犬恶丝虫病ELISA诊断方法的建立

Development of indirect ELISA for the detection of canine dirofilariasis

目的建立犬恶丝虫病的ELISA诊断方法。方法将犬恶丝虫成虫粗制抗原经过SephaexG-200凝胶柱层析后进行间接ELISA试验,确定最佳实验条件,建立犬恶丝虫病的同种纯化抗原ELISA诊断法。通过阻断性试验、交叉反应试验、敏感性和特异性试验、重复性试验及与病原学方法比较,评价其效果。结果纯化抗原ELISA方法与犬钩虫和蛔虫感染犬血清无交叉反应,检出的阳性血清最高抗体滴度为1∶25600,重复性好。结论成功建立犬恶丝虫病ELISA诊断方法,可替代虫检法对犬恶丝虫病进行早期诊断。

Objective To establish diagnostic method of ELISA on heartworm disease. Methods The raw protein of Dirofilaria irnrnitis was separated by Sephadex G-200. All peaks of protein were collected and detected as ELISA antigen. The best experiment condition was determined through estimating the opitimal working concentration. Effect of the ELISA was estimated by compared with interruption test, cross-reaction test, sensibility and specificity test, repetition test and nosetiology method. Results It has been proved that the antigen had higher sensitivity, specificity and stability. There was no cross-reaction with Toxocara and Ancylostoma. And the 1 ： 25 600 antibody titer could be detected with purified antigen. Conclusion The ELISA could replace the parasite examination in dirofilariasis diagnosis in earlier period.