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| 猪繁殖与呼吸综合征病毒重组N蛋白间接ELISA方法的建立 | |
| 引用本文： | 李小欢,郭万柱,左健,林华,韩存波,徐志文,王印,朱玲.猪繁殖与呼吸综合征病毒重组N蛋白间接ELISA方法的建立[J].畜牧与兽医,2011,43(8):17-21. |
| 作者姓名： | 李小欢 郭万柱 左健 林华 韩存波 徐志文 王印 朱玲 |
| 作者单位： | 1. 四川农业大学动物生物技术中心,四川雅安,625014 2. 四川农业大学动物生物技术中心,四川雅安625014;四川农业大学动物疫病与人类健康四川省重点实验室,四川雅安625014 3. 四川万众生物技术服务有限公司,四川成都,610041 |
| 基金项目： | 科技部国家科技支撑计划资助(2006BAD06A18); 教育部长江学者和创新团队发展计划项目资助(IRT0848) |
| 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13.3 μg/mL,37℃1h或4℃过夜；5％脱脂乳37℃封闭2h;血清1:40稀释,37℃作用90 min；酶标二抗1∶4000稀释,37... |
| 关 键 词： | 猪繁殖与呼吸综合征 重组N蛋白 ELISA |
An indirect ELISA based on recombinant nucleoprotein for detection of the antibodies to porcine reproductive and respiratory syndrome virus |
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| LI Xiao-huan,GUO Wan-zhu,ZUO Jian,Lin Hua,Han Cun-bo,XU Zhi-wen,WANG Yin,ZHU Ling.An indirect ELISA based on recombinant nucleoprotein for detection of the antibodies to porcine reproductive and respiratory syndrome virus[J].Animal Husbandry & Veterinary Medicine,2011,43(8):17-21. | |
| Authors: | LI Xiao-huan GUO Wan-zhu ZUO Jian Lin Hua Han Cun-bo XU Zhi-wen WANG Yin ZHU Ling |
| Abstract: | 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%. |
| Keywords: | porcine reproductive and respiratory syndrome recombinant nucleoprotein ELISA |
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