Evaluation of the Abbott RealTime HCV assay for quantitative detection of hepatitis C virus RNA

Michelin, B.; Müller, Zsófia; Stelzl, Evelyn; Marth, Egon; Kessler, Harald H.

doi:10.1016/j.jcv.2006.11.007

Cited by 55 publications

(39 citation statements)

References 9 publications

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“…However, for subtypes 1a and 2b, the mean difference in VL was not significantly different from non-1a or non-2b specimens, respectively, indicating that the underlying reason for the large discrepancy in VL results for these specimens was not related to subtype. The observed precision, linearity, and sensitivity of the ART assay reported here are similar to results from other laboratories and testing environments (2,5,15,(17)(18)(19)22). In addition, the negative bias of results from the CAP-CTM assay compared to those from the ART assay that we described is consistent with most (14-16), although not all (2, 17), published studies.…”

supporting

confidence: 90%

Performance of the Abbott RealTime and Roche Cobas TaqMan Hepatitis C Virus (HCV) Assays for Quantification of HCV Genotypes

et al. 2012

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We evaluated the Abbott RealTime (ART) and Roche Cobas TaqMan Hepatitis C virus (HCV) viral load assays for quantification of HCV genotypes in patient specimens. The ART HCV assay was a more sensitive and precise tool for accurate HCV viral load quantification across the HCV genotypes tested, especially genotype 1b. Hepatitis C virus (HCV) infection is associated with significant liver-related morbidity and mortality around the world. The World Health Organization estimates that 3% of the global population is infected with HCV and that there are approximately 170 million people at risk of developing cirrhosis or hepatocarcinoma (23,24). Treatment of HCV infection typically consists of pegylated interferon plus ribavirin or pegylated interferon, ribavirin, and a direct-acting antiviral (DAA) protease inhibitor (triple therapy) for non-genotype 1 and genotype 1, respectively (11,12,13). Depending on the particular treatment regimen and the genotype of HCV, treatment success as measured by failure to detect viral replication 24 weeks after cessation of treatment can be achieved in 50 to 80% of patients (12).Measurement of HCV viral load (VL) for the different HCV genotypes is crucial to clinical management of HCV-infected patients, both treated and not, for disease staging, decisions regarding treatment initiation, and individualization of treatment strategy (i.e., dosage and duration based on response kinetics) (1,6,7,13). Furthermore, with the advent of DAAs, VL monitoring may help prevent protease inhibitor resistance development by allowing for switching or stopping therapy if VL does not decrease or returns while on treatment.There are currently several commercially available HCV VL assays; real-time PCR assays are generally preferred because of their wide dynamic ranges and good sensitivities (1, 3). Two commercial real-time PCR platforms are available, the Cobas AmpliPrep/Cobas TaqMan HCV assay version 1.0 (CAP-CTM; Roche Molecular Systems, Pleasanton, CA) and the Abbott RealTime HCV assay (ART; Abbott Molecular, Des Plaines, IL). We characterized the performance of the ART assay and compared results obtained with both assays using clinical specimens of diverse HCV genotypes in a university hospital central testing laboratory.HCV VL was determined with the ART and the CAP-CTM as per the manufacturers' recommendations. A serum specimen volume of 500 l was required for ART, compared to 850 l for CAP-CTM. Sample preparation for the ART assay was performed using the Abbott m2000sp instrument. HCV genotypes were identified using the Versant Inno-LIPA HCV Genotype 2.0 assay (Siemens Healthcare Diagnostics, Deerfield, IL); specimens with indeterminate results were tested with the RealTime Genotyping II RUO assay (Abbott Molecular, Des Plaines, IL) or by direct sequencing and phylogenetic analysis of NS5B (performed by Siemens Clinical Laboratories, Berkeley, CA). Statistical tests were performed using Prism 5.0 (GraphPad Software, La Jolla, CA).To evaluate the sensitivity, linearity, and intra-and interrun precisio...

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confidence: 90%

Performance of the Abbott RealTime and Roche Cobas TaqMan Hepatitis C Virus (HCV) Assays for Quantification of HCV Genotypes

et al. 2012

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“…Previous studies have highlighted the challenge of unbiased detection of different HCV genotypes (3,6,8,10,13). However, these and other studies (including this study) have limited ability to measure genotype bias for several reasons.…”

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confidence: 92%

Evaluation of the Abbott Investigational Use Only RealTi m e Hepatitis C Virus (HCV) Assay and Comparison to the Roche TaqMan HCV Analyte-Specific Reagent Assay

et al. 2009

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The accurate and sensitive measurement of hepatitis C virus (HCV) RNA is essential for the clinical management and treatment of infected patients and as a research tool for studying the biology of HCV infection. We evaluated the linearity, reproducibility, precision, limit of detection, and concordance of viral genotype quantitation of the Abbott investigational use only RealTime HCV (RealTime) assay using the Abbott m2000 platform and compared the results to those of the Roche TaqMan Analyte-Specific Reagent (TaqMan) and Bayer Versant HCV bDNA 3.0 assay. Comparison of 216 samples analyzed by RealTime and TaqMan assays produced the following Deming regression equation: RealTime ‫؍‬ 0.940 (TaqMan) ؉ 0.175 log 10 HCV RNA IU/ml. The average difference between the assays was 0.143 log 10 RNA IU/ml and was consistent across RealTime's dynamic range of nearly 7 log 10 HCV RNA IU/ml. There was no significant difference between genotypes among these samples. The limit of detection using eight replicates of the World Health Organization HCV standard was determined to be 7.74 HCV RNA IU/ml by probit analysis. Replicate measurements of commercial genotype panels were significantly higher than TaqMan measurements for most samples and showed that the RealTime assay is able to detect all genotypes with no bias. Additionally, we showed that the amplicon generated by the widely used Roche COBAS Amplicor Hepatitis C Virus Test, version 2.0, can act as a template in the RealTime assay, but potential cross-contamination could be mitigated by treatment with uracil-N-glycosylase. In conclusion, the RealTime assay accurately measured HCV viral loads over a broad dynamic range, with no significant genotype bias.Methods for accurate quantitation of serum and plasma hepatitis C virus (HCV) RNA levels have become key tools both for understanding the biology of HCV infection and for the clinical management of patients under treatment. The ability to predict likelihood of response to combination interferon/ ribavirin therapy by assessing rates of HCV viral load decline has provided a more individualized treatment algorithm that can identify nonresponsive patients early in treatment, sparing them significant morbidity and cost. New algorithms that examine the kinetics of HCV viral decline provide an even more refined tool for management and complement the information provided by HCV genotype determination. Finally, HCV viral kinetics data are essential for the understanding of new therapeutics such as the class of protease inhibitors. For all applications, accurate quantitation of all HCV types over a broad dynamic range is critical. The advent of real-time PCR methods provides a powerful tool for a broad dynamic range of quantitation of viruses; however, targets such as HCV require careful assay design to avoid errors due to sequence variations intrinsic to RNA viruses. Abbott Molecular, Inc. (Des Plaines, IL), recently released the m2000 system and tests for human immunodeficiency virus type 1, HCV, and chlamydia/gonorrhea (Chlamydia trac...

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“…HCV RNA was isolated by the Abbott m2000sp instrument from 1,000-l aliquots (sample volume required, 500 l), as described previously (17). Extracted samples and controls were then amplified and detected with the Abbott m2000rt instrument, according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Differences between Two Real-Time PCR-Based Hepatitis C Virus (HCV) Assays (RealTime HCV and Cobas AmpliPrep/Cobas TaqMan) and One Signal Amplification Assay (Versant HCV RNA 3.0) for RNA Detection and Quantification

et al. 2008

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Hepatitis C virus (HCV) RNA detection and quantification are the key diagnostic tools for the management of hepatitis C. Commercially available HCV RNA assays are calibrated to the HCV genotype 1 (gt1)-based WHO standard. Significant differences between assays have been reported. However, it is unknown which assay matches the WHO standard best, and little is known about the sensitivity and linear quantification of the assays for non-gt1 specimens. Two real-time reverse transcriptase PCR-based assays (RealTime HCV and Cobas Ampliprep/Cobas TaqMan HCV [CAP/CTM]) and one signal amplification-based assay (the Versant HCV RNA, version 3.0, branched DNA [bDNA] assay) were compared for their abilities to quantify HCV RNA in clinical specimens (n ‫؍‬ 65) harboring HCV isolates of gt1 to g5. The mean differences in the amounts detected by RealTime HCV in comparison to those detected by the bDNA assay and CAP/CTM were ؊0.02 and 0.72 log 10 IU/ml HCV RNA, respectively, for gt1; ؊0.22 and 0.03 log 10 IU/ml HCV RNA, respectively, for gt2; ؊0.27 and ؊0.22 log 10 IU/ml HCV RNA, respectively, for gt3; ؊0.19 and ؊1.27 log 10 IU/ml HCV RNA, respectively, for gt4; and ؊0.03 and 0.09 log 10 IU/ml HCV RNA, respectively, for gt5. The lower limits of detection for RealTime HCV and CAP/CTM were 16.8 and 10.3 IU/ml, respectively, for the WHO standard and in the range of 4.7 to 9.0 and 3.4 to 44.4 IU/ml, respectively, for clinical specimens harboring gt1 to gt6. Direct comparison of the two assays with samples of the WHO standard (code 96/798) with high titers yielded slightly smaller amounts by RealTime HCV (؊0.2 log 10 at 1,500 IU/ml and ؊0.3 log 10 at 25,000 IU/ml) and larger amounts by CAP/CTM (0.3 log 10 at 1,500 IU/ml and 0.2 log 10 at 25,000 IU/ml). Finally, all three tests were linear between 4.0 ؋ 10 3 and 1.0 ؋ 10 6 IU/ml (correlation coefficient, >0.99). In conclusion, the real-time PCR based assays sensitively detected all genotypes and showed comparable linearities for the quantification of HCV RNA, with the exception of gt1 and gt4. The previously reported differences in the absolute quantification of samples harboring gt1 were confirmed and may be explained by different calibrations to the WHO standard.Hepatitis C virus (HCV) infection is a common cause of chronic liver disease that can lead to end-stage liver disease, including hepatocellular carcinoma.At present, combination therapy with pegylated interferon and ribavirin is the standard of care. However, this treatment regimen appears to be effective in only 40% to 50% of patients infected with genotype 1 and approximately 80% of patients infected with genotypes 2 and 3 (12, 13, 16).A sustained virologic response, defined as undetectable HCV RNA at least 6 months after the completion of therapy, may be predicted by a number of host and viral factors, including age, race, liver fibrosis, the HCV genotype, and the baseline viral load.Recent reports suggest that decisions on the optimal treatment duration may be made on the basis of the baseline viral load and the virologi...

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Evaluation of the Abbott RealTime HCV assay for quantitative detection of hepatitis C virus RNA

Cited by 55 publications

References 9 publications

Performance of the Abbott RealTime and Roche Cobas TaqMan Hepatitis C Virus (HCV) Assays for Quantification of HCV Genotypes

Performance of the Abbott RealTime and Roche Cobas TaqMan Hepatitis C Virus (HCV) Assays for Quantification of HCV Genotypes

Evaluation of the Abbott Investigational Use Only RealTi m e Hepatitis C Virus (HCV) Assay and Comparison to the Roche TaqMan HCV Analyte-Specific Reagent Assay

Differences between Two Real-Time PCR-Based Hepatitis C Virus (HCV) Assays (RealTime HCV and Cobas AmpliPrep/Cobas TaqMan) and One Signal Amplification Assay (Versant HCV RNA 3.0) for RNA Detection and Quantification

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