塞内卡谷病毒荧光RT-RPA快速检测方法的建立  

Development of a real-time RT-RPA assay for the rapid detection of Seneca valley virus

作　　者：林彦星[1] 曹琛福[1] 花群俊 黄超华[1] 杨俊兴[1] 徐铮[3] 史卫军[1] 阮周曦[1] 花群义[1] LIN Yan-xing;CAO Chen-fu;HUA Qun-jun;HUANG Chao-hua;YANG Jun-xing;XU Zheng;SHI Wei-jun;RUAN Zhou-xi;HUA Qun-yi(Inspection and Quarantine Center for Animals and Plants,Shenzhen Entry-exit Inspection and Quarantine Bureau,Shenzhen 518045,China;W uding Center for Animal Disease Control and Prevention,Wuding 651601,China;College of Animal Science,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518045 [2]武定县动物疫病预防控制中心,云南武定651601 [3]华南农业大学动物科学学院,广东广州510642

出　　处：《中国兽医科学》2018年第12期1497-1502,共6页Chinese Veterinary Science

基　　金：国家“十三五”重点研发计划项目(2016YFD0500708,2017YFD0501805)

摘　　要：本研究利用重组酶聚合酶扩增（RPA）技术建立快速检测塞内卡谷病毒（SVV）的实时荧光RT-RPA方法。试验结果表明,该方法特异性强,能准确检测SVV核酸,与水泡性口炎病毒（VSV-IND和VSV-NJ）、猪水泡病病毒（SVDV）、口蹄疫病毒（FMDV）、猪瘟病毒（CSFV）和猪繁殖与呼吸综合征病毒（PRRSV）等灭活抗原核酸无交叉反应;灵敏度高,本试验中最低可检测SVV核酸浓度为1.35×10^（-3）g/mL;简便快速,反应时间小于20 min;重复性好。利用所建立方法对116份临床样品进行检测,结果与实时荧光RT-PCR一致。本研究为SVV防控提供了一种快速检测的新方法,可用于基层实验室对SVV的快速检测。In this study,a real-time fluorescent reverse transcription-recombinase polymerase amplification（RT-RPA） assay was developed to detect the Seneca valley virus（SVV） using primers and probes specific for the gene of SVV.SVV was detected specifically by the assay without cross-reactions with the other inactivated virus antigens,including VSV-IND,VSV-NJ,SVDV,FMDV,CSFV or PRRSV RNA.The RT-RPA assay was rapid（20 min） and sensitive.The limit of detection of RT-RPA for SVV RNA was 1.35×10^（-3） g/mL in this test.A total of 116 clinical samples were tested by the RT-RPA assay and the reference method,and the test results were consistent between the developed RT-RPA and the reference method.In conclusion,the RT-RPA assay can be used for rapid detection of SVV in primary laboratories.

关 键 词：塞内卡病毒病 塞内卡谷病毒 反转录重组酶聚合酶扩增(RT-RPA) 检测 

分 类 号：S852.659.5[农业科学—基础兽医学]