title: Use of 16S rRNA Gene-Targeted Group-Specific Primers for Real-Time PCR Analysis of Predominant Bacteria in Mouse Feces.
abstract: Mouse models are widely used for studying gastrointestinal (GI) tract-related diseases. It is necessary and important to develop a new set of primers to monitor the mouse gut microbiota. In this study, 16S rRNA gene-targeted group-specific primers for Firmicutes, Actinobacteria, Bacteroidetes, Deferribacteres, "Candidatus Saccharibacteria," Verrucomicrobia, Tenericutes, and Proteobacteria were designed and validated for quantification of the predominant bacterial species in mouse feces by real-time PCR. After confirmation of their accuracy and specificity by high-throughput sequencing technologies, these primers were applied to quantify the changes in the fecal samples from a trinitrobenzene sulfonic acid-induced colitis mouse model. Our results showed that this approach efficiently predicted the occurrence of colitis, such as spontaneous chronic inflammatory bowel disease in transgenic mice. The set of primers developed in this study provides a simple and affordable method to monitor changes in the intestinal microbiota at the phylum level.
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing, China.
authors.forename: Yun-Wen
authors.initials: YW
authors.lastname: Yang
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing, China.
authors.forename: Mang-Kun
authors.initials: MK
authors.lastname: Chen
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing, China.
authors.forename: Bing-Ya
authors.initials: BY
authors.lastname: Yang
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing, China.
authors.forename: Xian-Jie
authors.initials: XJ
authors.lastname: Huang
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing, China.
authors.forename: Xue-Rui
authors.initials: XR
authors.lastname: Zhang
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing, China.
authors.forename: Liang-Qiang
authors.initials: LQ
authors.lastname: He
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing, China zchua@nju.edu.cn jzhang08@nju.edu.cn.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing, China zchua@nju.edu.cn jzhang08@nju.edu.cn.
authors.forename: Zi-Chun
authors.initials: ZC
authors.lastname: Hua
mesh_terms: D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology*; D000069196:Gastrointestinal Microbiome; D041981:Gastrointestinal Tract / Q000382:microbiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D012336:RNA, Ribosomal, 16S