猪细小病毒NS1非结构蛋白和VP2结构蛋白主要抗原区间接ELISA方法的建立与联合应用  

Establishment and joint application of indirect ELISA assay for the major antigenic domain of porcine parvovirus NSl non-structural protein and VP2 structural protein

作　　者：刘建[1] 汤德元[1] 李春燕[1] 曾智勇[1] 罗险峰[2] 郝飞[1] 姜德荣[1] 王洪光[1] 李达[1] 

机构地区：[1]贵州大学动物科学学院,贵阳550025 [2]贵州省动物疫病预防控制中心,贵阳550008

出　　处：《黑龙江畜牧兽医》2014年第11期41-45,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：贵州省科学技术基金项目"猪呼吸道病毒性疫病新型特异诊断技术多重PCR检测的研究"(黔科合J字(2013)2110号);贵州省2011年农业攻关项目"贵州省规模化猪场主要病毒性疫病的调查及综合防控对策的研究与示范"[黔科合NY字(2011)3103];贵州大学研究生创新基金项目"猪细小病毒NS1蛋白主要抗原区间接ELISA方法的建立"[研农2013020]

摘　　要：为了建立猪细小病毒野毒抗体的NS1-i ELISA和猪群PPV免疫抗体水平的VP2-i ELISA方法,试验采用猪细小病毒NS1和VP2基因主要抗原区纯化后的原核表达重组蛋白作为包被抗原。结果表明:检测灵敏度为1∶12 800;批内、批间重复性试验变异系数均小于10%,NS1-i ELISA方法与HI试验的符合率为100%;VP2-i ELISA方法与HI试验的符合率为94.7%,且比HI试验具有更高的敏感性。用这两种方法同时检测猪伪狂犬病毒(PRV)、猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪圆环病毒2型(PCV-2)、猪乙型脑炎病毒(JEV)和猪O型口蹄疫病毒(FMDV)6种常见猪病病毒的阳性血清结果均为阴性。说明所建立的NS1-i ELISA和VP2-i ELISA诊断方法具有良好的重复性、敏感性和特异性。这两种方法可联合应于PPV野毒感染的快速诊断、流行病学调查、猪群免疫疫苗后PPV抗体水平的检测以及猪群PPV的净化。To establish the methods of NS1 - iELISA for the wild virus antibody of porcine parvovirus （PPV） and VP2 - iELISA for the herd im- mune levels of PPV antibody in pigs. The recombinant protein with prokaryotic expression was used as the coated antigen after purified using the major antigenic domain of the NS1 and VP2 genes of porcine parvovirus. The results showed that the detection sensitivity of these two methods was 1 ： 12 800, and the intro - assay and inter - assay coefficients of variation for the reproducibility tests using these two methods were both less than 10%. The concordance rate of NS1 - iELISA was 100% with that of the HI assay; the concordance rate. of VP2 - iELISA was 94.7% with that of the HI assay, and VP2 - iELISA had higher sensitivity than the HI assay. These two methods were used to detect the positive sera of six common swine disease viruses including pseudorabies virus （PRV）, classical swine fever virus （CSFV） , porcine reproductive and respiratory syndrome virus （ PRRSV）, porcine circovirus type 2 （ PCV - 2 ）, Japanese encephalitis virus （ JEV ）, and foot - and - mouth disease virus type O （ FMDV Type O） , and their detection results were negative. The results indicate that the established NS1 -iELISA and VP2 -iELISA diagnostic methods have good reproducibility, sensitivity and specificity. These two methods can be combined to use for a rapid diagnosis in PPV wild virus infection, epidemiologic survey of PPV, detection of PPV antibody levels of herd immunity after vaccination, and the purification of PPV in swine herd.

关 键 词：猪细小病毒 NS1非结构蛋白 VP2结构蛋白 主要抗原区 间接ELISA 

分 类 号：S852.659.2[农业科学—基础兽医学] S784[农业科学—兽医学]