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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 quantitatively detect the amount of hepatitis B virus (HBV) and immune parameters in serum by fluorescence quantitative polymerase chain reaction (FQPCR) to guide the clinical practice. Methods The HBV FQPCR diagnostic kit was designed and synthesized. A total of 532 clinical serum samples were detected by a real-time PCR assay generated by a completely closed PCR and fluorescent probe hybridization. The results of FQ PCR detection, 158 copies of HBV s antigen, e antigen, c antibody (HBsAg, HBeAg, HBcAb) positive samples, the serum samples were all HBVDNA positive, the average copy number of HBV DNA was 1.1 × 108 / ml; the positive rate of 98 cases of HBsAg, HBeAb, HBcAb positive specimens was 73% (71 cases), the average copy number was 8.6 × 105 / ml, while the conventional PCR only 33% positive rate; 67 cases of HBsAb The average copy number of HBeAb and HBcAb positive specimens was 2.3 × 105 / ml. Conclusion The false positives caused by PCR can be avoided and accurate quantification can be achieved. FQ PCR can detect the true infection and replication of HBV, for the clinical diagnosis of hepatitis B, the choice of treatment options and curative effect of a great guiding significance.