Abstract
BackgroundThe infectivity of Plasmodium gametocytes is typically determined by microscopically examining the midguts of mosquitoes that have taken a blood meal containing potentially infectious parasites. Such assessments are required for the development and evaluation of transmission-reducing interventions (TRI), but are limited by subjectivity, technical complexity and throughput. The detection of circumsporozoite protein (CSP) by enzyme-linked immunosorbent assay (ELISA) and enhanced chemiluminescent slot-blot (ECL-SB) may be used as objective, scalable alternatives to microscopy for the determination of infection prevalence.MethodsTo compare the performance of the CSP ELISA and ECL-SB for the detection of mosquito infection, four groups of Anopheles stephensi mosquitoes were infected with cultured Plasmodium falciparum gametocytes. At day-8 post-infection (PI), parasite status was determined by microscopy for a sample of mosquitoes from each group. At days 8 and 10 PI, the parasite status of separate mosquito samples was analysed by both CSP ELISA and ECL-SB.ResultsWhen mosquito samples were analysed 8 days PI, the ECL-SB determined similar infection prevalence to microscopy; CSP ELISA lacked the sensitivity to detect CSP in all infected mosquitoes at this early time point. When mosquitoes were analysed 48 h later (10 days PI) both assays performed as well as microscopy for infection detection.ConclusionsWhilst microscopical examination of mosquito guts is of great value when quantification of parasite burden is required, ECL-SB and CSP ELISA are suitable alternatives at day 10 PI when infection prevalence is the desired endpoint, although CSP ELISA is not suitable at day 8 PI. These results are important to groups considering large-scale implementation of TRI.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-015-0954-2) contains supplementary material, which is available to authorized users.
Highlights
The infectivity of Plasmodium gametocytes is typically determined by microscopically examining the midguts of mosquitoes that have taken a blood meal containing potentially infectious parasites
104 test mosquitoes were analysed by microscopy on day 8 PI, 336 test mosquitoes were analysed in the enhanced chemiluminescent slot-blot (ECL-SB) on days 8 and 10 PI, and 336 test mosquitoes were analysed in the enzyme-linked immunosorbent assay (ELISA) on days 8 and 10 PI
This study describes the ability of two immuno-assays, the circumsporozoite protein (CSP)-ELISA and ECL-SB, to detect low-density mosquitostage P. falciparum infections based on the detection of CSP in the mosquito carcass
Summary
The infectivity of Plasmodium gametocytes is typically determined by microscopically examining the midguts of mosquitoes that have taken a blood meal containing potentially infectious parasites. Such assessments are required for the development and evaluation of transmission-reducing interventions (TRI), but are limited by subjectivity, technical complexity and throughput. Sporozoites that succeed in entering the salivary glands are detectable from about 11 days PI, and the number in the glands appears to plateau after approximately 14 days PI [11] where they remain viable for long periods [12, 13] Because of this stability and because very few sporozoites are egested during blood feeding [14, 15] it is generally accepted that a mosquito with any number of salivary gland sporozoites is infective to humans
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