A multicenter study evaluation of the Digene Hybrid Capture II signal amplification technique for detection of hepatitis B virus DNA in serum samplesand testing of EUROHEP standards

We have evaluated the new Digene Hybrid Capture If HBV DNA Test (HCII HBV), which is a 96-well microtiter plate-based signal amplification assay. This test uses hybrid capture technology that specifically detects RNA-DNA hybr ids. HCII HBV is able to quantify hepatitis B virus (HBV) DNA at between 1. 4 x 10(5) and 1.7 x 10(9) HBV copies per mi in a standard format. By using a modified sample preparation method, which allows the input of 30-fold mor e serum for an ultrasensitive format, the sensitivity of the assay can be i ncreased reproducibly to approximately 8,000 copies of HBV per ml, By using a combination of these two formats, the assay can quantify over a total ra nge of 6 logs. In our multicenter evaluation study, the mean laboratory-to- laboratory coefficients of variation were 22, 7, and 12% at the three sites , respectively, with a combined specificity of 98.4%. The linearities of bo th the standard test and the ultrasensitive test were excellent, with Spear man correlation coefficients of 0.997 and 0.999, respectively. Furthermore, the intra-assay reproducibility for the standard assay gave coefficients o f variation of from 13 to 33, 9 to 21, and 3 to 8% at the three sites, resp ectively, HCII HBV was shown to be genotype independent when the EUROHEP st andards for genotypes A and D were used. This assay allows the accurate mea surement of HBV DNA levels in serum and can be clinically used for the moni toring of responses to antiviral agents for patients chronically infected w ith HBV.