Comparison of Gastric Microbiota Between Gastric Juice and Mucosa by Next Generation Sequencing Method

Jihee Sung, Nayoung Kim, Jaeyeon Kim, Hyun Jin Jo, Ji Hyun Park, Ryoung Hee Nam, Yeong-Jae Seok, Yeon-Ran Kim, Dong Ho Lee, Hyun Chae Jung, Jihee Sung, Nayoung Kim, Jaeyeon Kim, Hyun Jin Jo, Ji Hyun Park, Ryoung Hee Nam, Yeong-Jae Seok, Yeon-Ran Kim, Dong Ho Lee, Hyun Chae Jung

Abstract

Background: Not much is known about the role of gastric microbiota except for Helicobacter pylori in human health and disease. In this study, we aimed to detect human gastric microbiota in both gastric mucosa and gastric juice by barcoded 454-pyrosequencing of the 16S rRNA gene and to compare the results from mucosa and juice.

Methods: Gastric biopsies and stomach juices were collected from 4 subjects who underwent standard endoscopy at Seoul National University Bundang Hospital. Gastric microbiota of antral mucosa, corpus mucosa samples, and gastric fluids were analyzed by barcoded 454-pyrosequencing of the 16S rRNA gene. The analysis focused on bacteria, such as H. pylori and nitrosating or nitrate-reducing bacteria.

Results: Gastric fluid samples showed higher diversity compared to that of gastric mucosa samples. The mean of operational taxonomic units was higher in gastric fluid than in gastric mucosa. The samples of gastric fluid and gastric mucosa showed different composition of phyla. The composition of H. pylori and Proteobacteria was higher in mucosa samples compared to gastric fluid samples (H. pylori, 66.5% vs. 3.3%, P = 0.033; Proteobacteria, 75.4% vs. 26.3%, P = 0.041), while Actinobacteria, Bacteroidetes, and Firmicutes were proportioned relatively less in mucosa samples than gastric fluid. However there was no significant difference. (Actinobacteria, 3.5% vs. 20.2%, P = 0.312; Bacteroidetes, 6.0% vs. 14.8%, P = 0.329; Firmicutes, 12.8% vs. 33.4%, P = 0.246).

Conclusions: Even though these samples were small, gastric mucosa could be more effective than gastric fluid in the detection of meaningful gastric microbiota by pyrosequencing.

Keywords: Gastric juice; Gastric mucosa; Helicobacter pylori; Microbiota.

Figures

Figure 1.
Figure 1.
(A) Rarefaction curve and composition of phylum in the gastric mucosa and gastric fluid samples. The rarefaction curve indicates the bacterial community in the gastric fluid samples and mucosa samples (1–3, Helicobacter pylori [HP] (+) sample; 4, HP (−) sample). OTUs stands for operational taxonomic units. (B) The phylum composition between gastric mucosa samples and gastric fluid samples.

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