| 应用间接竞争ELISA检测山羊痘抗体方法的建立 |
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| 引用本文： | 何光志,周颖,田维毅,王平,王文佳,韩洁,安永如,简昌友,崔小东.应用间接竞争ELISA检测山羊痘抗体方法的建立[J].安徽农业科学,2010,38(35):20125-20127. |
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| 作者姓名： | 何光志 周颖 田维毅 王平 王文佳 韩洁 安永如 简昌友 崔小东 |
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| 作者单位： | [1]贵阳中医学院,贵州贵阳550002 [2]贵州省安顺市职业技术学院,贵州安顺561000 [3]贵州省思南县动物疫病预防控制中心,贵州思南565100 [4]贵州省凤冈县人民医院,贵州凤冈564200 |
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| 摘 要： | 目的]为山羊痘疫苗接种山羊后产生的抗体效价检测和山羊痘的诊断奠定基础。方法]采用Vero-E6传代细胞分离山羊痘病毒抗原,建立检测山羊痘抗体的间接竞争ELISA方法。结果]检测山羊痘抗体的间接竞争ELISA的最佳工作条件：最佳抗原包被浓度为1μg/ml,兔抗高免血清最佳稀释度为1∶16 000,待检血清的工作浓度为1∶400,阳性血清的临界值为45.19%,阴性血清的临界值为38.89%。用该试验建立的ELISA方法检测3 000份接种山羊痘疫苗的山羊血清,阳性检出率为83.33%,琼脂扩散试验检出率为77.33%,竞争抑制ELISA方法较琼脂扩散试验方法敏感。结论]该方法具有特异性高、快速、敏感等特点,适合检测山羊痘抗体。
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| 关 键 词： | 山羊痘 间接竞争ELISA 山羊痘抗体 |
Establishment of Indirect Competitive Enzyme-linked Immunosorbent Assay of Goat Pox Antibody |
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| Institution: | HE Guang-zhi et al(Guiyang college of Traditional Chinese Medicine,Guizhou,Guiyang 550002) |
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| Abstract: | Using Vero-E6 cell separation goat pox virus as coating antigen,a indirect competitive enzyme-linked immunosorbent assay for detection of goat pox antibody was developed and the reaction condition was optimized in indirect competitive ELISA.The optimal concentration of coating antigen in indirect competitive ELISA was 1 μg/ml,the optimal dilution of the rabbit anti-high was 1∶16 000,the optimal dilution of inspection of serum was 1∶400,and the cut-off value of 45.19% was set for the positive sera,and the cut-off value of 38.89% was set for the known negative sera.Out of 3 000 goat sera,83.33% were positive GPV by a indirect competitive ELISA,77.33%were by agar gel precipitation(AGP),the method of indirect competitive ELISA was more sensitive than the AGP.This studies reveal that the method has a high specificity,rapid and sensitive. |
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| Keywords: | Goat pox indirect competitive enzyme-linked immunosorbent assay goat pox antibody |
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