Plasmodium vivax: isolation of mature asexual stages and gametocytes from infected human blood by colloidal silica (Percoll) gradient centrifugation.

The densities of human erythrocytes infected with P. vivax obtained from infected patients were determined by isopycnic centrifugation in continuous gradients of Percoll. The approximate densities of erythrocytes infected with rings were 1.086 to 1.1, trophozoites (amoeboid stages) 1.053 to 1.086, schizonts and gametocytes 1.053 to 1.056 and of the other cellular elements of blood, uninfected erythrocytes, 1.086 to 1.1, polymorphonuclear leucocytes 1.073 to 1.086 and mononuclear cells 1.062 to 1.073 g/ml. Based on these values, a one-step gradient of 47% Percoll was devised to separate erythrocytes infected with the more mature stages (trophozoites, schizonts and gametocytes) from uninfected erythrocytes. By this method it is possible to obtain parasitaemias of 88% to 98% from blood with starting parasitaemias of less than 0.7%. This method is therefore being routinely used for immunological, biochemical and molecular biological studies on P. vivax.