伊氏锥虫变异表面糖蛋白抗原双抗体夹心ELISA检测方法的建立及初步应用

Development and Preliminary Application of Double Antibody Sandwich ELISA for Detection of Trypanosoma evansi VSG Antigen

试验旨在建立一种能鉴定水牛临床感染伊氏锥虫的方法。试验以纯化的伊氏锥虫3D7腹水单抗作为包被抗体,HRP标记的伊氏锥虫5B9单抗作为第2抗体,建立了检测伊氏锥虫变异表面糖蛋白(variant surface glucoprotein,VSG)抗原的双抗体夹心ELISA方法,并进行了灵敏度、特异性试验及初步检测应用试验。结果显示,用该法检测阳性血清,1∶3200稀释时其D450nm值仍大于阴阳临界值;交叉反应试验结果显示,其不与泰勒虫、弓形虫、衣原体、大片吸虫等抗原发生交叉反应,表明该法具有良好的检测特异性;用该法检测了当地82份水牛血清,阳性率为9.76%。结果表明本研究建立的伊氏锥虫VSG抗原双抗体夹心ELISA方法检测灵敏度高、特异性好,能作为检测水牛临床感染伊氏锥虫的有效方法,对于临床伊氏锥虫病的诊断和防控具有重要意义。

A double antibody sandwich ELISA for detection of Trypanosoma evansi variant surface glucoprotein （VSG） an- tigen was established by using two monoclonal antibodies against Trypanosoma evansi VSG antigen. The result of sensitivity showed when positive serum was diluted to 3200 doubles, its D450 nm value was still greater than the critical value of positive and negative, and specificity test result showed that there was no cross reaction with the antigens of Theileria, Toacoplasma gondii, Chlamydia and Fasciola gigantica. 82 clinical sera were detected by this double antibody sandwich ELISA and its positive rate was 9. 76%. The experiment results demonstrated that this double antibody sandwich ELISA for detection of Trypanosorna evansi VSG antigen had good detecting sensitivity and specificity,and could be used as an effective method for clinical detecting buffalo Trypanosoma evansi disease.