提高乙型肝炎病毒P基因区聚合酶链反应检测阳性率的策略

The strategy to increase the sensitivity of polymerase chain reaction in the detection of hepatitis B virus P gene

目的 提高乙型肝炎病毒 (HBV)DNAP基因区聚合酶链反应 (PCR)检测的阳性率。方法 采用优化PCR反应条件 ,降低退火温度以及采用 3′末端碱基游移兼并引物 ,减少引物与模板引物结合区的错配对PCR的影响等多种方法 ,对常规PCR检测HBVDNAP基因区阴性的病人血清 ,再进行PCR检测。结果 通过降低退火温度及减少 3′末端错配 ,PCR检测阳性率由 81 7% ( 6 7/ 82 )增加至 92 7% ( 76 / 82 ,P <0 0 5 )。结论 综合采用优化PCR反应条件 ,降低退火温度和采用 3′末端碱基游移兼并引物等方法 ,可提高基因高突变区PCR检测阳性率。

Objective To increase the sensitivity of polymerase chain reaction(PCR) in the detection of hepatitis B virus(HBV) P gene. Methods To reduce the blight of the mismatch between primers and their targets, some methods were applied such as choosing primers, modifying primers, optimizing PCR conditions, decreasing the anneal temperature etc. Results The sensitivity of PCR in the detection of HBV P gene increased from 81。7%(67/82) to 92。7%(76/82, P<0。05) by using methods of amending primers and decreasing the anneal temperature. Conclusion Comprehensive methods including choosing primers, modifying primers, optimizing PCR conditions and decreasing the anneal temperature should be used to increase the sensitivity of PCR in the detection of variable region of HBV DNA。