Multicenter Evaluation of the COBAS AMPLICOR HCV Assay, an Integrated PCR System for Rapid Detection of Hepatitis C Virus RNA in the Diagnostic Laboratory

Abstract

The benefits shown by the recent introduction of PCR for the in vitro diagnosis of hepatitis C virus (HCV) infection has prompted the development of standardized, ready-to-use assays that can be implemented in routine clinical laboratories. We have evaluated the clinical performance of COBAS AMPLICOR HCV (COBAS), the first instrument system that allows the automation of HCV RNA amplification and detection, to determine its performance in the routine laboratory setting. More than 2,000 specimens collected at five centers were analyzed in parallel by the COBAS and the manual AMPLICOR HCV (AMPLICOR) tests, and the results were compared with the results for biochemical and serological markers of HCV. In this study the two PCR systems showed the same accuracy, with a concordance rate of 99.8%. As expected, the correlation between serology and PCR was not absolute because the presence of anti-HCV antibodies may be associated with a latent or past infection. On the other hand, if the presence of confirmed anti-HCV antibodies and elevated alanine aminotransferase levels are taken as the “gold standard,” indicating an active, ongoing infection, the COBAS and AMPLICOR tests show high and comparable sensitivities (100%) and specificities (98%), with positive and negative predictive values of 100 and 97%, respectively. During the study no false-positive reactions were detected. The use of an internal control allowed the identification of inhibitory substances that prevented amplification for 0.3 and 0.4% of samples tested by the COBAS and AMPLICOR tests, respectively. Compared to the manual system, the COBAS system allowed a significant reduction of hands-on time and could improve the overall laboratory work flow. In conclusion, these results support the use of the COBAS and AMPLICOR tests for the molecular diagnosis of active HCV infections.

Factors affecting efficacy in patients with genotype 2 chronic hepatitis C treated by pegylated interferon alpha-2b and ribavirin: reducing drug doses has no impact on rapid and sustained virological responses

“…Serum HCV RNA level was quantified by PCR assay (COBAS Amplicor HCV Test v2.0, Chugai-Roche Diagnostics, Tokyo, Japan), with a sensitivity limit of 5000 IU/mL and a dynamic range from 5000 to 5 000 000 IU/mL [11].…”

Evaluation of clinical usefulness of second‐generation HCV core antigen assay: comparison with COBAS AMPLICOR HCV MONITOR assay version 2.0

“…As shown in the present study, 204 (95.8%) of 213 cases were detectable by the second-generation assay, providing evidence that sensitivity was indeed significantly improved. The sensitivity of this assay was comparable with that of the COBAS v2.0 assay, a well-calibrated, sensitive and non-serotype-dependent assay (8,(17)(18)(19)(20)(24)(25)(26).…”

“…The quantification of HCV has become an important issue because low viral load is a strong predictor of sustained response to interferon (IFN) monotherapy for chronic hepatitis C (13-15). Among the assays for quantification of HCV load, combined reverse transcription PCR assay, the AM-PLICOR HCV MONITOR assay version 2.0 (CO-BAS v2.0) (Roche Molecular Systems, Branchburg, NJ), is now one of the most sensitive and reliable assays (16)(17)(18)(19)(20).…”

HCV RNA levels during therapy with amantadine in addition to interferon and ribavirin in chronic hepatitis C patients with previous nonresponse or response/relapse to interferon and ribavirin

“…In order to measure the HCV RNA levels, serum samples were obtained at week 0, 2, 4, 8, 12, 16, 20, 24 during treatment, and at week 48 at the end of follow-up. Sera were immediately frozen at )70°C, and later analysed by a quantitative PCR (Roche, Cobas Amplicor HCV Monitor R version 2.0), performed according to instructions of the manufacturer [13]. This assay has a sensitivity level of approximately 600 IU ml )1 .…”