多重PCR检测方法在鼠疫监测中的应用  

Multiplex polymerase chain reaction in Yersinia pestis surveillance

作　　者：张志凯[1] 海荣[1] 张忠兵[2] 赵斌[3] 史献明[4] 石国祥[5] 秦迎旭[6] 魏绍振[7] 黄富 唐永姣[9] 夏连续[1] 蔡虹[1] 张建华[1] 马凤琴[1] 张恩民[1] 俞东征[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室,北京102206 [2]内蒙古自治区地方病防治研究中心 [3]中国疾病预防控制中心鼠疫布氏菌病预防控制基地 [4]河北省鼠疫防治所 [5]浙江省疾病预防控制中心 [6]宁夏回族自治区疾病预防控制中心 [7]青海省地方病预防控制所 [8]内蒙古自治区乌兰察布盟地方病防治中心 [9]甘肃省疾病预防控制中心

出　　处：《中国地方病学杂志》2007年第5期475-477,共3页Chinese Jouranl of Endemiology

基　　金：国家“十五”攻关课题（2004BA718807-01）

摘　　要：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.

关 键 词：耶尔森菌 鼠疫 聚合酶链反应 内部对照 

分 类 号：R686[医药卫生—骨科学]