Levels of viraemia in subjects with serological markers of past or chronic hepatitis B virus infection.

Subjects with serological markers for a past HBV infection may still have HBV DNA in their serum, but the levels of viraemia in such cases are not known. In the present study, of 63 consecutive HBsAg-negative, anti-HBc-positive serum samples with or without anti-HBs, 20 were HBV DNA-positive as analysed by a highly sensitive quantitative PCR, the Cobas Amplicor HBV Monitor test. However, all of these 20 samples had viraemia levels below 1000 copies/ml, compared with median viraemia levels of 10(8.6) and 10(4.3) copies/ml, respectively, in 98 HBeAg-positive and 124 HBeAg-negative HBsAg carriers. There was no difference in viraemia between subjects with anti-HBc alone compared with both anti-HBs and anti-HBc, nor between those with or without hepatitis C virus antibodies. The findings indicate that HBsAg-negative subjects may retain a low infectivity. Their risk for progressive liver damage is probably low, but this deserves further study.

Automated quantitative analysis of hepatitis B virus DNA by using the Cobas Amplicor HBV monitor test.

A highly sensitive method of quantitative analysis of hepatitis B virus (HBV) DNA in serum, the Cobas Amplicor HBV Monitor (Cobas-AM) test, was evaluated. Following a manual extraction of viral DNA, amplification, colorimetric detection, and quantitative determination are all automatically performed in the Cobas analyzer. Serially diluted samples with known HBV DNA concentrations were analyzed blindly. All samples with a virus concentration of 400 copies/ml and 83% of samples with a virus concentration of 100 copies/ml could be detected. A linear correlation between input HBV DNA and measured HBV DNA was seen in the range from 100 to 10(5) copies/ml. The mean coefficient of variation was 29.6% for all input levels and 18.9% for HBV DNA concentrations above 400 copies/ml. Samples with an HBV DNA level above 10(9) copies/ml could be reproducibly measured after predilution to 10(-4) or 10(-6) in negative serum; however, the level was underestimated if target DNA after dilution was still above the linear range of the assay. Quantitative results of the Cobas-AM test were interchangeable with measurements by the manual microwell plate version of Amplicor HBV Monitor (MWP-AM); the mean ratio for log Cobas-AM results/log MWP-AM results was 0.97 (standard error of the mean, 0.007) when serum samples from 153 chronic carriers were analyzed. The test should be of value for clinical assessment of chronic carriers and for monitoring the response to antiviral treatment. A limitation is the relatively narrow linear range of the assay, requiring predilution of high-titer (mainly hepatitis B e-antigen-positive) samples.