猪人住肉孢子虫环介导等温核酸扩增检测技术的建立与应用  

Establishment and application of a loop-mediated isothermal amplification assay for detection of Sarcocystis suihominis in pigs

作　　者：刘丹丹[1,2] 禚振男 候照峰 陶建平 许金俊[1,2] LIU Dan-dan;ZHUO Zhen-nan;HOU Zhao-feng;TAO Jian-ping;XU Jin-jun(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou University,Yangzhou 225009,China;Animal Husbandry and Veterinary Bureau of Zhucheng City,Weifang 262200,China)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏高校动物重要疫病与人兽共患病防控协同创新中心,扬州大学,江苏扬州225009 [3]诸城市畜牧兽医局,山东潍坊262200

出　　处：《中国兽医科学》2022年第9期1102-1107,共6页Chinese Veterinary Science

基　　金：扬州大学“青蓝工程”资助项目(2018年度);江苏高校优势学科建设二期工程资助项目;江苏省人兽共患病学重点实验室开放课题(R1408);国家重点研发计划项目(2017YFD0500-402)。

摘　　要：为建立快速有效的猪人住肉孢子虫检测技术,保证猪及其肉产品的公共卫生安全,以感染猪人住肉孢子虫阳性猪的肌肉组织DNA为模板,扩增18S rRNA基因片段,针对该基因高保守区设计并筛选出1对外引物F3、B3与1对内引物FIP、BIP,建立环介导等温核酸扩增(LAMP)检测技术并初步应用。结果显示,建立的LAMP检测技术特异性强,与常见的猪寄生虫无交叉反应;且敏感性好,对虫体18S rRNA基因的最低检出限为10 copies。临床实际应用结果显示该检测方法效果良好,为临床上猪人住肉孢子虫的检测提供了一种简便快速的方法,值得进一步推广。The aim of this study was to establish an effective and rapid detection method,named loop-mediated isothermal amplification(LAMP),for the detection of Sarcocystis suihominis to protect the pigs and the public healthy safety of pork products.An LAMP method was established by using 18S r RNA gene of S.suihominis with outer primers F3 and B3,and inner primers FIP and BIP.And then,it was used to detect S.suihominis from DNA template extracted from muscle tissue of pigs infected with S.suihominis.As results,this LAMP method showed high specificity and sensitivity to detect S.suihominis with 10 copies of DNA at least,and had no cross reaction with other common parasites in pigs.It also showed expected effect in detecting clinical samples.In conclusion,the LAMP method established in this study may be a useful tool for rapid detection and identification of S.suihominis in pigs,and it should be promoted.

关 键 词：猪人住肉孢子虫 18S rRNA 环介导等温核酸扩增 

分 类 号：S852.723[农业科学—基础兽医学]