A scalable assessment of Plasmodium falciparum transmission in the standard membrane feeding assay using transgenic parasites expressing GFP-luciferase.


Stone, WJ; Churcher, TS; Graumans, W; van Gemert, GJ; Vos, MW; Lanke, KH; van de Vegte-Bolmer, MG; Siebelink-Stoter, R; Dechering, KJ; Vaughan, AM; Camargo, N; Kappe, SH; Sauerwein, RW; Bousema, T; (2014) A scalable assessment of Plasmodium falciparum transmission in the standard membrane feeding assay using transgenic parasites expressing GFP-luciferase. The Journal of infectious diseases, 210 (9). pp. 1456-63. ISSN 0022-1899 DOI: 10.1093/infdis/jiu271

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Abstract

BACKGROUND: The development of drugs and vaccines to reduce malaria transmission is an important part of eradication plans. The transmission-reducing activity (TRA) of these agents is currently determined in the standard membrane-feeding assay (SMFA), based on subjective microscopy-based readouts and with limitations in upscaling and throughput.<br/> METHODS: Using a Plasmodium falciparum strain expressing the firefly luciferase protein, we present a luminescence-based approach to SMFA evaluation that eliminates the requirement for mosquito dissections in favor of a simple approach in which whole mosquitoes are homogenized and examined directly for luciferase activity.<br/> RESULTS: Analysis of 6860 Anopheles stephensi mosquitoes across 68 experimental feeds shows that the luminescence assay was as sensitive as microscopy for infection detection. The mean luminescence intensity of individual and pooled mosquitoes accurately quantifies mean oocyst intensity and generates comparable TRA estimates. The luminescence assay presented here could increase SMFA throughput so that 10-30 experimental feeds could be evaluated in a single 96-well plate.<br/> CONCLUSIONS: This new method of assessing Plasmodium infection and transmission intensity could expedite the screening of novel drug compounds, vaccine candidates, and sera from malaria-exposed individuals for TRA. Luminescence-based estimates of oocyst intensity in individual mosquitoes should be interpreted with caution.<br/>

Item Type: Article
Faculty and Department: Faculty of Infectious and Tropical Diseases > Dept of Immunology and Infection
Research Centre: Malaria Centre
PubMed ID: 24829466
Web of Science ID: 344610600014
URI: http://researchonline.lshtm.ac.uk/id/eprint/1726146

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