猪流行性腹泻病毒SYBR Green I荧光定量RT-PCR检测方法的建立与应用  

Establishment and Application of SYBR Green I Fluorescent Quantitative RT-PCR for Detecting Porcine Epidemic Diarrhea Virus

作　　者：陈浩 鞠永政 王文文[2] 尹明荣 李阳 王一新[2] Chen Hao;Ju Yongzheng;Wang Wenwen;Yin Mingrong;Li Yang;Wang Yixin(College of Agricultural Science and Technology,Shandong Agriculture and Engineering University,Jinan,Shandong 250100,China;College of Animal Science and Technology,Shandong Agricultural University,Tai'an,Shandong 271018,China;China Animal Health and Epidemiology Center,Qingdao,Shandong 266032,China)

机构地区：[1]山东农业工程学院农业科技学院,山东济南250100 [2]山东农业大学动物科技学院,山东泰安271018 [3]中国动物卫生与流行病学中心,山东青岛266032

出　　处：《中国动物检疫》2022年第9期110-114,共5页China Animal Health Inspection

基　　金：三亚崖州湾科技城管理局科研项目(SKJC-2020-02-007);山东省农业良种工程项目(2021LZGC001)。

摘　　要：为快速、准确、灵敏检测猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV),以PEDV N基因为靶标设计引物,建立了一种PEDV SYBR Green I荧光定量RT-PCR检测方法。该方法最低可以检测到10个拷贝的病毒核酸,与猪传染性胃肠炎病毒、猪δ冠状病毒和猪轮状病毒等胃肠道病毒均无交叉反应,批内、批间重复试验的变异系数均在2%以内,表明该方法具有较好的敏感性、特异性和重复性。运用该方法对2018—2019年采集自山东省不同地区猪场的161份临床样品进行检测,检测结果与RT-PCR方法完全一致。以上结果说明,本试验所建立的SYBR Green I实时荧光定量RT-PCR检测方法为PEDV的快速检测提供了良好工具和技术支持。In order to develop a rapid,accurate and sensitive method for detecting porcine epidemic diarrhea virus(PEDV),specific primers were designed by taking PEDV N gene as a target,and a SYBR Green I fluorescent quantitative RT-PCR assay for PEDV was established,which could detect at least 10 copies of viral nucleic acid,without cross reaction with porcine infectious gastroenteritis virus,porcineδcoronavirus,porcine rotavirus and other gastrointestinal viruses,and both the variable coefficients of intra and inter batch repeated tests were within 2%,indicating a good sensitivity,specificity and repeatability.161 clinical samples collected from swine farms in different regions of Shandong Province from 2018 to 2019 were tested by the established method,and the results were completely consistent with the RT-PCR assay.In conclusion,the established method provided a good tool and technical support for rapid detection of PEDV.

关 键 词：猪流行性腹泻病毒 染料法 荧光定量RT-PCR 检测 应用 

分 类 号：S855.3[农业科学—临床兽医学]