猪捷申病毒荧光RT-RAA检测方法的建立及应用  

Establishment and Application of Fluorescent RT-RAA Method for Detecting Porcine Teschovirus

作　　者：孙志华 林青 康龙滨 王隆柏 周伦江[2] 俞道进[1] 陈秋勇[1,2] SUN Zhi-hua;LIN Qing;KANG Long-bin;WANG Long-bai;ZHOU Lun-jiang;YU Dao-jin;CHEN Qiu-yong(University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare of Fujian Province,College of Animal Sciences,Fujian Agriculture and Forestry University,Fuzhou,Fujian,350002,China;Institute of Animal Husbandry and Veterinary Medicine of Fujian Academy of Agricultural Sciences/Engineering and Technology of Fujian Livestock and Poultry Disease Prevention Research Center,Fuzhou,Fujian,350013,China)

机构地区：[1]福建农林大学动物科学学院,中西兽医结合与动物保健福建省高校重点实验室,福建福州350002 [2]福建省农业科学院畜牧兽医研究所/福建省畜禽疫病防治工程技术研究中心,福建福州350013

出　　处：《动物医学进展》2025年第3期130-133,共4页Progress In Veterinary Medicine

基　　金：地方畜禽优异性状基因挖掘应用与重要疫病绿色防控技术研究(XTCXGC2021008);畜病防控科技创新团队(CXTD2021007-2)。

摘　　要：为快速检测猪捷申病毒(PTV),根据PTV 5′UTR基因保守序列设计特异性引物和探针,优化荧光逆转录重组酶介导等温扩增(RT-RAA)反应条件和体系,并通过特异性、敏感性、重复性试验和临床应用对RT-RAA进行评价。结果表明,在37℃恒温反应22 min即可完成对PTV核酸扩增,最低检出限度为30.1 copies/μL;与猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒(TGEV)、猪轮状病毒(PoRV)、猪德尔塔冠状病毒(PDCoV)均无交叉反应;重复性试验显示,组内和组间变异系数均小于5%;23份临床样品检测显示PTV阳性率为21.74%(5/23),检测结果与RT-qPCR一致。说明建立的PTV荧光RT-RAA检测方法具有简便、高效、稳定等优点,为PTV的快速检测和流行病学调查提供了技术手段。In order to establish a rapid test method of PTV,we designed specific primers and probes based on the conserved sequence of PTV 5′UTR gene,optimized the conditions and system of the reaction of reverse-transcription recombinase-aid amplification.In addition,the study evaluated the method by specificity,sensitivity,repeatability tests and clinical application.The resuts indicated that this method can be applied under the coditions of 37℃constant temperature reaction for 22 minutes,and the minimum detection limit was 30.1 copies/μL,no cross-reaction with porcine epidemic diarrhea virus,porcine transmissible gastroenteritis virus,porcine rotavirus and porcine deltacoronavirus;the repeatability test showed that the variation coefficient was less than 5%.Moreover,23 clinical samples showed PTV positive rate of 21.74%(5/23),and the test results were consistent with RT-qPCR.The resuts showed that the PTV fluorescence RT-RAA detection method was established which has the advantages of simplicity,high efficiency and stability,and provided new technical support for the rapid detection and epidemiological investigation of PTV.

关 键 词：猪捷申病毒 荧光逆转录重组酶介导等温扩增 检测方法 

分 类 号：S852.65[农业科学—基础兽医学]