摘要
目的用荧光定量聚合酶链反应(FQPCR)方法准确地定量检测血清中乙型肝炎病毒(HBV)的数量和免疫指标,以指导临床。方法设计合成了HBVFQPCR诊断试剂盒,以一种完全闭管式的PCR和荧光探针杂交技术相结合所产生的实时检测定量PCR方法,检测了532份临床血清标本。结果经FQPCR检测,158份HBV的s抗原、e抗原、c抗体(HBsAg、HBeAg、HBcAb)都阳性的标本,其血清标本HBVDNA也全部阳性,平均HBVDNA拷贝数为1.1×108/ml;98例HBsAg、HBeAb、HBcAb都阳性的标本,其阳性率为73%(71例),平均拷贝数为8.6×105/ml,而常规PCR只有33%的阳性率;67例HBsAb、HBeAb、HBcAb都阳性标本的平均拷贝数为2.3×105/ml。结论能够避免PCR后处理导致的假阳性污染,并实现准确定量。FQPCR可以检测HBV的真实感染和复制情况,对于乙型肝炎的临床诊断,治疗方案的选择和疗效考察有较大的指导意义。
Objective To study the relationship of HBV in
serum, syndrome of patients, and immune indexes by FQPCR method. Method Fluorescence
quantitative PCR (FQPCR), which combines PCR and fluorescence probe hybridization, was
used to measure DNA/RNA. Results A clinical diagnostic kit for hepatitis B virus (HBV) was
developed. 532 clinical serum samples were tested with this kit, using ELISA as contrast. In 158
HBsAg+/HBeAg+/HBcAb+ samples, FQPCR results were positive, with 1.1108/ml of HBV on
average. In 98 HBsAg+/HBeAb+/HBcAb+ samples, the average was 8.6105/ml with a positive
rate of 73%; normal PCR only reached 30%. In 67 HBsAb+/HBeAb+/HBcAb+ samples, the
amount was 2.3105/ml. Conclusion Intube monitoring eliminates PCR crosscontamination
which causes false positive, and realtime detection ensures accurate quantity. So FQPCR can
be used to monitor the true state of HBV infection and amplification.
基金
1998年国家技术创新项目资助