绵羊肺炎支原体LAMP检测方法的建立及初步应用  

Development and application of a loop-mediated isothermal amplification assay for detection of Mycoplasma ovipneumoniae

作　　者：张双翔[1] 周碧君[1,2] 程振涛[1,2] 文明[1,2] 王开功[1,2] 李泽民[1] 王慧[1] 崔亚兰[1] 覃岚[1] 

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病研究室,贵州贵阳550025

出　　处：《中国兽医学报》2013年第3期362-366,共5页Chinese Journal of Veterinary Science

基　　金：贵州省重大科技专项计划资助项目(黔科合重大专项字[2011]6009号);贵州省农业科技攻关资助项目(黔科合NY字[2011]3105号);贵州省自然科学技术基金资助项目(黔科合J字[2011]2332号);贵州大学引进人才科研资助项目(贵大人基合字[2010]35号);贵州大学2012年研究生创新基金资助项目(校基金农科[2012]034号)

摘　　要：针对绵羊肺炎支原体(Mo)16SrRNA基因设计4条环介导等温扩增(LAMP)引物,通过优化反应条件与体系,建立了Mo检测LAMP方法,并对其进行特异性、敏感性及临床应用性检测。结果显示,于60℃保温60min即可最优化检测Mo,且所建方法仅能特异扩增Mo,最低检出分子拷贝数为1.0×102copies/μL,具有良好的特异性和敏感性。对4份已知阳性病料进行LAMP检测均为阳性,表明所建LAMP可初步应用于临床检测,为羊Mo感染病例诊断提供了新方法。Four primers of Loop-mediated isothermal amplification（LAMP） were designed to es- tablish an assay for detecting Mycoplasma Ovipneumoniae （Mo）, which was rapid, easy and low cost. The LAMP was developed after optimizing the reaction condition and system. The specifici- ty,sensitivity and clinical application of the assay were also verified. The results showed that mo- lecular copy count detection of Mo could reached to 1. 0× 10^2copies/μL at 60℃ for 60 min by LAMP,with good specificity. As well, detecting to 4 known Mo positive goat lungs was whole positive by LAMP. The LAMP detection of Mo could be used in clinical diagnosis.

关 键 词：绵羊肺炎支原体 环介导等温扩增 建立 应用 

分 类 号：S852.62[农业科学—基础兽医学]