2种PCR试剂盒定量检测血清HBV DNA含量的比较

Comparison of two PCR assays for serum HBV DNA quantification

目的探讨AMPLIPREP-COBAS TAQMAN法(罗氏COBAS法)和北京鑫诺美迪PCR-荧光探针"一管法"检测血清HBV DNA含量的性能差异。方法采用2种方法同步检测175例乙型肝炎患者血清样本,并将黄疸、溶血和脂血标本作为干扰样本,分析2种方法的相关性、一致性和抗干扰性。取一已知定量为2.24×109IU/ml的标本,用阴性血清做1+9(即1∶10)的稀释,并依次稀释至2.24×10 IU/ml,比较2种方法的线性范围和灵敏性的差异。结果 175份均有数值的标本中,2种试剂检测结果比较差异无统计学意义。2种试剂对黄疸、溶血和脂血标本的定量值影响不大。对于HBV DNA>1.70×108IU/ml的样本,罗氏COBAS法结果只显示>1.70×108IU/ml,而"一管法"无须稀释仍能准确定量;对于HBV DNA<1.00×102IU/ml的样本,"一管法"仅能检测到病毒,而罗氏COBAS法的稳定性和线性更好。结论 PCR-荧光探针"一管法"与进口罗氏COBAS法具有良好的一致性,且省时、省力,价格低廉,适合在我国推广应用。

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 2.24×10^9 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 ×10^8 IU/ml, Roche COBAS TaqMan HBV assay only showed HBV DNA level of 〉1.70×10^8 IU/ml, but HBV DNA could be accurately quantified without being diluted by one-tube nested PCR assay. For samples with HBV DNA〈1.00×10^2 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.