Avraham H, Golenser J, Gazitt Y, Spira D T, Sulitzeanu D
J Immunol Methods. 1982 Aug 27;53(1):61-8. doi: 10.1016/0022-1759(82)90240-x.
A highly sensitive radioimmunoassay for detection of P. falciparum antibodies and antigens is described. A partially purified P. falciparum antigen preparation is obtained from in vitro cultured parasites enriched after gelatin sedimentation by sonicating the infected red blood cells and precipitating the proteins with 50% saturated ammonium sulfate. The precipitate is dissolved in buffer, ultracentrifuged and used to coat wells of microtiter plates. Anti-P. falciparum antibodies are detected by incubating antiserum dilutions in the coated wells and detecting the bound IgG with radioiodinated staphylococcal protein A. P. falciparum antigens are detected by their ability to inhibit binding of antibodies to the coated wells. Sera of individuals with a history of P. falciparum infection contain antibodies detectable at a dilution of 1:75,000. P. falciparum RBC infected in vitro can be detected at levels of parasitemia of the order of 1 parasite or less per 10(6) RBC.
本文描述了一种用于检测恶性疟原虫抗体和抗原的高度灵敏的放射免疫测定法。通过对感染的红细胞进行超声处理并用50%饱和硫酸铵沉淀蛋白质，从经明胶沉淀富集后的体外培养寄生虫中获得部分纯化的恶性疟原虫抗体制剂。将沉淀物溶解在缓冲液中，进行超速离心，然后用于包被微量滴定板的孔。通过在包被孔中孵育抗血清稀释液并用放射性碘化葡萄球菌蛋白A检测结合的IgG来检测抗恶性疟原虫抗体。通过其抑制抗体与包被孔结合的能力来检测恶性疟原虫抗原。有恶性疟原虫感染史的个体血清中，抗体在1:75,000的稀释度下可被检测到。体外感染恶性疟原虫的红细胞在每10(6)个红细胞中有1个或更少寄生虫的寄生虫血症水平下即可被检测到。
