血清旋毛虫p49抗原IgG抗体的斑点金免疫渗滤法检测

Detection of the IgG antibodies against p49/GST antigen in sera of patients with Trichinella spiralis infection by dot-immunogold filtration assay

目的 建立一种快速诊断人体旋毛虫感染的新途径。方法 以基因工程表达的猪旋毛虫融合蛋白p49/GST为抗原,将其结合于硝酸纤维素膜上,以胶体金颗粒结合的羊抗人IgG作为标记抗体,建立斑点金免疫渗滤法(Dot-immunogoldfiltration assay,DIGFA),检测人血清抗旋毛虫p49抗原IgG抗体。共检测76份旋毛虫患者血清并与酶联免疫吸附试验(ELISA)作对比研究。结果 抗原抗体通过渗滤在膜上反应,10min肉眼即可观察到结果,在76份患者血清的检测中,阳性率为98.68％,显著高于ELISA法的检出阳性率86.84％,(X2=7.94,P<0.01)。交叉试验与重复试验结果显示DIGFA具有较好的特异性及稳定性。结论 以纯化的融合蛋白p49/GST为抗原建立的DIGFA可快速准确检测旋毛虫病人血清特异性旋毛虫IgG抗体,具有推广应用的价值。

In order to establish a rapid method to detect the antibodies against antigens in sera of patients with Trichinella spiralis infection, the genetically engineered fusion protein p49/GST used as antigen was coated on the nitrocellulose membrane, and combined with colloid gold labeled goat anti human IgG, thus to establish the dot -immunogold filtration assay(DIGFA). It was found the results of this method of assay could be observed within 10 minutes by naked eyes. By using this method to examine 76 clinical serum samples, the positive rate for the IgG antibodies against p49/GSF antigen was 98.68%, that was significantly higher than that of ELISA assay. It indicates that dot immunogold filtration assay is a rapid, simple , sensitive and specific method to detect the specific antibodies in sera of patients with Trichinella spiralis infection.