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Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples.
- Source :
-
BMC research notes [BMC Res Notes] 2018 Mar 27; Vol. 11 (1), pp. 206. Date of Electronic Publication: 2018 Mar 27. - Publication Year :
- 2018
-
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 <superscript>®</superscript> (Qiagen) and the manual QIAamp <superscript>®</superscript> 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.<br />Results: Whereas EZ1 <superscript>®</superscript> (Qiagen) and QIAamp <superscript>®</superscript> DNA Stool Mini Kit (Qiagen) yielded similar performances for the detection of Cryptosporidium spp. and D. fragilis, significant lower Ct values (pā<ā0.002) pointed out a better performance of EZ1 <superscript>®</superscript> on the five remaining pathogens. DNA extraction using the semi-automated EZ1 <superscript>®</superscript> 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.
- Subjects :
- Blastocystis genetics
Blastocystis pathogenicity
Cryptosporidium parvum genetics
Cryptosporidium parvum pathogenicity
Cyclospora genetics
Cyclospora pathogenicity
DNA, Protozoan genetics
Eukaryota classification
Eukaryota genetics
Giardia lamblia genetics
Giardia lamblia pathogenicity
Humans
Microsporidia genetics
Microsporidia pathogenicity
Molecular Diagnostic Techniques methods
Reproducibility of Results
Sensitivity and Specificity
DNA, Protozoan isolation & purification
Eukaryota pathogenicity
Feces parasitology
Polymerase Chain Reaction methods
Protozoan Infections diagnosis
Protozoan Infections parasitology
Subjects
Details
- Language :
- English
- ISSN :
- 1756-0500
- Volume :
- 11
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- BMC research notes
- Publication Type :
- Academic Journal
- Accession number :
- 29587846
- Full Text :
- https://doi.org/10.1186/s13104-018-3300-2