单克隆抗体夹心斑点免疫金银染色法诊断恶性疟的初步研究

PRELIMINARY STUDIES ON THE DIAGNOSIS OF PLASMODIVM FALCIPARVM MALARIA BY MONOCLONAL ANTIBODY SANDWICH DOT-IMMUNOGOLD SILVER STAINING ASSAY

用单克隆抗体(McAb)夹心斑点免疫金银染色法(Dot-IGSSA),对采自海南省的30价原虫血症范围为0.015-0.58％的恶性疟病人血和30份正常人血进行了检测。当用McAb 11G_5、13A_2、13A_1分别与金标记McAb 14D_9夹心时,其阳性检出率分别为96.7％、93.3％和93.3％,假阳性反应约为3.3％,最低可检测出约0.0001％的疟原虫血症。其中11G_5、13A_1分别与金标记14D_9夹心时,对体外培养的云南和安徽的恶性疟红内期抗原呈阳性反应,但其McAb与诺氏疟原虫和伯氏疟原虫均无交叉反应。McAb 13A_1与14D_9夹心时,对食蟹猴疟原虫和间日疟病人感染血中的疟原虫抗原有部分交叉反应。

In this report, the blood samples from 30 falciparum malaria patients with parasftemia 0.015-0.58% and the blood samples from 30 healthy persons were examined by monoclonal antibody (McAb) sandwich dot-immunogold silver staining assay (Dot-IGSSA). When the McAb 11G5, 13A2 and 13A1 were used for sandwith Dot-IGSSA with McAb 14D9 labeled with colloidal gold respectively, the 0.0001% of parasitemia could be detected and the McAb 11G5, 13A1 and 14D9 labeled with colloidal gold could also be used to detect the an-tigens of asexual blood stages of Yunnan and Anhui isolates of Plasmodium falciparum cultured in vitro. These McAbs did not cross-react with the antigens of Plos .Mum know-lesi and Plasmodium berghei, however, the McAbs 13A1 and 14D9 weakly cross-reacted with the antigens of Plasmodium cynomolgi and the antigens in the infected blood samples from patients with vivax malaria (Fig. 1).