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作者用纯化的抗原以ELISA法测定疟疾患者血清中的循环抗体,并与IFI比较。按下列程序制备抗原:将恶性疟原虫(Palo Alto株)感染松鼠猴(Saimiri sciureus),取被感染的猴血约10ml,其红细胞约80%被裂殖体寄生。经冻融处理使红细胞溶解,以1,400g×15分钟离心,用0.15M的NaCl洗涤,再以1,400g×15分钟离心,得沉淀物A(内含疟原虫和红细胞间质)及上清液B。将上清液B经Amicon YM10超滤膜过滤浓缩,然后再经CM葡聚糖凝胶A50柱分离得第1峰(作为抗Ⅱ)和经62%硫酸胺处理的第2峰(作为抗原Ⅲ)。将上述沉淀物A经超声粉碎并经40,000g×30分钟离心,得沉淀C和上清液D。将所得沉淀C再次超声
The authors used purified antigens to measure circulating antibodies in sera from malaria patients by ELISA and compared them to IFI. Antigen was prepared according to the following procedure: Plasmodium falciparum (Palo Alto strain) was infected with Saimiri sciureus, and about 10 ml of infected monkey blood was taken. About 80% of erythrocytes were parasitized by schizont. The erythrocytes were lysed by freeze-thawing treatment, centrifuged at 1,400 g for 15 minutes, washed with 0.15 M NaCl and then centrifuged at 1,400 g for 15 minutes to obtain a precipitate A (containing plasmodia and erythrocyte interstitium) and a supernatant B. The supernatant B was concentrated by filtration on an Amicon YM10 ultrafiltration membrane and then the first peak (as anti-II) and the second peak treated with 62% ammonium sulfate (as antigen III ). The above precipitate A was sonicated and centrifuged at 40,000 g for 30 minutes to precipitate C and supernatant D. The resulting precipitate C was sonicated again