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目的建立尖吻蝮蛇血凝酶(商品名苏灵,代号Saculin)多克隆抗体检测的间接ELISA法,并应用于其临床免疫学检测。方法免疫家兔获得多克隆抗体,Western blot鉴定其特异性,通过比较各种不同条件的检测结果来确立检测尖吻蝮蛇血凝酶多克隆抗体的间接ELISA法的最佳工作条件,并以此为参考检测临床血清。结果间接ELISA法测定兔抗血清效价高达1∶6 553 600,55例临床试验受试者(36例为Saculin实验组,19例为安慰剂组)给药3周后血清抗体检测结果均为阴性。结论本研究建立的抗尖吻蝮蛇血凝酶多克隆抗体间接ELISA法能够用于临床血清的判定。
Objective To establish an indirect ELISA for the detection of polyclonal antibodies against Agkistrodon halysii seroglutase (trade name Su Ling, code-named Saculin) and to apply it in clinical immunology tests. Methods The rabbit was immunized with polyclonal antibody and identified by Western blot. The optimal conditions of indirect ELISA for detection of polyclonal antibody against Agkistrodon halysii were established by comparing the results of different conditions. This is a reference test of clinical serum. Results The titer of rabbit antisera was up to 1: 555500 by indirect ELISA. In the 55 clinical trial subjects (36 for the Saculin experimental group and 19 for the placebo group), the serum antibody test results were negative. Conclusion The indirect ELISA method of polyclonal antibody against Agkistrodon halysii in this research can be used for clinical serum determination.