Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples
Abstract 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) va...
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| Format: | Article |
| Language: | English |
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BMC
2018-03-01
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| Series: | BMC Research Notes |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13104-018-3300-2 |
| _version_ | 1818034236610838528 |
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| author | Estelle Menu Charles Mary Isabelle Toga Didier Raoult Stéphane Ranque Fadi Bittar |
| author_facet | Estelle Menu Charles Mary Isabelle Toga Didier Raoult Stéphane Ranque Fadi Bittar |
| author_sort | Estelle Menu |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-10T06:35:57Z |
| format | Article |
| id | doaj.art-fb0d724cb853404da600dc32d1240c78 |
| institution | Directory Open Access Journal |
| issn | 1756-0500 |
| language | English |
| last_indexed | 2024-12-10T06:35:57Z |
| publishDate | 2018-03-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Research Notes |
| spelling | doaj.art-fb0d724cb853404da600dc32d1240c782022-12-22T01:58:57ZengBMCBMC Research Notes1756-05002018-03-011111610.1186/s13104-018-3300-2Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samplesEstelle Menu0Charles Mary1Isabelle Toga2Didier Raoult3Stéphane Ranque4Fadi Bittar5Aix Marseille Univ, IRD, APHM, MEPHI, IHU-Méditerranée InfectionAix Marseille Univ, IRD, APHM, VITROME, IHU-Méditerranée InfectionAix Marseille Univ, IRD, APHM, VITROME, IHU-Méditerranée InfectionAix Marseille Univ, IRD, APHM, MEPHI, IHU-Méditerranée InfectionAix Marseille Univ, IRD, APHM, VITROME, IHU-Méditerranée InfectionAix Marseille Univ, IRD, APHM, MEPHI, IHU-Méditerranée InfectionAbstract 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.http://link.springer.com/article/10.1186/s13104-018-3300-2Enteric parasitesProtozoaMicrosporidiaqPCRDNA extraction |
| spellingShingle | Estelle Menu Charles Mary Isabelle Toga Didier Raoult Stéphane Ranque Fadi Bittar Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples BMC Research Notes Enteric parasites Protozoa Microsporidia qPCR DNA extraction |
| 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 | Enteric parasites Protozoa Microsporidia qPCR DNA extraction |
| url | http://link.springer.com/article/10.1186/s13104-018-3300-2 |
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