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Updated: Jun 6, 2025

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Gut microbiota alterations and associations with nutrients in children with celiac disease
Yasemin Ertaş Öztürk1, Efsun Karabudak2, Ödül Eğritaş Gürkan3
1Department of Nutrition and Dietetics, Faculty of Health Sciences Ondokuz Mayıs University Samsun Türkiye.
Abstract:
Celiac disease is a chronic inflammatory condition that is not well understood in relation to the microbiome. Our objective was to demonstrate changes in the microbiota and the relationships between nutrients in children with celiac disease (CD) who followed a gluten-free diet (GFD). A group of 11 children who were recently diagnosed with CD, ranging in age from 3 to 12, were monitored for a period of 6 months. GFD is designed based on the individual's specific energy and nutrient needs, with strict control over dietary adherence. Food consumption, blood, and fecal samples were taken. Fecal samples were put through 16s rRNA sequencing. Microbial modifications were demonstrated using alpha diversity, beta diversity, nonmetric multidimensional scaling analysis (NDMS), t-test, and metastats. Mean age was 6.4 ± 2.66 years and 54.5% were male participants. Serological parameters were negative after 6 months. Both unweighted (p = .019) and weighted (p = .021) Unifrac distances were higher before GFD, and differences were reliable according to NDMS analysis (stress = 0.189). The abundance of Bacteroides ovatus was increased (p = .014), whereas unidentified Lachnospiraceae, Paeniclostridium, Paraclostridium Peptostreptococcus, and Dielma were decreased after GFD (p < .001). Associations between nutrients and several genera and species were identified. The presence of genus Bifidobacterium and Bifidobacterium adolescentis was inversely associated with fat intake after GFD (p < .01). Microbiota changes became evident over a period of 6 months. The presence or absence of small bacteria may play a role in the development of CD. Modifying the children's dietary intake can potentially influence the microbial composition.
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