猪轮状病毒TaqMan荧光定量检测方法的建立及初步应用  

Establishment and preliminary application of TaqMan fluorescence quantitative detection method for porcine rotavirus

作　　者：林正丹 涂军 詹存林 孙秀秀 冯贺龙 刘茜 胡薛英[1] 谷长勤[1] 张万坡[1] 陶攀 陈品[1] 余腾 钱平[1] 程国富[1] LIN Zhengdan;TU Jun;ZHAN Cunlin;SUN Xiuxiu;FENG Helong;LIU Xi;HU Xueying;GU Changqin;ZHANG Wanpo;TAO Pan;CHEN Pin;YU Teng;QIAN Ping;CHENG Guofu(College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;Guangxi Yangxiang Co.,Ltd.,Guigang,Guangxi 537000,China)

机构地区：[1]华中农业大学动物医学学院,湖北武汉430070 [2]广西扬翔股份有限公司,广西贵港537000

出　　处：《中国兽医学报》2024年第2期244-248,共5页Chinese Journal of Veterinary Science

基　　金：中央高校基本科研业务费专项基金资助项目(140422008);扬翔股份有限公司重大创新变革基金资助项目(JYKJ-R&D2022-01)。

摘　　要：为了对早期猪轮状病毒(porcine rotavirus,PoRV)感染情况进行监测,根据GenBank中已有的A型PoRV VP6序列,设计2对引物和荧光定量探针,建立了TaqMan荧光定量RT-PCR方法,并对该方法特异性、灵敏性和重复性进行评价。结果显示所建立的TaqMan荧光定量RT-PCR方法特异性强、重复性好、灵敏度高,可检测到101拷贝/μL。经对2366份腹泻仔猪的粪便样品进行检测,所建立的方法检出率(5.92%,140/2366)高于常规RT-PCR(4.95%,117/2366),两者的符合率为98.58%。结果表明,本研究建立的PoRV TaqMan荧光定量RT-PCR方法可用于猪轮状病毒病的临床检测。In order to monitor the early porcine rotavirus infection,according to the existing porcine rotavirus A VP6 sequence in GenBank,two pairs of primers and fluorescence probes were designed,and the TaqMan real-time RT-PCR method was established,and the specificity,sensitivity and repeatability of the method were evaluated.The results showed that the established TaqMan real-time RT-PCR method had strong specificity,good repeatability and high sensitivity,and could detect 10~1 copies/μL.After testing the fecal samples of 2366 diarrheal piglets,the detection rate of the established method(5.92%,140/2366)was higher than that of conventional RT-PCR(4.95%,117/2366),and the compliance rate of the two was 98.58%.The results showed that the TaqMan fluorescence RT-PCR method established in this study can be used for the clinical detection of porcine rotavirus disease.

关 键 词：猪轮状病毒 TAQMAN探针 荧光定量RT-PCR方法 

分 类 号：S858.28[农业科学—临床兽医学]