Zibaei Mohammad, Sadjjadi Seyed Mahmoud, Ishiyama Satoko, Sarkari Bahador, Uga Soji
Department of Parasitology and Mycology, School of Medicine, Lorestan University of Medical Sciences, Khorram Abad, Iran.
Hybridoma (Larchmt). 2010 Jun;29(3):217-20. doi: 10.1089/hyb.2009.0108.
Toxocariasis is an important zoonotic disease caused by the second-stage larvae of Toxocara species. Human toxocariasis is usually diagnosed by serological methods. We prepared monoclonal antibody specific for Toxocara cati (T. cati) and performed the following experiments using this antibody to increase the sensitivity of our capture ELISA for antigen detection. The isotype of monoclonal antibody was IgG3. Using this monoclonal antibody, a capture ELISA was developed. The produced monoclonal antibody reacted well with excretory-secretory (ES) antigen of T. cati with protein nature but not with other T. cati related antigens. A monoclonal antibody was raised against ES antigen of T. cati larval stage. No cross-reaction was found with antigens of Ascaris suum, Dirofilaria immitis, Trichostrongylus orientalis, encysted larvae of Trichinella spiralis, and metacercariae of Centrocestus armatus. The system had enough sensitivity to detect 5 ng/mL of circulating antigens in the serum. The results indicate that the new antigen detection system provides a useful method for the diagnosis of visceral and ocular larva migrans caused by T. cati and fulfils the requirements of a serological method for diagnosis of toxocariasis.
弓蛔虫病是一种由弓蛔虫属第二期幼虫引起的重要人畜共患病。人体弓蛔虫病通常通过血清学方法诊断。我们制备了针对猫弓首蛔虫(T. cati)的单克隆抗体,并使用该抗体进行了以下实验,以提高我们用于抗原检测的捕获ELISA的灵敏度。单克隆抗体的亚型为IgG3。利用该单克隆抗体,开发了一种捕获ELISA。所产生的单克隆抗体与具有蛋白质性质的猫弓首蛔虫排泄分泌(ES)抗原反应良好,但与其他猫弓首蛔虫相关抗原无反应。制备了一种针对猫弓首蛔虫幼虫期ES抗原的单克隆抗体。未发现与猪蛔虫、犬恶丝虫、东方毛圆线虫、旋毛虫包囊幼虫和有棘中央线虫囊蚴的抗原发生交叉反应。该系统具有足够的灵敏度来检测血清中5 ng/mL的循环抗原。结果表明,新的抗原检测系统为诊断由猫弓首蛔虫引起的内脏和眼部幼虫移行症提供了一种有用的方法,并满足了弓蛔虫病血清学诊断方法的要求。