| REV重组env蛋白间接ELISA诊断试剂盒的研制 |
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| 引用本文： | 李彦,牛钟相.REV重组env蛋白间接ELISA诊断试剂盒的研制[J].中国兽药杂志,2011,45(3):26-30. |
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| 作者姓名： | 李彦 牛钟相 |
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| 作者单位： | 1. 山东农业大学动物医学院,山东,泰安,271018;山东省动物生物技术与疫病防治重点实验室,山东,泰安,271018
2. 山东畜牧兽医职业学院,山东,潍坊,261061 |
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| 基金项目： | 国家十一.五科技支撑计划项目（2006BAD06A11） |
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| 摘 要： | 以纯化的env蛋白为包被抗原,建立了检测REV抗体的间接ELISA（ienv-ELISA）方法。ienv-ELISA工作条件优化结果表明,最佳包被液为CB;最佳封闭液浓度为1%BSA-PBS;最佳抗原包被量为8μg/mL;血清最佳稀释度为1∶300;血清最佳作用时间为1 h;二抗的最佳稀释度为1∶5000;二抗最佳作用时间为1 h;底物最佳显色时间为25 min。重复性试验、特异性试验和对比试验结果显示,建立的ienv-ELISA方法有良好的可重复性,标准差均小于15%;与鸡MDV、ALV、NDV、H5N1、H9N1等病毒阳性抗体均无交叉反应;与REV抗体检测试剂盒（以REV全病毒为包被抗原）对比,所建立的ienv-ELISA诊断敏感性、诊断特异性以及准确率分别为90.625%9、3.75%和92.7%。
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| 关 键 词： | 网状内皮组织增生症病毒 env蛋白 间接ELISA 试剂盒 抗体检测 |
| 收稿时间： | 2010/12/11 0:00:00 |
| 修稿时间： | 2011/1/21 0:00:00 |
Development of an Indirect ELISA Kit to Detect REV Sera with Recombinant Env Protein |
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| liyan and.Development of an Indirect ELISA Kit to Detect REV Sera with Recombinant Env Protein[J].Chinese Journal of Veterinary Drug,2011,45(3):26-30. |
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| Authors: | liyan and |
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| Institution: | Shandong Agricultural University, |
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| Abstract: | Using the recombinant and purified env protein as antigen to coat micro-plate of 96 wells,an indirect ELISA assay（ienv-ELISA）was developed to detect REV antibody.The optimum reaction conditions for ienv-ELISA were investigated,and the results showed that optimal coating buffer was CB;optimal confining liquid was 1% BSA-PBS;optimal concentration of antigen was 8 μg/mL;optimal dilution of sera was 1∶300;optimal reaction time of sera was 1 h;optimal dilution of HRP-labled goat-anti chicken IgG was 1∶5000;optimal reaction time of HRP-labled goat-anti chicken IgG was 1 h;optimal reaction time of substrate was 25 min.Repeatability tests revealed that the coefficients of variation of positive sera within and between runs were less than 15%.Cross-reactivity assay showed that this assay was REV-specific.This assay was validated by comparison with a REV-based ELISA.The diagnostic sensitivity（DSN）,specificity（DSP） and accuracy of the ienv-ELISA were 90.625%,93.75% and 92.7% respectively. |
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| Keywords: | REV env protein indirect ELISA kit antibody detection |
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