鹅星状病毒鹅细小病毒和鹅坦布苏病毒三重PCR检测方法的建立与应用  

Establishment and application of a triple PCR method for detection of astrovirus,parvovirus and Tembusu virus in goose

作　　者：张红 田秋丰 王志强 黄宇翔 邹跃 李鑫 吕明哲 杨昊天 马志刚 霍明东 董佳强 杨坤 魏念冬 苗艳 钟鹏 陈志峰 ZHANG Hong;TIAN Qiu-feng;WANG Zhi-qiang;HUANG Yu-xiang;ZOU Yue;LI Xin;Lü Ming-zhe;YANG Hao-tian;MA Zhi-gang;HUO Ming-dong;DONG Jia-qiang;YANG Kun;WEI Nian-dong;MIAO Yan;ZHONG Peng;CHEN Zhi-feng(Branch of A nimal Husbandry and Veterinary,Heilongjiang Academy of A gricultural Sciences,Qiqihar 161005,China)

机构地区：[1]黑龙江省农业科学院畜牧兽医分院,黑龙江齐齐哈尔161005

出　　处：《中国兽医科学》2023年第2期163-169,共7页Chinese Veterinary Science

基　　金：现代农业产业技术体系建设专项(CARS-42-34);黑龙江省农业科学院畜牧兽医分院2022年自拟项目(ZNKT-202203);黑龙江省农业科学院“农业科技创新跨越工程”专项(HNK2019CX-18)。

摘　　要：为建立一种可同时检测鹅星状病毒(GoAstV)、鹅细小病毒(GPV)和鹅坦布苏病毒(TMUV)的方法,根据GenBank上公布的GoAstV、GPV和TMUV的保守基因序列设计3对特异性引物,通过优化反应条件和反应体系,建立了能够同时检测GoAstV、GPV和TMUV的三重PCR方法,并初步应用于临床。结果显示:所建立的三重PCR检测方法能特异性扩增GoAstV、GPV和TMUV三种病毒的目的片段,对鹅副黏病毒、禽流感病毒、禽腺病毒和鹅圆环病毒的检测结果均为阴性,对GoAstV、GPV和TMUV的最低检出浓度分别为1.26×10^(5)、1.26×10^(3)和1.26×10^(5)copies/μL。利用三重PCR和单项PCR方法同时对黑龙江省部分鹅场的32份临床样品进行检测,3种病毒的阳性检出率分别为56.25%、6.25%和6.25%,2种方法的检测结果一致。结果表明,本试验中建立的三重PCR检测方法快捷、敏感、特异,能够用于GoAstV、GPV和TMUV的流行病学调查和临床检测。In order to establish one method for simultaneous detection of goose astrovirus(Go Ast V),goose parvovirus(GPV)and goose Tembusu virus(TMUV),three specific primer sets were designed by conserved sequence of Go Ast V,GPV and TMUV in Gen Bank.After the reaction conditions and reaction system were optimized,a triple PCR method for detection of Go Ast V,GPV and TMUV was established and applied to detect clinical samples.The results showed that the established triple PCR method could specifically amplify the target fragments of Go Ast V,GPV and TMUV,but goose paramyxovirus,avian influenza virus,fowl adenovirus,and goose circovirus as negative control could not.Sensitivity assay showed that the minimum concentrations of Go Ast V,GPV and TMUV were 1.26×10^(5)copies/μL,1.26×10^(3)copies/μL and 1.26×10^(5)copies/μL,respectively.Thirty-two clinical samples from several farms in Heilongjiang Province were simultaneously detected by this triple PCR and single PCR,and results showed that the positive rates of Go Ast V,GPV and TMUV were 56.25%,6.25%and 6.25%,respectively.The results of the two methods were consistent.These results indicated that the triple PCR method established in this study was rapid,sensitive and specific,and could be used for the clinical detection and epidemiological investigation of Go Ast V,GPV and TMUV.

关 键 词：鹅星状病毒 鹅细小病毒 鹅坦布苏病毒 三重PCR 

分 类 号：S852.659.2[农业科学—基础兽医学]