猫疱疹病毒Ⅰ型RAA检测方法的建立与应用  

Establishment and application of RAA detection method of feline herpesvirus-Ⅰ

作　　者：王颖 戚睿斌 崔赛赛 杨育鹏 冯可昕 刘家森[2] 姜骞[2] 杨鸣发 康洪涛 曲连东[1,2] WANG Ying;QI Rui-bin;CUI Sai-sai;YANG Yu-peng;FENG Ke-xin;LIU Jia-sen;JIANG Qian;YANG Ming-fa;KANG Hong-tao;QU Lian-dong(College of Veterinary Medicine,Northeast A gricultural University,Harbin 150030,China;State Key Laboratory of Veterinary Biotechnology/Harbin Veterinary Research Institute,China Academy of A gricultural Sciences,Harbin 150069,China)

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨宾150030 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150069

出　　处：《中国兽医科学》2023年第1期29-33,共5页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2019YFC1200400);“一带一路”沿线口岸病媒生物监测、快速检测和溯源技术研究(2019HK125)。

摘　　要：为建立一种快速检测猫疱疹病毒Ⅰ型(FHV-1)的方法,本研究根据GenBank上登录的FHV-1 UL55序列设计特异性探针及引物,筛选最佳引物对,建立了FHV-1的重组酶介导等温扩增(RAA)检测方法。该方法在39℃反应20 min即可快速、特异性地检测FHV-1。灵敏性试验结果显示,RAA最低检测限为7.60×101copies/μL,比普通PCR最低检测限高100倍。特异性试验结果显示,该RAA方法与猫杯状病毒、猫传染性腹膜炎病毒、猫细小病毒无交叉反应。对34份临床样本进行检测,其中13份为阳性,阳性检出率为38.23%。综上所述,本研究成功建立了猫疱疹病毒Ⅰ型RAA检测方法。In order to establish a method for rapid detection of feline herpesvirus-Ⅰ,this study designed specific probes and primers according to the FHV-1 UL55 sequence on GenBank,screened the best primer pairs,and established a recombinase-aided amplification of FHV.The method can rapidly and specifically detect FHV-1 at 39℃for 20 minutes.The sensitivity test results showed that the minimum detection limit of RAA was 7.60×10~1 copies/μL,which was 100 times higher than the minimum detection limit of ordinary PCR.The specificity test results showed that the RAA method did not cross-react with feline calicivirus,feline infectious peritonitis virus and feline parvovirus.We tested 34 clinical samples,13 of which were positive.The positive detection rate was 38.23%.In conclusion,this study successfully established a RAA detection method for FHV-1.

关 键 词：猫疱疹病毒 重组酶介导等温扩增 快速检测方法 

分 类 号：S852.659.6[农业科学—基础兽医学]