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A malaria gametocytocidal assay using oxidoreduction indicator, alamarBlue
Takeshi Q Tanaka1, Kim C Williamson
1Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20892, United States.
Molecular and Biochemical Parasitology
|February 15, 2011
Summary
Developing new anti-malarial drugs requires blocking malaria transmission. A new assay using alamarBlue fluorescent indicator screens for gametocyte viability, identifying compounds with gametocytocidal activity.
Area of Science:
- Medical Research
- Parasitology
- Drug Discovery
Background:
- Malaria eradication necessitates novel strategies beyond control.
- Blocking malaria transmission requires targeting gametocytes.
Purpose of the Study:
- To develop a novel assay for screening gametocytocidal compounds.
- To evaluate the viability of malaria gametocytes using a fluorescent indicator.
Main Methods:
- Utilized the alamarBlue fluorescent oxidoreduction indicator to assess gametocyte viability.
- Established a dose-response curve for epoxomicin to determine assay sensitivity and reproducibility (IC(50) 2.16±0.57 nM, Z'-factor 0.81±0.01).
- Screened six anti-malarial compounds for gametocytocidal activity at 10 µM.
Main Results:
- The fluorescent signal from alamarBlue correlated linearly with gametocyte number (R(2)=0.99).
- Primaquine and dihydroartemisinin (DHA) demonstrated significant gametocytocidal activity at 10 µM.
- The assay proved reproducible and sensitive, validated by epoxomicin testing.
Conclusions:
- A new, efficient fluorescent assay using alamarBlue can screen for compounds with gametocytocidal activity.
- This assay is a valuable tool for advancing malaria eradication efforts by identifying transmission-blocking drug candidates.
