Optimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.

<h4>Background</h4>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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Maurice Marcel Sandeu, Azizath Moussiliou, Nicolas Moiroux, Gilles G Padonou, Achille Massougbodji, Vincent Corbel, Nicaise Tuikue Ndam
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/11a8dd26ae92420d85873f4c92e40817
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:11a8dd26ae92420d85873f4c92e40817
record_format	dspace
spelling	oai:doaj.org-article:11a8dd26ae92420d85873f4c92e408172021-11-18T08:03:19ZOptimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.1932-620310.1371/journal.pone.0052719https://doaj.org/article/11a8dd26ae92420d85873f4c92e408172012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23285168/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>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.<h4>Methods</h4>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.<h4>Results</h4>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.<h4>Conclusion</h4>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.Maurice Marcel SandeuAzizath MoussiliouNicolas MoirouxGilles G PadonouAchille MassougbodjiVincent CorbelNicaise Tuikue NdamPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e52719 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maurice Marcel Sandeu Azizath Moussiliou Nicolas Moiroux Gilles G Padonou Achille Massougbodji Vincent Corbel Nicaise Tuikue Ndam Optimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.
description	<h4>Background</h4>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.<h4>Methods</h4>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.<h4>Results</h4>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.<h4>Conclusion</h4>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.
format	article
author	Maurice Marcel Sandeu Azizath Moussiliou Nicolas Moiroux Gilles G Padonou Achille Massougbodji Vincent Corbel Nicaise Tuikue Ndam
author_facet	Maurice Marcel Sandeu Azizath Moussiliou Nicolas Moiroux Gilles G Padonou Achille Massougbodji Vincent Corbel Nicaise Tuikue Ndam
author_sort	Maurice Marcel Sandeu
title	Optimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.
title_short	Optimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.
title_full	Optimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.
title_fullStr	Optimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.
title_full_unstemmed	Optimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.
title_sort	optimized pan-species and speciation duplex real-time pcr assays for plasmodium parasites detection in malaria vectors.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/11a8dd26ae92420d85873f4c92e40817
work_keys_str_mv	AT mauricemarcelsandeu optimizedpanspeciesandspeciationduplexrealtimepcrassaysforplasmodiumparasitesdetectioninmalariavectors AT azizathmoussiliou optimizedpanspeciesandspeciationduplexrealtimepcrassaysforplasmodiumparasitesdetectioninmalariavectors AT nicolasmoiroux optimizedpanspeciesandspeciationduplexrealtimepcrassaysforplasmodiumparasitesdetectioninmalariavectors AT gillesgpadonou optimizedpanspeciesandspeciationduplexrealtimepcrassaysforplasmodiumparasitesdetectioninmalariavectors AT achillemassougbodji optimizedpanspeciesandspeciationduplexrealtimepcrassaysforplasmodiumparasitesdetectioninmalariavectors AT vincentcorbel optimizedpanspeciesandspeciationduplexrealtimepcrassaysforplasmodiumparasitesdetectioninmalariavectors AT nicaisetuikuendam optimizedpanspeciesandspeciationduplexrealtimepcrassaysforplasmodiumparasitesdetectioninmalariavectors
_version_	1718422589221634048

Optimized Pan-species and speciation duplex real-time PCR assays for Plasmodium parasites detection in malaria vectors.

Ejemplares similares