A new quantitative 16S rRNA amplicon sequencing method.

Author: Na HAN ¹ ; Xianhui PENG ¹ ; Tingting ZHANG ¹ ; Yujun QIANG ¹ ; Xiuwen LI ¹ ; Wen ZHANG ¹
Author Information

1. Department of Bioinformatics, State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Publication Type:Journal Article
Keywords: 16S rRNA; amplicon; internal marker; quantitative; random tag
MeSH: Bacteria/genetics*; Gastrointestinal Microbiome/genetics*; High-Throughput Nucleotide Sequencing; Humans; Polymerase Chain Reaction; RNA, Ribosomal, 16S/genetics*
From: Chinese Journal of Biotechnology 2020;36(12):2548-2555
CountryChina
Language:Chinese
Abstract: In recent years, 16S rRNA amplicon sequencing technology has been widely used to study human gut microbiota and to detect unknown pathogens in clinical samples. However, its resolution to bacterial population can only reach the relative abundance of genus level, and different factors affect the final bacterial profile, such as sample concentrations, PCR cycle numbers and amplification primers. In order to solve these problems, we developed a quantitative 16S rRNA amplicon sequencing method by combining random tag and internal marker method. The new methods improved the accuracy of human gut microbiota, reduced the impact of experimental operation on the results, and improved the comparability between sequencing and other molecular biological methods.