猫泛白细胞减少症病毒环介导等温扩增检测方法的建立  

Establishment of Loop-mediated Isothermal Amplification for Detection of Feline Panleukopenia Virus

作　　者：马芹[1] 闫文卓 周洁 高诚 刘铁龙 赵丽丽[1] 陈洪岩[1] 陆涛峰[1] MA Qin;YAN Wen-zhuo;ZHOU Jie;GAO Cheng;LIU Tie-long;ZHAO Li-li;CHEN Hong-yan;LU Tao-feng(Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 100193,China;Shanghai Laboratory Animal Research Center,Shanghai 201203,China;Jiyuan Animal Health Inspection,Jiyuan 454650,China)

机构地区：[1]中国农业科学院哈尔滨兽医研究所,哈尔滨100193 [2]上海实验动物研究中心,上海201203 [3]河南省济源市动物卫生监督所,济源454650

出　　处：《实验动物与比较医学》2019年第1期34-38,共5页Laboratory Animal and Comparative Medicine

基　　金：上海市科委科研计划项目(16140900500)

摘　　要：目的建立一种快速、简便检测猫泛白细胞减少症病毒(FPV)的环介导等温扩增(LAMP)方法。方法针对Genbank中FPV基因组设计了4条LAMP扩增引物,通过优化反应温度和反应时间,确定高效的LAMP扩增条件。结果建立的LAMP方法在65℃水浴条件下反应60min可对FPV进行高效扩增,检测限量为5.01×10^2拷贝/μL,比常规PCR检测的敏感性高10倍,且与猫疱疹病毒1型(FHV-1),犬细小病毒(CPV)及犬腺病毒(CAV)均无交叉反应。结论建立的LAMP方法设备要求低、特异性好、灵敏度高,适用于FPV临床样品的快速检测。Objective To establish a rapid,convenient detection method of feline panleukopenia virus(FPV)by loop-mediated isothermal amplification(LAMP).Methods Two pairs of primers were designed according to the conserved sequences of FPV in Genbank.The efficient LAMP amplification conditions were determined by optimizing temperature and time.Results The results showed a highly efficient amplification for FPV nucleic acid which performed at 65℃for 60 min.It also showed a high sensibility with a detection limit of 5.01×10^2 copies/μL,10 times higher than the conventional PCR detection in the sensitivity.There was no cross reaction with feline herpesvirus type l(FHV-l),canine parvovirus(CPV)and vanine adenovirus(CAV).Conclusion The established LAMP method is good in duplication,stability,specificity,and sensitivity.This method can be used for the fast detection of FPV nucleic acid in clinical samples from cats.

关 键 词：猫泛白细胞减少症病毒(FPV) 环介导等温扩增(LAMP) 检测方法 

分 类 号：Q95-33[生物学—动物学]