A rapid, simple and sensitive flow cytometric system for detection of Plasmodium falciparum

Saito-Ito, Atsuko; Akai, Yasumasa; He, Shenyi; Kimura, Mikio; Kawabata, Masato

doi:10.1016/s1383-5769(01)00091-5

Cited by 31 publications

(35 citation statements)

References 14 publications

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“…In contrast with previous reports (Saito-Ito et al 2001), our results based on FCA data, showed that IC 50 's of each antimalarial compound were consistent with those obtained by microscopy, and 3 [H]-hypoxanthine uptake.…”

Section: Discussionsupporting

confidence: 58%

“…FCA have proved to be useful to study diverse features related with human and experimental malaria (Janse et al 1987, Pattanapanyasat et al 1993, 1999, Kumaratilake & Ferrante 2000, Saito-Ito et al 2001. In this study, we have described by flow cytometry the anti-malarial activity of four drugs using continuos cultures of P. falciparum geographically different strains.…”

Section: Discussionmentioning

confidence: 99%

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Stage-specific activity of potential antimalarial compounds measured in vitro by flow cytometry in comparison to optical microscopy and hypoxanthine uptake

Contreras

Rivas

Domínguez

et al. 2004

Mem. Inst. Oswaldo Cruz

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Key words: Plasmodium falciparum -antimalarial compounds -flow cytometry -hypoxanthine uptake -chloroquine -quinolone Drug resistance of Plasmodium falciparum, the most deadly human malaria parasite, is a major factor in the widespread persistence of malaria (Ouellette & Kunding 1997, Macreadi et al. 2000. Current efforts focus on research into novel compounds and on measures to prevent or delay resistance once new drugs are introduced. However, malaria therapy has generally not taken into consideration the stage-specificity of action of different drugs. This is an important consideration, since inappropriate timing of administration of antimalarial drugs might limit drug efficacy and favor the selection of drug-resistant parasites.Few studies have focused on the in vitro stage-specific efficacy of antimalarial compounds (Chimanuka et al. 2001). Most in vitro studies monitoring resistance and susceptibility to antimalarial compounds have been performed by microscopy and by uptake of a radiolabelled nucleic acid precursor 3 [H]-hypoxanthine (Desjardins et al. 1979). They are poorly suited to discriminate stagespecific activity. More recently, flow cytometry analysis (FCA) using different fluorescent dyes has been reported (van Vianen et al. 1990, Pattanapanyasat et al. 1996.We used propidium iodide (PI) whose incorporation as fluorescent molecule has been shown to be proportional to the DNA parasite content. In the present study, results of in vitro effect of antimalarial compounds on P. falciparum parasitized erythrocytes obtained from FCA were compared with other assays. FCA results were comparable to optical microscopy observation and to [ 3 H]-hypoxanthine uptake assay. Moreover, FCA offers advantages of high throughput, ready automation, simplified data processing, and easy determination of the stage-specific effectiveness of potential antimalarial compounds.

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Section: Discussionsupporting

confidence: 58%

Section: Discussionmentioning

confidence: 99%

Stage-specific activity of potential antimalarial compounds measured in vitro by flow cytometry in comparison to optical microscopy and hypoxanthine uptake

Contreras

Rivas

Domínguez

et al. 2004

Mem. Inst. Oswaldo Cruz

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“…FCM has found a broad range of applications in bacteriology and parasitology and has been used in studies of various parasitic protozoa such as Plasmodium, Toxoplasma, Eimeria, Leishmania, free living amoeba, Cryptosporidium, Giardia, and microsporidia (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). It has been used to quantitate and determine the viability of E. cuniculi spores from culture (5), to isolate E. bieneusi spores from stool samples (19), to compare the nucleic acid contents of microsporidian spores isolated from fish (20), and to discriminate between the spores of the three species of Encephalitozoon (22).…”

Section: Discussionmentioning

confidence: 99%

“…However, the use of FCM in parasitology has been largely restricted to the study of extracellular parasites. Exceptions are the Plasmodium spp., which can be specifically labeled with fluorescent DNA-binding dyes (18), and cytometric assays for the quantitation of intracellular Leishmania and Toxoplasma infections (12,15,23). In this report, we describe the use of FCM as a tool to allow the determination of intracellular microsporidia in various cell lines and humanderived macrophages.…”

Section: Discussionmentioning

confidence: 99%

Quantitation of microsporidia in cultured cells by flow cytometry

et al. 2004

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Background: Microsporidia are obligate intracellular protozoan parasites that emerged as major opportunistic pathogens in humans since the onset of the AIDS pandemic. In the present study, we investigated whether FCM is a useful method for the quantitation of intracellular microsporidian spores in cultured cells. Methods: Microsporidia (Encephalitozoon cuniculi) were grown in cell cultures and various cell-lines were coincubated with microsporidian spores at different multiplicities of infection, as well as for different periods of time. After permeabilization of the cells, intracellular spores were stained with a polyclonal anti-E. cuniculi serum and a FITC-labeled secondary antibody. Stained cells were analyzed on a flow cytometer and results were compared with those of fluorescence microscopy.

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“…Several methods of detection of malaria parasite by flow cytometry have been developed taking advantage of the absence of DNA in erythrocytes. Different dyes such as acridine orange (17,18), hydroethidine (19,20), SYTO 16 (21) and thiazole orange (22) were already employed for the determination of parasitemia in cultures of P. falciparum by FCM. Some previous studies using YOYO-1 showed that this dye possesses high affinity for DNA and discriminates infected RBC from the uninfected cells in a very effective way (14,23,24).…”

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confidence: 99%

Flow cytometry as a tool for analyzing changes in Plasmodium falciparum cell cycle following treatment with indol compounds

et al. 2011

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Melatonin and its derivatives modulate the Plasmodium falciparum and Plasmodium chabaudi cell cycle. Flow cytometry was employed together with the nucleic acid dye YOYO-1 allowing precise discrimination between mono-and multinucleated forms of P. falciparum-infected red blood cell. The use of YOYO-1 permitted excellent discrimination between uninfected and infected red blood cells as well as between early and late parasite stages. Fluorescence intensities of schizont-stage parasites were about 10-fold greater than those of ring-trophozoite form parasites. Melatonin and related indolic compounds including serotonin, N-acetyl-serotonin and tryptamine induced an increase in the percentage of multinucleated forms compared to non-treated control cultures. YOYO-1 staining of infected erythrocyte and subsequent flow cytometry analysis provides a powerful tool in malaria research for screening of bioactive compounds. ' 2011 International Society for Advancement of Cytometry Key termsPlasmodium falciparum; melatonin; cell cycle; flow cytometry MALARIA is caused by the Apicomplexan parasite Plasmodium falciparum. Its asexual replicative cycle inside red blood cell (RBC) is responsible for pathogenesis and induces several structural and biochemical changes within the host cell (1,2). One striking feature regarding P. falciparum infection is associated with 48-h fever peaks intervals, as a result of from synchronous release of merozoites into the blood stream (3).Melatonin, a hormone produced by the pineal gland of vertebrates, transmit the darkness signal based on circadian and seasonal time measurements (4). The presence of melatonin, however, is not restricted to vertebrates but is also found in phylogenetically divergent species including bacteria, plants and protozoa (4). We have previously shown that this hormone is able to synchronize Plasmodium falciparum and P. chabaudi cell infections (5-7). The synchronicity was lost in vitro when parasites had been incubated with the melatonin antagonist luzindole; the same effect was obtained in vivo in pinealectomized mice and after the injection of luzindole (5). In P. falciparum, melatonin induces a complex signaling pathway with the participation of IP3 generation (8) and subsequent transients in cytosolic calcium concentration, cAMP production and protein kinase A (PKA) (9,10) and protease activation (11).Flow cytometry (FCM) is a powerful method for evaluation of human erythrocyte infection rates as well as for discrimination of Plasmodium falciparum developmental stages (12-16). Several methods of detection of malaria parasite by flow cytometry have been developed taking advantage of the absence of DNA in erythrocytes. Different dyes such as acridine orange (17,18), hydroethidine (19,20), SYTO 16 (21) and thiazole orange (22) were already employed for the determination of parasitemia

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A rapid, simple and sensitive flow cytometric system for detection of Plasmodium falciparum

Cited by 31 publications

References 14 publications

Stage-specific activity of potential antimalarial compounds measured in vitro by flow cytometry in comparison to optical microscopy and hypoxanthine uptake

Stage-specific activity of potential antimalarial compounds measured in vitro by flow cytometry in comparison to optical microscopy and hypoxanthine uptake

Quantitation of microsporidia in cultured cells by flow cytometry

Flow cytometry as a tool for analyzing changes in Plasmodium falciparum cell cycle following treatment with indol compounds

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