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摘要抗蛇毒血清是蛇伤中毒解救中最有效的特异性治疗药物，抗蛇毒血清的生产也己常规化。目前使用的抗蛇毒血清大部分是由蛇毒免疫马匹而制得的抗体，而本研究以2种国产眼镜蛇（孟加拉眼镜蛇和舟山眼镜蛇）为对象，用其减毒后的蛇毒各自免疫大白兔，结合血清提取和纯化技术获取有大白兔血清，采用ELISA、Western blot方法检测大白兔抗血清及商用抗眼镜蛇毒血清与蛇毒之间的交叉反应强度和效价。ELISA结果显示，我们发现两种自制血清对孟加拉眼镜蛇蛇毒与舟山眼镜蛇蛇毒均有强烈的中和效果。在与商用血清对比中则发现，稀释比低于1:5000时，自制血清对蛇毒的中和效果好于商用血清，且抗孟加拉眼镜蛇血清效果最好。Western blot检测结果显示，这些自制血清均和商用血清一样能使两种蛇毒的主要条带呈现出强烈的免疫原性。自制血清具有特异性强、效价较高和成本低的特点，希望为蛇伤患者提供新的抗蛇毒血清。48332
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Abstract
Specific antivenoms are the most effective drug treatment for snakebite poisoning rescue, and antivenom production has also been normalized. At present, most of the anti snake venom serums are made of immune horse, and this research uses two kinds of homemade Cobras (Naja kaouthia and Naja atra). We used attenuated snake venoms to immunize rabbits, then used serum extraction and purification technology to obtain the serums of rabbits, and then used ELISA and Western blot methods to detect the titer and response intensity of between self-made serum and commercial antivenom. ELISA results showed that the two kinds of self-made serum have a strong neutralizing effect on Naja kaouthia and Naja atra venoms. In comparison with the commercial antivenom, it was found that when the dilution ratio was less than 1:5000, the neutralization effect of the self-made serum on snake venom was better than that of commercial antivenom. Western blot test results revealed that the homemade and commercial serum can make two kinds of snake venoms showing a strong immunogenicity. homemade serums have the characteristics of strong specificity, high titer and low cost, and we hope to provide a new anti snake venom serum for the patients with snake injury.
毕业论文关键词：抗眼镜蛇毒血清，孟加拉眼镜蛇，舟山眼镜蛇，血清效价，ELISA，Western blot
Keyword: anti cobra venom serum, Naja atra, Naja kaouthia, serum titer, ELISA,Western blot
目录
引言 4
1 材料与方法 5
1.1 实验动物 5
1.2 蛇毒采集 5
1.3 抗原制备及大白兔免疫 5
1.4 眼镜蛇毒抗血清提取和纯化 6
1.5 ELISA检测 6
1.6 Western blot检测 6
2 结果 8
2.1 ELISA结果 8
2.2 Western blot 8
3 讨论 10
参考文献 10
致谢 11
引言
世界上已知的蛇类有2700多种，其中毒蛇600多种。在有毒动物导致的人类中毒病例中，毒蛇咬伤占首要位置。2007年1月，世界卫生组织（World Health Organization，WHO）在瑞士日内瓦专门召开了抗蛇毒血清及抗狂犬病毒血清国际会议，会议上各国专家对本国蛇伤治疗情况进行了交流[1-2]。其中巴西、泰国和印度等国尽管都能生产抗蛇毒血清，但品种有限，且产量不多，无法应对其境内高发的毒蛇致伤事件。2010年5月4日，WHO向媒体发表简讯，强调急需拯救生命的抗蛇毒血清品种及许多国家所面临的抗蛇毒血清短缺问题，抗蛇毒血清及其市场需求无疑就是个世界性问题[13]。