融合蛋白斑点ELISA检测恶性疟抗体

FUSION PROTEIN Dot-ELISA FOR THE DETECTION OF PLASMODIVM FALCIPARVM ANTIBODIES

利用基因工程技术将恶性疟原虫基因组DNA与噬菌体λgt11重组,并在宿主菌E.Coli Y1090中表达出融合蛋白,以此蛋白作为抗原,用斑点ELISA法检测恶性疟抗体。当血清1:100稀释时,46份病人血清全为阳性,20份正常人血清全为阴性,与IFA检查结果一致。研究表明此种融合蛋白可替代疟疾全虫抗原,可解决抗原来源和纯化问题。

The feasibility of using fusion protein Dot-ELISA to detect the antibodies to Plas-modium falciparum was presented. Fusion protein expressed in the P. falciparum genomic DNA expression library with λgtll. was used as the antigen to coat the solid support nitrocellulose strips. Horseradish peroxidase conjugated with goat antihuman IgG was used as the second antibody. The enzyme activity was read by naked eyes according to the dot colour. The results by Dot-ELISA correlated well with those by IFA. The study indicated that this method was specific, sensitive, simple, and replicable. It would be valuable for further studying the usefulness of fusion protein in the immuncdiagnosis of malaria.