间日疟原虫、恶性疟原虫和伯氏疟原虫各期的提纯和分离

用纤维素(Cellulose,CF-11)过滤去除白细胞后再用 Percoll 密度梯度离心方法提纯分离间日疟(P.v.)、恶性疟(P.f.)和伯氏疟(P.b.)3种疟原虫。经纤维素过滤后,P.v.病人血和P.b.感染鼠血中白细胞去除率分别平均达 94.7％和 75.4％,而且不影响原虫密度和各期构成比。多个 Percoll 不连续梯度离心测试3种疟原虫裂殖体、滋养体、环状体和未感染红细胞的相对平均密度(kg/L):P.v.分别为 1.059、1.072、1.096 和 1.099;P.f.分别为 1.070、1.079、1.10 和 1.10,配子体 1.085;P.b.分别为 1.059、1.062、1.103 和 1.104。并据此选择各自合适的 Percoll 梯度分离3 种疟原虫,将裂殖体、滋养体同环状体分开,获取原虫纯度(原虫数/100 个红细胞),P.v.由分离前 0.8％至分离后 93.5％,P.f.由2.3％至100％,P.b.由 30.6％至93％,分别提高 113.8、43.4和 2倍。

PURIFICATION AND ISOLATION OF DIFFERENT STAGES OF PLASMODIUM VIVAX, P. FALCIPARUM AND P. BERGHEI

The present paper reported the results of purification and isolation of different stages of P. vivax, P. falciparum and P. berghei by Percoll gradient centrifugation after removal of leukocytes (WBCs) by passing the blood suspension through CF -11 cellulose columns. 94. 7% and 75. 4% of WBCs from the patients* blood of vivax malaria and the mice blood infected with P. berghei respectively were removed with no alteration of the parasite density and the malaria stage ratio after the cellulose filtration.The densities of the uninfected and the infected erythrocytes with different stages of 3 species of malaria parasites were determined by isopycnic centrifugation on multi continuous gradients of Percoll. The mean densities (kg/L) of schizonts, trophozoites, ring form and uninfected cells were approximately 1. 059,1. 072,1. 096 and 1. 099 in P. vivax) 1. 070,1. 079,1.10 and 1. 10, gametocytes 1. 085 in P. falciparum; 1. 059,1. 062,1. 103 and 1. 104 in P. berghei, respectively. Based on these values, 3 different density gradients of Percoll were devised to separate erythrocytes infected with the more mature stages (trophozoites and schizonts) from ring forms and uninfected cells. By these method, it is possible to obtain high purified parasites of 93. 5% from blood with starting parasitaemias of 0. 8% in P. vivax, 100% from 2. 3% in P. falciparum and 93. 0% from 30. 6% in P. berghei, being raised bv 113. 8, 43. 4 and 2. 0 times respectively.