Purification of Plasmodium vivax by Percoll Density Gradient

Park, Do Sim; Lee, Young Jin; Cho, Ji Hyun

Abstract

BACKGROUND
In Korea, reemerging of indigenous malaria (Plasmodium vivax) have been reported in 1993 and malarial patients were rapidly increased in number and made some social problems. For the effective control for malaria cost effective, high sensitive and specific diagnostic methods has been needed. We tried to purify trophozoites of P. vivax by Percoll density gradient for detecting malaria in P. vivax infection patients who were early stage or the state after taking anti-malarial medicines.
METHODS
Ten mL of EDTA venous bloods from malarial patients were collected and removed plasma after centrifugation. The packed cells were suspended by RPMI 1640 supplemented with 1% fetal bovine serum (1% FBS/RPMI) and overlaid onto the 47%, 45%, and 43% of Percoll solutions, respectively. The buffy coat and under layer were collected after centrifugation. Finally cell pellet suspension with 1% FBS/RPMI were prepared and cytospin slides were made and Wright-Giemsa stained.
RESULTS
The density gradient of 43% Percoll solution showed best result for purifying P. vivax under microscopic observation. Also infected RBC with malarial trophozoite phagocytosed by activated monocyte and neutrophil were noted in 47%, 45%, and 43% of Percoll solutions. CONCLUSTIONS: The Percoll density gradient is very useful laboratory method to detect malaria in P. vivax infection patients who were early stage or the state after taking anti-malarial medicines.