Abstract
Profiling of oral microbiota has traditionally been performed using conventional methods. These methods are relatively time-consuming and labor-intensive. Metagenomic analysis of oral microbiota using high-speed next-generation sequencing is a highly promising technology. However, it is expensive. This study sought to develop a simple and cost-effective profiling method for oral microbiota using 16S rRNA gene polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of PCR-amplified 16S ribosomal RNA genes. Oral isolates of 59 bacterial species from human saliva, including Streptococcus, Actinomyces, and Veillonella, were cultured anaerobically on CDC Anaerobe 5% sheep blood agar plates. Genomic DNA was extracted from single colonies and 16S rRNA genes were PCR-amplified using the 27F and 1492R universal primers. The PCR products were purified and characterized by single digestion with HpaII restriction endonuclease. 16S rRNA gene sequences were obtained from the GenBank database, and the expected restriction profiles were compared with the RFLP patterns obtained from agarose gel electrophoresis. Sixty-five RFLP patterns were obtained from 27 genera and 59 species. The expected fragment sizes of the...Continue Reading
References
Oct 23, 1997·International Journal of Systematic Bacteriology·T SatoE Hoshino
Feb 17, 1998·Applied and Environmental Microbiology·J R MarchesiW G Wade
May 9, 1998·Oral Microbiology and Immunology·T SatoE Hoshino
Jun 19, 1998·Archives of Oral Biology·T SatoE Hoshino
Apr 29, 1999·Oral Microbiology and Immunology·T Sato, H K Kuramitsu
Aug 22, 2003·Oral Microbiology and Immunology·T SatoN Takahashi
May 2, 2012·International Journal of Dentistry·Takuichi SatoNobuhiro Takahashi
Apr 30, 2013·Journal of Dental Research·Y W Han, X Wang
May 15, 2013·Journal of Alzheimer's Disease : JAD·Sophie PooleStJohn Crean
Jul 23, 2013·Applied and Environmental Microbiology·Dong-Lei SunNing-Yi Zhou
Sep 10, 2013·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Heba S SaidMasahira Hattori
Jul 28, 2015·Nature Medicine·Xuan ZhangJun Wang
Sep 13, 2016·Molecular Metabolism·Siegfried UssarC Ronald Kahn
Oct 6, 2016·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Rosario Lucas LópezRubén Pérez Pulido