论文部分内容阅读
本文报道ELISA法检测抗恶性疟原虫裂殖子主要表面蛋白gp190特异性抗体的一种新方法(β-Gal桥连ELISA),可用于诊断人体初次感染急性疟疾。 F2,M6和F2.7在大肠杆菌中的表达:M6含有MAD_(20)株的gp190的384～595氨基酸序列,F2和F2.7分别含有K_1株的gP190 105～321和216～321氨基酸序列。M6DHFR(二氢叶酸还原梅),F2-CAT(氯胺苯醇乙酰转移梅)和F2.7-CAT融合蛋白的克隆和表达按Crisanti等(1988)方法进行。β-Gal桥连ELISA法:将500ng的M6-DHFR和100ng的F2-CAT溶于50μ10.1mol/L NaHCO_3中,分别包被平底微量滴定板。非特异性结合位点用BSA封闭。用TBST液洗5次,每井加入50μl抗血清,孵育过
This article reports a new method (β-Gal bridging ELISA) to detect gp190-specific antibody against major surface protein of Plasmodium falciparum meconium by ELISA, which can be used to diagnose acute malaria infection in humans. Expression of F2, M6 and F2.7 in Escherichia coli: M6 contains the amino acid sequence of 384-595 of gp190 of MAD 20 strain, F2 and F2.7 contain the gP190 105-312 and 216-221 amino acid sequences of K 1 strain respectively . Cloning and expression of M6DHFR (plum dihydrofolate reductase), F2-CAT (chloramphenicol acetyltransfera) and F2.7-CAT fusion protein were performed according to Crisanti et al. (1988). β-Gal Bridging ELISA: 500ng of M6-DHFR and 100ng of F2-CAT were dissolved in 50μ10.1mol / L NaHCO_3 and coated with flat-bottomed microtiter plates respectively. Non-specific binding sites are blocked with BSA. Wash with TBST 5 times, add 50μl antiserum to each well and incubate