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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Optimized Pan-species and Speciation Duplex Real-time PCR Assays for Plasmodium Parasites Detection in Malaria Vectors
PLoS ONE, Volume 7, No. 12, Article e52719, Year 2012
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Description
Background: An accurate method for detecting malaria parasites in the mosquito's vector remains an essential component in the vector control. The Enzyme linked immunosorbent assay specific for circumsporozoite protein (ELISA-CSP) is the gold standard method for the detection of malaria parasites in the vector even if it presents some limitations. Here, we optimized multiplex real-time PCR assays to accurately detect minor populations in mixed infection with multiple Plasmodium species in the African malaria vectors Anopheles gambiae and Anopheles funestus. Methods: Complementary TaqMan-based real-time PCR assays that detect Plasmodium species using specific primers and probes were first evaluated on artificial mixtures of different targets inserted in plasmid constructs. The assays were further validated in comparison with the ELISA-CSP on 200 field caught Anopheles gambiae and Anopheles funestus mosquitoes collected in two localities in southern Benin. Results: The validation of the duplex real-time PCR assays on the plasmid mixtures demonstrated robust specificity and sensitivity for detecting distinct targets. Using a panel of mosquito specimen, the real-time PCR showed a relatively high sensitivity (88.6%) and specificity (98%), compared to ELISA-CSP as the referent standard. The agreement between both methods was "excellent" (κ = 0.8, P<0.05). The relative quantification of Plasmodium DNA between the two Anopheles species analyzed showed no significant difference (P = 0, 2). All infected mosquito samples contained Plasmodium falciparum DNA and mixed infections with P. malariae and/or P. ovale were observed in 18.6% and 13.6% of An. gambiae and An. funestus respectively. Plasmodium vivax was found in none of the mosquito samples analyzed. Conclusion: This study presents an optimized method for detecting the four Plasmodium species in the African malaria vectors. The study highlights substantial discordance with traditional ELISA-CSP pointing out the utility of employing an accurate molecular diagnostic tool for detecting malaria parasites in field mosquito populations. © 2012 Sandeu et al.
Authors & Co-Authors
Sandeu, Maurice Marcel
Benin, Cotonou
University of Abomey-calavi
France, Montpellier
Maladies Infectieuses et Vecteurs : Écologie, Génétique, Évolution et Contrôle
Benin
Centre D'etudes et de Recherche Sur le Paludisme Associe a la Grossesse et a L'enfant Cerpage, Faculté Des Sciences de la Santé Fss
Moussiliou, Azizath
Benin
Centre D'etudes et de Recherche Sur le Paludisme Associe a la Grossesse et a L'enfant Cerpage, Faculté Des Sciences de la Santé Fss
Benin
Institut de Recherche Pour le Développement Ird 216
Moiroux, Nicolas
Benin, Cotonou
University of Abomey-calavi
France, Montpellier
Maladies Infectieuses et Vecteurs : Écologie, Génétique, Évolution et Contrôle
Padonou, Gil Germain
Benin, Cotonou
University of Abomey-calavi
Massougbodji, Achille
Benin
Centre D'etudes et de Recherche Sur le Paludisme Associe a la Grossesse et a L'enfant Cerpage, Faculté Des Sciences de la Santé Fss
Corbel, Vincent
Benin, Cotonou
University of Abomey-calavi
France, Montpellier
Maladies Infectieuses et Vecteurs : Écologie, Génétique, Évolution et Contrôle
Tuikue-Ndam, Nicaise
Benin
Centre D'etudes et de Recherche Sur le Paludisme Associe a la Grossesse et a L'enfant Cerpage, Faculté Des Sciences de la Santé Fss
Benin
Institut de Recherche Pour le Développement Ird 216
France, Paris
Université Paris Cité
Statistics
Citations: 33
Authors: 7
Affiliations: 5
Identifiers
Doi:
10.1371/journal.pone.0052719
e-ISSN:
19326203
Research Areas
Genetics And Genomics
Infectious Diseases
Study Locations
Benin