云南省玉龙县一起鼠间鼠疫的实验室判定  

Laboratory determination of a rat plague epidemic in Yulong county of Yunnan province

作　　者：王鹏[1] 张福新[2] 郭英[1] 谭红丽[1] 张正飞[2] 宋志忠[1] 

机构地区：[1]云南省地方病防治所云南省自然疫源性疾病防控技术重点实验室,云南大理671000 [2]丽江市疾病预防控制中心,云南丽江674100

出　　处：《中国卫生检验杂志》2015年第8期1105-1107,共3页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金(81160354);卫生部行业基金(201-202021);云南省应用基础研究(2011FB134)

摘　　要：目的对云南省玉龙县一起鼠间鼠疫进行实验室判定。方法取6份干燥自毙鼠标本的肌肉及骨骼,用生理盐水进行浸泡,然后取浸泡液进行免疫胶体金(GICA)及鼠疫反相间接血凝抑制试验(RIHA)检测;采用DNA提取试剂盒对以上6份标本的骨骼及肌肉进行了全DNA的提取;采用聚合酶链式反应(PCR)检测caF1及pla基因;采用实时荧光定量PCR(real^time PCR)检测caf1、pla及YPO0392基因;采用LB平板对肌肉及骨髓标本进行细菌分离培养。结果经GICA初筛,6份标本中4份为阳性;RIHA检测4份阳性标本的效价分别是1:320、1:640、1:1 280及1:640;4份免疫学阳性标本的普通PCR检测,其中3份标本为阳性,1份为阴性;实时荧光PCR检测4份免疫学阳性标本DNA,其中3份标本为阳性,1份为阴性;所有6份标本细菌培养皆阴性。结论判定玉龙县此次事件为鼠间鼠疫疫情,同时建立了一套对干燥标本进行鼠疫实验室诊断的技术流程。Objective To determine a rat plague epidemic in Yulong county of Yunnan province. Methods Muscle and bone from 6 dry mice of were soaked with saline, and the above immersion liquid was detected for GICA and RIHA. Total bone and muscles DNA of above 6 specimens were extracted using DNA extraction kit. PCR was used to detect cafl and pla, and real - time PCR was used to detect cafl, pla and YPO0392 genes. Muscle and bone marrow specimens were cultured for bacterial iso- lation by LB medium. Results After the primary GICA screening, 4 of 6 samples were positive. RIHA titers of 4 positive sam- ples were 1 ： 320, 1 ： 640, 1 ： 1 280 and 1 ： 640 respectively. For PCR and real - time PCR detecting, 3 of above 4 speci- mens were positive, while 1 of them was negative. Bacterial culture to all 6 samples was negative. Conclusion It was con- firmed that a rat plague epidemic occurred in Yulong county. Moreover, a set of process for detection plague to dry specimen was established.

关 键 词：鼠疫 实时荧光定量PCR 自毙鼠 玉龙县 

分 类 号：R516.8[医药卫生—内科学]