青环海蛇毒素抗毒血清的制备和效价的实验研究

Study on the preparation of antivenin and potency test against Hydrophis cynocinctus venom

目的 制备我国沿海常见的青环海蛇毒素抗毒血清 ,为治疗海蛇咬伤提供特效解毒剂。方法 青环海蛇毒素经减毒灭菌处理后 ,对 10只新西兰兔进行免疫 ,免疫过程中通过免疫双向扩散试验和酶联免疫吸附试验进行效价评价 ,达到最高效价后动物取血 ,进行试血试验。结果 动物经 19周免疫后 ,最终免疫效价为免疫双向扩散试验 1∶ 8可见明显沉淀线 ,酶联免疫吸附试验效价达到 12 80 0 0 0 ,试血实验 1ml血清可中和 0 .4 0 m g青环海蛇毒素 ,0 .34m g平颏海蛇毒素。结论 确定了一套快速、高效的抗青环海蛇毒素血清方案 ,研制出高效价的抗毒血清。为治疗海蛇咬伤提供了生产特效抗毒血清的实验基础。

Objective To investigate the preparation of rabbit antivenin against the venom of Hydrophis cynocinctus and provide antidote for the sea snake bites. Methods The detoxified and sterilized venom was used for immunization of 10 rabbits, in the process of which the potency of antivenin was traced through the Immunodiffusion and ELISA Tests. When the potency was at its peak, blood was collected from the immunized rabbits, and then the neutralization test for the prepared antivenin was carried out. Results Potency test of the antivenin showed that the precipitate lines could be clearly observed in the 1∶8 group of Immunodiffusion Test, while the potency was determined to be 1 280 000 in ELISA test. In the neutralization test, 1 ml of the antivenin could neutralize 0.4 mg (24.5 LD50) and 0.34 mg (18.8 LD50) of Hydrophis cynocinctus and Lapemis hardwickii venin respectively. Conclusions A rapid, highly effective method for the preparation of antivenin against the venom of Hydrophis cynocinctus has been developed, and a high potency antivenin has been prepared. This paper has provided a good experimental basis for the preparation of antivenin, which can be used for the treatment of sea snake bites.