猪繁殖与呼吸综合征病毒M基因的截短表达及胶体金免疫层析法的建立  

Prokaryotic Expression of Truncated M Gene and Development of Colloidal Gold Immunochromatographic Method of Porcine Reproductive and Respiratory Syndrome Virus

作　　者：胡冉冉 赵云环 刘莹[1] 张帅[1] 郭禹 左玉柱[1,2] 范京惠 HU Ranran;ZHAO Yunhuan;LIU Ying;ZHANG Shuai;Guo Yu;ZUO Yuzhu;FAN Jinghui(College of Veterinary Medicine,Hebei Agricultural University,Baoding 07001,China;Hebei Veterinary Biotechnology Innovation Center,Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]河北农业大学动物医学院,保定071001 [2]河北省兽医生物技术创新中心,保定071001

出　　处：《中国动物传染病学报》2023年第6期94-100,共7页Chinese Journal of Animal Infectious Diseases

基　　金：河北省重点研发计划项目(19226622D);河北省农业产业技术体系生猪创新团队(HBCT2018110207)。

摘　　要：为建立一种快速、便捷的检测猪繁殖与呼吸综合征病毒(PRRSV)抗体的胶体金免疫层析方法,本试验通过原核表达截短的M基因作为检测线,葡萄球菌A蛋白(SPA)和鸡抗SPA作为金标抗原和质控线。结果显示,该试纸条与猪伪狂犬病毒(PRV)、猪传染性胃肠炎病毒(TGEV)、猪瘟病毒(CSFV)、猪流行性腹泻病毒(PEDV)和猪圆环病毒2型(PCV2),均不存在交叉反应,特异性强;最低检测限度为1∶6400,灵敏性高;批间、批内无差异性;4℃可保存30 d,稳定性好;临床样品检测结果与爱德士试剂盒符合率达到94.82%。综上所述,本试验建立了一种敏感性高、特异性好的PRRSV抗体检测的免疫层析试纸条,为PRRSV的监测、预防及诊断提供了技术支持。In order to develop a rapid and convenient method for detecting porcine reproductive and respiratory syndrome virus(PRRSV)antibodies,the truncated M protein was expressed in prokaryotic system and used as the test line for development of a colloidal gold immunochromatography,in which SPA and chicken anti-SPA served as the gold-labeled antigen and control line.The results showed that the test strips did not cross-react with PRV,TGEV,CSFV,PEDV and PCV2.The minimum detection limit was 1∶6,400.In addition,no difference occurred between batches and within batches;The prepared strips were stable at 4℃ for 30 days.The coincidence reached 98.2%when clinical sample were tested using both colloidal gold immunochromatography and IDEXX kit.In summary,the colloidal gold immunochromatography developed here had high sensitivity and good specificity,which might be used for the detection of PRRSV antibodies for the monitoring,prevention and diagnosis.

关 键 词：猪繁殖与呼吸综合征病毒 M基因 原核表达 胶体金免疫层析法 

分 类 号：S852.651[农业科学—基础兽医学]