志贺氏菌属环介导恒温扩增快速检测方法的建立及初步应用  

Rapid detection of Shigella by loop-mediated isothermal amplification

作　　者：徐义刚 崔丽春[2] 张昕哲[3] 李苏龙 于恒纯 张子群 李丹丹[4] 

机构地区：[1]黑龙江出入境检验检疫局检验检疫技术中心,黑龙江哈尔滨150001 [2]东北林业大学,黑龙江哈尔滨150040 [3]东北农业大学,黑龙江哈尔滨150030 [4]海南出入境检验检疫局,海南海口570125

出　　处：《中国预防兽医学报》2010年第9期691-694,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家质检总局科技项目(2008IK162);黑龙江省博士后基金(LBH-Z09);黑龙江省青年基金(QC2009C90)

摘　　要：为建立快速检测志贺氏菌属的方法,本研究针对其ipaH基因,建立了志贺氏菌属环介导恒温护增(LAMP)快速检测方法。利用该方法对50株细菌进行检测,结果显示7株志贺氏菌均为阳性,其它菌均为阴性,表明该方法具有高度特异性。灵敏度试验显示,对志贺氏菌纯培养菌的检测灵敏度为28cfu/mL,对污染食品中志贺氏菌的检测灵敏度为35cfu/mL。利用该方法对195份涉及肉类、奶类、豆制品及人工污染样品进行检测,共检出29份阳性样品,与细菌分离鉴定检测结果一致。该LAMP方法操作简便、特异性强、灵敏度高且成本低廉,具有良好的应用前景。In this experiment,a rapid method for detecting Shigella was established with loop-mediated isothermal amplification （LAMP） using the ipaH gene of Shigella as target genes. The specificity of the LAMP method was confirmed by detecting fifty bacteria strains of thirty-two species where seven Shigella strains were tested positive. The assay had a detection limit of 28 cfu/mL for cultured Shigella and 35 cfu/mL in contaminated food. It was easy to perform and could be completed in 40 to 60 min utes. Tests on 195 samples （meat,milk,bean products and artificial contamination） using the LAMP method showed that 29 were Shigella positive,which was consistent with the bacterial culture method. This method could provide a sensitive,rapid and simple tool for the detection of Shigella.

关 键 词：志贺氏菌属 IPAH基因 环介导恒温扩增 快速检测 

分 类 号：S852.61[农业科学—基础兽医学]