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 > 猪繁殖与呼吸综合征病毒重组N蛋白间接ELISA方法的建立 畜牧与兽医 2011,43 (8) 17-21
Position: Home > Articles > An indirect ELISA based on recombinant nucleoprotein for detection of the antibodies to porcine reproductive and respiratory syndrome virus Animal Husbandry & Veterinary Medicine 2011,43 (8) 17-21

猪繁殖与呼吸综合征病毒重组N蛋白间接ELISA方法的建立

作  者：
李小欢;郭万柱;左健;林华;韩存波;徐志文;王印;朱玲
单  位：
四川万众生物技术服务有限公司;四川农业大学动物生物技术中心
关键词：
猪繁殖与呼吸综合征;重组N蛋白;ELISA
摘  要：
用纯化的猪繁殖与呼吸综合征病毒(PRRSV)重组核衣壳(N)蛋白作为包被抗原,建立了PRRSV抗体检测的间接ELISA方法,并优化确定了ELISA最佳工作条件:重组抗原包被浓度为13.3μg/mL,37℃1 h或4℃过夜;5%脱脂乳37℃封闭2 h;血清1∶40稀释,37℃作用90 min;酶标二抗1∶4 000稀释,37℃作用40 min;底物TMB溶液37℃显色15 min;统计学分析后确定阴阳性的临界值为0.434。与美国IDEXX公司PRRSV抗体检测试剂盒相比较,其相对特异性和敏感性分别为93.0%和95.6%,符合率达到94.7%,检测结果无显著性差异。
译  名：
An indirect ELISA based on recombinant nucleoprotein for detection of the antibodies to porcine reproductive and respiratory syndrome virus
作  者：
LI Xiao-huan1,GUO Wan-zhu1,3,ZUO Jian2,Lin Hua1,Han Cun-bo2,XU Zhi-wen1,WANG Yin1,ZHU Ling1(1.Animal Biotechnology Center of Sichuan Agricultural University,Ya′an 625014,China; 2.Sichuan Peoples Biological Technology Service CO.LTD,Chengdu 610041,China; 3.Animal Disease and Human Health Key Laboratory of Sichuan Province of Sichuan Agricultural University,Ya′an 625014,China)
关键词：
porcine reproductive and respiratory syndrome;recombinant nucleoprotein;ELISA
摘  要：
An indirect ELISA for detection of the antibodies to porcine reproductive and respiratory syndrome virus(PRRSV) was developed using the purified nucleoprotein,and its optimal reaction conditions were determined: coating antigen at 37 ℃ 1 h or 4 ℃ overnight at a concentration of 13.3 μg/mL,5% defatted milk powder as blocking agent at 37 ℃ 2 h,serum sample(1∶40) at 37 ℃ for 90 min,HRP labeled rabbit anti-pig IgG(1∶4000) at 37 ℃ for 40 min,the substrate TMB for ELISA being incubated at room temperature for 15 min.The shreshold of positive and negative sera was 0.434 by statistical analysis.The ELISA assay was confirmed to have a good repeatability,sensitivity and specificity by repeatability test,cross test and blocking test.And compared with the IDEXX Test kit,the sensitivity and specificity of the ELISA were 95.6% and 93.0%,respectively,which showed no significant difference between the two assays.The coincidence rate of the two assays was up to 94.7%.

相似文章

导出引用
计量
文章访问数: 1
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