检测日本血吸虫抗体的胶体金-葡萄球菌A蛋白试剂盒研制

Development of a colloidal gold-SPA assay kit for detection of Schistosoma japonicum antibody

目的研制胶体金-葡萄球菌A蛋白试剂盒用于检测日本血吸虫抗体。方法通过不同筛目尼龙绢和低频超声等方法,制备、纯化日本血吸虫可溶性虫卵抗原,研制胶体金-葡萄球菌A蛋白试剂盒。用该试剂盒检测人血清中日本血吸虫特异性抗体,并用ELISA方法进行平行比较。结果胶体金-葡萄球菌A蛋白试剂盒检测慢性血吸虫病的敏感性为93%,检测急性血吸虫病的敏感性为100%,检测健康人血清的特异性为96%,检测肺吸虫病的交叉反应为40%,对其他寄生虫和乙肝未见有交叉反应。与ELISA检测结果比较,两种方法具有良好的一致性。结论胶体金-葡萄球菌A蛋白试剂盒具有快速、简便、不需要仪器、敏感性高和特异性强等优点,可广泛用于血吸虫病流行区现场查病。

Objective To develop a colloidal gold-SPA assay kit for detection of Schistosoma japonicum antibody.Methods The soluble egg antigen of schistosome japonicum was obtained and purified by using nylon mesh filters with different pore sizes and ultrasound treatment.The colloidal gold-SPA assay kit consisted of purified soluble egg antigen(SEA)of Schistosoma japonicum coated onto nitrocellulose membrane, mounted on a lateral flow test device to provide the assay capture matrix.SPA absorbed to a colloidal gold served as detecting reagents.Several different kinds of sera were examined with the kit,and the results were compared with those tested with a ELISA kit.Results The sensitivity of the colloidal gold-SPA assay kit was 93% for chronic schistosomiasis and 100% for acute ones.The specificity was 96%.The cross-reaction was 40% for Paragonimiasis,and no possitive was observed in peoples infected each with clonorchiasis sinensis,hook-worm, or Hepatitis Type B infection.There were no significant difference between the colloidal gold-SPA assay kit and the ELISA kit.Conclusion The results indicated that the colloidal gold-SPA assay kit may provide a simple,rapid test for the detection of schistosome infection with high sensitivity and specificity,suitable for field surveys without any complex instrument.