摘要
有效防治猪流感需要借助于准确而快速的检测方法,为了建立一种高效、快速的多重检测方法,本研究应用H1和H3亚型猪流感病毒(SIV)的血凝素(HA)蛋白抗原偶联不同编码的荧光微球,应用间接法检测模式,建立了H1、H3亚型猪流感病毒IgG抗体荧光微球免疫学检测方法,并将该方法与血凝抑制试验(HI)、病毒中和试验(VN)和酶联免疫吸附试验(ELISA)进行比较分析。结果表明,该方法的抗体检测灵敏度比ELISA方法高100～1 000倍;与猪繁殖与呼吸综合征病毒、猪圆环病毒2型、猪瘟病毒、伪狂犬病病毒及日本乙型脑炎病毒的阳性血清均无交叉反应,且两种亚型血清之间也不存在交叉反应。重复性试验表明,批内和批间精密度测试结果分别为10.2%和12.8%。与HI、VN和ELISA方法比较,总符合率分别为97.82%、98.27%和97.83%。综上,本研究建立的检测方法具有双重检测特性,特异性好、灵敏性高、重复性强,可用于H1、H3亚型SIV的鉴别诊断。
Effective prevention and control of swine influenza requires the help of accurate and rapid detection methods.In order to establish a highly efficient and rapid multiple detection method,this study applied H1 and H3 subtypes swine influenza virus(SIV) hemagglutinin(HA) protein antigen coupled with different encoded fluorescent microspheres,and used the indirect method for detectingthe IgG antibodies against H1 and H3 subtypes SIV.The immunological microspheres of IgG antibodies against swine influenza virus were detected and compared with the methods of blood coagulation inhibition test(HI),virus neutralization test(VN) and enzyme linked immunosorbent assay(ELISA).The results showed that the detection sensitivity of the antibodies was 100─1 000 times higher than that of the ELISA method.There was no cross-reactivity with the positive serum with porcine reproductive and respiratory syndrome virus,type II porcine circovirus,swine fever virus,pseudorabies virus and Japanese encephalitis virus and there was no cross-reaction between the H1 and H3 subtype SIV sera.Repeatability experiments showed that the intra and inter batch precision test indexes were 10.2% and12.8% respectively.Compared with HI,VN and ELISA,the total coincidence rates were 97.82%,98.27%and 97.83% respectively.In conclusion,the detection method established in this study has dual detection characteristics,good specificity,high sensitivity and strong repeatability,which can be used for the differential diagnosis of the antibodies of H1 and H3 subtype SIV.
作者
冀池海
朱婉君
盛金良
韦应芳
曾梦
韩晓亮
朱棣华
麦湛卓
王衡
张桂红
JI Chi-hai;ZHU Wan-jun;SHENG Jin-liang;WEI Ying-fang;ZENG Meng;HAN Xiao-liang;ZHU Di-hua;MAI Zhan-zhuo;WANG Heng;ZHANG Gui-hong(College of Veterinary Medicine,South ChinaAgricultural University,Guangzhou 510642,China;GuangdongKey Laboratory of Zoonosis and Prevention of Zoonosis ,Guangzhou 510642,China;Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases,Guangzhou 510642,China;College of A nimal Science and Technology, Shihezi University,Shihezi 832003,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2019年第2期159-168,共10页
Chinese Veterinary Science
基金
国家重点研发计划项目(2016YFD0500707)
现代农业产业技术体系建设专项资金项目(CARS-35)
国家自然科学基金项目(31272564)
广东省高等学校优秀青年教师培养计划项目(YQ201530)
