| 猪瘟抗体间接ELISA检测方法的建立和优化 |
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| 引用本文： | 徐璐,范学政,徐和敏,赵启祖,邹兴启,朱元源,王琴,宁宜宝,王芳.猪瘟抗体间接ELISA检测方法的建立和优化[J].中国兽医杂志,2012,48(3):15-17,97. |
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| 作者姓名： | 徐璐 范学政 徐和敏 赵启祖 邹兴启 朱元源 王琴 宁宜宝 王芳 |
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| 作者单位： | 1. 中国兽医药品监察所,北京海淀,100081
2. 中国牧工商(集团)总公司研究院,北京丰台,100070 |
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| 基金项目： | 农业部"948"项目，国家科技支撑计划 |
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| 摘 要： | 采用真核系统表达纯化的猪瘟病毒E2蛋白作为包被物,通过对各个反应条件的摸索和优化,建立了检测猪瘟病毒抗体的猪瘟抗体间接ELISA检测方法。研究结果表明,E2蛋白的最佳包被浓度为0.25μg/mL,最佳包被条件为4℃16h;待检血清的最佳稀释倍数为100倍;特异性检测试验证明,该方法与猪常见病原的阳性血清没有交叉反应。通过对大量猪瘟抗体阴、阳性血清的检测,最终确定了本检测方法的判定标准。
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| 关 键 词： | 猪瘟 ELISA 判定标准 |
Development and optimization of the indirect ELISA method to detect antibody against classical swine fever virus |
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| XU Lu
,
FAN Xue-zheng
,
XU He-min
,
ZHAO Qi-zu
,
ZOU Xing-qi
,
ZHU Yuan-yuan
,
WANG Qin
,
NING Yi-bao
,
WANG Fang.Development and optimization of the indirect ELISA method to detect antibody against classical swine fever virus[J].Chinese Journal of Veterinary Medicine,2012,48(3):15-17,97. |
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| Authors: | XU Lu FAN Xue-zheng XU He-min ZHAO Qi-zu ZOU Xing-qi ZHU Yuan-yuan WANG Qin NING Yi-bao WANG Fang |
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| Institution: | 1(1.China Institute of Veterinary Drug Control,Beijing 100081,China; 2.Zhongmu Institute of China Animal Hunsbandry Group,Beijing 100070,China) |
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| Abstract: | Classical swine fever virus(CSFV) E2 protein was expressed in BAC-To-BAC system.After purification,E2 protein was coated to the ELISA plate.Through a series of optimization tests,the indirect ELISA for CSFV antibody was established.The results showed that the optimal concentration of E2 protein for coating was 0.25 μg/mL and coating condition was 4℃ for 16 h.The proper dilution of serum sample was 1∶100.And the specificity tests showed that the purified E2 protein had higher specificity for CSFV antibody in serum.Hundreds of samples were detected by the ELISA and standard of criterion was established by data analysis. |
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| Keywords: | classical swine fever ELISA standard of criterion |
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