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Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples
OBJECTIVE: Efficient and easy-to-use DNA extraction and purification methods are critical in implementing PCR-based diagnosis of pathogens. In order to optimize the routine clinical laboratory diagnosis of eukaryotic enteric pathogens, we compare, via quantitative PCR cycle threshold (Ct) values, th...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5869780/ https://www.ncbi.nlm.nih.gov/pubmed/29587846 http://dx.doi.org/10.1186/s13104-018-3300-2 |
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author | Menu, Estelle Mary, Charles Toga, Isabelle Raoult, Didier Ranque, Stéphane Bittar, Fadi |
author_facet | Menu, Estelle Mary, Charles Toga, Isabelle Raoult, Didier Ranque, Stéphane Bittar, Fadi |
author_sort | Menu, Estelle |
collection | PubMed |
description | OBJECTIVE: Efficient and easy-to-use DNA extraction and purification methods are critical in implementing PCR-based diagnosis of pathogens. In order to optimize the routine clinical laboratory diagnosis of eukaryotic enteric pathogens, we compare, via quantitative PCR cycle threshold (Ct) values, the efficiency of two DNA extraction kits: the semi-automated EZ1(®) (Qiagen) and the manual QIAamp(®) DNA Stool Mini Kit (Qiagen), on six protozoa: Blastocystis spp., Cryptosporidium parvum/hominis, Cyclospora cayetanensis, Dientamoeba fragilis, Giardia intestinalis and Cystoisospora belli and one microsporidia: Enterocytozoon bieneusi. RESULTS: Whereas EZ1(®) (Qiagen) and QIAamp(®) DNA Stool Mini Kit (Qiagen) yielded similar performances for the detection of Cryptosporidium spp. and D. fragilis, significant lower Ct values (p < 0.002) pointed out a better performance of EZ1(®) on the five remaining pathogens. DNA extraction using the semi-automated EZ1(®) procedure was faster and as efficient as the manual procedure in the seven eukaryotic enteric pathogens tested. This procedure is suitable for DNA extraction from stools in both clinical laboratory diagnosis and epidemiological study settings. |
format | Online Article Text |
id | pubmed-5869780 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-58697802018-03-29 Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples Menu, Estelle Mary, Charles Toga, Isabelle Raoult, Didier Ranque, Stéphane Bittar, Fadi BMC Res Notes Research Note OBJECTIVE: Efficient and easy-to-use DNA extraction and purification methods are critical in implementing PCR-based diagnosis of pathogens. In order to optimize the routine clinical laboratory diagnosis of eukaryotic enteric pathogens, we compare, via quantitative PCR cycle threshold (Ct) values, the efficiency of two DNA extraction kits: the semi-automated EZ1(®) (Qiagen) and the manual QIAamp(®) DNA Stool Mini Kit (Qiagen), on six protozoa: Blastocystis spp., Cryptosporidium parvum/hominis, Cyclospora cayetanensis, Dientamoeba fragilis, Giardia intestinalis and Cystoisospora belli and one microsporidia: Enterocytozoon bieneusi. RESULTS: Whereas EZ1(®) (Qiagen) and QIAamp(®) DNA Stool Mini Kit (Qiagen) yielded similar performances for the detection of Cryptosporidium spp. and D. fragilis, significant lower Ct values (p < 0.002) pointed out a better performance of EZ1(®) on the five remaining pathogens. DNA extraction using the semi-automated EZ1(®) procedure was faster and as efficient as the manual procedure in the seven eukaryotic enteric pathogens tested. This procedure is suitable for DNA extraction from stools in both clinical laboratory diagnosis and epidemiological study settings. BioMed Central 2018-03-27 /pmc/articles/PMC5869780/ /pubmed/29587846 http://dx.doi.org/10.1186/s13104-018-3300-2 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Note Menu, Estelle Mary, Charles Toga, Isabelle Raoult, Didier Ranque, Stéphane Bittar, Fadi Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples |
title | Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples |
title_full | Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples |
title_fullStr | Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples |
title_full_unstemmed | Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples |
title_short | Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples |
title_sort | evaluation of two dna extraction methods for the pcr-based detection of eukaryotic enteric pathogens in fecal samples |
topic | Research Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5869780/ https://www.ncbi.nlm.nih.gov/pubmed/29587846 http://dx.doi.org/10.1186/s13104-018-3300-2 |
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