乙型肝炎病毒基因分型的双通道荧光PCR检测及其在未成年乙型肝炎病毒感染者中的临床应用

Detection of HBV genotypes by dual-color qPCR and its clinical application for juveniles infected with HBV

目的评价双通道荧光PCR(d-qPCR)法检测乙型肝炎病毒(HBV)基因型的方法学性能,并在未成年HBV感染者中进行临床应用。方法对基于d-qPCR的HBV基因型检测方法进行性能验证,并将该方法用于未成年HBV感染者的基因型检测,分析其临床意义。结果用于HBV基因分型的dd-qPCR法能检测B型、C型和D型,准确度为100%;最低检测限为1×10^(3 )IU/ml,批内和批间精密度高,抗干扰能力强,特异性高。在本地区200例未成年慢性HBV型携带者基因型中,以B型和C型为主,B型、C型和D型分别占34.0%、65.5%和0.5%,未发现混合型和不能分型的标本。B型和C型HBV DNA定量、HBeAg阳性率差异无统计学意义(P>0.05),肝功能异常率(ALT>50 U/L)差异有统计学意义(P<0.05)。结论 d-qPCR法检测HBV基因型具有分型全面、性能可靠和操作简便等优点。本院就诊的HBV C型未成年HBV感染者存在较高比例的肝功能损害,临床应结合HBV基因分型结果,尽早进行治疗干预。

Objective To evaluate the method of HBV genotypes by dual-color qPCR(d-qPCR)and apply it for the juveniles infected with HBV.Methods The performance of HBV genotype detection by d-qPCR was validated.The d-qPCR was applied to detect the juveniles infected with HBV and its clinical significance was analyzed.Results d-qPCR for HBV genotyping can be used to detect type B,type C and type D,and the accuracy was 100%.The limit of detection reaches 1×10^3 IU/ml.The method also had high precision of within-run and between-run.Moreover,the assay had a strong anti-jamming ability and high specificity.The major HBV genotypes in this region included type B and type C in 200 cases of HBV infection,and the proportion of type B,type C and type D was 34.0%,65.5% and 0.5%,respectively.However,no mixed and undivided cases were found.There was no statistical significance on the difference in the HBV DNA load and positive rate of HBeAg between type B and type C(P>0.05),while the difference in abnormal rate of liver function(ALT>50 U/L)was statistically significant(P<0.05).Conclusion d-qPCR for HBV genotyping has the advantages of more subtypes detection,reliable performance and easy operation.There is a certain proportion of liver function damage in juveniles infected with HBV type C.Therefore,combined with the results of HBV genotyping,the juveniles infected with HBV should be monitored and appropriate treatment measures should be applied as soon as possible.