ABC-ELISA法检测犊弓首蛔虫IgG抗体实验研究

DETECTION OF TOXOCARA VITULORUM IgG ANTIBODY BY USING ABC-ELISA

首次以动物为模型,应用 ABC-ELISA 法对家兔血清中犊弓首蛔虫 IgG 抗体进行了测试研究。实验中对犊弓首蛔虫的成虫体壁抗原、体腔液抗原、生殖器官抗原、虫卵抗原、幼虫抗原进行了比较研究,结果显示:幼虫抗原对血清检测的 P/N 值及对被检血清的检出率最高,而检测对照组血清未出现假阳性反应.由此表明,幼虫抗原是最敏感和特异的抗原.在检测方法上,ABC-ELISA 法对被检血清的检出率为96.62%。常规 ELISA 法和 IHA 法的检出率分别为90%和56.67%。3种方法检测对照组血清未出现假阳性反应;在抗体检测滴度上,ABC-ELISA 法比常规 ELISA 法高1～4倍,比 IHA 法高4～128倍.经统计处理,ABC-ELISA 法比常规 ELISA 法敏感1.21倍,比 IHA 法敏感33.76倍.在特异性实验中,用 ABC-ELISA 法检测家兔血清中犊弓首蛔虫 IgG 抗体,与实验家兔乳突类圆线虫和肝片吸虫阳性血清均未出现交叉反应,表明该法检测犊弓首蛔虫抗体特异性良好.另外,对4份血清多次重复检测结果,批内重复和批间试验 OD 值变异系数不超过10%,表明该法稳定性良好.实验结果表明,ABC-ELISA 法具有敏感性高,特异性强,并有较好的稳定性,可用于犊弓首蛔虫抗体的检测,故该法的建立无疑具有理论和实践意义.

Firstly using rabbit as cxperimental model,in the detection of Toxocara viluloum IgG antibody by ABC—ELISA.Comparisons were made of nive antigens of Toxocara vitulorum,and this providcd cvidence that larvae antigen has the highest P/N response and with highcst positivc pcrccntagc among these fivc antigcns.Among the detcc- tivc mcthods,the positive percentage od ABC—ELISA is 96.67%,convcntional ELISA is 90%and IHA is 56.67%.Comparisons of antibody titers dcteetcd,ABC—ELTSA is 1—4 and 4—128 times higher than that of convcntional ELISA and IHA rcspectively.The results of statistical treatment showcd that the scnsitivity of ABC—ELTSA is 1.21 and 33.76 times higher than that of conventional ELISA and IHA.No false positivc reaction was obscrvcd when used ABC—ELISA to cxamine the sera of control group.Furthermore,when ABC—ELISA was uscd to dcteet the Toxocara vitlourum TgG antibody,it showed no cross reaction with positive scra of Fasciola hepatic and Strongyloides papillosus in cxperi- mcntal rabbits.It is vcry stablc,its rcsults were almost idcntical when the same sample was examincd scvcral times in different periods.The results indicate that ABC—ELISA can be uscd to deteet the Toxocara vitulorum antibody cffectively,so the establishmcnt of this method must has grcat significance in thcory and practiee.