荧光探针定量PCR在检测HBV-DNA的应用

Application of Fluorescence Quantitative PCR(FQ-PCR) in Detection of Hepatitis B Virus

目的 用荧光定量聚合酶链反应 (FQ PCR)方法准确地定量检测血清中的HBV DNA数量和免疫指标 ,以指导临床。方法 用FQ PCR方法和酶联免疫吸附试验 (ELISA)方法分别检测 184份临床血清标本的HBVDNA和免疫指标。结果 用 (ELISA)作对照 ,经FQ PCR检测 ,HBeAg(+)组 (82份 )的阳性率为 10 0 % ,HBV DNA阳性拷贝数范围在 10 5～ 10 10 /ml;HBeAb(+)组 (5 2份 )的阳性率为 5 5 8% ,HBV DNA阳性拷贝数范围在 10 4～ 10 7/ml;HBsAb(+)组 (2 0份 )的阳性率为 15 % ,HBV DNA阳性拷贝数范围在 10 4～ 10 5/ml;对照组 (30份 )的阳性率为 0。HBeAg(+)组血清中HBV DNA含量 (10 7 2± 1 13 )显著高于HBeAb(+)组 (10 5 8± 1 4)和HBsAb(+)组 (10 4 67± 0 58)。结论 FQ PCR检测HBV DNA具有较高的灵敏度和特异性 ,且能准确定量 ,FQ PCR可以检测HBV的真实感染和复制情况 ,对乙型肝炎临床诊断 ,治疗及疗后观察有重要意义。

Objective To detect the number HBV in serum by FQ PCR method.Methods The amount of HBV DNA in the serum of 184 patients were estimated by FQ PCR.The serum HBV immune markers of these patients were also tested with ELISA. Results ELISA indexes are taken for comparison. In 82 HBeAg+ samples, FQ PCR results are all positive, with a HBV range of 10 5 to 10 10 /ml.In 52 HBeAb+samples,the range is 10 4 to 10 7/ml with a positive rate of 55 8%.In 20 HBsAb+ samples,the range is 10 4 to 10 5/ml with a positive rate of 15%.The amount of HBV in HBeAg+samples(10 7 2±1 13 )is significantly higher than that in HBeAb+samples(10 5.8±1.4 ) and HBsAb+samples (10 4.67±0.58 )samples.Conclusions FQ PCR has very high sensitivity and specificity.It can be used for accurate measurement.FQ PCR can be for monitoring and prognosis of the HBV infection.