荧光定量分型PCR法在HBV-DNA检测中的应用

目的：探讨荧光定量分型PCR法在乙型肝炎病毒（HBV）-DNA检测中的作用，为HBV基因的分型提供参考。方法对120份 HBV-DNA阳性样本分别采用荧光定量分型PCR法和直接测序法检测，比较两种方法的检测效果。结果120份样本均被荧光定量分型PCR 法和直接测序法成功分型，荧光定量分型PCR 检出29（24．17％）例混合感染样本，直接测序法检出7（5．83％）例，前者检出率高于后者（χ^2＝15．817，P＝0．000）；两种方法的一致性较好（Kappa 值为81．67％）；检测结果不一致的样本经TA 克隆后，证实与荧光定量分型PCR 法的检测结果一致。结论荧光定量分型PCR 法是一种简便、快速高效的HBV 基因分型法，对基因型混合感染样本的检出率高于直接测序法，且正确率高。

Application of fluorescence quantitative PCR method for detection of HBV-DNA

Objective To investigate the role of fluorescent quantitative PCR method for detection of HBV-DNA,and to provide the reference for the HBV genotyping. Methods 120 HBV-DNA positive samples were detected by fluorescence quantitative PCR typing method and direct sequencing respectively,and the results of two methods were compared. Results 120 samples were suc- cessfully typed with fluorescent quantitative PCR method and direct sequencing method, 29 （24.17%） samples with mixed geno- types were detected by fluorescence quantitative PCR method,7 （5.83%） cases with mixed genotypes were detected by direct se- quencing method, the detection rate of the former was higher than that of the latter （χ^2=5. 817, P= 0. 000）. The consistency of two methods was better （Kappa= 81. 67 %0）. Conclusion Fluorescence quantitative PCR method is a simple, rapid and efficient HBV genotyping method, detection rate for the mixed genotypes of samples is higher than that of direct sequencing, and the accura- cy is high.