重组ASl6蛋白抗原检测猪蛔虫抗体间接ELISA方法的建立

目的]建立检测猪蛔虫抗体的间接ELISA方法。方法]用猪蛔虫重组ASl6蛋白作为包被抗原，通过方阵滴定试验确定ELISA的最佳工作条件，包括抗原最佳包被浓度与血清最佳稀释度、最佳包被液、抗原封闭时间、血清反应时间、酶标二抗工作时间和间接EusA判定标准，并通过交叉试验现场对1200份猪血清样本采用该试验建立的EusA和AGP同时进行蛔虫抗体的检测。结果]ELISA的最佳工作条件是：抗原包被浓度为10μg／ml，37℃1h，4℃过夜；血清稀释度为1：80；酶标SPA稀释度为1：t0000，37℃孵育45min。该试验制备的ELISA酶标板的临界值为0．196。交叉试验检测表明，ELISA方法阳性检出率为45％，而AGP阳性检出率为30％。该方法具有特异高、快速、敏感等特点。结论]研究结果证实重组ASl6蛋白具有良好的反应原性，可以用来建立ELISA方法诊断猪蛔虫病。

Establishment of An Indirect ELISA Method Based on Recombinant AS16 Protein Antigen for Detection of Antibodies to Ascaris suum

Objective] The study aimed to establishment of an indirect ELISA method for detection of antibodies to Ascaris suum.Method] With recombinant AS16 protein of Ascaris suum as the coating antigen,the optimal reaction conditions of ELISA including the optimal coating concn.of antigen and dilution degrees of serum,the optimal coating liquid,antigen closing time,reaction time of serum,working time of enzyme-labeled second antibody and the judging standard of indirect ELISA were determined through the matrix tit...