环介导等温扩增技术结合横向流动试纸条检测猪链球菌2型的应用研究  

Rapid detection of Streptococcus suis serotype 2 with loop mediated isothermal amplification combined with a lateral flow dipstick

作　　者：张莉[1] 王利丽[1] 魏财文[2] 杨春蕾[1] 路超[1] 池晶晶[1] 鄢明华[1] ZHANG Li WANG Li-li WEI Cai-wen YANG Chun-lei LU Chao CHI Jing-jing YAN Ming-hua(Tianjin Veterinary Research Insititution, Tianjin 300384, China China Institute of Veterinary Drug Control, Beijing 100081, China)

机构地区：[1]天津市畜牧兽医研究所,天津300384 [2]中国兽医药品监察所,北京100081

出　　处：《中国人兽共患病学报》2016年第12期1077-1082,1090,共7页Chinese Journal of Zoonoses

基　　金：国家高技术研究发展计划资助项目(No.2012AA101605);天津市农业科技成果转化与推广项目(No.201301030);天津市农业科学院基金项目(No.16003)联合资助~~

摘　　要：目的建立一种猪链球菌2型的快速检测方法。方法以猪链球菌2型cps2J基因序列为检测靶标设计6条环介导等温扩增(LAMP)特异性引物,其中2条促环引物分别标记异硫氰酸荧光素(FITC)和地高辛(Dig),用横向流动试纸条(LFD)检测扩增产物,建立猪链球菌2型LAMP-LFD检测方法。结果所建立的检测方法能够特异性的检测出猪链球菌2型,与试验对照菌株不存在交叉反应,其最低检测限为76cfu/mL,在对100份样品的检测中,LAMP-LFD方法检测出14份阳性样品,培养法检测出12份阳性样品,LAMP-LFD方法检测阳性率略高于病原培养法,两者间的P>0.05无统计学差异。结论所建立LAMP-LFD检测方法具有特异性好、敏感性高、操作简单、快速等特点,适合于动物食品中猪链球菌2型的检测。The aim of this study is to establish a rapid detection method for Streptococcus suis serotype 2.Six specific primers of transcription loop method were designed based on the conservative regions of cps2 Jgene of S.suis serotype 2,two loop primers were respectively labeled by fluorescein isothiocyanate（FITC）and digoxin（Dig）.After amplification,the products of LAMP were detected by lateral flow devices（LFD）,then the LAMP-LFD was established.The assay was highly specific for the S.suis serotype 2.No cross-reaction was observed with any of the other pathogenic bacteria.The sensitivity of the assay was 76cfu/mL.For the 100 suspected samples,14 positive samples were identified by the LAMP-LFD method,while 12 positive samples were identified by culture method.The LAMP-LFD method had higher sensitivity than that of the culture method,but Pvalue0.05,that was not in significant level.The LAMP-LFD method established in this study was a simple and rapid operation with good sensitivity and specificity.It can be used as reliable method for animal produce detection of S.suis serotype2.

关 键 词：猪链球菌2型 cps2J基因 环介导等温扩增 横向流动试纸条 

分 类 号：R379[医药卫生—病原生物学]