卵黄抗体用于蛇毒抗原检测的初步研究  

A preliminary study of egg yolk antibody for kingcobra venom antigens detection

作　　者：王桂平[1] 余清声[1] 刘新艳[1] 朱柳[1] 覃媛[2] 黄劭[2] 

机构地区：[1]广州医学院药物研究中心 [2]广州医学院广州蛇毒研究所,广东广州510182

出　　处：《中国药理学通报》2005年第11期1351-1354,共4页Chinese Pharmacological Bulletin

基　　金：广州市科技局应用基础研究基金资助项目(No2002J1-C0171)

摘　　要：目的在常规ELISA检测中,以卵黄抗体取代哺乳动物来源的IgG类抗体,以探讨卵黄抗体应用于蛇毒检测的可行性.方法以甲醛减毒的眼镜王蛇毒抗原免疫莱航母鸡,制备抗眼镜王蛇毒抗体;采用过碘酸钠法对特异卵黄抗体进行酶标;将卵黄抗体及相应酶标抗体应用于常规ELISA中,对国内常见蛇毒样品抗原(包括眼镜王蛇毒、眼镜蛇毒、金环蛇毒、银环蛇毒、五步蛇毒及广东园斑蝰蛇毒)进行检测,并进行灵敏度、精确度、特异性及交叉反应等分析.结果检测的灵敏度可达32 μg·L-1,并且眼镜王蛇毒抗原浓度在32～750 μg·L-1的范围内,线性关系较好(r=0.963);初步应用表明卵黄抗体对眼镜王蛇毒具有较高的特异性,与蝰蛇科的五步蛇毒、园斑蝰蛇毒无交叉反应,与眼镜蛇科的金环及银环蛇毒仅在高剂量(剂量大于500 μg·L-1)时存在轻度交叉反应,不干扰实验检测,但与眼镜蛇毒之间交叉反应明显,干扰实验检测;应用本方法对低(100 μg·L-1)、中(300 μg·L-1)、高(1 000 μg·L-1)不同浓度的眼镜王蛇毒样品检测,结果平均板内变异系数均在1%～3%之内,板间变异系数在8%以内.结论 特异的卵黄抗体应用于微量蛇毒检测研究是可行的,具有灵敏度高、特异性较强等优点.该研究将为蛇伤诊断试剂盒的研制提供相应的理论依据.Aim To discuss whether specific Egg yolk antibody （IgY） can be used for snake venom antigens detection. Methods Chickens （ white Leghorn） were immunized with detoxicated king cobra venom by form- aldehyde. Egg yolk antibody were isolated from egg yolk, and labeled with horse radish peroxidase （HRP）. Snake venom antigens samples, including king cobra venom, cobra venom, bungrarus fasciatus venom, bun- grams muhicinctus venom, agkistrodon actus venom and Guangdong viper venom, were detected using ELISA , and sensitivity,precision and specificity were tested, respectively. Results At about 32 μg · L^-1, the king cobra antigens were detected using the method ; Linear relation was better （ r = 0. 963 ） when the concentration of kingcobra venom was within 32-75μg · L^-1. The method had good specificity and no cross reactivity was observed among the reagents and agkistrodon acutus guenther venom and vipera russelli siamensis smith venom; little cross reactivity was shown with bungarus muhicinctus blyth venom and bungarus fasciatus chmeider venom;cross reactivity was obvious with cobra venom; the average intra-assay coefficient of variation （CV） was 1% - 3% ,and the inter-assay CV was within 8%. Conclusions The study indicates that IgY can be good reagents for snake venom antigens detecton,and the study provides foundation for the development of the diagnosis kits of snakebites.

关 键 词：眼镜王蛇毒 卵黄抗体(IgY) 酶联免疫吸附试验(ELISA) IGG类抗体 

分 类 号：R392.11[医药卫生—免疫学] R446.61[医药卫生—基础医学]