多重PCR检测方法在鼠疫监测中的应用

Multiplex polymerase chain reaction in Yersinia pestis surveillance

目的 验证在鼠疫疫源地调查中应用多重PCR方法快速检测鼠疫菌的实用性。方法在14个省的17个鼠疫监测点采集标本2524份。选用针对鼠疫菌特异序列的引物,并加入内部对照模板．直接从鼠肝、脾等脏器标本中进行鼠疫核酸检测，与细菌分离培养结果相比较并进行统计学分析。结果 两种检测方法的符合率为92．67％。PCR方法阳性检出率为10．38％，较细菌培养阳性检出率（4．48％）高（Х^2=682．25，P〈0．01）。结论 多重PCR方法可应用于鼠疫监测中，比传统方法迅速、灵敏，但还有待于优化。

Objective To verify the feasibility of multiple locus polymerase chain reaction（PCR） to rapidly detect Yersinia pestis in the surveillance. Methods 2524 specimen were collected in 17 monitoring spots among 14 provinces. Nucleotide sequences were identified in the rat liver or spleen. Using the specific primers targeting at Yersinia pestis which was co-amplified by PCR with an internal control. The PCR results were compared with that of the isolation of bacteria and statistically analyzed. Results The accordant rate of two methods was 92.67%. The positive ratio revealed by PCR method was 10.38%, higher than that of 4.48% of bacteria culture（Х^2 = 682.25, P 〈 0.01 ）. Conclusion Faster and more sensitve than the traditional method, multiplex polymerase chain reaction can be used to identify Yersinia pestis, yet it needs further improvement.