副猪嗜血杆菌环介导等温扩增检测方法的建立及应用  

Establishment and Application of Loop-Mediated Isothermal Amplification Method for Detection of Haemophilus parasuis

作　　者：李璐璐[1] 骆延波[1] 张庆[1] 赵效南 任金瑞 胡明[1] 张印[1] 齐静[1] 刘玉庆[1] Li Lulu;Luo Yanbo;Zhang Qing;Zhao Xiaonan;Ren Jinrui;Hu Ming;Zhang Yin;Qi Jing;Liu Yuqing(Institute of Animal Science and Veterinary Medicine,Shandong Academy of Agricultural Sciences/Key Laboratory of Animal Disease Prevention and Breeding of Shandong Province,Jinan 250100,China;College of Life Sciences,Shandong Normal University,Jinan 250014,China)

机构地区：[1]山东省农业科学院畜牧兽医研究所/山东省畜禽疫病防控与繁育重点实验室,山东济南250100 [2]山东师范大学生命科学学院,山东济南250014

出　　处：《山东农业科学》2020年第10期163-167,共5页Shandong Agricultural Sciences

基　　金：国家重点研发计划项目(2016YFD0501301,2018YFD0500500);国家科技重大专项(2018ZX10733402-013);传染病预防控制国家重点实验室开放课题(2019SKLID317);山东省现代农业产业技术体系家禽创新团队环境控制岗位专家项目(SDAIT-13-011-09);山东省农业科学院农业科技创新工程项目(CXGC2018E10,CXGC2017A01)。

摘　　要：本试验针对副猪嗜血杆菌的16S rRNA基因设计环介导等温扩增(LAMP)引物,建立副猪嗜血杆菌LAMP方法,并进行特异性和灵敏度试验。结果显示,建立的LAMP检测方法特异性良好,与其它5种细菌无交叉反应,灵敏度为10^(-4)ng/μL。分别使用LAMP和PCR方法对30份临床采集样品进行检测,阳性检出率均为40%,符合率100%。本试验建立的LAMP检测方法不需要特殊的仪器设备,可用于副猪嗜血杆菌的鉴定。In order to establish a rapid and accurate method for detecting Haemophilus parasuis,loopmediated isothermal amplification(LAMP)primers were designed according to 16S rRNA gene of H.parasuis,and the specificity and sensitivity tests were performed.The results of specificity assay demonstrated that there was no cross reaction with the other five bacteria and the sensitivity was 10-4 ng/μL.The LAMP method and PCR assay were used for detecting 30 clinical samples,and the positive detection rates were all 40%and the coincidence rate was 100%.This LAMP assay could be used for the identification of H.parasuis without special equipment.

关 键 词：副猪嗜血杆菌 环介导等温扩增 快速检测 

分 类 号：S858.281.2-0[农业科学—临床兽医学]