Counting the Countless: Bacterial Quantification by Targeting rRNA Molecules to Explore the Human Gut Microbiota in Health and Disease

Frontiers in Microbiology
Hirokazu TsujiKoji Nomoto

Abstract

Over the past decade, the advent of next-generation-sequencing tools has revolutionized our approach to understanding the human gut microbiota. However, numerical data on the gut bacterial groups-particularly low-cell-count microbiota, such as indigenous pathobionts, that are otherwise important components of the microbiota-are relatively limited and disparate. As a result, the comprehensive quantitative structure of the human gut microbiota still needs to be fully defined and standardized. With the aim of filling this knowledge gap, we have established a highly sensitive quantitative analytical system that is based on reverse transcription-quantitative PCR and targets microbial rRNA molecules. The system has already been validated in the precise, sensitive, and absolute quantification of more than 70 target bacterial groups belonging to various human gut bacterial clades, including predominant obligate and facultative anaerobes. The system demonstrates sensitivity several hundred times greater than that of other rRNA-gene-targeting methods. It is thus an efficient and valuable tool for exhaustive analysis of gut microbiota over a wide dynamic range. Using this system, we have to date quantified the gut microbiota of about 2,00...Continue Reading

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Citations

Dec 16, 2020·Biochimica Et Biophysica Acta. Reviews on Cancer·Yinghui ZhaoAjay Goel
Sep 9, 2021·PLoS Computational Biology·Chenhao LiNiranjan Nagarajan
Dec 25, 2021·Frontiers in Cellular and Infection Microbiology·Alaa YaseenMalik Sallam

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Methods Mentioned

BETA
PCR
amplicon sequencing
PCA

Software Mentioned

YIF
SCAN

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