Evaluation of Two DNA Extraction Methods for the PCR-based Detection of Eukaryotic Enteric Pathogens in Fecal Samples

Overview

Journal BMC Res Notes

Publisher Biomed Central

Specialties Biology
General Medicine

Date 2018 Mar 29

PMID 29587846

Citations 21

Authors

Estelle Menu

Charles Mary

Isabelle Toga

Didier Raoult

Stephane Ranque

Fadi Bittar

Affiliations

Soon will be listed here.

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.

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