Mackey L J, McGregor I A, Paounova N, Lambert P H
Bull World Health Organ. 1982;60(1):69-75.
An ELISA method has been developed for the diagnosis of Plasmodium falciparum infection in man. Parasites from in vitro cultures of P. falciparum were used as source of antigen for the solid phase and the source of specific antibody was immune Gambian sera; binding of antibody in antigen-coated wells was registered by means of alkaline phosphatase-conjugated anti-human IgG. Parasites were detected on the basis of inhibition of antibody-binding. The test was applied to the detection of parasites in human red blood cells (RBC) from in vitro cultures of P. falciparum and in RBC from infected Gambians; RBC from 100 Geneva blood donors served as normal, uninfected controls. In titration experiments, the degree of antibody-binding inhibition correlated with the number of parasites in the test RBC. Parasites were detected at a level of 8 parasites/10(6) RBC. Samples of RBC were tested from 126 Gambians with microscopically proven infection; significant antibody-binding inhibition was found in 86% of these cases, where parasitaemia ranged from 10 to 125 000/mul of blood. The presence of high-titre antibody in the test preparations was found to reduce the sensitivity of parasite detection in infected RBC from in vitro cultures mixed with equal volumes of different antibody-containing sera. The sensitivity was restored in most cases by recovering the RBC by centrifugation before testing. In a preliminary experiment, there was no significant difference in antibody-binding inhibition using fresh infected RBC and RBC dried on filter-paper and recovered by elution, although there was greater variation in the latter samples.
已开发出一种酶联免疫吸附测定(ELISA)方法用于诊断人类恶性疟原虫感染。来自恶性疟原虫体外培养物的寄生虫用作固相抗原的来源,特异性抗体的来源是冈比亚免疫血清;通过碱性磷酸酶偶联的抗人IgG来记录抗原包被孔中抗体的结合情况。基于抗体结合的抑制来检测寄生虫。该试验应用于检测来自恶性疟原虫体外培养物的人红细胞(RBC)以及感染冈比亚人的红细胞中的寄生虫;来自100名日内瓦献血者的红细胞用作正常、未感染的对照。在滴定实验中,抗体结合抑制程度与测试红细胞中的寄生虫数量相关。在每10⁶个红细胞中有8个寄生虫的水平下可检测到寄生虫。对126名经显微镜证实感染的冈比亚人的红细胞样本进行了检测;在这些病例中,86%发现有明显的抗体结合抑制,其中疟原虫血症范围为每微升血液10至125000个。发现测试制剂中高滴度抗体的存在会降低在与等体积不同含抗体血清混合的体外培养感染红细胞中检测寄生虫的敏感性。在大多数情况下,通过在测试前离心回收红细胞可恢复敏感性。在一项初步实验中,使用新鲜感染的红细胞以及在滤纸上干燥并通过洗脱回收的红细胞,抗体结合抑制没有显著差异,尽管后一种样本的变化更大。