Comparison of two PCR assays for serum HBV DNA quantification

ZHANG Ji-wan; ZHOU Jian-li; ZHANG Xiao-fang; LUO Li-sha; WANG Hai-bin

Abstract：Objective To investigate the performance differences in detecting serum HBV DNA quantification between Roche COBAS TaqMan HBV assay and one-tube nested PCR assay. Methods The serum samples from 175 patients with hepatitis B were detected by Roche COBAS TaqMan HBV assay and one-tube nested PCR assay, with the jaundice, hemolytic and lipemic samples as interference samples, so as to analyze the correlation, consistency and interference performance of the two assays. A known sample with 2.24×109 IU/ml of HBV DNA was diluted by 1∶10 in negative serum in turn, and finally to an HBV DNA level of 2.24×10 IU/ml. The linear range and sensitivity of the two assays were analyzed. Results There was no significant difference in HBV DNA quantification of the 175 samples detected by the two assays. HBV DNA quantification in the jaundice, hemolytic and lipemic samples detected by the two assays was not significantly different. For samples with HBV DNA>1.70 ×108 IU/ml, Roche COBAS TaqMan HBV assay only showed HBV DNA level of >1.70×108 IU/ml, but HBV DNA could be accurately quantified without being diluted by one-tube nested PCR assay. For samples with HBV DNA<1.00×102 IU/ml, one-tube nested PCR assay could only detect the virus, but Roche COBAS TaqMan HBV assay had better stability and linearity. Conclusions The detection result of HBV DNA quantification by one-tube nested PCR assay is consistent with that by Roche COBAS TaqMan HBV assay, and one-tube nested PCR assay, as a timesaving, laborsaving, and cheap method for quantifying HBV DNA levels, is suitable for application in China.