Multicenter Evaluation of the VERSANT HCV RNA Qualitative Assay for Detection of Hepatitis C Virus RNA

Hendricks, David A.; Friesenhahn, Michel; Tanimoto, Lorine; Goergen, Bernd; Dodge, Deborah E.; Comanor, Lorraine

doi:10.1128/jcm.41.2.651-656.2003

Cited by 40 publications

(34 citation statements)

References 10 publications

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“…[11][12][13] Positive TMA results at the end of treatment (week 48, or W48) have improved the identification of patients who subsequently relapse. 9,[14][15][16] However, the clinical usefulness of the TMA assay over less sensitive PCR-based qualitative assays in monitoring responsiveness to interferon alphabased treatment regimens at other time points is less clear.…”

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confidence: 99%

HCV RNA detection by TMA during the hepatitis C antiviral long-term treatment against cirrhosis (Halt-C) trial

et al. 2006

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For making treatment decisions related to chronic hepatitis C, the utility of HCV RNA tests with increased sensitivity has not been defined. Prior interferon nonresponders with advanced fibrosis (n ‫؍‬ 1,145) were retreated with peginterferon alpha-2a and ribavirin. Patients who were HCV RNA-negative by a polymerase chain reaction (PCR)-based assay (Roche COBAS Amplicor ™ HCV Test, v. 2.0; lower limit of detection [LOD] 100 IU/mL) at week 20 (W20) received treatment for 48 weeks. Stored specimens were tested using the Bayer VERSANT HCV RNA Qualitative (TMA) Assay (LOD 9.6 IU/mL) and compared to PCR results for the ability to predict sustained virological response (SVR; defined as undetectable HCV RNA by PCR at W72). Nearly all PCR-positive samples (1006/1007, 99.9%) were positive as assessed by TMA. Among 1,294 PCR-negative samples, 22% were TMApositive. Negative TMA results were more predictive of SVR than were negative PCR results at W12 (82% vs. 64%, P < .001) and at W20 (66% vs. 52%, P ‫؍‬ 0.001). SVR was more likely the earlier TMA had become negative during treatment (82% at W12, 44% at W20, 20% at W24). Among 45 patients who were TMA-positive but were PCR-negative at W20 and W24, none achieved SVR (95% CI: 0%-8%). Approximately 10% of patients with a Abbreviations: HCV, hepatitis C virus; SVR, sustained virological response. From the

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confidence: 99%

HCV RNA detection by TMA during the hepatitis C antiviral long-term treatment against cirrhosis (Halt-C) trial

et al. 2006

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“…The bDNA assay can be confidently used as a comparator, since it has been shown to be precise and accurate and to equally quantify HCV genotypes 1 to 6, due to the use of a set of 6 capture and 17 extender oligonucleotide probes spanning the full-length 5Ј-noncoding region and the 5Ј third of the core coding region for hybridization of the HCV genome (1,8,9,12,13,(17)(18)(19)24).…”

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Performance of the Abbott Real-Time PCR Assay Using m 2000 _sp and m 2000 _rt for Hepatitis C Virus RNA Quantification

2009

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Quantification of hepatitis C virus (HCV) RNA is essential for the everyday management of chronic hepatitis C therapy. "Real-time" PCR techniques are potentially more sensitive than classical PCR techniques, are not prone to carryover contamination, and have a consistently wider dynamic range of quantification. Thus, they are rapidly replacing other technologies for routine quantification of HCV RNA. We extensively evaluated the intrinsic characteristics and clinical performance of the m2000 sp -m2000 rt Abbott real-time PCR platform for HCV RNA quantification. The study shows that the m2000 sp -m2000 rt platform is sensitive, specific, and precise; that the results are reproducible; and that the platform has a broad dynamic range of quantification. When comparing HCV RNA levels measured in the same individuals with the m2000 sp -m2000 rt platform and the third-generation branched-DNA assay, a trend toward a modest overestimation of HCV RNA levels was observed in the m2000 sp -m2000 rt platform in all genotypes except genotype 5. The differences, however, were unlikely to have any impact in clinical practice. In conclusion, our study shows that the Abbott m2000 real-time PCR system for HCV RNA quantification is sensitive, specific, and precise; that the results are reproducible; and that the platform's broad dynamic range of quantification is well suited to HCV RNA monitoring in the clinical setting.Monitoring of hepatitis C virus (HCV) RNA levels is essential for the management of chronic hepatitis C therapy with the combination of pegylated alpha interferon and ribavirin. Indeed, the rapid (undetectable HCV RNA at week 4 of therapy) and early (more than 2 log 10 drop or undetectable HCV RNA at week 12 of therapy) virologic response is a strong predictor of the likelihood of sustained viral eradication and is used to tailor the treatment duration and to improve cure rates (2,6,7,10,14,16,23,26,27). HCV RNA quantification will continue to be of major importance in the management of therapy with the new HCV drugs in development to assess the virologic response and to detect virologic breakthroughs related to viral resistance early enough to alter therapy and rescue antiviral efficacy (20).Ideally, HCV RNA quantification assays should be sensitive, specific, accurate, and precise, and the results should be reproducible. They should have a broad range of linear quantification that fully covers the HCV RNA levels observed in clinical practice in both untreated and treated patients, and quantification should be independent of the HCV genotype. Real-time PCR techniques currently are replacing classical PCR methods and the branched-DNA (bDNA) technology, which did not fulfill all of these criteria. Real-time PCR methods are sensitive, with lower limits of detection/quantification on the order of 10 to 15 HCV RNA international units (IU)/ml, and are not prone to carryover contamination. They benefit from a broad dynamic range of quantification of 7 to 8 log 10 units that covers most of the HCV RNA levels encountered in ...

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“…13,14 Qualitative HCV RNA detection assays have been approved for the diagnosis of HCV infection, but these tests are also used to assess response to antiviral therapy, due to their low LODs compared to assays that quantitate HCV RNA. The utility of highly sensitive qualitative assays such as the modified COBAS AMPLICOR version 2.0 and Versant HCV RNA Qualitative Assay ([TMA, LOD ϭ 2 to 10 IU/ml] 15,16 in the setting of therapeutic monitoring is not yet clear. However, data in the literature suggest potential roles for these types of tests.…”

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confidence: 99%

Increased Sensitivity of the Roche COBAS AMPLICOR HCV Test, Version 2.0, Using Modified Extraction Techniques

Forman

Valsamakis

2004

The Journal of Molecular Diagnostics

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Processing modifications were made to the COBAS AMPLICOR HCV version 2.0 assay to enhance sensitivity. Two methods of specimen concentration, centrifugation ("ultraspin") and cationic detergent plus silica membrane ("ultracolumn"), were compared to the standard method. The effect of these changes on assay sensitivity and specificity was examined using commercial hepatitis C virus (HCV) preparations. The limits of detection (LOD, defined as detection of HCV RNA in > 95% of replicates) of genotype 1a were 50, 12, and 6 by standard method, ultraspin and ultracolumn, respectively. For genotype 1b, the LOD was 25 IU/ml, 12 IU/ml, and 3 IU/ml; for 2b, it was 50, 12, and 3; for 3a, it was 25, 12, and 1.5; for 4 it was 18, 4, and 2; for 5a, it was 38, 9, and 2; and for 6a it was 47, 6, and 3. No false positives were detected after ultraspin when controls containing high or low HCV concentrations were alternated with normal human plasma. Plasmas in which HCV RNA was not detected by the standard assay were re-tested with modified methods to assess the effect of altered processing in clinical specimens. Three of 152 specimens with no detectable HCV RNA by the standard method were positive by ultraspin and 2 of 109 were positive by ultracolumn, suggesting that these methods may increase assay sensitivity in clinical specimens. Accurate, sensitive detection of hepatitis C virus (HCV) in serum or plasma is essential for diagnosis of HCV infections and assessing response to anti-HCV therapy. The standard, FDA approved COBAS AMPLICOR HCV version 2.0 assay has a reported LOD of 50 IU/ml for HCV genotype 1b. 1 This assay detects HCV RNA during active HCV infection. 2 It has been used to diagnose viremia 3,4 and to assess treatment response. 5 Therapy for chronic HCV infection has evolved from interferon alfa (IFN-alfa) alone, to second generation combination therapy with IFN-alfa plus ribavirin for either 24 or 48 weeks, to the latest generation of pegylatedinterferon ␣ with ribavirin (pegIFN-alfa/riba) for either 24 or 48 weeks. The lowest rates of response and highest rates of relapse were observed with IFN-␣ monotherapy. 6 Treatment with pegIFN-␣/riba resulted in the highest rates of response and the lowest rates of relapse. 5,7 Variables that potentially affect rates of virologic relapse include treatment regimen, genetic variability of the virus, and HCV RNA detection assay sensitivity. The ability to detect very low levels of virus may prove useful in identifying patients who may relapse at end of treatment. Transcription-mediated amplification (TMA) has been used to assess the presence of residual HCV RNA in plasma from patients treated with pegINF-alfa-2a and INF-alfa-2a. 8 In patients who relapsed after therapy, residual HCV RNA was detected in 7% of plasma samples at end of treatment with pegINF-alfa-2a and 33% of samples following therapy with INF-alfa-2a.The aim of this study was to increase the sensitivity of COBAS AMPLICOR HCV version 2.0 through alteration of extraction methods. Two modifications were used to extra...

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Multicenter Evaluation of the VERSANT HCV RNA Qualitative Assay for Detection of Hepatitis C Virus RNA

Cited by 40 publications

References 10 publications

HCV RNA detection by TMA during the hepatitis C antiviral long-term treatment against cirrhosis (Halt-C) trial

HCV RNA detection by TMA during the hepatitis C antiviral long-term treatment against cirrhosis (Halt-C) trial

Performance of the Abbott Real-Time PCR Assay Using m 2000 _sp and m 2000 _rt for Hepatitis C Virus RNA Quantification

Increased Sensitivity of the Roche COBAS AMPLICOR HCV Test, Version 2.0, Using Modified Extraction Techniques

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Multicenter Evaluation of the VERSANT HCV RNA Qualitative Assay for Detection of Hepatitis C Virus RNA

Cited by 40 publications

References 10 publications

HCV RNA detection by TMA during the hepatitis C antiviral long-term treatment against cirrhosis (Halt-C) trial

HCV RNA detection by TMA during the hepatitis C antiviral long-term treatment against cirrhosis (Halt-C) trial

Performance of the Abbott Real-Time PCR Assay Using m 2000 sp and m 2000 rt for Hepatitis C Virus RNA Quantification

Increased Sensitivity of the Roche COBAS AMPLICOR HCV Test, Version 2.0, Using Modified Extraction Techniques

Contact Info

Product

Resources

About

Performance of the Abbott Real-Time PCR Assay Using m 2000 _sp and m 2000 _rt for Hepatitis C Virus RNA Quantification