie Shen, and Yiqi Chen
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Centrifugal force ,Fabrication ,Microfluidics ,Biomedical Engineering ,Mechanical engineering ,Centrifugation ,Bioengineering ,02 engineering and technology ,01 natural sciences ,Biochemistry ,law.invention ,Mice ,law ,medicine ,Animals ,Immunoassay ,Electromagnet ,medicine.diagnostic_test ,010401 analytical chemistry ,food and beverages ,Serum Albumin, Bovine ,Rotational speed ,General Chemistry ,Microfluidic Analytical Techniques ,Mycotoxins ,021001 nanoscience & nanotechnology ,Chip ,0104 chemical sciences ,Magnet ,Magnets ,Cattle ,0210 nano-technology - Abstract
The lab-on-a-disc is a powerful microfluidic platform that skillfully takes advantage of centrifugal force to controllably drive liquids with the assistance of passive or active valves. However, the passive valves are mainly triggered by the rotation speed and can be easily influenced by the surface chemistry of the channel, while the active valves usually require a complicated fabrication or actuation procedure. In this study, a novel active valve that can be easily triggered by an electromagnet was proposed and applied on the centrifugation platform. This valve, named the electromagnet-triggered pillar (ETP) valve, consisted of a metal pin and pressure sensitive adhesive (PSA) tape, and is closed until the pin is lifted up by an electromagnet to partially separate the PSA tape from the substrate. As a typical application, this valve is utilized to construct a centrifugal chip for mycotoxin detection. With four ETP valves in a unit, the sample and liquid reagents can be sequentially released into the reaction chamber that was spotted with mycotoxin conjugates to accomplish the whole immunoassay. Four mycotoxins (aflatoxin B1, ochratoxin A, T-2 toxin, and zearalenone) were simultaneously detected on this chip with limits of detection lower than the permissible limits set by the regulatory agencies of China, demonstrating the practicability of this easy-to-use active valve.
- Published
- 2019
7. Rapid and efficient isolation and detection of extracellular vesicles from plasma for lung cancer diagnosis
- Author
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Xun Wang, Fan Yang, Youchun Xu, Junge Chen, Ying Lu, Wanli Xing, Xiurui Zhu, and Dongchen Liu
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Lung Neoplasms ,Protein biomarkers ,Isolation (health care) ,Chemistry ,Vesicle ,Biomedical Engineering ,Cancer ,Bioengineering ,Equipment Design ,General Chemistry ,respiratory system ,Cell Fractionation ,medicine.disease ,Biochemistry ,Extracellular vesicles ,Extracellular Vesicles ,MicroRNAs ,Plasma ,A549 Cells ,Lab-On-A-Chip Devices ,Biomarkers, Tumor ,Cancer research ,medicine ,Humans ,Lung cancer - Abstract
Extracellular vesicles (EVs) are cell-derived nanoscale vesicles that provide promising biomarkers for the non-invasive diagnosis of cancer because they carry important cancer-related DNA, RNA and protein biomarkers. However, the clinical application of EVs is limited by tedious and non-standardized isolation methods that require bulky instrumentation. Here, we propose an easy-to-operate, simple dielectrophoretic (DEP) method for EV isolation with higher recovery efficiency (>83%) and higher purity than ultracentrifugation (UC). The DEP chip reduces the isolation procedure from 8 h to 30 min. To facilitate subsequent analysis, our DEP chip achieved integration of EV isolation and in situ lysis of EVs for the first time. Our chip also achieved on-chip siRNA delivery to EVs isolated by DEP. We found that EVs isolated from the plasma of lung cancer patients contained higher levels of miR-21, miR-191 and miR-192 compared to those from healthy people. With on-chip detection, EGFR in EVs could distinguish lung cancer patients from healthy people. Overall, this study provides an efficient and practical approach to the isolation and detection of EVs, which could be used for the early diagnosis of lung cancer.
- Published
- 2019
8. A self-contained and fully integrated fluidic cassette system for multiplex nucleic acid detection of bacteriuria
- Author
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Youchun Xu, Ying Lu, Jing Cheng, and Nan Li
- Subjects
DNA, Bacterial ,Salmonella typhimurium ,Staphylococcus aureus ,Bacteriuria ,Biomedical Engineering ,Bioengineering ,02 engineering and technology ,medicine.disease_cause ,Biochemistry ,Microbiology ,03 medical and health sciences ,Lab-On-A-Chip Devices ,medicine ,Escherichia coli ,Humans ,Multiplex ,Proteus mirabilis ,030304 developmental biology ,Detection limit ,0303 health sciences ,biology ,Chemistry ,Pathogenic bacteria ,General Chemistry ,021001 nanoscience & nanotechnology ,biology.organism_classification ,medicine.disease ,Molecular diagnostics ,DNA extraction ,0210 nano-technology ,Bacteria - Abstract
The gold standard for diagnosing infectious diseases is culture-based identification of bacterial pathogens, which is time-consuming and labour-intensive. Current advances in molecular diagnostics and microfluidic technologies have made the rapid detection of bacteria or viruses in clinical specimens possible. However, the need for rapid, sensitive and multiplex detection of pathogens in a "sample-in and answer-out" manner has not been fully satisfied. In this study, a self-contained and fully integrated fluidic cassette and its supporting analyser were constructed for multiplex detection of bacteria to accelerate the diagnosis of urinary tract infections (UTIs). The fully integrated cassette contains all the necessary components and reagents for bacterial analysis. All of the bacterial analysis processes, including bacterial lysis, magnetic silica bead-based DNA extraction, DNA elution and multiplex loop-mediated amplification (LAMP), are automatically conducted in the cassette. This cassette was successfully applied for the detection of four major pathogenic bacteria in UTIs, i.e., Escherichia coli, Proteus mirabilis, Salmonella typhimurium and Staphylococcus aureus. The first three were successfully detected with a limit of detection (LoD) of 1 colony-forming unit (CFU) μL-1 and the last was with a LoD of 10 CFU μL-1 in urine samples, demonstrating that the cassette has similar sensitivity compared to that of the manual protocol, which is lower than that required by UTIs. The turnaround time for this cassette-based sample-to-answer system was approximately 100 minutes, and the detection is sensitive, fully automated, and accurate, demonstrating the potential to be a useful diagnostic tool for UTIs.
- Published
- 2019
9. A portable urine analyzer based on colorimetric detection
- Author
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Ying Lu, Nan Li, Jing Cheng, Youchun Xu, and Jiancheng Ye
- Subjects
Spectrum analyzer ,Computer science ,General Chemical Engineering ,010401 analytical chemistry ,General Engineering ,Analytical chemistry ,02 engineering and technology ,Urine ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,Analytical Chemistry ,Matrix (chemical analysis) ,Consistency (statistics) ,Urine examination ,Chromaticity ,0210 nano-technology ,Biomedical engineering - Abstract
Urine examination is a basic program in routine check-ups that can reveal multiple parameters about our health and thus has significant meaning. However, instruments for urine analysis in hospitals are usually bulky and expensive. In this study, we have constructed a pocket-sized and CMOS imaging-based analytical device for urine diagnosis at home or on the spot. The shape of traditional dipsticks was transformed into matrix arrays and pasted on a reaction unit for colorimetric arrays. Chromaticity values were used to establish the functional relationship between the red-green-blue intensities of images and the concentrations of different indices. The size of the entire system was reduced to 30 × 30 × 45 mm, and once charged this device can continuously work for >4 h accommodating more than 1000 tests. To validate the performance of our device, more than 200 real human urine samples were tested and the results generated by our portable device and a commercial instrument show a great consistency (the consistency for most results is >90%) between the two systems.
- Published
- 2017
10. A sheath-less electric impedance micro-flow cytometry device for rapid label-free cell classification and viability testing
- Author
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Xinwu Xie, Zhen Cheng, Youchun Xu, Jing Cheng, Ran Liu, and Qi Li
- Subjects
education.field_of_study ,Materials science ,medicine.diagnostic_test ,General Chemical Engineering ,010401 analytical chemistry ,Microfluidics ,Population ,General Engineering ,Nanotechnology ,02 engineering and technology ,021001 nanoscience & nanotechnology ,Chip ,01 natural sciences ,0104 chemical sciences ,Analytical Chemistry ,Flow cytometry ,medicine ,Miniaturization ,0210 nano-technology ,education ,Sensitivity (electronics) ,Electrical impedance ,Susceptance ,Biomedical engineering - Abstract
Microfluidic electric impedance flow cytometry (IFC) chips have strong advantages over the traditional flow cytometry system because they are self-contained, disposable, economic in reagent consumption, and easier to operate. However, the throughput, sensitivity, and simplicity of the microfluidic IFC chips are inversely related to one another, and their reported impedance-based cell differentiation capability is in general limited. In this paper, we designed a sheath-less microfluidic IFC chip with a constriction structure between the detection electrodes to enhance the particle sensing performance, and built the entire sensing system around it, including the sensing circuit and data processing software. The measurement of the volume, limit of detection (∼3 μm), coefficient variation (6.83%) and other characteristics of the device was performed using the standard polymer beads. This sheath-less polydimethylsiloxane microfluidic device had a simple structure, which could maintain a single-cell sequence flow at the detection area, and displayed high signal-to-noise ratio (23.5–32.6 dB) and low coincidence ratio signals for further analysis. The throughput of the chip for single-cell screening can reach up to 172 cells per s, and thus, 10 000 cells could be analyzed in a few minutes for statistical analysis. Moreover, the electrical conductance and susceptance were found to be good at differentiating the bead/cell sizes and membrane/surface characteristics of cells/beads, respectively. These parameters were used to classify the population of a large amount of drug-treated cells (>10 000 cells per sample), displaying good performance in distinguishing apoptotic/necrotic cells from live cells. The ratios of apoptotic, necrotic, and live cells analyzed using our system were consistent with the traditional flow cytometry results (R2 = 0.9796). Along with the miniaturization of the electric sensing circuit, our system can be applicable for novel, compact and easy operative (single) cell analysis systems in the future.
- Published
- 2017
11. Enhancing the Sensitivity of Lateral Flow Immunoassay by Centrifugation-Assisted Flow Control
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Minjie Shen, Mangsuo Zhao, Yiqi Chen, Yunzeng Zhu, and Youchun Xu
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Centrifugal force ,Detection limit ,Immunoassay ,medicine.diagnostic_test ,Chemistry ,010401 analytical chemistry ,Collodion ,Rotational speed ,Centrifugation ,Prostate-Specific Antigen ,010402 general chemistry ,Rotation ,01 natural sciences ,Sensitivity and Specificity ,0104 chemical sciences ,Analytical Chemistry ,Volumetric flow rate ,Standard curve ,Flow control (fluid) ,medicine ,Humans ,Biomedical engineering - Abstract
Lateral flow immunoassay (LFIA) is widely used but is limited by its sensitivity. In this study, a novel centrifugation-assisted lateral flow immunoassay (CLFIA) was proposed that had enhanced sensitivity compared to traditional LFIA based on test strips. For CLFIA, a vaulted piece of nitrocellulose membrane was prepared and inserted into a centrifugal disc. Powered by the centrifugal force, the sample volume on the disc was not limited and the flow rate of the reaction fluid was steady and adjustable at different rotation speeds. It was found that lower rotation speeds and larger sample volumes resulted in greater signal intensity in the nitrocellulose membrane as well as higher sensitivity, indicating that the actively controlled flow on the disc allowed for sensitivity enhancement of CLFIA. To operate CLFIA on the centrifugal disc, a portable and cost-effective operating device was constructed to rotate the disc with a stepper motor and collect the results with a smartphone. The proposed method was successfully applied to detect prostate specific antigen (PSA) in human serum. Standard curves were established for CLFIA and LFIA, and both had correlation coefficients of up to 0.99. Under optimal conditions (1500 rpm rotation speed, 120 μL sample volume), the detection limit of CLFIA reached 0.067 ng/mL, showing a 6.2-fold improvement in sensitivity compared to that of LFIA. With clinical serum samples, a good correlation was observed between PSA concentrations measured by CLFIA and by a bulky commercial instrument in hospital. In summary, this portable, cost-effective, and easy-to-use system holds great promise for biomarker detection with enhanced sensitivity compared to traditional LFIA.
- Published
- 2019
12. Self-served and fully automated biochemical detection of finger-prick blood at home using a portable microfluidic analyzer
- Author
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Jie Li, Pan Liangbin, Youchun Xu, Xiangrui Meng, Yiqi Chen, Jing Cheng, Yunzeng Zhu, Ying Lu, and Shao Haoying
- Subjects
Spectrum analyzer ,Microfluidics ,02 engineering and technology ,010402 general chemistry ,Biochemical detection ,01 natural sciences ,Materials Chemistry ,Medicine ,Electrical and Electronic Engineering ,Instrumentation ,Finger prick ,business.industry ,Blood biochemistry ,Metals and Alloys ,food and beverages ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,0104 chemical sciences ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials ,Microfluidic chip ,Fully automated ,Medical training ,0210 nano-technology ,business ,Biomedical engineering - Abstract
The burden of chronic diseases is unquestionably a predominant challenge to global health. Chronic diseases are the long-term diseases and their intervention requires frequent monitoring of disease development, which makes the self-checkup of blood biochemistry at home highly desired. However, there is few devices that can be used at home for simultaneous test of multiple finger-prick blood biochemicals. In this study, a fully integrated microfluidic analyzer which can perform multiple blood biochemical analysis is proposed. The finger-prick blood can be directly aspirated into the chip containing diluent and lyophilized reagents, and the following processes, including serum/plasma separation and metering, diluent release and metering, quantified serum/plasma and diluent mixing, the diluted serum/plasma distribution to the reaction chambers, and the biochemical reaction and results measurement, were sequentially and automatically performed on the self-contained microfluidic chip. Using this one key operated portable analyzer, users can easily accomplish their own biochemical detection without medical training, making self-served health monitoring for chronic diseases at biochemical level possible. In the current report, fully automated detection for three blood biochemical indexes, including blood glucose, total cholesterol and triglycerides, were achieved in 15 min. The finger-prick blood samples from volunteers were measured by this microfluidic analyzer, and the results were comparable to those obtained from a commercial biochemical analyzer used in hospital.
- Published
- 2020
13. Sensitive and rapid detection of pathogenic bacteria from urine samples using multiplex recombinase polymerase amplification
- Author
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Lei Wang, He Yan, Wanli Xing, Youchun Xu, Yunzeng Zhu, Yan Zhang, Ying Lu, and Junge Chen
- Subjects
Salmonella ,Time Factors ,Biomedical Engineering ,Recombinase Polymerase Amplification ,Bioengineering ,Centrifugation ,02 engineering and technology ,Bacteriuria ,DNA-Directed DNA Polymerase ,Biology ,Urinalysis ,medicine.disease_cause ,01 natural sciences ,Biochemistry ,Microbiology ,Recombinases ,Limit of Detection ,medicine ,Humans ,Escherichia coli ,Bacteria ,010401 analytical chemistry ,Pathogenic bacteria ,General Chemistry ,021001 nanoscience & nanotechnology ,medicine.disease ,biology.organism_classification ,Proteus mirabilis ,0104 chemical sciences ,Staphylococcus aureus ,Urinary Tract Infections ,0210 nano-technology ,Nucleic Acid Amplification Techniques ,Filtration - Abstract
Bacterial infections may cause severe diseases such as tuberculosis, sepsis, nephritis and cystitis. The rapid and sensitive detection of bacteria is a prerequisite for the treatment of these diseases. The current gold standard for bacterial identification is bacteriological culture. However, culture-based identification takes 3-7 days, which is time-consuming and laborious. In this study, bacteria in urine samples were enriched using a portable filter-based pipette. Then, a centrifugal chip was constructed to detect multiple pathogenic bacteria from urine samples by integrating the DNA extraction, multiplex recombinase polymerase amplification (RPA) and fluorescent detection together. This eliminated the time-consuming cultivation step, and thus accelerated the diagnosis of the urinary tract infections (UTIs). The five major pathogenic bacteria in UTIs were detected in this study, which are Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella typhimurium. Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa and Staphylococcus aureus were successfully detected with limits of detection of 100 CFU mL-1 from urine samples within 40 min. Salmonella typhimurium was successfully detected with a limit of detection of 1000 CFU mL-1 from urine samples. The chip-based bacteria detection proposed in this study is a promising tool for sensitive, accurate, and multiplex identification of bacteria in clinical urine samples of UTIs and bacteriuria.
- Published
- 2018
14. In-vitro hyperthermia studied by cellular impedance sensors
- Author
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Xinwu Xie, Ran Liu, Weixing Chen, Youchun Xu, and Jing Cheng
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Hyperthermia ,Materials science ,medicine ,Impedance sensor ,Thermal therapy ,medicine.disease ,Electrical impedance ,Cell damage ,In vitro ,Biomedical engineering - Abstract
In this paper, we used an experimental platform based on cellular impedance sensors to study cellular changes after hyperthermia treatments. Impedance profiling showed high correlation with traditional MTT results, thus the impedance data represent cell viabilities very well. Then, status of cells under six typical hyperthermia treatments with different temperatures between 41–46 °C and different treating durations (0–30 min, 45 °C) were examined by the platform separately. The results showed increasing cell damage occurred when temperature or duration increased, and indicated the thresholds of temperature (∼43 °C). Moreover, it is possible that a thermal isoeffect dose (TID) threshold exists in the two cell lines (equivalent time at 43 °C EM43=30∼40 min). These results fit previous theories of hyperthermia, demonstrating that the cellular impedance sensor is a useful analytical tool for the in vitro research of thermal therapy.
- Published
- 2015
15. A review of impedance measurements of whole cells
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Xinwu Xie, Yong Duan, Jing Cheng, Youchun Xu, Zhen Cheng, and Lei Wang
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Fabrication ,Computer science ,Microfluidics ,Biomedical Engineering ,Biophysics ,Nanotechnology ,02 engineering and technology ,Biosensing Techniques ,01 natural sciences ,Cell Physiological Phenomena ,Environmental safety ,Lab-On-A-Chip Devices ,Electrochemistry ,Microelectronics ,Impedance sensing ,Animals ,Humans ,Electrical impedance ,Label free ,business.industry ,010401 analytical chemistry ,General Medicine ,Equipment Design ,021001 nanoscience & nanotechnology ,Flow Cytometry ,0104 chemical sciences ,Equipment Failure Analysis ,Dielectric Spectroscopy ,Electric impedance spectroscopy ,0210 nano-technology ,business ,Biotechnology - Abstract
Impedance measurement of live biological cells is widely accepted as a label free, non-invasive and quantitative analytical method to assess cell status. This method is easy-to-use and flexible for device design and fabrication. In this review, three typical techniques for impedance measurement, i.e., electric cell-substrate impedance sensing, Impedance flow cytometry and electric impedance spectroscopy, are reviewed from the aspects of theory, to electrode design and fabrication, and applications. Benefiting from the integration of microelectronic and microfluidic techniques, impedance sensing methods have expanded their applications to nearly all aspects of biology, including living cell counting and analysis, cell biology research, cancer research, drug screening, and food and environmental safety monitoring. The integration with other techniques, the fabrication of devices for certain biological assays, and the development of point-of-need diagnosis devices is predicted to be future trend for impedance sensing techniques.
- Published
- 2015
16. A fully sealed plastic chip for multiplex PCR and its application in bacteria identification
- Author
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Wang Hui, Wanli Xing, He Yan, Shan Wang, Yan Zhang, Ying Lu, Yonghong Ren, Youchun Xu, and Kewei Jiang
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In situ ,DNA, Bacterial ,Materials science ,Chromatography ,Thermal cycler ,Bacteria ,Biomedical Engineering ,Analytical chemistry ,Nucleic Acid Hybridization ,Bioengineering ,General Chemistry ,Microfluidic Analytical Techniques ,Chip ,Biochemistry ,genomic DNA ,Multiplex polymerase chain reaction ,Drug Resistance, Bacterial ,Sample preparation ,Multiplex ligation-dependent probe amplification ,Primer (molecular biology) ,Multiplex Polymerase Chain Reaction ,Plastics ,DNA Primers - Abstract
Multiplex PCR is an effective tool for simultaneous multiple target detection but is limited by the intrinsic interference and competition among primer pairs when it is performed in one reaction tube. Dividing a multiplex PCR into many single PCRs is a simple strategy to overcome this issue. Here, we constructed a plastic, easy-to-use, fully sealed multiplex PCR chip based on reversible centrifugation for the simultaneous detection of 63 target DNA sequences. The structure of the chip is quite simple, which contains sine-shaped infusing channels and a number of reaction chambers connecting to one side of these channels. Primer pairs for multiplex PCR were sequentially preloaded in the different reaction chambers, and the chip was enclosed with PCR-compatible adhesive tape. For usage, the PCR master mix containing a DNA template is pipetted into the infusing channels and centrifuged into the reaction chambers, leaving the infusing channels filled with air to avoid cross-contamination of the different chambers. Then, the chip is sealed and placed on a flat thermal cycler for PCR. Finally, amplification products can be detected in situ using a fluorescence scanner or recovered by reverse centrifugation for further analyses. Therefore, our chip possesses two functions: 1) it can be used for multi-target detection based on end-point in situ fluorescence detection; and 2) it can work as a sample preparation unit for analyses that need multiplex PCR such as hybridization and target sequencing. The performance of this chip was carefully examined and further illustrated in the identification of 8 pathogenic bacterial genomic DNA samples and 13 drug-resistance genes. Due to simplicity of its structure and operation, accuracy and generality, high-throughput capacity, and versatile functions (i.e., for in situ detection and sample preparation), our multiplex PCR chip has great potential in clinical diagnostics and nucleic acid-based point-of-care testing.
- Published
- 2015
17. An individually addressable suspended-drop electroporation system for high-throughput cell transfection
- Author
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Ying Lu, Wanli Xing, and Youchun Xu
- Subjects
Materials science ,Electrical Equipment and Supplies ,Biomedical Engineering ,Bioengineering ,Nanotechnology ,Transfection ,Biochemistry ,Cell Line ,Printed circuit board ,Mice ,Suspensions ,Animals ,Humans ,Primary cell ,RNA, Small Interfering ,Throughput (business) ,business.industry ,Electroporation ,General Chemistry ,Cell culture ,Tissue Array Analysis ,Electrode ,Optoelectronics ,Drop (telecommunication) ,Printing ,RNA Interference ,business ,Plasmids - Abstract
High-efficiency transfection of genes, proteins, or drug compounds into cells without causing permanent damage is a prerequisite for many cell biology experiments. Here, we report a printed circuit board (PCB)-based electroporation device for high-throughput delivery of exogenous molecules into cells in an individually addressable manner. This device incorporates an array of 96 through-holes on the PCB with a pair of gold coated symmetric electrodes plated on the wall of each through-hole. A mixture of cell suspension and exogenous molecules is top-loaded, electroporated, and bottom-ejected with cell culture medium into the corresponding well of a 96-well plate placed under the PCB. One group of electrodes on the same side of the 96 through-holes were connected to eight top pads via connecting row wires plated on the top surface of the PCB and the other group of electrodes were connected in a column format to twelve bottom pads. Therefore, each pair of electrodes can be selectively energized, guaranteeing that appropriate electroporation parameters can be applied to different holes. We demonstrate the use of the present device to effectively introduce plasmid DNA and synthetic interfering RNA into cultured and primary cells with high cell viability and transfection efficiency. The high performance and low cost features make our device an ideal platform for cellular and molecular research and high-throughput screening.
- Published
- 2013
18. A microfluidic device with passive air-bubble valves for real-time measurement of dose-dependent drug cytotoxicity through impedance sensing
- Author
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Lei Wang, Wanli Xing, Jing Cheng, Yi Lv, and Youchun Xu
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Drug ,Materials science ,media_common.quotation_subject ,Microfluidics ,Biomedical Engineering ,Biophysics ,Dose dependence ,Drug Evaluation, Preclinical ,Nanotechnology ,Antineoplastic Agents ,chemistry.chemical_compound ,Cell Line, Tumor ,Neoplasms ,Electrochemistry ,Electric Impedance ,Impedance sensing ,Humans ,Cytotoxicity ,media_common ,Polydimethylsiloxane ,Drug discovery ,General Medicine ,Equipment Design ,Microfluidic Analytical Techniques ,chemistry ,Electrode ,Cisplatin ,Biotechnology ,Biomedical engineering - Abstract
The monitoring and evaluation of cell behaviors under various concentrations of diffusible molecules or drugs are important in drug screening and in many other types of biological studies. In the current study, a novel polydimethylsiloxane (PDMS)-based microfluidic device was established for the real-time monitoring of drug-induced cytotoxicity using electric cell-substrate impedance sensing (ECIS). This device consists of the following three components: a drug gradient generator, planar air-bubble valves, and parallel cell culture cavities that are combined with impedance-sensing electrodes. The gradient generator allows for the simultaneous administration of multiple drug doses to test the functional cytotoxicity, and the incorporated impedance sensing enables the dynamic, automatic and quantitative measurement of in vitro dose-dependent drug responses. The air-bubble valve presented here allows the automatic closure of the valve without the need for any external valve-control instrument. As a proof-of-concept demonstration, this device was applied to dynamically monitor the effects of the anticancer drug cisplatin on apoptosis in four cancer cell lines, which may be useful for drug discovery and other biological studies that require automated analysis combined with concentration gradients.
- Published
- 2011
19. The construction of an individually addressable cell array for selective patterning and electroporation
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Lei Wang, Youchun Xu, Huanfen Yao, Wanli Xing, and Jing Cheng
- Subjects
Materials science ,Cellular array ,Drug discovery ,Electroporation ,High-throughput screening ,Biomedical Engineering ,Tin Compounds ,Bioengineering ,Nanotechnology ,General Chemistry ,Dielectrophoresis ,Microarray Analysis ,Biochemistry ,Indium tin oxide ,Microelectrode ,Electrode ,Humans ,Microelectrodes ,HeLa Cells - Abstract
In basic cell biology research and drug discovery, it is important to rapidly introduce genes, proteins or drug compounds into cells without permanent damage. Here, we report a three dimensional SU-8 micro-well structure sandwiched with an indium tin oxide (ITO) electrode-covered slide from the top and an individually addressable array of microelectrodes on the bottom to allow parallel delivery of exogenous molecules into various cells in a spatially specific manner. A positive dielectrophoretic force was selectively applied by energizing appropriate electrodes to capture the dispersed cells at the bottom electrode, while the micro-wells were designed to confine cells in situ when the positive dielectrophoretic force is removed. The combination of spatial positive dielectrophoresis (pDEP) and micro-wells made it possible to construct cell microarrays with specific patterns. Once the cells become attached to the electrodes, different plasmids can be introduced sequentially for selective electroporation. The present cell arraying-assisted electroporation chip integrates a pDEP-assisted cell positioning function with selective electroporation to provide a simple and efficient method for gene transfer. This platform is ideal for high throughput screening of compounds in parallel and thus holds promise for applications in cellular and molecular research.
- Published
- 2011
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