A rapid, simple and sensitive flow cytometric system for detection of Plasmodium falciparum.
We have established a rapid, simple and sensitive flow cytometric system for the detection of Plasmodium falciparum that involves lysing erythrocytes and staining parasites at the same time using a newly developed hemolysing and staining solution containing dodecyl methyl ammonium chloride and 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 min per sample to obtain the scattergram and analyze the data, including the time of sample preparation for flow cytometric analysis. Sample preparation with this method does not require any 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 also almost identical to those obtained using microscopic examination. In this system, parasites at different erythrocytic stages could be easily distinguished. This system must prove useful and practical for basic laboratory studies of P. falciparum including those requiring the differential measurement of parasites at specific erythrocytic stages.
Saito-Ito
A
A
Department of Medical Zoology, Kobe University School of Medicine, 650-0017, Kobe, Japan. atsuko@med.kobe-u.ac.jp
Akai
Y
Y
He
S
S
Kimura
M
M
Kawabata
M
M
D000165:Acridine Orange / Q000737:chemistry; D000368:Aged; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000469:parasitology; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D020128:Inhibitory Concentration 50; D016778:Malaria, Falciparum / Q000097:blood / Q000469:parasitology*; D008297:Male; D018512:Parasitemia / Q000097:blood / Q000469:parasitology; D010963:Plasmodium falciparum / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D000165:Acridine Orange