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为了解孢子体与肝细胞相互作用的情况,作者报道1种新的检查方法——酶联免疫和免疫荧光双染色测定法来鉴定孢子体是寄生于肝细胞膜表面,还是侵入肝细胞内。该法首先将有代谢功能的肝细胞体外培养,再接种一定量的疟原虫孢子体继续培养数小时使其感染,然后用这些培养物分二步进行染色观察。第1步酶联免疫测定:培养物洗3次——4%聚甲醛固定5分钟孢子体表面抗原单克隆抗体37℃孵育30分钟——洗3次——过氧化物酶标记的第2抗体(羊抗鼠Ig)洗3次加底物显色——洗3次——无水乙醇处理20分钟——洗3次——置37℃过夜。第2步免疫荧光测定:次日将上述处理的培养物洗3次——单克隆抗体孵育洗3次——荧光
To understand the interaction of sporozoites with hepatocytes, the authors reported a new assay, enzyme-linked immunosorbent assay and immunofluorescence double staining, to identify sporozoites that are parasitic on the surface of hepatocyte membranes or invade hepatocytes. The method first of all the metabolic function of liver cells cultured in vitro, and then inoculated with a certain amount of Plasmodium sporozoites continued to culture a few hours to make it infected, and then use these cultures in two steps for staining. Step 1 Enzyme-linked immunoassay: Cultures were washed three times - 4% POM fixed for 5 minutes Spore body surface antigen Monoclonal antibodies incubated at 37 ° C for 30 minutes - washed three times - Peroxidase-labeled second antibody (Goat anti-mouse Ig) 3 times plus substrate color - wash 3 times - ethanol treatment for 20 minutes - wash 3 times - set 37 ℃ overnight. Step 2 Immunofluorescence Assay: The next day the culture of the above treatment was washed three times - Monoclonal antibody incubation wash three times - Fluorescence