新型鸭源小RNA病毒荧光定量PCR检测方法的建立  

Development of A Real-time PCR Assay for Novel Duck-derived Picornavirus

作　　者：李继桐 胡峰[2] 姜亦飞[2] 宋敏训[2] 李玉峰[2] 王建琳 LI Jitong;HU Feng;JIANG Yifei;SONG Minxun;LI Yufeng;WANG Jianlin(School of Animal Medicine,Qingdao Agricultural University,Qingdao 266109,China;Institute of Poultry,Shandong Academy of Agricultural Sciences,Jinan 250100,China)

机构地区：[1]青岛农业大学动物医学院,青岛266109 [2]山东省农业科学院家禽研究所,济南250100

出　　处：《病毒学报》2023年第5期1377-1384,共8页Chinese Journal of Virology

基　　金：山东省农业良种工程(2022LZGC014),题目:优质肉鸭育种技术创新与新品种培育;山东省家禽产业技术体系(SDAIT-11-01);韩国农林部国际合作(122014-2),题目:鸭病毒病国际联合监测与控制技术体系的建立~~。

摘　　要：为建立一种快速诊断新型鸭源小RNA病毒的检测方法,根据NCBI已公布的病毒基因组序列设计特异性引物,建立了新型鸭源小RNA病毒荧光定量PCR检测方法。结果显示:本方法能特异性扩增新型鸭源小RNA病毒基因序列;对质粒标准品的最低检测限为98拷贝/反应;组内与组间的重复试验变异系数均小于2%;对新分离的小RNA病毒株进行检测,均能扩增出目的基因。综上,本研究建立的检测方法特异性强、灵敏性高、重复性和实用性好,可用于新型鸭源小RNA病毒的快速鉴别诊断。To establish a rapid method for diagnosing novel duck-derived picornavirus,a fluorescent quantitative PCR assay was developed based on NCBI published viral genomic sequences with specific primers.Results showed that this method was able to specifically amplify the gene sequences of duck-derived picornavirus.The minimum detection limit for plasmid standards was 98 copies/reaction.The coefficient of variation of intra-and inter-group replicates was less than 2%.The target gene was amplified in all newly isolated picornavirus strains.In conclusion,this assay is highly specific,sensitive,reproducible and practical,and can be used for rapid differential diagnosis of novel duck-derived picornavirus.

关 键 词：鸭源小RNA病毒 荧光定量PCR 检测方法 

分 类 号：S834.89[农业科学—畜牧学] S855.3[农业科学—畜牧兽医]