摘要
为建立猪呼肠孤病毒(PRV)抗体间接ELISA检测方法,本研究采用PCR方法扩增PRV GD-1株S1基因,并以原核系统表达其编码的σ1蛋白。以纯化的σ1蛋白作为包被抗原,鹅抗猪Ig G-HRP为检测抗体,建立了PRV血清型3型间接ELISA血清学的检测方法。该方法对其他几种常见猪腹泻病毒均无交叉反应,具有较强的特异性。组内和组间变异系数均低于10%,重复性好。利用该方法和e Bioscience同类ELISA进口抗体检测试剂盒对606份临床样品进行检测,两者符合率为96.2%。本研究建立的方法为进一步开发商品化试剂盒奠定了基础。
To establish a rapid method for detecting antibody against the porcine reovirus （PRV）, the S1 gene of serotype 3 PRV GD-1 strain was amplified by PCR and inserted into pET-32a vector to express the σ1 protein in E.coli, and the indirect ELISA was developed with the purified crl protein as coating antigen and the goose anti-pig IgG-conjugated horseradish peroxidase （HRP） as the detection antibody. The results showed the assay was specificity for detecting serotype 3 PRV without any cross-reaction to the positive sera of other related viruses causing diarrheal diseases in swine. The coefficients of variation in both intra- and inter-assay were less than 10%. Furthermore, a total of 606 clinical serum samples were detected by this assay and the agreement was 96.2% with commercial ELISA kit （eBioscience）. The establishment of this assay provided the basis for further developing PRV commercial kits.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2016年第1期63-67,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
江西省教育厅科学技术研究项目(GJJ14298)
