Abstract
In preclinical development, the efficacy of agents with putative effects on Plasmodium transmission is determined using the standard membrane feeding assay (SMFA). Because the end-point of the SMFA is normally the enumeration of oocysts on the mosquito midgut, the assays reliance on mosquito dissections and microscopy makes it slow, labor-intensive, and subjective. Below, we describe a novel method of assessing the transmission of a Plasmodium falciparum strain expressing the firefly luciferase protein in the SMFA. The use of a transgenic parasite strain allows for the elimination of mosquito dissections in favor of a simple approach where whole mosquitoes are homogenized and examined directly for luciferase activity. Measuring the mean luminescence intensity of groups of individual or pooled mosquitoes provides comparable estimates of transmission reducing activity at 5–10-fold the throughput capacity of the standard microscopy based SMFA. This high efficiency protocol may be of interest to groups screening novel drug compounds, vaccine candidates, or sera from malaria exposed individuals for transmission reducing activity (TRA).
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References
Moonen B, Cohen JM, Snow RW et al (2010) Operational strategies to achieve and maintain malaria elimination. Lancet 376:1592–1603
Vannice K, Brown G, Akanmori B, Moorthy V (2012) MALVAC 2012 scientific forum: accelerating development of second-generation malaria vaccines. Malar J 11:372
Okell L, Drakeley C, Ghani A, Bousema T, Sutherland C (2008) Reduction of transmission from malaria patients by artemisinin combination therapies: a pooled analysis of six randomized trials. Malar J 7:125
Shekalaghe S, Drakeley C, Gosling R et al (2007) Primaquine clears submicroscopic Plasmodium falciparum gametocytes that persist after treatment with sulphadoxine-pyrimethamine and artesunate. PLoS One 2:e1023
Adjalley SH, Johnston GL, Li T et al (2011) Quantitative assessment of Plasmodium falciparum sexual development reveals potent transmission-blocking activity by methylene blue. Proc Natl Acad Sci 108:E1214–E1223
The malERA Consultative Group on Drugs (2011) A research agenda for malaria eradication: drugs. PLoS Med 8:e1000402
Delves M, Plouffe D, Scheurer C et al (2012) The activities of current antimalarial drugs on the life cycle stages of Plasmodium: a comparative study with human and rodent parasites. PLoS Med 9:e1001169
Delves MJ, Ruecker A, Straschil U et al (2013) Male and female Plasmodium falciparum mature gametocytes show different responses to antimalarial drugs. Antimicrob Agents Chemother 57:3268–3274
Bousema T, Churcher TS, Morlais I, Dinglasan RR (2013) Can field-based mosquito feeding assays be used for evaluating transmission-blocking interventions? Trends Parasitol 29:53–59
Outchkourov NS, Roeffen W, Kaan A et al (2008) Correctly folded Pfs48/45 protein of Plasmodium falciparum elicits malaria transmission-blocking immunity in mice. Proc Natl Acad Sci 105:4301–4305
Chowdhury DR, Angov E, Kariuki T, Kumar N (2009) A potent malaria transmission blocking vaccine based on codon harmonized full length Pfs48/45 expressed in Escherichia coli. PLoS One 4:e6352
Wu Y, Ellis RD, Shaffer D et al (2008) Phase 1 trial of malaria transmission blocking vaccine candidates Pfs25 and Pvs25 formulated with montanide ISA 51. PLoS One 3:e2636
Ouédraogo AL, Roeffen W, Luty AJF et al (2011) Naturally acquired immune responses to Plasmodium falciparum sexual stage antigens Pfs48/45 and Pfs230 in an area of seasonal transmission. Infect Immun 79(12):4957–4964
Bousema T, Sutherland CJ, Churcher TS et al (2011) Human immune responses that reduce the transmission of Plasmodium falciparum in African populations. Int J Parasitol 41:293–300
Roeffen W, Mulder B, Teelen K et al (1996) Association between anti-Pfs48/45 reactivity and P. falciparum transmission-blocking activity in sera from Cameroon. Parasite Immunol 18:103–109
Bousema T, Drakeley C (2011) Epidemiology and Infectivity of Plasmodium falciparum and Plasmodium vivax gametocytes in relation to malaria control and elimination. Clin Microbiol Rev 24:377–410
Churcher TS, Blagborough AM, Delves M et al (2012) Measuring the blockade of malaria transmission—an analysis of the standard membrane feeding assay. Int J Parasitol 42:1037–1044
van der Kolk M, de Vlas SJ, Saul A, van de Vegte-bolmer M, Eling WM, Sauerwein W (2005) Evaluation of the standard membrane feeding assay (SMFA) for the determination of malaria transmission-reducing activity using empirical data. Parasitology 130:13–22
Miura K, Deng B, Tullo G et al (2013) Qualification of standard membrane-feeding assay with Plasmodium falciparum malaria and potential improvements for future assays. PLoS One 8:e57909
Delves M, Sinden R (2010) A semi-automated method for counting fluorescent malaria oocysts increases the throughput of transmission blocking studies. Malar J 9:35
Bell AS, Ranford-Cartwright LC (2004) A real-time PCR assay for quantifying Plasmodium falciparum infections in the mosquito vector. Int J Parasitol 34:795–802
Janse CJ, Franke-Fayard B, Mair GR et al (2006) High efficiency transfection of Plasmodium berghei facilitates novel selection procedures. Mol Biochem Parasitol 145:60–70
Delves MJ, Ramakrishnan C, Blagborough AM, Leroy D, Wells TNC, Sinden RE (2012) A high-throughput assay for the identification of malarial transmission-blocking drugs and vaccines. Int J Parasitol 42:999–1006
Vaughan AM, Mikolajczak SA, Camargo N et al (2012) A transgenic Plasmodium falciparum NF54 strain that expresses GFP–luciferase throughout the parasite life cycle. Mol Biochem Parasitol 186:143–147
Stone WJR, Churcher TS, Graumans W et al (2014) A scalable assessment of Plasmodium falciparum transmission in the standard membrane feeding assay using transgenic parasites expressing GFP-luciferase. J Infect Dis. doi:10.1093/infdis/jiu271
Stone WJR, Eldering M, van Gemert G-J et al (2013) The relevance and applicability of oocyst prevalence as a read-out for mosquito feeding assays. Sci Rep 3
Zollner GE, Ponsa N, Garman GW et al (2006) Population dynamics of sporogony for Plasmodium vivax parasites from western Thailand developing within three species of colonized Anopheles mosquitoes. Malar J 5:68
Ponnudurai T, Lensen AHW, Van Gemert GJA, Bensink MPE, Bolmer M, Meuwissen JHET (1989) Infectivity of cultured Plasmodium falciparum gametocytes to mosquitoes. Parasitology 98:165–173
Ponnudurai T, Lensen AHW, Leeuwenberg ADEM, Meuwissen JHET (1982) Cultivation of fertile Plasmodium falciparum gametocytes in semi-automated systems. 1. Static cultures. Trans R Soc Trop Med Hyg 76:812–818
Molina-Cruz A, Garver LS, Alabaster A et al (2013) The human malaria parasite Pfs47 gene mediates evasion of the mosquito immune system. Science 340:984–987
Lensen AHW, van Druten J, Bolmer M, van Gemert GJA, Eling WM, Sauerwein R (1996) Measurment by membrane feeding of reduction in Plasmodium falciparum transmission induced by endemic sera. Trans R Soc Trop Med Hyg 90:20–22
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Stone, W.J.R., Bousema, T. (2015). The Standard Membrane Feeding Assay: Advances Using Bioluminescence. In: Vaughan, A. (eds) Malaria Vaccines. Methods in Molecular Biology, vol 1325. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2815-6_9
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DOI: https://doi.org/10.1007/978-1-4939-2815-6_9
Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-2815-6
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