隐孢子虫LAMP检测方法的建立及其在牦牛中的应用  

Establishment of LAMP Method for Detecting Cryptosporidium and Its Application in Yaks

作　　者：王丹[1,2,3] 任玫 吴飞 常建军 李璐瑶[1] 康明 林青[1,2,3] WANG Dan;REN Mei;WU Fei;CHANG Jian-jun;LI Lu-yao;KANG Ming;LIN Qing(College of Veterinary Medicine,Northwest A g~ F University,Yangling,Shaanxi,712100,China;College of Agriculture and Animal Husbandry,Qinghai University,Xining,Qinghai,810016,China;State Key Laboratory of Plateau Ecology and Agriculture,Qinghai University,Xining,Qinghai,810016,China 4.College of Animal Sciences,Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine,Zhejiang University,Hangzhou,Zhejiang,310058,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]青海大学农牧学院,青海西宁810016 [3]青海大学省部共建三江源生态与高原农牧业国家重点实验室,青海西宁810016 [4]浙江大学动物科学学院/浙江省动物预防医学重点实验室,浙江杭州310058

出　　处：《动物医学进展》2018年第10期13-17,共5页Progress In Veterinary Medicine

基　　金：青海省科技厅项目(2016-ZJ-754;2016-ZJ-Y01);青海大学省部共建三江源生态与高原农牧业国家重点实验室自主课题项目(2017-ZZ-08)

摘　　要：为了探索和建立隐孢子虫LAMP检测方法,以便能快速、简便检测牦牛隐孢子虫的感染情况,基于GenBank中牦牛源隐孢子虫18SrRNA基因序列,设计1组LAMP引物,优化并建立LAMP反应体系。对所建立的LAMP方法进行特异性、重复性、敏感性试验,并用该方法对牦牛的临床样本进行检测。结果显示,建立的LAMP方法特异性、重复性良好,其敏感性是套式PCR的8倍。LAMP方法与套式PCR临床检出率对比,符合率100%。表明成功建立了隐孢子虫的LAMP检测方法,该方法特异性和灵敏性良好,可以应用于牦牛粪便临床样本的检测。In order to detect Cryptosporidium infected yaks quickly and easily,the LAMP method was explored and established.A series of LAMP primers were designed based on the sequences of 18 S rRNA gene of Cryptosporidium of yaks in GenBank to optimize and establish a LAMP reaction system.The specificity, repeatability and sensitivity tests were performed on establishing the LAMP method, which was used to detect the clinical samples of yaks.The results showed that the LAMP method was specific and reproducible, and the sensitivity was 8 times than that of nested PCR method.The LAMP method was compared with nested PCR in clinical detection of yak samples,and the coincidence rate was 100%.The LAMP method for detecting yak Cryptosporidium was successfully established with good specificity and sensitivity, which can be applied to clinical detection.

关 键 词：牦牛 隐孢子虫 环介导等温扩增 18S RRNA基因 

分 类 号：S852.723[农业科学—基础兽医学] Q789[农业科学—兽医学]