mirage

Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp

Repositorio ABACUS/Manakin

Mostrar el registro sencillo del ítem

dc.contributor.author Paulos Viñas, Silvia
dc.contributor.author Mateo, Marta
dc.contributor.author Lucio, Aída de
dc.contributor.author Hernández de Mingo, Marta
dc.contributor.author Bailo, Begoña
dc.contributor.author Saugar, José M.
dc.contributor.author Cardona, Guillermo A.
dc.contributor.author Fuentes, Isabel
dc.contributor.author Mateo Barrientos, María
dc.contributor.author Carmena, David
dc.date.accessioned 2016-12-15T16:09:50Z
dc.date.available 2016-12-15T16:09:50Z
dc.date.issued 2016
dc.identifier.citation Paulos, S., Mateo, M., de Lucio, A., Hernández-de Mingo, M., Bailo, B., Saugar, J. M., Cardona, G. A., Fuentes, I., Mateo, M., & Carmena, D. (2016). Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp. Journal of Microbiological Methods, 127, 38-73. spa
dc.identifier.issn 01677012
dc.identifier.uri http://hdl.handle.net/11268/6101
dc.description.abstract High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n = 29), giardiasis (n = 47) and amoebiasis by Entamoeba histolytica (n = 3) or E. dispar (n = 10) and apparently healthy subjects (n = 24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56 °C were proven more efficient for the release of DNA from Cryptosporidium oocysts. spa
dc.description.sponsorship SIN FINANCIACIÓN spa
dc.language.iso eng spa
dc.subject.other Protozoan parasites spa
dc.subject.other DNA spa
dc.subject.other ADN spa
dc.title Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp spa
dc.type article spa
dc.description.impact 1.790 JCR (2016) Q3, 55/78 Biochemical Research Methods, 86/125 Microbiology spa
dc.identifier.doi 10.1016/j.mimet.2016.05.020
dc.rights.accessRights closedAccess spa
dc.subject.uem Genética humana spa
dc.subject.unesco Biotecnología spa
dc.subject.unesco Genética spa
dc.description.filiation UEM spa
dc.peerreviewed Si spa


Ficheros en el ítem

Ficheros Tamaño Formato Ver

No hay ficheros asociados a este ítem.

Este ítem aparece en la(s) siguiente(s) colección(ones)

Mostrar el registro sencillo del ítem