scholarly journals Performances of NeuMoDx, a random-access system for HBV-DNA and HCV-RNA quantification

2020 ◽  
Author(s):  
Juliette Besombes ◽  
Charlotte Pronier ◽  
Charles Lefevre ◽  
Gisèle Lagathu ◽  
Anne Maillard ◽  
...  
Keyword(s):  
Internal Control ◽  
Random Access ◽  
Turnaround Time ◽  
Correlation Factor ◽  
Hbv Dna ◽  
Clinical Samples ◽  
Hcv Rna ◽  
Rna Quantification ◽  
Kappa Agreement ◽  
Serial Dilutions

AbstractViral loads (VL) monitoring for hepatitis B and C is essential to evaluate disease progression and treatment response. Automated, random-access rapid systems are becoming standard to provide reliable VL to clinicians. The aim of this study was to evaluate the analytical performances of the recently launched NeuMoDx™ for HBV-DNA and HCV-RNA quantification. Clinical samples routinely quantified on the Beckman-Veris system were either retrospectively (frozen samples; HBV n=178, HCV n=249), or in parallel (fresh primary tubes; HBV n=103, HCV n=124) tested using NeuMoDx™. Linearity range was assessed on serial dilutions of high tittered plasmas containing different genotypes for HBV (A-E, n=10) and HCV (1a-b, 2-5, n=12). Overall test failure, mostly internal control amplification failure, was 2.3% and was not influenced by matrix types. For HBV-VL, Kappa agreement was 74%, with 27 (12.6%) discrepancies. Correlation between HBV assays on 72 quantified samples by both methods was excellent (r=0.963) with a mean bias (NeuMoDx™-Veris) of 0.21 log IU/mL. For HCV-VL, Kappa agreement reached 94%, with 9 (2.8%) discrepancies. The r-correlation factor between assays on 104 samples was 0.960 with a mean bias of −0.14 log IU/mL (NeuMoDx™-Veris). Serial dilutions confirmed the claimed linear ranges for all HBV and HCV genotypes. The mean turnaround time was 72’ [55-101] for HBV and 96’ [78-133] for HCV. These results obtained on the NeuMoDx™ confirmed the overall good functionality of the system with a short turn-around-time, full traceability and easy handling. These results on HBV- and HCV-VL look promising and should be challenged with further comparisons.

scholarly journals Performances of NeuMoDx™, a random-access system for HBV-DNA and HCV-RNA quantification

2021 ◽  
Author(s):  
Juliette Besombes ◽  
Charlotte Pronier ◽  
Charles Lefevre ◽  
Gisèle Lagathu ◽  
Anne Maillard ◽  
...  
Keyword(s):  
Random Access ◽  
Hbv Dna ◽  
Hcv Rna ◽  
Access System ◽  
Rna Quantification

scholarly journals Effect of Multiple Freeze-Thaw Cycles on Hepatitis B Virus DNA and Hepatitis C Virus RNA Quantification as Measured with Branched-DNA Technology

1999 ◽  
Vol 37 (6) ◽  
pp. 1683-1686 ◽  
Author(s):  
Mel Krajden ◽  
James M. Minor ◽  
Oretta Rifkin ◽  
Lorraine Comanor
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hbv Dna ◽  
Hcv Rna ◽  
Freeze Thaw ◽  
Rna Quantification ◽  
B Virus ◽  
Rna Levels

Quantification of hepatitis B virus (HBV) DNA and hepatitis C virus (HCV) RNA often is performed in specimens that have been frozen and thawed more than once. To ensure optimal therapeutic and prognostic value, it is important to establish whether viral load measurements are affected by repeated freeze-thaw (FT) cycles. We therefore evaluated the effect of multiple FT cycles on HBV DNA and HCV RNA quantification by testing serum specimens subjected to one (baseline), two, four, and eight FT cycles with the appropriate Chiron Quantiplex assay. Linear regression analysis showed minor increases of 1.7% per FT cycle for both HBV DNA and HCV RNA. The rise in HCV RNA levels was more pronounced among low-concentration samples, since further analysis revealed an increase of 3.2% per FT cycle among samples with 0.2 to 3.86 Meq of HCV RNA per ml. Given that the coefficient of variation for the Quantiplex assays is generally 10 to 15%, the minor increases in HBV DNA and HCV RNA levels with progressive FT cycles for the specimens tested were recognized only because analysis of variance revealed a statistically significant trend (P < 0.05). Due to the minor statistical trend, the clinical impact for individual patient specimens is likely to be limited, but it may deserve further study. In conclusion, the concentration of HBV DNA and HCV RNA in serum specimens subjected to up to eight short-term FT cycles was stable.

scholarly journals NeuMoDx random access molecular diagnostic system for detection and quantification of hepatitis B virus in clinical samples

2020 ◽  
Vol 14 (10) ◽  
pp. 1197-1203
Author(s):  
Ayse Arikan ◽  
Murat Sayan ◽  
Osman Doluca
Keyword(s):  
Statistical Significance ◽  
Random Access ◽  
Absolute Error ◽  
Hbv Dna ◽  
Molecular Diagnostic ◽  
Clinical Samples ◽  
External Quality ◽  
Chi Square ◽  
Deming Regression ◽  
The Mean

Introduction: Currently, several molecular assays are available to detect and quantify HBV DNA in clinical samples. We aimed to characterize and compare the clinical performance of newly designed NeuMoDx PCR to the existing artus PCR. Methodology: The plasma HBV DNA levels of 96 clinical and 5 external quality control samples were measured by NeuMoDx and artus assays simultaneously in Kocaeli University, Turkey. The linearity, agreement and the correlation between two assays were determined by Deming regression analysis, Bland-Altman plotting, the chi-square and the relative absolute error statistical analyzes. For all statistical analyzes, the XLSTAT statistical program was used. Results: The mean (standard deviation; SD) age was 45.07 ± 12.29. HBsAg S/Co median (range) was 4,273.4 ± 1,138.1 and ALT U/L median (range) was 27 ± 16. The mean (SD) of HBV DNA was 1.46+E6 ± 1.0+E4 for NeuMoDx and 1.54+E5 ± 4.7 + E4 for artus assays. The Deming regression indicates a linear correlation (95% confidence). The chi-square test indicates strong correlation (p < 0.001). Bland-Altman analysis confirms that the measurement difference is acceptable. The relative absolute error analysis for artus showed relatively less and more consistent error rate. With 5 external quality check samples, the statistical significance was low (p = 0.566). Conclusions: The NeuMoDx HBV assay showed an excellent analytical performance by providing a rapid, high throughput technology in a random-access testing system in clinical samples and may be a new solution for viral load quantification in the management of HBV infections.

scholarly journals Comparison of a novel real-time PCR method (RTA) and Artus RG for the quantification of HBV DNA and HCV RNA

2017 ◽  
Vol 11 (07) ◽  
pp. 543-548
Author(s):  
Sibel Aydogan ◽  
Ziya Cibal Acikgoz ◽  
Aysegul Gozalan ◽  
Fisun Kirca ◽  
Tuba Muderris ◽  
...  
Keyword(s):  
Quality Control ◽  
Real Time ◽  
Hbv Dna ◽  
Clinical Samples ◽  
Hcv Rna ◽  
Rt Pcr ◽  
B Virus ◽  
Pcr Method ◽  
Polymerase Chain

Introduction: The purpose of this study was to evaluate and compare two manual isolation and real-time (RT) polymerase chain reaction (PCR) kits (RTA RT-PCR with RTA isolation kit and Artus RG RT-PCR with QIAamp isolation kit) for molecular diagnosis of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. Methodology: The study was conducted on 121 and 54 clinical samples for the detection of HBV DNA and HCV RNA, respectively, with an additional 8 HCV RNA external quality control samples. Results: Though a high correlation was observed between the two kits for the HBV DNA (r = 0.955, p = 0.001) and HCV RNA quantifications (r = 0.828, p = 0.001), discordant results were found in nine of the HBV DNA and in six of the HCV RNA samples. The mean difference between the two systems was found to be 0.4 log IU/mL in Quality Control for Molecular Diagnostics (QCMD) HCV RNA samples by Bland-Altman analysis. Conclusions: Although there was a high correlation between HBV DNA and HCV RNA tests according to the results of the study, the RTA system requires improvement for the determination of HCV RNA.

scholarly journals Validation of a Novel IoT and AI based Point-of-Care Testing Laboratory: Analytical Accuracy and Clinical Agreement

2021 ◽  
Author(s):  
Lucca Malucelli ◽  
Gabriele Luise Neves Alves ◽  
Carolina Melchioretto dos Santos ◽  
Matheus Severo ◽  
Victor Henrique Alves Ribeiro ◽  
...  
Keyword(s):  
Influenza A ◽  
Point Of Care ◽  
Pearson Correlation ◽  
Turnaround Time ◽  
Clinical Samples ◽  
Point Of Care Testing ◽  
Reliable Performance ◽  
25 Hydroxy Vitamin D ◽  
Analytical Accuracy ◽  
Kappa Agreement

Point-of-care testing (POCT) offers several advantages over traditional laboratory testing. Offering less invasive testing with a faster turnaround time is not enough if not associated with an acceptable level of accuracy. Here, we show the analytical validation behind the multi-analyte POCT immunochromatography analyser, Hilab Flow (HiF). Analyses from 4,518 clinical samples were compared to College of American Pathologists accredited laboratories for ten quantitative and thirteen qualitative exams. Compatibility between methods was evaluated in terms of association/correlation and clinical agreement. Strong correlation/ concordance was observed between quantitative (CHOL, HDL-c, TG, HbA1c, Glycemia, 25-Hydroxy Vitamin D, TSH, Uric Acid, Creatinine, Urea) and qualitative methods (COVID-19 IgG/ IgM, Beta-hCG, Syphilis, Anti-HBsAg, Zika IgG/ IgM, Influenza A/B, HIV, HCV, HBsAg, Dengue NS1, COVID-19 Ag, Dengue IgG/ IgM, PSA). Approval criteria was obtaining a kappa agreement > 0.8 or a Pearson correlation > 0.9 depending on the exam. Overall percentage agreement was greater than 95% for all exams, indicating a good clinical agreement to gold-standard laboratory-based tests. Results indicate all exams are suitable for POCT and present a reliable performance. Data support the analyser is a useful tool to aid decision-making at the clinical setting, with potential to contribute with healthcare solutions in diagnostic medicine worldwide.

Naked-eye colorimetric detection of HCV RNA mediated by a 5′ UTR-targeted antisense oligonucleotide and plasmonic gold nanoparticles

The Analyst ◽  
2021 ◽  
Author(s):  
Almas Shamaila Mohammed ◽  
Aniket Balapure ◽  
Mahammad Nanne Khaja ◽  
Ramakrishnan Ganesan ◽  
Jayati Ray Dutta
Keyword(s):  
Gold Nanoparticles ◽  
Antisense Oligonucleotide ◽  
Early Stage ◽  
Colorimetric Detection ◽  
Sensitive Detection ◽  
Clinical Samples ◽  
Hcv Rna ◽  
Naked Eye

An Au NP based facile strategy for the rapid, early-stage, and sensitive detection of HCV RNA in clinical samples which avoids thiol tagging to the antisense oligonucleotide and expensive infrastructure is presented.

scholarly journals Essential properties and pitfalls of colorimetric Reverse Transcription Loop-mediated Isothermal Amplification as a point-of-care test for SARS-CoV-2 diagnosis

2021 ◽  
Vol 27 (1) ◽  
Author(s):  
Bruna de Oliveira Coelho ◽  
Heloisa Bruna Soligo Sanchuki ◽  
Dalila Luciola Zanette ◽  
Jeanine Marie Nardin ◽  
Hugo Manuel Paz Morales ◽  
...  
Keyword(s):  
Viral Load ◽  
Internal Control ◽  
Reverse Transcription ◽  
Color Change ◽  
Point Of Care ◽  
Colorimetric Detection ◽  
False Negative ◽  
Isothermal Amplification ◽  
Clinical Samples ◽  
Loop Mediated Isothermal Amplification

Abstract Background SARS-CoV-2 Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) colorimetric detection is a sensitive and specific point-of-care molecular biology technique used to detect the virus in only 30 min. In this manuscript we have described a few nuances of the technique still not properly described in the literature: the presence of three colors clusters; the correlation of the viral load with the color change; and the importance of using an internal control to avoid false-negative results. Methods To achieve these findings, we performed colorimetric RT-LAMP assays of 466 SARS-CoV-2 RT-qPCR validated clinical samples, with color quantification measured at 434 nm and 560 nm. Results First we determinate a sensitivity of 93.8% and specificity of 90.4%. In addition to the pink (negative) and yellow (positive) produced colors, we report for the first time the presence of an orange color cluster that may lead to wrong diagnosis. We also demonstrated using RT-qPCR and RT-LAMP that low viral loads are related to Ct values > 30, resulting in orange colors. We also demonstrated that the diagnosis of COVID-19 by colorimetric RT-LAMP is efficient until the fifth symptoms day when the viral load is still relatively high. Conclusion This study reports properties and indications for colorimetric RT-LAMP as point-of-care for SARS-CoV-2 diagnostic, reducing false results, interpretations and optimizing molecular diagnostics tests application.

scholarly journals Evaluation of mobile real-time polymerase chain reaction tests for the detection of severe acute respiratory syndrome coronavirus 2

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chukwunonso Onyilagha ◽  
Henna Mistry ◽  
Peter Marszal ◽  
Mathieu Pinette ◽  
Darwyn Kobasa ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Real Time ◽  
Point Of Care ◽  
Middle Eastern ◽  
Turnaround Time ◽  
Cross Reactivity ◽  
Clinical Samples ◽  
Diarrhea Virus ◽  
Highly Sensitive ◽  
Transmissible Gastroenteritis

AbstractThe coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), calls for prompt and accurate diagnosis and rapid turnaround time for test results to limit transmission. Here, we evaluated two independent molecular assays, the Biomeme SARS-CoV-2 test, and the Precision Biomonitoring TripleLock SARS-CoV-2 test on a field-deployable point-of-care real-time PCR instrument, Franklin three9, in combination with Biomeme M1 Sample Prep Cartridge Kit for RNA 2.0 (M1) manual extraction system for rapid, specific, and sensitive detection of SARS-COV-2 in cell culture, human, and animal clinical samples. The Biomeme SARS-CoV-2 assay, which simultaneously detects two viral targets, the orf1ab and S genes, and the Precision Biomonitoring TripleLock SARS-CoV-2 assay that targets the 5′ untranslated region (5′ UTR) and the envelope (E) gene of SARS-CoV-2 were highly sensitive and detected as low as 15 SARS-CoV-2 genome copies per reaction. In addition, the two assays were specific and showed no cross-reactivity with Middle Eastern respiratory syndrome coronavirus (MERS-CoV), infectious bronchitis virus (IBV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, and other common human respiratory viruses and bacterial pathogens. Also, both assays were highly reproducible across different operators and instruments. When used to test animal samples, both assays equally detected SARS-CoV-2 genetic materials in the swabs from SARS-CoV-2-infected hamsters. The M1 lysis buffer completely inactivated SARS-CoV-2 within 10 min at room temperature enabling safe handling of clinical samples. Collectively, these results show that the Biomeme and Precision Biomonitoring TripleLock SARS-CoV-2 mobile testing platforms could reliably and promptly detect SARS-CoV-2 in both human and animal clinical samples in approximately an hour and can be used in remote areas or health care settings not traditionally serviced by a microbiology laboratory.

scholarly journals A211 SPONTANEOUS HBV REMISSION AFTER HBV REACTIVATION FOLLOWING TREATMENT WITH SOFOSBUVIR AND VELPATASVIR IN A PATIENT WITH HCV AND HBV CO-INFECTION: CASE REPORT

2021 ◽  
Vol 4 (Supplement_1) ◽  
pp. 242-243
Author(s):  
A Chiang ◽  
K Tsoi
Keyword(s):  
Hepatitis C ◽  
Hepatitis B ◽  
Antiviral Agents ◽  
Clinical Signs ◽  
Hbv Dna ◽  
Hcv Rna ◽  
Hbv Reactivation ◽  
Genotype 3 ◽  
Direct Acting Antiviral ◽  
Direct Acting

Abstract Background In co-infected patients with hepatitis B (HBV) and hepatitis C (HCV), the treatment of HCV with direct-acting antiviral agents (DAA) can cause HBV reactivation. However, there are no clear guidelines on the timing of treatment initiation, especially in the absence of clinical signs of flare. Aims Here we discuss the case of a 34-year-old female with HBV and HCV genotype 3 who had HBV reactivation following HCV treatment, but did not require nucleos(t)ide therapy. Methods She initially presented with chronic inactive hepatitis B and chronic hepatitis C with HBV DNA level of 67.5 IU/mL and HCV RNA level of 3.33 x 106 IU/mL. She completed a 12 week course of sofosbuvir and velpatasvir for HCV and achieved sustained virologic remission, but subsequently developed reactivation of her HBV with HBV DNA peaking at 3.41 x 104 IU/mL twelve weeks post-treatment. She did not develop any signs of hepatitis and a decision was made to monitor her clinically. Results Two years later, she spontaneously went into remission with her HBV DNA levels being &lt;10 IU/mL. Conclusions The significance of this case is to illustrate HBV reactivation following treatment of HCV with DAAs may not necessitate immediate treatment, especially if there are no signs of flare. There have been similar reported cases, but larger prospective studies are required to determine the appropriate clinical context where monitoring may be acceptable instead of immediate treatment. Funding Agencies None

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