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 > 基因C型鸭甲肝病毒竞争ELISA抗体检测方法的建立 中国预防兽医学报 2015,37 (3) 211-215
Position: Home > Articles > Development of a competitive ELISA for detecting antibodies against genotype C duck hepatitis A virus based on the monoclonal antibody Chinese Journal of Preventive Veterinary Medicine 2015,37 (3) 211-215

基因C型鸭甲肝病毒竞争ELISA抗体检测方法的建立

作  者：
何美琳;张焕容;程方明;杨晓农;岳华
单  位：
西南民族大学生命科学与技术学院
关键词：
基因C型鸭甲肝病毒;单克隆抗体;竞争ELISA;抗体检测
摘  要：
为建立检测基因C型鸭甲肝病毒(DHAV-C)血清抗体的方法,本研究以纯化的DHAV-C作为包被抗原,抗DHAV-C单克隆抗体(MAb)5E3-9与待检血清竞争结合抗原,采用方阵法确定包被抗原、MAb及待检血清的最佳工作浓度,经过对反应条件的优化,建立了DHAV-C抗体的竞争ELISA检测方法。该方法仅对DHAV-C血清检测为阳性,与DHAV-A、鸭圆环病毒、鸭乙型肝炎病毒等相关病原的鸭阳性血清无交叉反应。敏感性试验表明,标准阳性血清1:128倍稀释时,检测结果仍为阳性,比中和试验灵敏性更高。批内批间变异系数分别为4.64%~5.21%和6.02%~8.68%,具有良好的重复性。与中和试验比较,阳性符合率为100%(15/15),阴性符合率为77.8%(14/18)。本研究基于纯化的DHAV-C及其特异性MAb建立的MAb竞争ELISA检测方法可以用于DHAV-C流行病学调查以及抗体的检测。
译  名：
Development of a competitive ELISA for detecting antibodies against genotype C duck hepatitis A virus based on the monoclonal antibody
作  者：
HE Mei-lin;ZHANG Huan-rong;CHENG Fang-ming;YANG Xiao-nong;YUE Hua;College of Life Science and Technology, Southwest University for Nationalities;Key Laboratory of Veterinary Medicine of University in Sichuan Province, Southwest University for Nationalities;
关键词：
duck hepatitis A virus genotype C;;monoclonal antibody;;competitive ELISA;;antibody detection
摘  要：
The objective of this study is to develop a competitive ELISA to detect the antibody against duck hepatitis A virus genotype C(DHAV-C). The purified DHAV-C was used as coating antigen, and DHAV-C specific MAb 5E3-9 as competitive antibody against DHAV-C antibody. Under the optimized reacting conditions, including 10 μg/m L purified DHAV-C coating antigen, 1∶800 dilution of MAb 5E3-9 diluting the test serum to 1 ∶2 ratio, and 1 ∶5,000 dilution of HRP conjugated goat anti-mice Ig G with the positive cut-off value of inhibition ratio 36.86%. The specificity test showed that there were no cross reaction with duck anti-sera of all tested relative pathogens, such as duck hepatitis A virus genotype A(DHAV-A), duck circovirus, duck hepatitis B virus and other 6 pathogens. The sensitivity test showed that 1 ∶64 diluted DHAV-C positive sera could be detected positive with this developed competitive ELISA. The 15 neutralization test detected positive sera were all positive when they were detected by the developed competitive ELISA, while among 18 neutralization tested negative sera, 14 of which were negative when they were detected by the developed competitive ELISA. Therefore, the developed competitive ELISA in this study was more sensitive than neutralization test. The intra- and inter-batch assay demonstrated that the co-efficient of maximum variation were 5.21% and 8.68%, respectively. The results revealed that the developed competitive ELISA could be used for seroepidemiological survey and infection or immune antibody detection of DHAV-C.

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