胶体染料斑点免疫试验检测血吸虫病人抗体方法的建立及初步应用

本研究首次从国产染料中筛选出一种能用于免疫反应显色的蓝色染料(D-1),并对用此染料标记羊抗人IgG的最适条件进行了探讨。这种标记抗体的胶体染料能与相关抗原反应并直接显色,双抗体夹心法检测人IgG蛋白的最低检出量为25—10ng/ml,胶体染料斑点免疫试验(DIA)检测62份血吸虫病人血清,61份阳性,阳性率为98.4％,40份姜片虫病人血清均为阴性,30份非血吸虫病流行区的健康人血清有1份出现阳性,假阳性率为3.3％,与Dot-ELISA检测结果相似。标记的胶体染料在室温下保存1wk仍保持活性不变,冻干能保存更长时间,与酶联免疫斑点法相比似有更高的实用价值。

ESTABLISHMENT AND PRELIMINARY APPLICATION OF DOT COLLOIDAL DYE IMMUNOASSAY FOR ANTIBODY DETECTION IN PATIENTS OF SCHISTOSOMIASIS JAPONICA

A kind of colloidal blue dye(D-l)used as a staining reagent for immunoassay was first selected from the dyes produced in China in this study. The optimun condition for labelling the dye onto sheep antihuman IgG antibody was explored. The dye-labelled antibody could react and stain with relative antigens. The minimal concentration of human IgG protein could be detected with the labelled dye by Dot Double Antibody Sandwich Assay was 20-10ng/ ml. 61 of 62 sera samples of schistosomiasis japonica were positive,the positive rate was 98.4%. Just 1 out of 30 healthy people's sera sambles was positive,the false positive rate was 3.3%. All of 40 Fasciolopsiasis sera samples were negative. The results were similar to that of Dot-ELISA. The activity of antibody labelled with colloidal dye could be maintained for 1 week in room temperature and be presered in lyophilized condition for a longer period. The assay was less expensive,simple to conduct and required no special equipment. It suggested that the Dot Colloidal Dye Immunoassay(DIA)might have potential value for use in schisto-somiasis endemic areas.