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Malaria Panel Assay versus PCR: detection of naturally infected Anopheles melas in a coastal village of Equatorial Guinea

BACKGROUND: A study was carried out in a village of the mainland region of Equatorial Guinea in order to ascertain a) which members of Anopheles gambiae complex could be involved in malaria transmission and b) the rate of infectivity for Anopheles melas comparing two different methods, a PCR able to...

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Autores principales: Moreno, Marta, Cano, Jorge, Nzambo, Sisinio, Bobuakasi, Leonardo, Buatiche, Jesús N, Ondo, Melchor, Micha, Francisco, Benito, Agustín
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2004
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC487905/
https://www.ncbi.nlm.nih.gov/pubmed/15238168
http://dx.doi.org/10.1186/1475-2875-3-20
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author Moreno, Marta
Cano, Jorge
Nzambo, Sisinio
Bobuakasi, Leonardo
Buatiche, Jesús N
Ondo, Melchor
Micha, Francisco
Benito, Agustín
author_facet Moreno, Marta
Cano, Jorge
Nzambo, Sisinio
Bobuakasi, Leonardo
Buatiche, Jesús N
Ondo, Melchor
Micha, Francisco
Benito, Agustín
author_sort Moreno, Marta
collection PubMed
description BACKGROUND: A study was carried out in a village of the mainland region of Equatorial Guinea in order to ascertain a) which members of Anopheles gambiae complex could be involved in malaria transmission and b) the rate of infectivity for Anopheles melas comparing two different methods, a PCR able to detect sporozoite-DNA and an immunochromatographic assay MPR (Malaria Rapid Dipstick Panel Assay). METHODS: Mosquitoes were sampled at night by indoor captures in two houses of a coastal village in Equatorial Guinea (Ayantang). Collected mosquitoes were identified as An. gambiae s.l. These were individually dried into silica-gel. The head-thorax of the An. gambiae s.l. mosquitoes were analysed by PCR to verify that the species was of the gambiae complex. Individual head-thorax and pools (5 pools) of homogenized mosquitoes employed in Malaria Rapid Panel assay (MRP assay) were lysed and DNA was extracted. PCR was designed from the 753 base pair insert of pBRKl-14 and DNA was amplified. The relationship between dipstick and PCR to detect Plasmodium falciparum sporozoites was measured in terms of sensitivity, specificity and test association (Cohen's kappa value). RESULTS: Two hundred and sixty-four An. gambiae s.l. females were studied (214 individually and five pools with 10 mosquitoes in each). PCR analysis showed that 207 mosquitoes were An. melas, 3 An. gambiae s.s. and 4 could not be identified. By using PCR as the gold standard method when dipstick assay was compared, matching results were obtained for 6 mosquitoes and, in one case MRP was positive while PCR was not reactive. MRP assay showed a low sensitivity (3.3%) when compared with falciparum-DNA detection (17,7% and 14,3%, series A and B respectively). Agreement between the two test formats was low (κ = 0,224). CONCLUSION: It was determined that An. melas is the main anopheline vector involved in malaria transmission in Ayantang, a coastal village in mainland Equatorial Guinea. A comparison of PCR and Vec-Test Assay(®), concluded that the PCR method proved to be a more sensitive and useful tool than the dipstick assay to determine the malarial infection rate in mosquitoes in an area of stable and high malaria transmission like Equatorial Guinea.
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spelling pubmed-4879052004-07-25 Malaria Panel Assay versus PCR: detection of naturally infected Anopheles melas in a coastal village of Equatorial Guinea Moreno, Marta Cano, Jorge Nzambo, Sisinio Bobuakasi, Leonardo Buatiche, Jesús N Ondo, Melchor Micha, Francisco Benito, Agustín Malar J Research BACKGROUND: A study was carried out in a village of the mainland region of Equatorial Guinea in order to ascertain a) which members of Anopheles gambiae complex could be involved in malaria transmission and b) the rate of infectivity for Anopheles melas comparing two different methods, a PCR able to detect sporozoite-DNA and an immunochromatographic assay MPR (Malaria Rapid Dipstick Panel Assay). METHODS: Mosquitoes were sampled at night by indoor captures in two houses of a coastal village in Equatorial Guinea (Ayantang). Collected mosquitoes were identified as An. gambiae s.l. These were individually dried into silica-gel. The head-thorax of the An. gambiae s.l. mosquitoes were analysed by PCR to verify that the species was of the gambiae complex. Individual head-thorax and pools (5 pools) of homogenized mosquitoes employed in Malaria Rapid Panel assay (MRP assay) were lysed and DNA was extracted. PCR was designed from the 753 base pair insert of pBRKl-14 and DNA was amplified. The relationship between dipstick and PCR to detect Plasmodium falciparum sporozoites was measured in terms of sensitivity, specificity and test association (Cohen's kappa value). RESULTS: Two hundred and sixty-four An. gambiae s.l. females were studied (214 individually and five pools with 10 mosquitoes in each). PCR analysis showed that 207 mosquitoes were An. melas, 3 An. gambiae s.s. and 4 could not be identified. By using PCR as the gold standard method when dipstick assay was compared, matching results were obtained for 6 mosquitoes and, in one case MRP was positive while PCR was not reactive. MRP assay showed a low sensitivity (3.3%) when compared with falciparum-DNA detection (17,7% and 14,3%, series A and B respectively). Agreement between the two test formats was low (κ = 0,224). CONCLUSION: It was determined that An. melas is the main anopheline vector involved in malaria transmission in Ayantang, a coastal village in mainland Equatorial Guinea. A comparison of PCR and Vec-Test Assay(®), concluded that the PCR method proved to be a more sensitive and useful tool than the dipstick assay to determine the malarial infection rate in mosquitoes in an area of stable and high malaria transmission like Equatorial Guinea. BioMed Central 2004-07-06 /pmc/articles/PMC487905/ /pubmed/15238168 http://dx.doi.org/10.1186/1475-2875-3-20 Text en Copyright © 2004 Moreno et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Research
Moreno, Marta
Cano, Jorge
Nzambo, Sisinio
Bobuakasi, Leonardo
Buatiche, Jesús N
Ondo, Melchor
Micha, Francisco
Benito, Agustín
Malaria Panel Assay versus PCR: detection of naturally infected Anopheles melas in a coastal village of Equatorial Guinea
title Malaria Panel Assay versus PCR: detection of naturally infected Anopheles melas in a coastal village of Equatorial Guinea
title_full Malaria Panel Assay versus PCR: detection of naturally infected Anopheles melas in a coastal village of Equatorial Guinea
title_fullStr Malaria Panel Assay versus PCR: detection of naturally infected Anopheles melas in a coastal village of Equatorial Guinea
title_full_unstemmed Malaria Panel Assay versus PCR: detection of naturally infected Anopheles melas in a coastal village of Equatorial Guinea
title_short Malaria Panel Assay versus PCR: detection of naturally infected Anopheles melas in a coastal village of Equatorial Guinea
title_sort malaria panel assay versus pcr: detection of naturally infected anopheles melas in a coastal village of equatorial guinea
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC487905/
https://www.ncbi.nlm.nih.gov/pubmed/15238168
http://dx.doi.org/10.1186/1475-2875-3-20
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