Naslov
Evaluation of human faecal microbiota content by 16S rRNA analysis using different collection, storage and DNA extraction methods – OMNIgene.GUT case study

Autori
Perić, Mihaela ; Panek, Marina ; Barešić, Anja ; Čipčić Paljetak, Hana ; Matijašić, Mario ; Ljubas Kelečić, Dina ; Brinar, Marko ; Urek, Marija ; Turk, Nikša ; Verbanac, Donatella ; Krznarić, Željko

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, ostalo

Izvornik
Journal of International Society of Microbiota / - Pariz, 2015, 107-107

Skup
3rd World Congress on Targeting Microbiota

Mjesto i datum
Pariz, Francuska, 21.10.2015. - 23.10.2015

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
human faecal samples; microbiota; DNA extraction; 16S rRNA gene sequencing

Sažetak
Methodologies for collection and storage of human faecal samples as well as DNA extraction govern the quality of DNA obtained and present crucial steps in obtaining representative models of bacterial community structure after 16S rRNA gene sequencing. Collection of faeces in the clinical settings necessitates the need for uniform, reliable, robust and patient compliant procedures. In this study we compared two different approaches for faeces collection and storage and three commercially available DNA extraction kits. Faeces from 6 healthy donors were collected fresh and in OMNIgene.GUT system, then stored for 14 days at -20°C and at room temperature, respectively. DNA yield and quality varied between DNA extraction kits in the order MP>QIAGEN>MoBio with OMNIgene.GUT preserving integrity of the obtained DNA. Donor-specific bacterial diversity was maintained irrespective of the collection, storage or extraction method used. However, kit-based trends were observed displaying different levels of abundance among the most prevalent families, notably Bacteroidaceae and Lachnospiraceae. Although OMNIgene.GUT faeces collection system slightly decreased the richness of detected bacteria, the change was not statistically significant. Overall, OMNIgene.GUT was proven as convenient and reliable human faecal collection, transport and storage system for human faecal microbiota analyses.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

POVEZANOST RADA