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Cytomegalovirus (CMV) nucleic acid amplification testing is important for CMV infection diagnosis and management. CMV DNA is found in plasma and various other fluids, including urine. If CMV can be reliably detected in urine, it may be considered a non-invasive alternative to blood tests. The cobas 6800 system (Roche Diagnostics, Mannheim, Germany) is a Food and Drug Administration-approved testing platform for measuring CMV DNA in plasma.To evaluate the analytical performance of the cobas 6800 system and compare the clinical feasibility of CMV detection in plasma and urine samples.Imprecision, linearity, limit of quantitation (LOQ), and cross-reactivity of the cobas 6800 system were assessed, and reference interval verification was performed. Plasma CMV DNA quantification was compared to CMV DNA values in urine samples obtained from 129 pediatric patients (18 years of age) from March 2020 to May 2020 at a tertiary hospital.The assay precision was within the acceptable range. Linearity was observed within the tested concentration range (2.36-6.33 log IU/mL) with a coefficient of determination of 0.9972. The LOQ was 34.5 IU/mL. The assay did not show cross-reactivity with 15 other viruses. Plasma and urine detection results were stratified into three categories: negative,LOQ, and positive to analyze the degree of agreement with the results. The quadratic weighted kappa value was 0.623 (P = 0.000), showing substantial concurrence.The cobas 6800 system offers good sensitivity, precision, and linearity and is suitable for monitoring CMV viral loads in the plasma and urine samples.

We compared the performance of the Roche Diagnostics Elecsys immunoassay for the detection of Treponema pallidum specific antibodies in patient serum samples with that of the Abbott Laboratories Architect chemiluminescent microparticle immunoassay and the InTec and KHB enzyme-linked immunosorbent assays, which are commonly used in China. We tested 13,767 serum samples collected from 13 independent laboratories throughout China, which included samples from 999 previously confirmed syphilis cases and 158 ‘borderline’ samples previously identified using the Architect, InTec, and KHB tests. The Mikrogen Syphilis Immunoblot was used to confirm positive test results. The consistency between the four different assays was 100%. The sensitivity of Elecsys immunoassay was 100% versus 98.26% for Architect, 99.11% for InTec; and 98.56% for KHB. The specificity of the Elecsys immunoassay was 99.81% versus 99.74% for Architect; 99.93% versus 99.80% for InTec; and 99.85% versus 99.77% for KHB. For borderline samples, the Elecsys immunoassay yielded no false-negative results and fewer false-positive results, compared to the other tests. Considering the ease-of-use, automation, high speed, and high throughput capacity of the Elecsys assay, the higher sensitivity and specificity indicate it is superior for routine screening of serum samples for syphilis diagnosis.

Background In 2016, the Food and Drug Administration (FDA) proposed to enhance performance expectations for point-of-care testing (POCT) international normalized ratio (INR) devices relative to International Organization for Standardization (ISO) 17593:2007. The objective of the study was to estimate the frequency of warfarin dosing errors associated with a central laboratory INR method, a POCT INR method, and the proposed FDA performance goals. Methods A data set of INR results (n = 51912) from adult patients with INR ≤4 was used to assess the influence of adding assay imprecision and bias on warfarin dose decisions. The frequency of error in warfarin dose and size of error (≥1 or ≥2 dose categories) was compared using published assay specifications for the Instrumentation Laboratory ACL TOP® and the Roche Diagnostics CoaguChek® XS relative to the proposed FDA guidelines. Results The frequency of warfarin dose misclassification was largely influenced by bias and was not sensitive to assay imprecision. The central laboratory and POCT INR methods met the FDA performance specifications, had equal rates of ≥2 warfarin dose category error, and had statistically different rates of ≥1 warfarin dose category error in large samples (n >250). Conclusions Simulation models are useful tools for evaluating POCT INR assay performance criteria required to achieve the proposed FDA guidelines. This simulation depicted how the Roche Diagnostics CoaguChek XS instrument meets the guideline.

Screening of blood for human T-cell lymphotropic virus type 1 and type 2 (HTLV-1 and -2, respectively) is important to diagnose and prevent infection and ensure the safety of blood supplies. The Elecsys HTLV-I/II assay is a newly developed, electrochemiluminescence screening assay for the detection of HTLV-1/2 infection. The sensitivity and specificity of the Elecsys HTLV-I/II assay were determined using well-characterized HTLV-1/2-positive serum and plasma samples and routine diagnostic and blood donor samples expected to be HTLV negative, respectively. These results were compared with those for at least one of the following CE-marked assays at seven independent laboratories and the Roche Diagnostics facility in Penzberg, Germany: Abbott Architect rHTLV-I/II, Ortho Avioq HTLV-I/II Microelisa system, Abbott Prism HTLV-I/HTLV-II, and DiaSorin Murex HTLV I+II. Fujirebio INNO-LIA HTLV-I/II Score was used as a confirmatory assay. The Elecsys HTLV-I/II, Abbott Architect rHTLV-I/II, and Abbott Prism HTLV-I/HTLV-II assays detected all HTLV-1/2-positive samples (sensitivity, 100%). Sensitivity for Ortho Avioq HTLV-I/II was 98.63%. The Elecsys HTLV-I/II assay had a specificity of 99.95% in blood donor samples, which was comparable to results for the other assays (range, 99.91 to 100%). In routine diagnostic samples, the specificity of the Elecsys HTLV-I/II assay was 99.83%, compared with 99.70% for Abbott Architect rHTLV-I/II. Specificity for the Elecsys HTLV-I/II assay in potentially cross-reactive samples was 100%, compared with 99.0% for Ortho Avioq HTLV-I/II and 99.2% for DiaSorin Murex HTLV I+II. The Elecsys HTLV-I/II assay has the sensitivity and specificity to support its use as a routine screening assay for detecting HTLV infection.

Objectives Free light chain (FLC) measurement gained a lot of interest for diagnostic workup of monoclonal gammopathy. Methods We evaluated the performance of turbidimetric polyclonal Freelite (The Binding Site, Birmingham, UK) assays on Cobas 6000 (Roche Diagnostics, Rotkreuz, Switzerland) and nephelometric monoclonal N Latex (Siemens Healthcare Diagnostics, Marburg, Germany) assays on BN ProSpec (Dade Behring, Deerfield, IL) vs established nephelometric Freelite assays on BN ProSpec. Results Analytical performance was acceptable. Method comparison (n = 118) showed significant proportional FLC differences for N Latex assays. However, good correlation and clinical concordance were shown. Recovery study in the low concentration range demonstrated consistent over- and underrecovery for Freelite reagents, hampering future research on prognostic value of suppressed noninvolved FLC. Antigen excess detection was successful for κ FLC in three-fourths of cases with Freelite reagents and in all cases with N Latex reagents. However, the latter resulted in underestimated κ FLC concentrations. Conclusions FLC analysis requires continuous awareness of analytical limitations. Monitoring of disease response requires FLC analysis on the same platform using the same reagents.

The aim of this study was to determine the reference values for thyrotropin (TSH), thyroid hormones (total and free thyroxine, T4 and fT4; total and free triiodothyronine, T3 and fT3), thyroglobulin (Tg) and thyroid antibodies (thyroid peroxidase, TPOAb and thyroglobulin antibody, TgAb) in the population of the Republic of Srpska.A total of 250 euthyroid subjects were enrolled in this study. A direct method for choosing reference subjects was used to establish reference intervals. The hormones and thyroid antibodies were measured by an electrochemiluminescence immunoassay method (ECLIA, Roche Diagnostics, Mannheim, Germany). We calculated the reference intervals by MedCalc, version 12.1.4.0 (MedCalc software, Belgium) as recommended by the IFCC (CLSI C28-A3).Using guidelines recommended by the National Academy of Clinical Biochemistry (NACB) and based on standard statistical approaches, the reference intervals derived for TSH, fT4, T4, fT3, T3 were 0.75-5.32 mIU/L, 12.29-20.03 pmol/L, 73.49-126,30 nmol/L, 4.11-6.32 pmol/L, 1.15-2.32 nmol/L and for Tg, TPOAb, TgAb were 3.63-26.00 μg/L,18.02 mIU/L,98.00 mIU/L, respectively. We found a significant difference (p0.05) in TSH and fT3 values between different age groups as well as in T4, fT4 and fT3 values between ge nder groups.The established reference values for the population of the Republic of Srpska were significantly different from the values recommended by the manufacturer of reagents (Roche Diagnostics). Our results showed that a laboratory needs to establish its own reference values in order to set up a proper diagnosis, as well as to treat patients successfully.Cilj ovog rada bio je da se utvrde referentne vrednosti tireotropnog hormona (TSH), tireoidnih hormona (ukupni tiroksin, T4 i slobodni tiroksin, fT4; ukupni trijodtironin, T3 i slobodni trijodtironin, fT3), tireoglobulina (Tg) i anti tela štitne žlezde (antitela na tireoperoksidazu, TPO-At i tireoglobulinska antitela, Tg-At) kod stanovništva Republike Srpske.Ukupno 250 eutireoidnih ispitanika bilo je uključeno u ovo istraživanje. Korišćen je direktan način izbora referentnih osoba za utvrđivanje referentnog intervala. Hormoni i antitela štitne žlezde određivani su u serumu elektrohemiluminiscentnom metodom (ECLIA, Roche Diagnostics, Mannheim, Germany). Referentne granice su određene korištenjem programa MedCalc, verzija 12.1.4.0 (MedCalc software, Belgium) po preporuci IFCC (CLSI C28-A3).Koristeći smernice Nacionalne akademije za kliničku biohemiju (NACB) i na temelju standardnih statistiČkih pristupa utvrđeni referentni intervali za TSH, fT4, T4, fT3, T3 bili su 0,75–5,32 mIU/L, 12,29–20,03 pmol/L, 73,49– 126,30 nmol/L, 4.11–6.32 pmol/L, 1,15–2,32 nmol/L kao i za Tg, TPOAt, TgAt 3, 63–26,00 μg/L,18,02 mIU/L,98,00 mIU/L. Takođe, postojala je i značajna razlika između različitih starosnih grupa za TSH i fT3 vrednosti, kao i razlika za T4, fT4 i fT3 vrednosti između žena i muškaraca.Dobijene referentne vrednosti za populaciju Republike Srpske znatno su se razlikovale od vrednosti preporučenih od strane proizvođača reagensa (Roche Diagnostics). Rezultati su ukazali da laboratorije moraju da izrade sopstvene referentne vrednosti sa ciljem postavljanja prave dijagnoze, kao i uspešnog lečenja pacijenata.

Post-vaccination testing after hepatitis B vaccination is indispensable to evaluate long-term immunological protection. Using a threshold level of antibodies against hepatitis B surface antigen (anti-HBs) to define serological protection, implies reproducible and valid measurements of different diagnostic assays.In this study we assess the performance of currently used anti-HBs assays.In 2013, 45 laboratories participated in an external quality assessment program using pooled anti-HBs serum samples around the cutoff values 10IU/l and 100IU/l. Laboratories used either Axsym (Abbott Laboratories), Architect (Abbott Laboratories), Access (Beckman-Coulter), ADVIA Centaur anti-HBs2 (Siemens Healthcare Diagnostics), Elecsys, Modular or Cobas (Roche Diagnostics) or Vidas Total Quick (Biomerieux) for anti-HBs titre quantification. We analysed covariance using mixed-model repeated measures. To assess sensitivity/specificity and agreement, a true positive or true negative result was defined as an anti-HBs titre respectively above or below the cutoff value by ≥4 of 6 assays.Different anti-HBs assays were associated with statistically significant (P0.05) differences in anti-HBs titres in all dilutions. Sensitivity and specificity ranged respectively from 64%-100% and 95%-100%. Agreement between assays around an anti-HBs titre cutoff value of 10IU/l ranged from 93%-100% and was 44% for a cutoff value of 100IU/l.Around a cutoff value of 10IU/l use of the Access assay may result in false-negative results. Concerning the cutoff value of 100IU/l, a sample being classified below or above this cutoff relied heavily on the specific assay used, with both the Architect and the Access resulting in false-negative results.

BACKGROUNDB-type natriuretic peptides (BNPs) are used clinically to diagnose and monitor heart failure and are present in the circulation as multiple proBNP-derived fragments. We investigated the specificity of BNP immunoassays with glycosylated and nonglycosylated BNP, N-terminal proBNP (NT-proBNP), and proBNP peptides to probe the cross-reactivity of each assay.METHODSNine B-type natriuretic peptides were studied,including synthetic and recombinant BNP (Shionogi, Scios, Mayo), human and synthetic glycosylated and nonglycosylated NT-proBNP (HyTest, Roche Diagnostics), and human glycosylated and nonglycosylated proBNP (HyTest, Scios). Five BNP [Abbott, Abbott POC, Alere, Beckman Coulter, Siemens (Centaur)], 9 NT-proBNP [Ortho-Clinical Diagnostics, Roche, Response, bioMerieux, Siemens (Dimension, Immulite, Stratus CS), Mitsubishi] and 3 research-use-only proBNP immunoassays [Biosite (Alere), Bio-Rad, Goetze] were evaluated. Specificity was assessed by calculating the recovery between baseline and peptide-spiked human plasma pools at target concentrations of 100 ng/L BNP, 300 ng/L proBNP, or 450 ng/L NT-proBNP. All assays were performed in duplicate.RESULTSBNP and NT-proBNP assays demonstrated substantial cross-reactivity with proBNP peptides. NT-proBNP assays do not detect glycosylated forms of either NT-proBNP or proBNP. proBNP assays preferentially detect the BNP 1–32 peptide and have minimal cross-reactivity with BNP peptides and glycosylated proBNP.CONCLUSIONSBNP or NT-proBNP results are not transferable among the current existing immunoassays owing to their differences in cross-reactivity and ability to detect various glycosylated forms of proBNP-derived fragments. Opportunities remain to standardize and harmonize BNP and NT-proBNP assays, as well as to develop specific proBNP assays, to widen their clinical scope of use.

There has been increasing interest in standardized and quantitative Epstein-Barr virus (EBV) DNA testing for the management of EBV disease. We evaluated the performance of the Real-Q EBV Quantification Kit (BioSewoom, Korea) in whole blood (WB). Nucleic acid extraction and real-time PCR were performed by using the MagNA Pure 96 (Roche Diagnostics, Germany) and 7500 Fast real-time PCR system (Applied Biosystems, USA), respectively. Assay sensitivity, linearity, and conversion factor were determined by using the World Health Organization international standard diluted in EBV-negative WB. We used 81 WB clinical specimens to compare performance of the Real-Q EBV Quantification Kit and artus EBV RG PCR Kit (Qiagen, Germany). The limit of detection (LOD) and limit of quantification (LOQ) for the Real-Q kit were 453 and 750 IU/mL, respectively. The conversion factor from EBV genomic copies to IU was 0.62. The linear range of the assay was from 750 to 10⁶ IU/mL. Viral load values measured with the Real-Q assay were on average 0.54 log₁₀ copies/mL higher than those measured with the artus assay. The Real-Q assay offered good analytical performance for EBV DNA quantification in WB.

Bloodstains are a widespread kind of biological evidence at the crime scene and one of the most used reagents for the presumptive identification of blood for forensic purposes is tetramethyl-benzidine. We have introduced and validated the tetramethylbenzidine-based Combur 3 Test® E (Roche Diagnostics Corporation, Basel, Switzerland), a colorimetric catalytic test based upon the detection of the peroxidase-like activity of the hemoglobin, due to its high sensitivity, easiness of use and capability to maintain the complete structural and morphological integrity of the bloodstain. Analytical performances related to a forensic use of the test and the suitable applicability to the presumptive detection of bloodstains when extremely diluted, aged, mixed with several substances and deposited over a plethora of substrates was reliably proved. In addition, possible positive interferences of the test chemicals on the subsequent Short Tandem Repeats (STRs) DNA typing analyses, especially in Low-Template DNA (LT DNA) conditions, was evaluated. While the Combur 3 Test® E showed the same chemical interference drawbacks as other presumptive tests for blood as for the low specificity, we demonstrated that its format and our suggested protocol of use make it appropriate for the forensic presumptive detection of blood, better performing and much easier to use than other analogous presumptive tests and usually compatible with the following STRs DNA typing analyses.

Background The comprehensive genomic profile (CGP) by next generation sequencing (NGS) ctDNA can identify a wide spectrum of GA that range from drivers with approved targeted therapies for routine use to other GA with lack of evidence for actionability. We aimed to assess the clinical utility of NGS-ctDNA based on ESCAT in a large cohort of NSCLC patients. Methods Advanced NSCLC patients were prospectively enrolled between 11.2015-05.2019 in the Liquid Biopsy Program in our institution. Plasma ctDNA was collected at diagnosis, under therapy or at progressive disease (PD) and analyzed by InVisionFirstTMLung. We evaluated the detection of driver GA on ctDNA and the clinical utility for accessing targeted therapies approved for routine use (EGFR mutation (m), ALK rearrangement (r), BRAFV600Em, ROS1r), according to ESCAT tiers. Results Preliminary results are available for 308 patients/547 samples (n = 117 untreated; 217 at PD). 58% were females, 42% nonsmokers, with median age of 61 (24-90) and 87% had adenocarcinoma. At diagnosis, ≥1 ctDNA GA was found in 79% (91/115; 2 failed analyses): 29% (26/91) were ESCAT tier I (16 EGFRm ex19/21, 1 ALKr, 1 ROS1r, 8 BRAFV600Em), 2% ESCAT tier II (2 METa ; 1 case co-driver with EGFRm) and 31% tier III (21 KRASm with 8 cases G12C; 5 HER2m, 2EGFRm ex20). ctDNA provided clinically informative results for 33% (38/115). At PD, ≥1 ctDNA GA was found in 75% (163/217); 2 failed analyses): 65% (106/163) in ESCAT tier I (66 EGFRm ex19/21, 7 ALKr, 32 BRAFV600Em, 1 ROS1r), Conclusions ctDNA proved clinically informative results for 33% in untreated patients, most of them ESCAT tier I GA (22%) directing targeted therapies in routine. At time of TKI failure, ctDNA was clinically informative assessing resistance in 50% of EGFR/ALK patients. Legal entity responsible for the study Gustave Roussy. Funding Has not received any funding. Disclosure L. Mezquita: Advisory / Consultancy: Roche Diagnostics; Honoraria (self): Bristol-Myers Squibb; Honoraria (self): Tecnofarma; Honoraria (self): Roche; Honoraria (self): AstraZeneca; Travel / Accommodation / Expenses: Chugai. D. Planchard: Advisory / Consultancy: Eli Lilly, Merck, Novartis, Pfizer, prIME Oncology, Peer CME, Roche; Honoraria (self): AstraZeneca, Bristol-Myers Squibb, Boehringer Ingelheim, Celgene, Eli Lilly, Merck, Novartis, Pfizer, prIME Oncology, Peer CME, Roche; Research grant / Funding (institution): AstraZeneca, Bristol-Myers Squibb, Boehringer Ingelheim, Eli Lilly, Merck, Novartis, Pfizer, Roche, Medimmun, Sanofi-Aventis, Taiho Pharma, Novocure, Daiichi Sankyo; Travel / Accommodation / Expenses: AstraZeneca, Roche, Novartis, prIME Oncology, Pfizer; Advisory / Consultancy: AstraZeneca, Bristol-Myers Squibb, Boehringer Ingelheim, Celgene, Daiichi Sankyo, Eli Lilly, Merck, Novartis, Pfizer, prIME Oncology, Peer CME, Roche. F. de Kievit: Shareholder / Stockholder / Stock options: INIVATA Ltd. L. Lacroix: Advisory / Consultancy: Abbott, Astrazeneca, Boehringer Ingelheim, Bristol Myers Squibb, Ca Bayer Healthcarer, Illumina, Genomic Health, Myriad, Novartis, Pfizer, Roche, Siemens, Thermofisher, VelaDx. J. Remon Masip: Advisory / Consultancy: Pfizer, MSD, BMS, Astrazeneca, Boehringer Ingelheim; Travel / Accommodation / Expenses: OSE Immunotherapeutics, BMS, Astrazeneca, Roche; Honoraria (self): OSE Immunotherapeutics. P. Lavaud: Travel / Accommodation / Expenses: Astellas-Pharma, AstraZeneca, Ipsen, Janssen Oncology, Mundi Pharma. A. Gazzah: Travel / Accommodation / Expenses: Boehringer Ingelheim, Novartis, Pfizer, Roche; Advisory / Consultancy: Novartis; Research grant / Funding (institution), Clinical trials at Gustave Roussy: Aduro Biotech, Agios Pharmaceuticals, Amgen, Argen-X Bvba, Arno Therapeutics, Astex Pharmaceuticals, AstraZeneca, Aveo, Bayer Healthcare Ag, Bbb Technologies Bv, Beigene, Bioalliance Pharma, Biontech Ag, Blueprint Medicines, Boehringer Ingelheim, Bristol; Research grant / Funding (institution): AstraZeneca, BMS, Boehringer Ingelheim, Janssen Cilag, Merck, Novartis, Pfizer, Roche, Sanofi; Non-remunerated activity/ies, Drug supply: Astrazeneca, Bayer, BMS, Boringher Ingelheim, Johnson & Johnson, Lilly, Medimmune, Merck, NH TherAGuiX, Pfizer, Roche. C. Morris: Shareholder / Stockholder / Stock options: INIVATA Ltd. K. Howarth: Shareholder / Stockholder / Stock options: INIVATA Ltd. E. Green: Shareholder / Stockholder / Stock options: INIVATA Ltd. C. Massard: Advisory / Consultancy: Amgen, Astellas, AstraZeneca, Bayer, BeiGene, BMS, Celgene, Debiopharm, Genentech, Ipsen, Janssen, Lilly, MedImmune, MSD, Novartis, Pfizer, Roche, Sanofi, Orion; Research grant / Funding (institution), Clinical Trials at Gustave Roussy: AbbVie, Aduro, Agios, Amgen, Argen-x, Astex, AstraZeneca, Aveopharmaceuticals, Bayer, Beigene, Blueprint, BMS, Boehringer Ingelheim, Celgene, Chugai, Clovis, Daiichi Sankyo, Debiopharm, Eisai, Eos, Exelixis, Forma, Gamamabs, Genentech, Gortec, GSK, H3 biomed. B. Besse: Research grant / Funding (institution): AbbVie, Amgen, AstraZeneca, Biogen, Blueprint Medicines, BMS, Celgene, Eli Lilly, GSK, Ignyta, IPSEN, Merck KGaA, MSD, Nektar, Onxeo, Pfizer, Pharma Mar, Sanofi, Spectrum Pharmaceuticals, Takeda, Tiziana Pharma; Research grant / Funding (institution), Clinical Trials at Gustave Roussy: Nerviano, GSK, Pfizer, Roche-Genentech, Lilly, OSE Pharma, MSD, Celgene, Stemcentrx, Ignyta, AbbVie, Loxo Oncology, AstraZeneca, Blueprint Medicines. All other authors have declared no conflicts of interest.

Background The addition of S100B protein to guidelines for the management of mild traumatic brain injury (mTBI) decreases the amount of unnecessary computed tomography (CT) scans with a significant decrease in radiation exposure and an increase in cost savings. Both DiaSorin and Roche Diagnostics have developed automated assays for S100B determination. Recently, bioMérieux developed a prototype immunoassay for serum S100B determination. For the first time, we present the evaluation of the S100B measurement using a bioMérieux Vidas® 3 analyzer. Methods We evaluated the matrix effects of serum and plasma, and their stability after storage at 2–8 °C, −20 °C and −80 °C. The new measurement prototype (bioMérieux) was compared with an established one (Roche Diagnostics), and a precision study was also conducted. Lastly, clinical diagnostics performance of the bioMérieux and Roche Diagnostics methods were compared for 80 patients referred to the Emergency Department for mTBI. Results Stability after storage at 2–8 °C, −20 °C, and −80 °C and validation of the serum matrix were demonstrated. The bioMérieux analyzer was compared to the Roche Diagnostics system, and the analytical precision was found to be efficient. Clinical diagnosis performance evaluation confirmed the predictive negative value of S100B in the management of mTBI. Conclusions The study’s data are useful for interpreting serum S100B results on a bioMérieux Vidas® 3 analyzer.

Background The detection of hepatitis C virus antibody (anti-HCV) is known to have high false-positive rates. Using signal-to-cutoff (S/Co) ratios in reflex supplemental testing, however, could reduce false-positive rates. Here, we analyzed the 2-year data of an anti-HCV assay to understand the significance of the S/Co ratio and make a new algorithm by confirming with a second anti-HCV assay. Methods We reviewed 32,573 samples of the Architect assay (Abbott Diagnostics) from a tertiary hospital. Retests with the Elecsys (Roche Diagnostics) and Vitros (Ortho Clinical Diagnostics) assays were performed in 346 anti-HCV-positive samples. HCV RNA PCR and recombinant immunoblot assay (RIBA) were performed in 147 and 11 anti-HCV-positive samples, respectively. Results Among 32,573 samples, 446 (1.37%) yielded positive results and 32,127 (98.6%) yielded negative results. Concordance rates in low S/Co samples (0.9–10.0) were 35.2%, 43.8%, and 81.9% for the Architect-Elecsys, Architect-Vitros, and Elecsys-Vitros comparisons, respectively. Correlation coefficients of S/Co ratios were as follows: Architect-Elecsys, 0.20; Architect-Vitros, 0.42; and Elecsys-Vitros, 0.46. In logistic regression, the S/Co value for predicting positivity with 95% probability was 3.13, while that for predicting 50% probability was 8.85. S/Co ratios of 1.70–3.34 showed one reactive antigen out of five antigens, and S/Co ratios of 13.54–17.72 showed three to five positive reactions out of five antigens used in the RIBA. Conclusions Supplementary testing of anti-HCV screening results is necessary to distinguish between true positivity and biological false positivity for anti-HCV. In this study, we presented an algorithm of supplementary testing by a retest with a second reagent, which could be useful in clinical laboratories.

The Diabetes Remission Clinical Trial (DiRECT) demonstrated initial remission of type 2 diabetes in about half of a large primary care population, and this was associated with decrease in intrapancreatic fat content and return of beta cell function. We have carried out detailed metabolic studies on a subgroup at 2 years, comparing those who remained in remission (HbA1c Disclosure A. Al-Mrabeh: None. S.V. Zhyzhneuskaya: None. A.C. Barnes: None. K.G. Hollingsworth: None. N. Sattar: Advisory Panel; Self; Amgen Inc., AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Novo Nordisk A/S. Consultant; Self; NAPP Pharmaceuticals Limited. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc. Speaker's Bureau; Self; Amgen Inc., Eli Lilly and Company, Mitsubishi Tanabe Pharma Corporation, Novo Nordisk A/S, Roche Diagnostics France, Sanofi. M.E. Lean: Consultant; Self; Counterweight Ltd., Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S. R. Taylor: Advisory Panel; Self; Wilmington Healthcare. Speaker's Bureau; Self; Lilly Diabetes, Novartis AG. Funding Diabetes UK

Background: We previously developed a clinical/proteomics panel for predicting procedural acute kidney injury (AKI) and now examine its performance in those with diabetes (DM). Methods: We measured 109 biomarkers in blood from patients undergoing coronary angiography. Procedural AKI defined as increase in serum creatinine ≥0.3 mg/dL, increase in serum creatinine of ≥50%, or documented oliguria ≤7 days after procedure. Clinical and biomarker predictors of AKI identified using least-angle regression; a final prognostic model was developed with LASSO; the model was then measured in those with DM. Results: Besides DM, 5 predictors were in the final model: 3 (blood urea nitrogen to creatinine ratio, C-reactive protein and osteopontin) had positive association, while 2 (CD5 antigen-like and Factor VII) had negative association with AKI risk. Among 217 patients with DM, 18 (8.3%) developed AKI. The final model had an AUC of 0.87 for predicting procedural AKI (P Conclusions: We describe a clinical and proteomics-supported biomarker model with high sensitivity/NPV for AKI in patients with DM following coronary angiography. Disclosure N.E. Ibrahim: Other Relationship; Self; Novartis Pharmaceuticals Corporation. C.P. McCarthy: None. S. Shrestha: None. H. Gaggin: Other Relationship; Self; Dr. Gaggin has received research grant support from Roche Diagnostics, Jana Care, Ortho Clinical, Novartis; consulting income from Merck, Roche Diagnostics; research payments for clinical endpoint com. R. Mukai: None. C.A. Magaret: Consultant; Self; Prevencio. R.F. Rhyne: Board Member; Self; Prevencio. Employee; Self; Prevencio. Stock/Shareholder; Self; Prevencio. J. Januzzi: Consultant; Self; Abbott, Roche Diagnostic USA. Research Support; Self; Abbott, Prevencio, Quest Diagnostics, Roche Diagnostic USA. Funding Prevencio, Inc.

Diphtheria is caused by toxigenic strains ofCorynebacterium diphtheriae, Corynebacterium ulceransandCorynebacterium pseudotuberculosis. For diagnostic purposes, species identification and detection of toxigenic strains (diphtheria toxin (tox)-positive strains) is typically performed using end-point PCR. A faster quadruplex real-time PCR (qPCR) was recently developed (De Zoysaet al. J Med Microbiol. 2016 65(12):1521-1527). Here, we present an improvement of the quadruplex method, in which a 16S rRNA gene target was added as an internal processing control, providing confirmation of the presence of bacterial DNA in the assays. This improved qPCR method was validated using 36 bacterial isolates and 16 clinical samples. The method allows detection of thetoxgene and distinguishingC. diphtheriae(including the newly described speciesC. belfantii) fromC. ulceransandC. pseudotuberculosis. Complete diagnostic specificity, sensitivity and experimental robustness of the method to temperature and reagent concentration variations were demonstrated. The lower limit of detection forC. diphtheriae, C. ulceransandtoxtargets was 1.86 genome copies per 5 μL reaction volume. Finally, the method was successfully used on two distinct qPCR technologies (LightCycler 480, Roche Diagnostics and Rotor-Gene Q, Qiagen) and in two laboratories (Institut Pasteur, Paris, France and Public Health England – National Infection Service, London, UK). This work describes validation of the improved qPCR quadruplex method and supports its implementation for the biological diagnosis of diphtheria.

Clinical practice guidelines for the management of germ cell tumors recommend the measurements of human chorionic gonadotropin (hCG) and/or free hCGβ subunit for earlier diagnosis/recognition of the residual disease, for the prognostic evaluation and for the post-chemotherapy surveillance. However, the marketed hCG assays are validated and approved only for pregnancy purposes, with the sole exception of the Elecsys ‘hCG+β’ assay (Roche Diagnostics), cleared in Europe for oncological application. Theoretically, the hCG assay design for oncological purposes should fulfil the recommendations of the International Society of Oncology and Biomarkers requiring the use of antibodies displaying an equimolar recognition of both intact hCG and hCGβ monomer. Further analytical requirements should also be considered, such as optimal analytical sensitivity to allow an early tumor detection and low cross-reactivity for luteinizing hormone (LH). For the Elecsys assay, the detection limit (0.2 U/L) and the reported cross-reactivity for LH (0.12%) may be considered adequate if compared with the recommended requirements. Another issue is the definition of decision limits for oncologic purposes. After 3 years of clinical experience using the Elecsys assay in the oncology setting, we were able to define limits partitioned by sex and age as follows: males 50 years, 2.3 U/L; female 50 years, 5.6 U/L. There is an urgent need to disseminate appropriate educational information and to boost the clinical use of selective, highly sensitive and precise assays, specifically manufactured for cancer application.

Coagulopathies in horses are common and potentially life-threatening. In equine field medicine, a portable point-of-care (POC) prothrombin time (PT) testing device could be useful to identify early changes in extrinsic clotting. The CoaguChek-XS (Roche Diagnostics) is a small, portable POC PT analyzer used in human medicine. Our preliminary study assessed the suitability of CoaguChek-XS for testing PT in horses and established the PT reference interval (PT RI) in healthy horses using this instrument. Blood samples collected from 102 healthy and ill horses were analyzed with the CoaguChek-XS and compared to a semi-automated coagulometric analyzer (SACA) as the gold standard. There was a significant positive correlation between the 2 measurement methods ( r = 0.765, p < 0.01), and very good agreement, with 97% of the samples falling within limits of agreement. The mean CoaguChek-XS PT coefficient of variation was 0.8%, indicating high precision. With high precision and good agreement with the coagulometric PT, the CoaguChek-XS should be further validated for PT measurement in horses.

Seasonal influenza virus causes significant morbidity and mortality each year. Point-of-care (POC) testing using rapid influenza diagnostic tests (RIDTs), immunoassays that detect viral antigens, are often used for diagnosis by physician offices and urgent care centers. These tests are rapid but lack sensitivity, which is estimated to be 50 to 70%. Testing by PCR is highly sensitive and specific, but historically these assays have been performed in centralized clinical laboratories necessitating specimen transport and increasing the time to result. Recently, Clinical Laboratory Improvement Amendments (CLIA)-waived, POC PCR influenza assays have been developed with >95% sensitivity and specificity compared to centralized PCR assays. To determine the clinical impact of a POC PCR test for influenza, we compared antimicrobial prescribing patterns of one urgent care location using the Cobas LIAT Influenza A/B assay (LIAT assay; Roche Diagnostics, Indianapolis, IN) to other urgent care centers in our health system using traditional RIDT, with negative specimens being reflexed to PCR. Antiviral prescribing was lower in patients with a negative LIAT PCR result (2.3%) than in patients with a negative RIDT result (25.3%; P < 0.005). Antivirals were prescribed more often in patients that tested positive by LIAT PCR (82.4%) than in those testing positive by either RIDT or reflex PCR (69.9%; P < 0.05). Antibacterial prescriptions for patients testing negative by LIAT PCR were higher (44.5%) than for those testing negative by RIDT (37.7%), although the difference was not statistically significant. In conclusion, having results from a PCR POC test during the clinic visit improved antiviral prescribing practices compared to having rapid results from an RIDT.

Study objectiveTo investigate the performance of a rapid RT-PCR assay to detect influenza A/B at emergency department admission.MethodsThis single-center prospective study recruited adult patients attending the emergency department for influenza-like illness. Triage nurses performed nasopharyngeal swab samples and ran rapid RT-PCR assays using a dedicated device (cobas Liat, Roche Diagnostics, Meylan, France) located at triage. The same swab sample was also analyzed in the department of virology using conventional RT-PCR techniques. Patients were included 24 hours-a-day, 7 days-a-week. The primary outcome was the diagnostic accuracy of the rapid RT-PCR assay performed at triage.ResultsA total of 187 patients were included over 11 days in January 2018. Median age was 70 years (interquartile range 44 to 84) and 95 (51%) were male. Nine (5%) assays had to be repeated due to failure of the first assay. The sensitivity of the rapid RT-PCR assay performed at triage was 0.98 (95% confidence interval (CI): 0.91-1.00) and the specificity was 0.99 (95% CI: 0.94-1.00). A total of 92 (49%) assays were performed at night-time or during the weekend. The median time from patient entry to rapid RT-PCR assay results was 46 [interquartile range 36-55] minutes.ConclusionRapid RT-PCR assay performed by nurses at triage to detect influenza A/B is feasible and highly accurate.

Introduction: Standardized pre-analytical blood sample procedures for the analysis of circulating cell-free DNA (ccfDNA) are still not available. Therefore, the present study aimed at evaluating the impact of storage conditions related to different times (24 and 48 h) and temperatures (room temperature (RT) and 4 - 8 °C) on the plasma ccfDNA concentration of blood samples drawn into Cell-Free DNA collection tubes (Roche Diagnostics GmbH, Mannheim, Germany). Materials and methods: Venous blood from 30 healthy individuals was collected into five 8.5 mL Cell-Free DNA Collection Tubes (Roche Diagnostics GmbH) each. Plasma samples were processed at time point of blood collection (tube 1), and after storage under the following conditions: 24 h at RT (tube 2) or 4-8 °C (tube 3), and 48 h at RT (tube 4) or 4 - 8 °C (tube 5). Circulating cell-free DNA concentrations were determined by EvaGreen chemistry-based droplet digital PCR (ddPCR). Results: No statistically significant differences between median (interquartile range) plasma ccfDNA concentrations (ng/mL) at time point of blood collection (3.17 (2.13 – 3.76)) and after storage for 24 h (RT: 3.02 (2.41 – 3.68); 4-8 °C: 3.21 (2.19 – 3.46)) and 48 h (RT: 3.13 (2.10 – 3.76); 4-8 °C: 3.09 (2.19 – 3.50)) were observed (P values from 0.102 – 0.975). Conclusions: No unwanted release of genomic DNA from white blood cells could be detected in plasma samples after tube storage for 24 and 48 h regardless of storage temperature.

Background Since more sensitive immunoassays have been introduced, false positive Hepatitis B surface antigen (HBsAg) results are increasing. This study was carried out to propose a process to reduce the burden of the laboratory while increasing positive predictive value in HBsAg. Methods Samples with Elecsys HBsAg II (Roche Diagnostics, Germany) between cutoff index (COI) 0.9 and 10.0 were tested with Elecsys HBsAg Confirmatory Test (Roche Diagnostics). If the COI value after neutralization is less than or equal to 60% of the COI treated with control reagent, the sample was determined as HBsAg positive. Results A total of 133 samples were analyzed and 70.7% were confirmed positive. The highest COI of negatively confirmed sample was 5.6. Receiver operating characteristic curve analysis of HBsAg assay showed an area under curve of 0.761. Specificity was 100% at COI 6.0. Conclusions Based on this finding, the authors propose that only samples with HBsAg COI less than 6.0 need confirmatory tests.

Biomarkers for diseases like cancer, heart failure, viral infections or metabolic disorders are present in body fluids like whole blood or serum. By using an immunoassay with specific labelled antibodies these biomarkers can be selectively bound to a solid phase and detected. For electrochemiluminescent (ECL) detection, the label contains an organometallic compound like Ru(2,2′-bipyridine)32+ capable of emitting light after undergoing a series of electrochemically induced redox reactions with a coreactant like tripropylamine. ECL efficiency can be improved by adding ECL enhancing compounds like carbonic acid amides or by optimization of the ECL label for example by substitution of the bipyridine rings, by polymerization or by exchanging ruthenium by iridium. Furthermore, also the linker connecting the ECL label and the antibody and the conjugation method has an influence on assay performance. Roche Diagnostics cobas e analyzers automate immunoassays based on ECL detection and together with the Elecsys® assays, which are available for more than 100 different biomarkers, they have become widely used for in-vitro diagnosis, because they allow high throughput, sensitive and precise quantification of biomarkers in small volumes of patient samples.

The global dissemination of carbapenemase-producing Enterobacteriaceae (CPE) is a major concern in public health. Due to the existence of the diversity of carbapenemases, development of an easily available, cost-effective multiplex detection assay for CPE is required worldwide. Using clinically available and reliable equipment, COBAS® z480 (Roche Diagnostics K.K., Tokyo, Japan), we developed a multiplex real-time PCR assay for the detection of two combinations of carbapenemases; first, blaNDM, blaKPC, and blaIMP (Set 1), and second, blaGES, blaOXA-48, and blaVIM (Set 2). We constructed standard curves for each carbapenemase gene using serial dilutions of DNA standards, then applied reference or clinical isolates with each carbapenemase gene to this assay. The multiplex assay showed satisfactory accuracy to detect CPE genes, with the correlation coefficients of greater than 0.99 for all genotypes. The assay appropriately differentiated the reference or clinical strains harboring each carbapenemase gene without cross reactivity. Lastly, the assay successfully detected multiple genes without false-positive reactions by applying six clinical isolates carrying both NDM and OXA-48-like carbapenemase genes. Major advantages of our assay include multiplicity, simple operation, robustness, and speed (1 h). We believe that the multiplex assay potentially contributes to early diagnosis of CPE with a diverse genetic background.

Highlights • Choice of test does not compromise SARS-CoV-2 detection. • Commercial assays and lab developed tests for detection of SARS-CoV-2 exhibited 100 % positive and negative agreement. • Nucleic acid extraction method influenced the analytical sensitivity (limit of detection) of the lab developed RT-PCR test., The global COVID-19 pandemic has led to the rapid development of tests for detection of SARS-CoV-2. Studies are required to assess the relative performance of different assays. Here, we compared the performance of two commercial assays, the cobas® SARS-CoV-2 (Roche Diagnostics) and Xpert® Xpress SARS-CoV-2 (Cepheid®) tests, and a laboratory developed RT-PCR test adapted for use on the Hologic® Panther Fusion® (Hologic®) instrument as well as Bio-Rad and QIAGEN real-time PCR detection systems. Performance characteristics for each test were determined by testing clinical specimens and reference material. All assays detect the pan-Sarbecovirus E (envelope structural protein) gene plus a SARS-CoV-2-specific target. The limit of detection for the E gene target varied from ∼2 copies/reaction to >30 copies/reaction. Due to assay-specific differences in sample processing and nucleic acid extraction, the overall analytical sensitivity ranged from 24 copies/mL specimen to 574 copies/mL specimen. Despite these differences, there was 100 % agreement between the commercial and laboratory developed tests. No false-negative or false-positive SARS-CoV-2 results were observed and there was no cross-reactivity with common respiratory viruses, including endemic coronaviruses.

Haemoglobinopathies may interfere with the haemoglobin A1c (HbA1c) measurement, leading to incorrect diagnosis and inappropriate treatment. It is essential that HbA1c assays are capable of identifying haemoglobinopathies. We report two cases of haemoglobin New York (HbNY) discovered through HbA1c analysis using capillary electrophoresis (Capillarys 2 Flex Piercing [C2FP], Sebia). We used these samples to evaluate the ability of three other HbA1c assays to identify this variant: ion-exchange high-performance liquid chromatography (Variant II Turbo [VII-T], Bio-Rad); boronate affinity high-performance liquid chromatography (Ultra2, Trinity Biotech) and immunoassay (Cobas c501 Tina-quant Generation 3, Roche Diagnostics). Each method was used for HbA1c assay of in samples from two cases of heterozygous haemoglobinopathy: β0-thalassemia/HbNY (Case 1) and HbA/NY (Case 2). Only the C2FP system detected HbNY (an additional peak appeared between HbA1c and HbA0). Clinical laboratories should be aware of the limitations of their HbA1c assay methods especially in geographic areas, where haemoglobinopathy prevalence is high.

Background: Anti-hepatitis C virus antibody (anti-HCV) assays are recommended for screening HCV-infected persons. The VIDAS Anti-HCV Assay (bioMerieux, France), based on the enzyme-linked fluorescence test principle, was recently introduced in Korea. We evaluated the clinical performance of the VIDAS assay. Methods: One hundred HCV-positive and 1,002 HCV-negative blood samples confirmed by Architect anti-HCV (Abbott Laboratories, USA) and COBAS TaqMan HCV real-time PCR (Roche Diagnostics, USA) or the Procleix Ultrio Plus Assay (Gen-Probe Incorporated, USA) were obtained from the Human Serum Bank (HSB) and tested by VIDAS. In case of discrepant results, we conducted a recombinant immunoblot assay (RIBA). Results: The agreement rates for known HCV-positive and HCV-negative samples between the VIDAS assay and the HSB testing were 100% (95% confidence interval [CI]: 96.4-100%) and 99.5% (95% CI: 98.8-99.8%), respectively. One of the five discrepant samples was positive for Core 2+ and NS3-2 2+ reactivity, two samples were negative, and the other two were indeterminate regarding NS4 2+ reactivity in RIBA. We observed a significant but weak positive correlation between the titers of VIDAS and Architect assays (r=0.315, P

Background A simple high-performance liquid chromatography (HPLC) method was developed for the determination of albumin in patients' urine samples without coeluting proteins and was compared with the immunoturbidimetric determination of albumin. Urine albumin is important biomarker in diabetic patients, but part of it is immuno-nonreactive. Methods Albumin was determined by high-performance liquid chromatography (HPLC), UV detection at 280 nm, Zorbax 300SB-C3 column. Immunoturbidimetric analysis was performed using commercial kit on automatic biochemistry analyzer COBAS INTEGRA® 400, Roche Diagnostics GmbH, Manheim, Germany. Results The HLPC method was fully validated. No significant interference with other proteins (transferrin, α-1-acid glycoprotein, α-1-antichymotrypsin, antitrypsin, hemopexin) was found. The results from 301 urine samples were compared with immunochemical determination. We found a statistically significant difference between these methods (P = 0.0001, Mann–Whitney test). Conclusion New simple HPLC method was developed for the determination of urine albumin without coeluting proteins. Our data indicate that the HPLC method is highly specific and more sensitive than immunoturbidimetry.

BackgroundHere we describe the determination of upper reference limits (URL) for galectin-3, high-sensitivity cardiac troponin I (hs-cTnI) and high-sensitivity cardiac troponin T (hs-cTnT) in a single cohort of healthy blood donors using routine assays.MethodsFor this reference value study, we used a cohort of 402 consecutive blood donors (64% were male and 36% were female). The median individuals' age was 35.0years (range, 18.0–64.4). Individuals of this reference population were free of cardiovascular disease, diabetes mellitus, renal disease, cancer, current infection and chronic inflammatory disease. Plasma concentrations of galectin-3 were measured with the “routine Galectin-3” assay (Abbott Diagnostics), of hs-cTnI with the “STAT High Sensitive Troponin-I” assay (Abbott Diagnostics), and of hs-cTnT with the “Troponin T hs” assay (Roche Diagnostics). URLs were calculated by using a non-parametric percentile method.ResultsThe 97.5th percentile URL for galectin-3 was 16ng/mL in males and 17ng/mL in females; the 99th percentile URL for hs-cTnI was 39ng/L in males and 24ng/L in females; and the 99th percentile URL for hs-cTnT was 14ng/L in males and 11ng/L in females. Those individuals with hs-cTnI values ≥15ng/L (n=8) were different from those individuals with hs-cTnT values ≥10ng/L (n=7). Of the 402 individuals, none had galectin-3 values below the limit of detection (LOD

Human insulin is a polypeptide hormone produced, stored, and secreted by the s-cells in the pancreatic islets of Langerhans. Its secretion is stimulated by an increase of the glucose concentration in circulation. Non-radioactive assays are frequently used in many laboratories to measure hormone concentrations, as an alternative to the traditional "gold standard" radioimmuno- and immunoradiometric assays. The precise and reliable determination of the insulin concentration is an important concern in numerous diagnostic procedures. The aim of this study was to compare two commercially available assays (manual and automated) for measurement of serum insulin concentrations. Aliquots of the 86 randomly selected serum samples were measured by Elecsys Insulin Assay (cobas e411 immunoassay analyzer, Roche Diagnostics GmbH, Mannheim, Germany) and DIAsource INS-IRMA Kit (DIAsource ImmunoAssays S.A., Louvain-la-Neuve, Belgium). Compared assays exhibit good correlation (r = 0.996). Insulin concentrations were on average 4.2 μ IU/mL lower (p < 0.05) with the cobas e411 immunoassay analyzer when compared to those measured with DIAsouce Immunoassay. Our findings suggest that electrochemiluminescence method on the cobas e411 analyzer and manual IRMA method offered by the DIAsource for the serum insulin determination could be considered interchangeable.

Objectives: Lipoprotein-associated phospholipase A2 (Lp-PLA 2 ) is increased in circulation in patients at higher risk of coronary heart disease (CHD) events and stroke. Therefore, measurement of Lp-PLA 2 can be used as an adjunct to traditional cardiovascular risk factors for identifying individuals at higher risk of cardiovascular events. Recently, a reagent for measuring Lp-PLA 2 activity (diaDexus, San Francisco, CA) received FDA approval. Here we evaluate the assay performance of the Lp-PLA 2 activity assay. Methods: Lp-PLA 2 activity assay reagent performance was evaluated on an open user-defined channel on a Cobas 6000/c501 (Roche Diagnostics, Indianapolis, IN) using a 5-point calibration curve (0–400 nmol/min/mL). Analytical performance was established for the following parameters: precision, linearity, accuracy, analytical sensitivity, analytical specificity, reference interval, reagent lot-to-lot comparison, specimen type, on-board reagent stability, and sample stability. Results: Assay limit of detection was determined to be 7.8 nmol/min/mL with an average %CV of 2.8%. Precision studies revealed a coefficient of variation ≤ 1.6% between 79 and 307 nmol/min/mL and accuracy was demonstrated between 4.8–368.7 nmol/min/mL. Comparable results were generated in paired SST serum and EDTA plasma. No age association was found with Lp-PLA 2 activity at the 95th percentile however a gender association was identified resulting in gender-specific 95th percentile limits in a healthy reference population. No bias was found when comparing results from several different lots of assay reagent. Lp-PLA 2 activity results are extremely stable in both serum and EDTA plasma under refrigerate and frozen storage conditions up to 31 days. Conclusions: Lp-PLA 2 activity assay displays accurate and precise performance characteristics on the Cobas c501 platform. The assay performance is significantly improved over the predecessor immunoassay allowing for adoption of Lp-PLA 2 activity in clinical practice.

Parathyroid hormone (PTH) determination is essential for the diagnosis of renal osteodystrophy, but differences between the laboratory assays can lead to different therapies. This study compared the new Tosoh ST AIA-Pack Intact PTH assay (Tosoh Bioscience, San Francisco, CA, USA) with the Elecsys Intact PTH Roche assay (Roche Diagnostics GmbH, Mannheim, Germany), currently considered the gold standard. Nineteen chronic stable hemodialysis patients were enrolled to check PTH levels with the two assays. Median age was 71 years (range 26-84), M/F = 10/9. Blood samples were taken before the start of the same midweek dialysis session. Two blood vacuettes were collected and immediately transported to the central laboratory. The median PTH value was 268 (range 35-901 pg/dL) with the AIA-Pack versus 184 (range 39-552 pg/dL) with Elecsys. The Wilcoxon test showed a significant difference between the two methods (P 300 pg/dL four patients were detected with AIA-Pack (21.0%) versus six patients with Elecsys (31.6%). The two assays showed no differences for each of the three PTH ranges considered. The two PTH assays tested are different and the attending physician should be aware of the differences when patients change their dialysis facility.

Ethylene glycol (EG) is a common cause of toxic ingestions. Gas chromatography (GC)-based laboratory assays are the gold standard for diagnosing EG intoxication. However, GC requires specialized instrumentation and technical expertise that limits feasibility for many clinical laboratories. The objective of this retrospective study was to determine the utility of incorporating a rapid EG assay for management of cases with suspected EG poisoning. The University of Iowa Hospitals and Clinics core clinical laboratory adapted a veterinary EG assay (Catachem, Inc.) for the Roche Diagnostics cobas 8000 c502 analyzer and incorporated this assay in an osmolal gap-based algorithm for potential toxic alcohol/glycol ingestions. The main limitation is that high concentrations of propylene glycol (PG), while readily identifiable by reaction rate kinetics, can interfere with EG measurement. The clinical laboratory had the ability to perform GC for EG and PG, if needed. A total of 222 rapid EG and 24 EG/PG GC analyses were documented in 106 patient encounters. Of ten confirmed EG ingestions, eight cases were managed entirely with the rapid EG assay. PG interference was evident in 25 samples, leading to 8 GC analyses to rule out the presence of EG. Chart review of cases with negative rapid EG assay results showed no evidence of false negatives. The results of this study highlight the use of incorporating a rapid EG assay for the diagnosis and management of suspected EG toxicity by decreasing the reliance on GC. Future improvements would involve rapid EG assays that completely avoid interference by PG.

Background The objective of this study was to quantify the sensitivity of very low concentrations of high‐sensitivity cardiac troponin T (hsTnT) at ED arrival for acute myocardial infarction (AMI) in a large cohort of chest pain patients evaluated in real‐world clinical practice. Methods This retrospective study included consecutive ED patients with suspected cardiac chest pain evaluated in four urban EDs, excluding those with ST‐elevation AMI, cardiac arrest or abnormal kidney function. The primary outcomes were AMI at 7, 30, and 90 days. Secondary outcomes included major adverse cardiac events (MACE; all‐cause mortality, AMI, and revascularization) and the individual MACE components. Test characteristics were calculated for hsTnT values from 3 to 200 ng/L . Results A total of 7,130 patients met inclusion criteria. AMI incidences at 7, 30, and 90 days were 5.8, 6.0, and 6.2%. When the hsTnT assay was performed at ED arrival, the limit of blank of the assay (3 ng/L) ruled out 7‐day AMI in 15.5% of patients with 100% sensitivity and negative predictive value (NPV). The limit of detection of the assay (5 ng/L) ruled out AMI in 33.6% of patients with 99.8% sensitivity and 99.95% NPV for 7‐day AMI. The limit of quantification (the Food and Drug Administration [FDA]‐approved cutoff for lower the reportable limit) of 6 ng/L ruled out AMI in 42.2% of patients with 99.8% sensitivity and 99.95% NPV. The sensitivities of the cutoffs of

Background: SARS-CoV-2 outbreak has impacted on the management of oncological p, leading to treatment delays in a considerable number of cases The aim of this study was to evaluate if oncological T affected negatively COVID-19 outcome Methods: We retrospectively analyzed clinical data from p with solid tumors under active systemic T (received in the last 6 months) that were diagnosed with SARS-CoV-2 infection (defined as positive PCR) between March and 11th May 2020 in our center Study endpoint was death due to COVID-19 We divided the patients in two groups;those who had received treatment in the last 4 weeks and those who had not Descriptive and univariate analysis were performed to detect the effect of T type and other variables on COVID-19 related mortality Results: A total of 70 p were included with a median follow-up of 28 days (10-47) and active oncological T had been administered in the past 4 weeks to 44 p Median age was 66 (IQR 56-74), 23 p (52 27%) were female and 41 (93 2%) had a baseline ECOG≤1 The most frequent primary site was lung tumor (12 p [27 3%]), followed by breast (11 p [25%]) and gastrointestinal (5 p [11 4%]) Thirty-one p (70 5%) had metastatic disease and 13 (29 5%) were included in clinical trials Twenty-four p (54 5%) received chemotherapy (CT), 14 (31 8%) targeted therapies, 9 (20 4%) immunotherapy (IT), 5 (11 4%) radiotherapy and 6 (13 6%) hormonotherapy A total of 13 p (29 5%) received different combinations of oncological T Death due to COVID-19 occurred in 5/22 (22 7%) p receiving CT and 6/21 (28 5%) p in the non-CT (p>0 05) Only 1/9 (11 1%) p treated with IT died compared to 11/35 (31 4%) p in the rest of the cohort (p>0 05) Age>71, comorbidities such as chronic obstructive pulmonary disease and ECOG status>2 were associated to a higher mortality The distribution of these variables between the anticancer T groups was not different Conclusions: Our results suggest that CT and other anticancer T might not worsen COVID-19 related mortality;nevertheless, the number of patients was small and decision making has to be individualized Our findings may warrant further investigation in larger studies Legal entity responsible for the study: The authors Funding: Has not received any funding Disclosure: E Felip: Advisory/Consultancy, Speaker Bureau/Expert testimony: AbbVie;AstraZeneca;Blueprint medicines;Boehringer Ingelheim;Bristol-Myers Squibb;Celgene;Eli Lilly;Guardant Health;Janssen;Medscape;Merck KGaA;Novartis;Pfizer;Roche;Takeda;Touchtime;Research grant/Funding (self), Research grant/Funding (institution): Fundacion Merck Salud;Oncology Innovation EMD Serono J Carles: Advisory/Consultancy, Speaker Bureau/Expert testimony: Johnson & Johnson;Bayer;Advisory/Consultancy, Speaker Bureau/Expert testimony, Research grant/Funding (self): Astellas Pharma;Advisory/Consultancy: Pfizer;Sanofi;MSD Oncology;Advisory/Consultancy, Research grant/Funding (self): Roche;Advisory/Consultancy, Research grant/Funding (self), Travel/Accommodation/Expenses: AstraZeneca;Speaker Bureau/Expert testimony: Asofarma;Research grant/Funding (self), Travel/Accommodation/Expenses: BMS;ravel/Accommodation/Expenses: Ipsen;Roche;Research grant/Funding (self): AB Science;Aragon Pharmaceuticals;Pharmaceuticals;INC;Blueprint Medicines Corporation;N Immunotherapeutics INC;Boehringer Ingelheim Espana, S A ;Clovis Oncology;Cougar Biotechnology INC;Deciphera Pharmaceuticals LLC;Exelixis INC;F Hoffmann-La Roche LTD;Genentech INC;Glaxosmithkline;Incyte Corporation;Janssen-Cilag International NV;Karyopharm Therapeutics INC;Laboratoires Leurquin Mediolanum SAS J Tabernero: Honoraria (self): Array Biopharma;AstraZeneca;Bayer;BeiGene;Boehringer Ingelheim;Chugai;Genentech;Genmab A/S;Halozyme;Imugene Limited;Inflection Biosciences Limited;Ipsen;Kura Oncology;Lilly;MSD;Merck Serono;Menarini;Merrimack;Merus;Molecular Partners;Novartis;Peptomyc;Pfizer;Pharmacyclics;ProteoDesign SL;Rafael Pharmaceuticals;F Hoffmann-La Roche Ltd;): Sanofi;eaGen;Seattle Genetics, Servier, Symphogen, Taiho, VCN Biosciences, Biocartis, Foundation Medicine, HalioDX SAS and Roche Diagnostics All other authors have declared no conflicts of interest

We appreciate the responses[1][1],[2][2] to our article published in CMAJ entitled, “Health Canada needs to act on laboratory-developed diagnostics.”[3][3] Michele D’Elia from Roche Diagnostics offered an update, indicating that the company’s noninvasive prenatal test was approved by Health

Background: This study compared the analytical performance of the Elecsys 602 (Roche Diagnostics) system with the I2000 (Abbott laboratories) system for the quantitative measurement of squamous cell carcinoma antigen (SCCA) to assess its role as an indicator in pan squamous cell carcinoma. Methods: 435 serum samples included pan squamous cell cancer group (n = 318) and healthy subjects (n = 52) and non-squamous cell group (n = 41) and benign diseases group (n = 24) were measured by 2 systems and compared. Results: The within-run precision coefficient of variation (CV) for Abbott and Roche systems were 3.34-4.88% and 0.95 -1.96%, and the total precision CV were 2.89-9.48% and 3.97-5.38%, respectively. Good correlation was showed in Abbott and Roche systems (slopes = 0.749, r = 0.9658). Serum SCCA in the groups of nasopharyngeal carcinomas, lung squamous cell carcinoma, esophageal squamous cell carcinoma, bladder cancer and cervical squamous cell carcinoma under the curve area (AUC) was more than 0.5, while the AUC in the non- nasopharyngeal carcinomas head and neck squamous cell carcinoma was less than 0.5. The AUC of 2 systems was statistically different in lung squamous cell carcinoma and nasopharyngeal carcinomas (P < 0.05). The levels of SCCA of 2 systems were similarities in esophageal squamous cell carcinoma(stage IV vs. stage 0a-II)and bladder cancer(stage I vs. stage Oa)and cervical squamous cell carcinoma(stage IIB-III vs. stage I-IIA), which advanced stage had higher level of SCCA than early stage. But the SCCA levels of 2 systems were inconsistent in bladder cancer (stage II-IV vs. stage Oa in Abbott), head and neck squamous cell carcinoma (stage IV vs. stage Oa-I in the Roche) and lung squamous cell carcinoma (stage III vs. stage I-II in the Roche). (P < 0.05) Conclusions: 2 systems correlated well in SCCA detection of squamous cell carcinoma, but there were individual differences. Serum SCCA may also contribute to the diagnosis of bladder cancer.

Introduction Haemoglobin A1c (HbA1c) is considered to be the gold standard for the follow-up of glycaemic control in patients with diabetes mellitus and is also a diagnostic tool. Accordingly, reliable and efficient methods must be used for its quantification. Roche Diagnostics have recently adapted the Tina-quant® HbA1c Third Generation immunoassay on a fully dedicated analyser, the Cobas c513, which allows a high throughput of up to 400 samples per hour. The present article deals with the evaluation of the analytical performances of this system which has been recently introduced to the market. Materials and methods Precision, comparison with two ion-exchange high-performance liquid chromatography (HPLC) methods (Variant II and D-100 systems, BioRad Laboratories) using Passing Bablok and Bland-Altman analyses, accuracy and interference of the most frequent haemoglobin (Hb) variants on HbA1c measurement were evaluated. Results Precision was high, with coefficients of variation lower than 1.1% (HbA1c values expressed in National Glycohemoglobin Standardization Program units, 1.7% for values expressed in International Federation of Clinical Chemistry and Laboratory Medicine [IFCC] units). The comparison study showed similar results with the two HPLC systems. The analysis of samples with IFCC-assigned values showed high methodological accuracy. Finally, no interference of bilirubin, triglycerides and common Hb variants (Hb AC, AD, AE, AS) was observed. Conclusions This evaluation showed that the analytical performance of the Cobas c513 analyser for HbA1c assay makes it suitable for a routine use in clinical chemistry laboratories.

Anti-cyclic citrullinated peptide antibodies (anti-CCPs) are important diagnostic markers for rheumatoid arthritis (RA). We evaluated the analytical and clinical performance of the QUANTA Flash CCP3 (INOVA Diagnostics, USA), a fully automated third-generation anti-CCP assay, in comparison with three second-generation anti-CCP (CCP2) assays. A total of 300 sera (67 from RA patients, 64 from other rheumatic diseases, 43 from osteoarthritis [OA], and 126 from other conditions) were tested with QUANTA Flash CCP3, Kallestad Anti-CCP II (Bio-Rad, USA), Elecsys Anti-CCP (Roche Diagnostics GmbH, Germany), and ARCHITECT Anti-CCP (Abbott Diagnostics, USA). Within-run and total imprecision (% coefficient of variation) of the QUANTA Flash CCP3 were

Chest pain is one of the most common reasons for patients attending the Emergency Department (ED). Accurately assessing for Acute Coronary Syndromes (ACS) remains a challenge. There is strong evidence supporting use of the Troponin-only Manchester Acute Coronary Syndrome (T-MACS) risk prediction model. How clinicians perform compared to these models is unknown.We aimed to externally validate the diagnostic accuracy of clinicians’ estimated probability of ACS (gestalt) compared to the T-MACS calculated probability of ACS.The Bedside Evaluation of Sensitive Troponin prospective multi-centre diagnostic accuracy study included adults presenting to the ED with potential ACS. Alongside clinical, ECG and blood sample data, the emergency clinician recorded their estimated probability of ACS (%) following review. The probability of ACS was also calculated using T-MACS. The primary outcome was Major Adverse Cardiac Events (MACE) within 30-days. For this planned secondary analysis, patients from sites using the high-sensitivity cardiac troponin T (Roche Diagnostics Elecsys) were eligible.Of 782 included, 116 (14.8%) had MACE. The C-statistic for clinician gestalt and T-MACS were 0.76 (95% CI 0.71–0.81) and 0.93 (0.90–0.95) respectively (pClinician gestalt has inferior diagnostic accuracy to T-MACS. T-MACS requires a clinician’s skill for appropriate application. Our conclusion is therefore not that computers are better, but that clinician performance can be augmented using T-MACS.

Aim: The PDM-ProValue study program showed the benefit of iPDM for people with type 2 diabetes mellitus (T2D) on insulin therapy. Here, we analyze which treatment process conditions may be key for this success. Methods: The study program was conducted over 12 months in a prospective, controlled, cluster-randomized setting. 101 medical practices were randomized to the iPDM arm (n=53) or the control (CNL) arm (n=48). In addition to medical and patient related outcomes, parameters of process quality and data sources for therapy decisions were monitored e.g., by means of Likert-scaled requests. Results: After 12 months, HbA1c reduction vs. baseline was higher for patients in iPDM (0.5%, p Discussion: A more holistic and beneficial use of data sources by implementing iPDM may be key for the improvements observed in this difficult to treat patient group. A more structured treatment process applied by the physicians and better personalization of therapy seem to enhance physician-patient interaction. Physicians’ positive perception of iPDM may facilitate more focused treatment decisions, thereby overcoming clinical inertia. Disclosure L. Heinemann: Stock/Shareholder; Self; Profil Institute for Metabolic Research, ProSciento. Consultant; Self; Roche Diabetes Care Health and Digital Solutions. D. Messinger: Other Relationship; Self; Roche Diagnostics Corporation, Roche Diagnostics Corporation, Roche Pharma, AbbVie Inc., Merck KGaA. W. Schramm: Advisory Panel; Self; Roche Diabetes Care Health and Digital Solutions. Research Support; Self; Roche Diabetes Care Health and Digital Solutions. I. Vesper: None. J. Weissmann: Employee; Self; Roche Diabetes Care Deutschland GmbH. B. Kulzer: Research Support; Self; Berlin-Chemie AG. Speaker's Bureau; Self; Berlin-Chemie AG, Novo Nordisk Inc.. Advisory Panel; Self; Roche Diabetes Care Health and Digital Solutions. Speaker's Bureau; Self; Roche Diabetes Care Health and Digital Solutions. Research Support; Self; Abbott. Speaker's Bureau; Self; Abbott, Eli Lilly and Company. Advisory Panel; Self; Novo Nordisk Inc., Medtronic, Ascensia Diabetes Care. Speaker's Bureau; Self; Ascensia Diabetes Care.

Introduction: In patients with type 1 diabetes mellitus (T1D) the relationship between low number of self-monitoring of blood glucose (SMBG) measurements and elevated hemoglobin A1c (HbA1c) levels has been reported. However, the optimal frequency of SMBG has not been defined. Flash glucose monitoring (FGM) is an emerging technology allowing patient with diabetes to measure interstitial glucose concentrations in a non-invasive manner, with patients obtaining more measurements than with SMBG. Materials and Methods: We collected data from 103 adult T1D patients using FGM technology in real life for at least 28 days. We recorded patients demographics, diabetes duration, type of treatment (CSII or MDI) and we downloaded FGM data to the Abbott Freestyle Libre software, which provides the number of daily FGM scans and estimates of HbA1c based on continuous glucose profile readings. The correlation between median FGM scan frequency over 28 days and the estimated HbA1c was analyzed using Spearman rho. We also computed HbA1c median levels for quintiles of scan frequency and compared them using Kruskal-Wallis test. Results: A significant correlation was found between median scan Spearman rho was -0.48 (p Conclusions: In T1D patients we confirmed that frequent measurements of interstitial glucose benefits glucose control in real life conditions. The maximum benefit in term of HbA1c decrease is obtained between 10 to 14 scans daily, a number of glucose measurements that would unlikely be sustainable with SMBG. Disclosure A. Laurenzi: Other Relationship; Self; Animas Corporation, LifeScan, Inc..A. Caretto: None.M. Barrasso: None. A. Bolla: Other Relationship; Self; Roche Diabetes Care Health and Digital Solutions. C. Molinari: Other Relationship; Self; Eli Lilly and Company, Sanofi. A. Zanardini: Other Relationship; Self; Roche Diagnostics Corporation.N.C. Dozio: None.E. Meneghini: None. E. Bosi: Consultant; Self; Abbott, AstraZeneca, Lexicon Pharmaceuticals, Inc., LifeScan, Inc.. Employee; Spouse/Partner; LifeScan, Inc.. Consultant; Self; Medtronic, Merck Sharp &Dohme Corp., Novartis AG, Roche Diabetes Care Health and Digital Solutions, Sanofi. M. Scavini: Consultant; Self; Sanofi, Roche Diabetes Care Health and Digital Solutions.

Tubeless patch pumps may offer unique benefits enhancing long-term glycemic benefits seen in adults with diabetes starting continuous subcutaneous insulin infusion (CSII) therapy. We retrospectively evaluated adults with diabetes >6 months to assess changes in A1c and weight 1 year after initiating the Omnipod® Insulin Management System (Insulet Corp., Billerica, MA). Changes were assessed by baseline A1c category ( Disclosure L.J. Tinsley: None. L.M. Laffel: Consultant; Self; Eli Lilly and Company, Novo Nordisk Inc., Sanofi US, MannKind Corporation, Roche Diagnostics Corporation, Dexcom, Inc., Insulet Corporation, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Johnson & Johnson Diabetes Institute, LLC.. Z. Guo: None. N. Naik: None. J.E. Layne: Employee; Self; Insulet Corporation. T.T. Ly: Employee; Self; Insulet Corporation. S.N. Mehta: None.

The Elecsys Troponin T Gen 5 STAT test (distributed in the United States (US) by Roche Diagnostics, Indianapolis, IN) is the first high-sensitivity cardiac troponin test approved for use by the FDA in the US (2017). Areas covered: The test offers clinicians the opportunity for more rapid decision-making for diagnosing myocardial infarction (MI) in the emergency department (ED). The Troponin T Gen 5 STAT test (labeled as TNT-G5ST on the reagent pack) is similar to the Troponin T hs STAT (TNT-HSST) and Troponin T hs (TNT-HS) tests that have been available outside the US since 2009. Collectively, these tests can all be considered as high-sensitivity cardiac troponin T (hs-cTnT) assays. Expert commentary: Studies performed in the US and throughout the world using 0 and 3 h blood draws for hs-cTnT testing in patients with possible MI have reliably achieved a sensitivity of94% and negative predictive value of ≥99% for MI in the ED setting.

Context This commentary discusses the implications of disease-modifying treatments for Alzheimer’s disease which seem likely to appear in the next few years and results from a meeting of British experts in neurodegenerative diseases in Edinburgh. The availability of such treatments would help change public and professional attitudes and accelerate engagement with the prodromal and preclinical populations who might benefit from them. However, this would require an updated understanding of Alzheimer’s disease, namely the important distinction between Alzheimer’s disease and Alzheimer’s dementia. Consensus Since treatments are likely to be most effective in the early stages, identification of clinically relevant brain changes (for example, amyloid burden using imaging or cerebrospinal fluid biomarkers) will be crucial. While current biomarkers could be useful in identifying eligibility for new therapies, trial data are not available to aid decisions about stopping or continuing treatment in clinical practice. Therefore, effective monitoring of safety and effectiveness when these treatments are introduced into clinical practice will be necessary to inform wide-scale use. Equity of access is key but there is a tension between universal access for everyone with a diagnosis of Alzheimer’s disease and specifying an eligible population most likely to respond. We propose the resources necessary for an optimal care pathway as well as the necessary education and training for primary and secondary care. Conclusion The majority of current services in the UK and elsewhere would not be able to accommodate the specialist investigations required to select patients and prescribe these therapies. Therefore, a stepped approach would be necessary: from innovating sentinel clinical-academic centres that already have capacity to deliver the necessary phase IV trials, through early adoption in a hub and spoke model, to nationwide adoption for true equity of access. The optimism generated by recent and anticipated developments in the understanding and treatment of Alzheimer’s disease presents a great opportunity to innovate and adapt our services to incorporate the next exciting development in the field of dementia.

Severe, early onset fetal growth restriction (FGR) is characterised by a fetus presenting at a gestational age at the borderline of viability which is extremely small for gestational age by ultrasonic biometry and exhibits other signs of utero-placental insufficiency, such as abnormal patterns of uterine or umbilical blood flow. Following diagnosis, there is, currently, no disease modifying therapy other than medically indicated delivery, which carries a high risk of neonatal death or, if the infant survives, severe neurodisability in later life. Conversely, expectant management carries a substantial risk of intra-uterine fetal death. Balancing these conflicting risks is a day to day element of practice in Maternal-Fetal Medicine. Practitioners looks to a future of disease modifying therapies other than delivery. Unfortunately, the STRIDER trial suggests that one of the promising candidates, sildenafil citrate, is very unlikely to offer hope in this dismal situation. The study was powered to detect a 7 day prolongation in pregnancy. In reality, the trend was towards a reduction in the duration of pregnancy and the 95% confidence interval indicates that the best one could expect is a 1.3 day prolongation. In reality, it is more likely that the drug hastens delivery rather than postpones it.

Objectives The new immunochemistry cobas e 801 module (Roche Diagnostics) was developed to meet increasing demands on routine laboratories to further improve testing efficiency, while maintaining high quality and reliable data. Design and methods During a non-interventional multicenter evaluation study, the overall performance, functionality and reliability of the new module was investigated under routine-like conditions. It was tested as a dedicated immunochemistry system at four sites and as a consolidator combined with clinical chemistry at three sites. Results We report on testing efficiency and analytical performance of the new module. Evaluation of sample workloads with site-specific routine request patterns demonstrated increased speed and almost doubled throughput (maximal 300 tests per h), thus revealing that one cobas e 801 module can replace two cobas e 602 modules while saving up to 44% floor space. Result stability was demonstrated by QC analysis per assay throughout the study. Precision testing over 21 days yielded excellent results within and between labs, and, method comparison performed versus the cobas e 602 module routine results showed high consistency of results for all assays under study. In a practicability assessment related to performance and handling, 99% of graded features met (44%) or even exceeded (55%) laboratory expectations, with enhanced reagent management and loading during operation being highlighted. Conclusion By nearly doubling immunochemistry testing efficiency on the same footprint as a cobas e 602 module, the new module has a great potential to further consolidate and enhance laboratory testing while maintaining high quality analytical performance with Roche platforms.

Background: Standard antimicrobial susceptibility testing (AST) approaches lead to delays in the selection of optimal antimicrobial therapy. We sought to determine the accuracy of antimicrobial resistance (AMR) determinants identified by Nanopore whole genome sequencing in predicting AST results. Methods: Using a cohort of 40 clinical Klebsiella pneumoniae isolates, three separate sequencing and analysis pipelines were performed: (1) a real-time Nanopore analysis approach that identified acquired AMR genes, (2) an assembly-based Nanopore approach that identified acquired AMR genes and chromosomal mutations, and (3) a Pilon-corrected Nanopore-Illumina hybrid approach. The Pilon-corrected assemblies served as the reference standard to determine the accuracy of Nanopore sequencing results. Findings: With the real-time analysis approach, full annotation of acquired AMR genes occurred within 8 hours of subcultured isolates. Assemblies sufficient for full resistance gene and single nucleotide polymorphism annotation were available within 14 hours from subcultured isolates. The overall agreement of genotypic results and anticipated AST results for the 40 K. pneumoniae isolates was 75% (range 30-95%) and 91% (range 80-98%) for the real-time approach and the assembly approach, respectively. Evaluating the patients contributing the 40 isolates, the real-time approach and assembly approach could substantially shorten the median time to effective antibiotic therapy by 20 hours and 26 hours, respectively, compared to standard AST. Interpretations: Nanopore sequencing offers a rapid approach to both accurately identify resistance mechanisms as well as to predict AST results. Bioinformatics improvements enabling real-time alignment coupled with rapid extraction and library preparation will further enhance the accuracy and workflow of the Nanopore real-time approach. Funding Statement: The work was supported by funding from the National Institutes of Health K23-AI127935 awarded to PDT, a Canadian Institutes of Health Research fellowship awarded to YF, an R01-HG009190 awarded to WT, an R01-HG006677 awarded to MCS, and an R21-AI130608 awarded to PJS. Declaration of Interests: PDT has received research funding from Merck, outside of the submitted work. KCC has received research funding from GenMark, Inc. and Singulex outside of the submitted work. WT has two patents (8,748,091 and 8,394,584) licensed to Oxford Nanopore. WT, YF, PJS have received travel funds from Oxford Nanopore Technologies. PJS has received grants and personal fees from Accelerate Diagnostics, grants from BD Diagnostics, Inc., grants from bioMerieux, Inc., grants from Check-Points Diagnostics, BV, grants from Hardy Diagnostics, personal fees from Roche Diagnostics, personal fees from Opgen Inc, outside the submitted work. All other authors declare no competing interests. Ethics Approval Statement: The Johns Hopkins University School of Medicine Institutional Review Board approved this study, with a waiver of informed consent.

International audience; Direct-acting antiviral (DAA) drug performances depend on the viral genotype. So international recommendations give typing of the virus a prerequisite for treatment choice and patient management. Commercially available HCV genotyping kits are scarce and this analysis is often in-house using tedious PCRs and Sanger sequencing, leading to a lack of standardization. A newly commercialized HCV genotyping assay based on real-time PCR has been developed by Roche Diagnostics (Mannheim, Germany). We compared this new assay with our in-house PCRs -sequencing technique on 101 regular samples and 81 LiPA failures or low viral load samples. No genotype or 1a/1b subtype mismatch was observed. Two samples were misidentified at the subtype level without clinical impact. Three genotype 1b and two genotype 1a samples with low viral load could not be subtyped. Nevertheless, 13 (13%) samples from the regular panel and 35 (43%) from the more difficult-to-type panels failed to give results on first pass with the Roche kit. Failures were mostly associated with genotype 3 subtype a, with genotype 4 subtype non-a, or with viral loads