双抗体夹心ELISA法检测广州管圆线虫循环抗原的研究

目的建立双抗体夹心ELISA法检测广州管圆线虫循环抗原(Cag)。方法采用两株抗广州管圆线虫成虫可溶性抗原单克隆抗体,应用双抗体夹心ELISA法检测广州管圆线虫CAg,并摸索该法的最佳实验条件。对感染大鼠、小鼠、疑似和确诊广州管圆线虫病病例血清进行检测,评价方法的敏感性;对日本血吸虫、肺吸虫、旋毛虫、囊虫病人血清进行交叉反应试验,评价方法的特异性。结果10μg/ml单抗包被,酶标记单抗1∶1 600稀释为最佳工作浓度。利用该法检测不同血清广州管圆线虫循环抗原的敏感性为84.2%~87.2%,特异性为100%。结论建立的双抗体夹心ELISA法检测广州管圆线虫循环抗原具有敏感性高、特异性强和经济、简便易行等优点。

Application of the double antibody sandwich ELISA assay for the detection of circulating antigens of Angiostrongylus cantonensis

By means of two monoclonal antibodies against soluble antigens of adults worms of Angiostrongylus cantonensis,a new method of double antibody sandwich ELISA for detection of circulating antigens was established with the best experimental conditions for performance.The circulating antigens in sera of the rats and mice infected with A.cantonensis as well as in sera of suspected and definitely confirmed patients with A.cantonensis infection were detected with this method to evaluate its sensitivity,and the cross-reaction testing of sera with schistosomiasis,paragonimiasis,trichinellosis and cysticercus cellolosae infection was performed to determine its specificity.It was proved that the optimal working concentration to coat the monoclonal antibody 3F1 was about 10 μg/ml,while the best working concentration of the enzyme-conjugated monoclonal antibody 4H2 was 1∶1 600 for this double antibody sandwich ELISA assay.The sensitivity of this method to detect the circulating antigens of A.cantonensis was 84.2% to 87.2%,and its specificity was 100%.It is evident that the advantages of this method include a very good specificity and moderate degree of sensitivity,easy to handle without need of high technique requirement and instrument and low expense of performance