聚合酶链反应荧光偏振技术在乙型肝炎病毒DNA检测中的应用及价值

Detecting HBV DNA in serum by PCR-FP

目的 探讨聚合酶链反应荧光偏振技术 (PCR FP)检测乙型肝炎病毒 (HBV)DNA的临床应用价值。方法 将PCR扩增得到的HBVDNA特定片段 ,与荧光素标记探针和PCR产物中的一条DNA链杂交 ,形成特异杂交体 ,用荧光偏振光检测仪检测特异杂交体 ,最终判断HBVDNA是否存在。并对对 10 2份乙型肝炎患者和 30份非乙型肝炎患者血清进行检测结果比较。结果 PCR FP检测HBVDNA的最低检测限为 1× 10 1拷贝 ,在 4 0份乙型肝炎表面抗原 (HBsAg)、乙型肝炎e抗体 (HBeAg)和抗乙型肝炎核心抗原 (抗HBc)均阳性患者血清标本中检出HBVDNA阳性率为 10 0 % ;33份HBsAg、抗乙型肝炎e抗体 (抗Hbe)和抗HBc均阳性患者血清HBVDNA阳性率为 81 8% ;2 9份HBsAg和抗HBc阳性患者血清HBVDNA阳性率为 72 4 % ;14份乙型肝炎患者和 30份非乙型肝炎患者血清均为阴性。方法的灵敏性和特异性分别为 86 %和 10 0 %。结论 PCR FP操作简单、灵敏、特异 ,适用于临床HBV基因检测。

Objective To evaluate the clinical application value of PCR-FP for detecting HBV DNA in serum. Methods Specific segment of HBV DNA was amplified by PCR, Fluorescence-labeled probes were hybridized specifically with the products of PCR. Then the hybridized products were detected by FP instrument to identify HBV DNA.Results The lowest detecting limit of PCR-FP was 1×10 1 copy HBV DNA. The positive rates for 40 serum samples with HBsAg(+),HBeAg(+),anti-HBc(+)were 100%(40/40), 33 serum samples with HBsAg(+),anti-HBe(+),anti-HBc(+)were 81.8%(27/33),29 serum samples with HBsAg(+)、anti-HBc(+) were 72.4%(21/29). The rest 30 normal serum were negative. The sensitivity and the specificity were 86% and 100% respectively. Conclusion The method of PCR-FP was simple, rapid, sensitive , specific, and could be used for detecting pathogen DNA in clinic.