中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > PRRSV E蛋白的表达及检测抗体胶体金试纸的研制 黑龙江畜牧兽医 2014 (9) 19-21+26
Position: Home > Articles > Expression of the E protein of porcine reproductive and respiratory syndrome virus and development of colloidal gold strip for the detection of antibodies against PRRSV Heilongjiang Animal Science and Veterinary Medicine 2014 (9) 19-21+26

PRRSV E蛋白的表达及检测抗体胶体金试纸的研制

作  者：
杨欢;吴天成;刘禹;郑鑫;崔焕忠
单  位：
吉林农业大学动物科技学院
关键词：
猪繁殖与呼吸综合征病毒(PRRSV);糖蛋白E;表达;胶体金试纸;抗体
摘  要：
为了建立快速检测猪繁殖与呼吸综合征病毒(PRRSV)抗体的方法,试验采用RT-PCR扩增ORF5基因,克隆至原核表达载体pET-28a上,诱导重组质粒转化菌表达,SDS-PAGE分析目的蛋白表达情况,Western-blot检测表达蛋白免疫活性,亲和层析纯化表达蛋白。用SPA标记胶体金,分别以纯化E融合蛋白和羊抗猪抗体为检测线和质控线制备胶体金试纸,用所研制的试纸检测猪血清样品40份,并与IDEXX ELISA试剂盒的检测结果进行比较。结果表明:成功表达E融合蛋白,表达蛋白具有良好的反应原性,纯度为90%;制备的胶体金试纸与IDEXX ELISA试剂盒检测符合率为92%。说明该种方法可检测PRRSV抗体。
译  名：
Expression of the E protein of porcine reproductive and respiratory syndrome virus and development of colloidal gold strip for the detection of antibodies against PRRSV
作  者：
YANG Huan;WU Tian-cheng;LIU Yu;ZHENG Xin;CUI Huan-zhong;College of Animal Science and Technology,Jilin Agricultural University;
关键词：
porcine reproductive and respiratory syndrome virus(PRRSV);;glycoprotein E;;expression;;colloidal gold strip;;antibodies
摘  要：
To establish a method of rapid detection of antibodies against porcine reproductive and respiratory syndrome virus( PRRSV),the ORF5 gene was amplified by RT- PCR and cloned into prokaryotic expression vector pET28a,and then the recombinant plasmid was transformed into Escherichia coli BL21( DE3) and the expression was induced by IPTG. SDS- PAGE and Western- blot were used to analyze the expression of the target protein and detect the immune activity of the expressed protein,respectively,and the expressed protein was purified by affinity chromatography. The colloidal gold strip for detecting antibodies against PRRSV was developed using Staphylococcal protein A( SPA) labeled with colloidal gold,taking the purified E fusion protein and goat anti- pig antibodies as test line and control line,respectively. 40 pig serum samples were tested by colloidal gold strip,and the detection results were compared with the results using IDEXX ELISA kit. The result showed that the E fusion protein was successfully expressed,and the expressed protein had good reactogenicity,with the purity of 90%. The coincidence rate between the colloidal gold strip and IDEXX ELISA kit for the detection was 92%. The results indicate that the method with colloidal gold strip can be used for the detection of antibodies against PRRSV.

相似文章

导出引用
计量
文章访问数: 8
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46