The rapid isolation from human blood of concentrated, white-cell-free preparations of Plasmodium falciparum.
The sucrose gradient centrifugation method has been applied to representative samples of human (Gambian) blood infected with ring, trophozoite, schizont and gametocyte stages of P. falciparum in order to assess quantitatively the efficiency of recovery, white cell removal and the degree of enrichment of the infected cell fraction. Maximal white cell removal was 90%. (a.v.) Average infected cell recoveries varied with the level of white cell contamination, namely 47% (2-5% WBC), 34% (1-2% WBC) and 24% (smaller than 1% WBC); infected cells were enriched 2-9-fold on average, and up to 22-fold in the case of gametocytes. Preliminary attempts to prepare free parasites by nitrogen cavitation of infected cells showed that disruption took place at much lower pressures than those required to break normal red cells. Gel diffusion analyses showed that the most highly purified infected cell preparations retained the full precipitinogenic spectrum of the original crude preparation.
D000941:Antigens / Q000302:isolation & purification; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000379:methods; D004912:Erythrocytes / Q000469:parasitology / Q000648:ultrastructure; D005714:Gambia; D006801:Humans; D007962:Leukocytes / Q000032:analysis*; D008288:Malaria / Q000097:blood* / Q000276:immunology; D008855:Microscopy, Electron, Scanning; D010963:Plasmodium falciparum / Q000302:isolation & purification*
D000941:Antigens