Multicenter comparative study of Enterocytozoon bieneusi DNA extraction methods from stool samples, and mechanical pretreatment protocols evaluation - Microbes, Intestin, Inflammation et Susceptibilité de l'Hôte Accéder directement au contenu
Article Dans Une Revue Scientific Reports Année : 2024

Multicenter comparative study of Enterocytozoon bieneusi DNA extraction methods from stool samples, and mechanical pretreatment protocols evaluation

Résumé

Nowadays, the use of qPCR for the diagnosis of intestinal microsporidiosis is increasing. There are several studies on the evaluation of qPCR performance but very few focus on the stool pretreatment step before DNA extraction, which is nevertheless a crucial step. This study focuses on the mechanical pretreatment of stools for Enterocytozoon bieneusi spores DNA extraction. Firstly, a multicenter comparative study was conducted evaluating seven extraction methods (manual or automated) including various mechanical pretreatment. Secondly, several durations and grinding speeds and types of beads were tested in order to optimize mechanical pretreatment. Extraction methods of the various centers had widely-varying performances especially for samples with low microsporidia loads. Nuclisens® easyMAG (BioMérieux) and Quick DNA Fecal/Soil Microbe Microprep kit (ZymoResearch) presented the best performances (highest frequencies of detection of low spore concentrations and lowest Ct values). Optimal performances of mechanical pretreatment were obtained by applying a speed of 30 Hz during 60 s with the TissueLyser II (Qiagen) using commercial beads of various materials and sizes (from ZymoResearch or MP Biomedicals). Overall, the optimal DNA extraction method for E. bieneusi spores contained in stool samples was obtained with a strong but short bead beating using small-sized beads from various materials.
Fichier principal
Vignette du fichier
s41598-024-66154-2.pdf (978.05 Ko) Télécharger le fichier
41598_2024_66154_MOESM1_ESM.pdf (256 Ko) Télécharger le fichier
41598_2024_66154_MOESM2_ESM.pdf (274.35 Ko) Télécharger le fichier
41598_2024_66154_MOESM3_ESM.pdf (376.8 Ko) Télécharger le fichier
Origine Fichiers éditeurs autorisés sur une archive ouverte
Commentaire The datasets generated during and/or analysed during the current study are all presented in supplementary tables.

Dates et versions

hal-04653289 , version 1 (18-07-2024)

Licence

Identifiants

Citer

Céline Nourrisson, Maxime Moniot, Maxime Tressol, Céline Lambert, Emilie Fréalle, et al.. Multicenter comparative study of Enterocytozoon bieneusi DNA extraction methods from stool samples, and mechanical pretreatment protocols evaluation. Scientific Reports, 2024, 14 (1), pp.15404. ⟨10.1038/s41598-024-66154-2⟩. ⟨hal-04653289⟩
0 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Mastodon Facebook X LinkedIn More