Comparación de dos protocolos de extracción de ADN para la detección de infección por Plasmodium spp. en Anopheles spp.

ABSTRACT: Detection of anophelines infected with Plasmodium spp. has traditionally been conducted by optical microscopy and immuno-assays. Currently, methodologies based on Polymerase Chain Reaction (PCR), provide high sensitivity and specificity; however, some authors have reported that inhibitors present in the mosquito’s body can decrease PCR sensitivity for Plasmodium spp. detection and that their effect could be reduced using DNA extraction protocols that efficiently remove those potential inhibitors. Objective To compare two DNA extraction protocols for the detection of Plasmodium spp. infecting Anopheles spp. Materials and methods Ten Anopheles stephensi mosquitoes infected with Plasmodium falciparum were dissected to separate head-thorax and abdomen and processed by one of the extraction protocols, dilutions of the DNA were evaluated by nested PCR. Differences between amplification frequencies were analyzed by the t-Student’s test and the hypotheses were analyzed by the test of difference between proportions of two populations. Results In specimens with DNA extracted by the protocol without chelating resin amplification of parasite DNA was achieved in 11.6% of the samples and with the protocol with chelating resin, in 6.6%. From all of the amplifications, 20% corresponded to undiluted DNA and 7.5% to DNA diluted 1:10; the difference between amplification proportions was statistically significant (p <0.05). Conclusions The protocol that more efficiently detected parasite DNA in mosquitoes was the one without chelating resin; with it, amplification was approximately two fold to the one obtained with the protocol with resin.

• Autores: Monsalve Restrepo, María Isabel; Ortega Arellano, Héctor Flavio; Gutiérrez Builes, Lina Andrea; Correa Ochoa, Margarita María; Zapata Tamayo, Mario Augusto
• Tipo de recurso: Article of investigation
• Fecha de publicación: 2011
• Institución: Universidad de Antioquia
• Repositorio: Repositorio UdeA
• Idioma: spa
• OAI Identifier: oai:bibliotecadigital.udea.edu.co:10495/10442
• Acceso en línea: http://hdl.handle.net/10495/10442
• Palabra clave: Anopheles; Extracción de ADN; Malaria; PCR (Reacción en cadena de la polimerasa); Plasmodium; DNA extraction; Plasmodium
• Rights: openAccess
• License: Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO)