Abstract
The gastrointestinal tract harbors the gut microbiota, structural alterations of which (dysbiosis) are linked with an increase in gut permeability (“leaky gut”), enabling luminal antigens and bacterial products such as nanosized bacterial extracellular vesicles (BEVs) to access the circulatory system. Blood-derived BEVs contain various cargoes and may be useful biomarkers for diagnosis and monitoring of disease status and relapse in conditions such as inflammatory bowel disease (IBD). To progress this concept, we developed a rapid, cost-effective protocol to isolate BEV-associated DNA and used 16S rRNA gene sequencing to identify bacterial origins of the blood microbiome of healthy individuals and patients with Crohn’s disease and ulcerative colitis. The 16S rRNA gene sequencing successfully identified the origin of plasma-derived BEV DNA. The analysis showed that the blood microbiota richness, diversity, or composition in IBD, healthy control, and protocol control groups were not significantly distinct, highlighting the issue of ‘kit-ome’ contamination in low-biomass studies. Our pilot study provides the basis for undertaking larger studies to determine the potential use of blood microbiota profiling as a diagnostic aid in IBD.
Highlights
Introduction iationsThe human body harbors on average 3.8 × 1013 microbes, termed the microbiota [1], the largest community of which resides in the gastrointestinal tract (GIT)
Using 16S rRNA gene sequencing, we found that there was no difference in the BEVassociated blood microbiota richness, diversity, or composition between inflammatory bowel disease (IBD), Healthy control (HC), and protocol control groups, highlighting the issue of ‘kit-ome’ contamination in low-biomass studies
Using 16S rRNA gene sequencing, we found that the bacterial extracellular vesicles (BEVs)-associated blood microbiota composition was not distinct between IBD, HC, or protocol control groups
Summary
The human body harbors on average 3.8 × 1013 microbes, termed the microbiota [1], the largest community of which resides in the gastrointestinal tract (GIT). Comprising bacteria, viruses, fungi, protozoa, and archaea, the intestinal microbiota contributes to the life-long health of the host, playing key roles in digestion, the provision of essential amino acids and micronutrients, and in the maturation and maintenance of the host immune system as well as maintaining intestinal epithelial barrier integrity [2]. Associations have been demonstrated between an altered structural composition of the microbiota (dysbiosis) and many human diseases [3], in particular, those with a concomitant “leaky gut” and increased permeability of the intestinal barrier as seen in inflammatory bowel disease (IBD) [4]. IBD is a debilitating group of chronic diseases which causes relapsing–remitting inflammation of the GIT and affects millions of people worldwide [5].
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