HBV基因变异对荧光定量PCR系统检测HBV-DNA准确性的影响

Influence of HBV gene mutation on the accuracy of HBV-DNA detection by fluorescence quantitative PCR system

目的探讨乙型肝炎病毒(HBV)基因型和病毒变异对Roche公司COBAS AmpliPrep/COBAS TaqMan 48系统和荧光定量PCR系统检测HBV-DNA的影响。方法收集医院肝病中心2014年3月-2015年10月HBV感染患者100例,明确HBV基因型和变异情况,同时采用Roche公司COBAS AmpliPrep/COBAS TaqMan48系统和荧光定量PCR系统检测HBV-DNA,比较针对不同HBV基因型和病毒变异时两种方法检测结果的差异和相关性。结果两种方法检测B型、C型患者的病毒载量差异均无统计学意义,对未发生病毒变异的患者,两种方法的检测结果差异无统计学意义;对发生病毒变异的患者,两种方法的检测结果差异有统计学意义(t=2.29,P=0.028);两种方法检测B、C型HBV-DNA时的相关系数r分别为0.82、0.86,差异无统计学意义;两种方法检测未发生病毒变异患者HBV-DNA的相关系数r为0.91,显著高于发生病毒变异时的相关系数(r=0.70)(Z=3.13,P<0.01)。结论对于不同的HBV基因型,Roche公司COBAS AmpliPrep/COBAS TaqMan系统和国产荧光定量PCR系统在检测HBV-DNA方面有较好的相关性,但HBV基因变异可能会影响后者的准确性。

OBJECTIVE To investigate the influence of HBV genotypes and viral variation on HBV DNA detection by Roche COBAS AmpliPrep/COBAS TaqMan 48 system and domestic fluorescence quantitative PCR system.METHODS A total of 100 patients with HBV infection in liver disease center from Mar.2014 to Oct.2015 were enrolled.The HBV genotype and variation of all patients were detected,and HBV DNA loads were detected using Roche COBAS AmpliPrep/COBAS TaqMan 48 system and domestic fluorescence quantitative PCR system at the same time.The differences and correlations between the results of the two methods for different HBV genotypes and viral variation were compared.RESULTS The differences of viral loads by the two methods in genotype B and C were not significant.The results of the two methods in patients without virus mutation had no significant difference,but the differences between the two methods for patients with virus mutation were significant（t=2.29,P=0.028）.The correlation coefficients of the two methods for HBV DNA viral load detection in genotype B and C patients were 0.82 and 0.86,and the difference was not significant.The correlation coefficient of the two method for HBV DNA viral load detection in patients without virus mutation was 0.91,which was significantly higher than that with virus mutation（r=0.70）（Z=3.13,P〈0.01）.CONCLUSION For different HBV genotypes,Roche COBAS AmpliPrep/COBAS TaqMan 48 system and domestic fluorescence quantitative PCR system have a good correlation in terms of detection of HBV DNA,but HBV gene mutation may affect the accuracy of domestic fluorescence quantitative PCR system.