同时检测猪瘟病毒、圆环病毒2型抗体的液相芯片法  

Simultaneous detection of classical swine fever virus and circovirus type 2 antibody by liquid chip method

作　　者：肖丽 石欣鹭 练月晓 饶丹[3] 黄碧洪 黄韧 郭鹏举 陈梅丽 XIAO Li;SHI Xin-lu;LIAN Yue-xiao;RAO Dan;HUANG Bi-hong;HUANG Ren;GUO Peng-ju;CHEN Mei-li(Guangdong Key Laboratory of Laboratory Animals,Guangzhou 510663,China;Rural Economic Bureau of Xiuyan County,Anshan,Liaoning 114300,China;Guangdong Laboratory Animals Monitoring Institute ,Guangzhou 510663,China)

机构地区：[1]广东省实验动物重点实验室,广东广州510663 [2]辽宁省岫岩县农村经济局,辽宁鞍山114300 [3]广东省实验动物监测所,广东广州510663

出　　处：《中国兽医学报》2018年第12期2267-2270,共4页Chinese Journal of Veterinary Science

基　　金：广东省科技计划资助项目(2015B070701003,2016A040403060);广州市科技计划资助项目(201707010440,2016201604030059)

摘　　要：通过液相芯片新技术建立了一种同时检测猪瘟病毒(classical swine fever virus,CSFV)、圆环病毒2型(porcine circovirus typeⅡ,PCV2)抗体的方法。研究方法及步骤包括:分别原核表达CSFV E2蛋白和PCV2Cap蛋白,与荧光微球偶联;偶联后的微球与各个标准血清反应,利用Luminax系统检测藻红蛋白的荧光信号强弱,从而建立抗体检测方法。该方法经过验证后,结果显示特异性良好,与其他猪病原抗体无交叉反应;批内及批间变异系数均在10%以内,稳定性好。与ELISA方法对比发现,液相芯片法的灵敏度均高于ELISA方法,CSFV和PCV2分别提高4,8倍。临床样品符合率分别是90.48%,60.32%。本研究为同时检测CSFV和PCV2抗体提供一种新方法。Through liquid chip technology to establish a new method for simultaneous detection of classical swine fever virus(CSFV),porcine circovirus type 2(PCV2)antibody.Respectively,the prokaryotic expressed CSFV E2 protein and PCV2 Cap protein were coupled with fluorescent microspheres;reacted to various standard serum after coupling,using Luminax detection system to captrue the phycoerythrin fluorescence signal,finallly establishing antibody detection method.The experimental method showed good specificity,no cross reaction to other swine pathogenic antibody;experimental coefficient of variation of intra batch and inter batch was less than 10%.The sensitivity of the liquid chip method was 4 times for CSFV and 8 times for PCV2 higher than that of ELISA.The coincidence rate of clinical samples was 90.48%(CSFV)and 60.32%(PCV2)respectively.This study provides a new technical basis for simultaneous detection of antibodies against classical swine fever virus,circovirus type 2 and transmissible gastroenteritis virus.

关 键 词：液相芯片 CSFV PCV2 

分 类 号：S852.65[农业科学—基础兽医学]