Performance characteristics of a transcription-mediated nucleic acid amplification assay for qualitative detection of hepatitis C virus RNA

The detection of hepatitis C virus (HCV) RNA by nucleic acid amplification techniques is the method of choice to differentiate between ongoing and pas t infection, and can be used to monitor the course of HCV infection. In thi s study, we evaluated the performance characteristics of a newly developed transcription-mediated amplification (TMA)-based assay, the VERSANT (R) HCV RNA qualitative assay, which was designed to qualitatively detect HCV RNA. Samples tested by the TMA assay included 100 HCV antibody negative sera; s erial dilutions of an HCV genotype 1 a panel; the WHO HCV RNA standard 76/7 90; an HCV genotyping panel; and 150 clinical specimens, including sera fro m patients who had received alpha interferon (IFN) treatment or liver trans plants. TMA test results were compared with the Cobas Amplicor (R) HCV poly merase chain reaction (PCR) assay. The analytical specificity of the HCV TM A assay was > 98%. No carry-over contaminations were observed. The assay de monstrated an analytical sensitivity of 100% at 41 HCV RNA copies/mL (genot ype 1 a panel) and 5 IU/mL (WHO standard), respectively. HCV genotypes and subtypes did not affect the results. Qualitative RNA detection by diagnosti c Amplicor (R) PCR and TMA was in agreement in > 97% of all 150 clinical sa mples tested. In our study, the TMA-based assay proved to be a specific and sensitive method for qualitative HCV RNA detection. The test may turn out to be an attractive alternative to already established techniques for HCV R NA amplification in routine clinical laboratories. J. Clin. Lab. Anal. 15:3 08-313, 2001. (C) 2001 Wiley-Liss, Inc.