沙门菌invA基因LAMP快速检测法的建立和初步应用  

Establishment and preliminary application of LAMP invA gene assay for rapid detection of Salmonella

作　　者：王敏雅[1] 徐明汉[2] 潘宏伟[3] 王志刚[4] 

机构地区：[1]浙江省台州市路桥区疾病预防控制中心,浙江台州318050 [2]浙江省温州医学院,浙江温州325035 [3]浙江省台州医院,浙江临海317000 [4]浙江省疾病预防控制中心,杭州310009

出　　处：《中国卫生检验杂志》2008年第10期1971-1973,1981,共4页Chinese Journal of Health Laboratory Technology

摘　　要：目的:建立适合基层检验部门使用的快速检测沙门菌LAMP法。方法:在65℃普通恒温水浴中、60 min扩增沙门菌invA基因片段,目测或电泳快速检测沙门菌。分别对环介导等温扩增(LAMP)反应影响较大的温度、Betaine浓度等进行优化,并对实验室保存的28种沙门菌不同血清型共34株和其他9种肠杆菌科细菌进行检测;对部分沙门菌进行污染血清直接LAMP模拟检测。结果:28种不同血清型的沙门菌LAMP检测都呈阳性,其它9种肠杆菌科细菌均为阴性;血清模拟实验与菌株直接DNA提取LAMP检测结果一致。结论:LAMP法能够快速、特异地检测沙门菌,且不需要昂贵的仪器,适合基层检验部门使用。Objective：To establish a method of loop-mediated isothermal amplification（LAMP） for the rapid detection of Salmonella in basic clinical and epidemilogicl laboratory.Methods：Optimizing the temperature and Betaine concentration which are sensitive for the LAMP.Estimating by eyeball or electrophoresis the amplification characteristics of the salmonella invA gene segment in thermostatic water-bath under 65℃ and 60 min.The 34 strains of 28 serotypes of Salmonella and 9 other kinds of enterbacteria were tested.Some human serums experimentally infected by Salmonella were directly amplified by LAMP method.Results：All LAMP reactions of 28 serotypes of salmonella were positive,and others are negative.The results of the infected serum directly amplified by LAMP were consisted with the results of common strain test.Conclusion：This LAMP-based assay is rapid and specific,and no expensive apparatus is needed.Therefore,it is especially suitable for basic clinical and epidemilogicl lab.

关 键 词：沙门菌 LAMP法 invA基因 快速测定 

分 类 号：R378.22[医药卫生—病原生物学]