Development of new malaria parasite detection system : Establishment of automatic and simple system

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We established a simple and sensitive flow cytometric detection system for Plasmodium falciparum that involves lysing erythrocytes and staining parasites at the same time using a newly developed hemolysing solution containing acridine orange. In this system, freed parasites of P.falciparum could be plotted separately from erythrocyte ghosts, white blood cells and platelets on the two-dimensional scattergram of forward angle light scatter and green fluorescence by flow cytometry with an argon laser. It took only 2-3 minutes per sample to obtain the scattergram and analyze the data, including the time of sample preparation for the flow cytometric analysis. Sample preparation does not require any particularly difficult handling procedures. The threshold of parasite detection was almost equal to that of microscopic examination for cultured P.falciparum. The results of drug susceptibility assays using this system were quite similar to those obtained using microscopic examination. In this system, parasites at different erythrocytic stages could be easily distinguished. Moreover, using a flow cytometer with imaging device, clear images of gametocytes were obtained. This system has not only the potential to be useful for laboratory studies, but also a high potential of becoming an ideal new diagnostic system for malaria in the near future.