MICROBIAL INVESTIGATION OF INFECTED ROOT CANAL OF SYMPTOMATIC TEETH WITH PERIAPICAL LESION BY CULTURE, CLONING AND 16S rRAN SEQUENCING

• Grant number: 09/07760-5
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date until: August 01, 2009
• End date until: September 30, 2012
• Field of knowledge: Health Sciences - Dentistry - Dental Clinics
• Principal Investigator: Brenda Paula Figueiredo de Almeida Gomes
• Grantee: Letícia Maria Menezes Nóbrega
• Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil

Abstract

Microorganisms are important for the establishment of endodontic diseases. Different from phenotypic identification, 16S rRNA gene sequencing analysis has the potential to give a more reliable bacterial identification. It makes possible the identification of some species that is believed not to be cultivated, or to find species that had never been isolated before. Cloning and 16S rRNA sequencing do not depend on culture and identify non-viable microorganisms. The aim of this study is to identify cultivable bacteria isolated from symptomatic root canals with periapical lesions using culture and identification by both biochemical methods and 16S rRNA sequencing, and also identify uncultivable bacteria from the same samples by cloning and 16S rRNA sequencing. Samples will be collected from 20 root canals, using sterile paper points that will be immediately transferred to the transport medium VMGA III. Serial dilution will be plated out and incubated in anaerobic atmosphere. Microbial colony will be isolated, featured and identified by biochemical methods. The V6-V8 region of 16S rRNA gene will be amplified by PCR and the product will be selected for sequencing. Moreover, 10 samples will be cloned for 16S rRNA sequencing. The resulting sequences will be edited and aligned using the BioEdit and ClustaW. The Blast n algorithm will compare the obtained nucleic acid sequence against a nucleotide sequence database at NCBI. Data will be statistically analized using SPSS for Windows.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MENEZES NOBREGA, LETICIA MARIA; MONTAGNER, FRANCISCO; RIBEIRO, ADRIANA COSTA; PINTO MAYER, MARCIA ALVES; FIGUEIREDO DE ALMEIDA GOMES, BRENDA PAULA. Bacterial diversity of symptomatic primary endodontic infection by clonal analysis. Brazilian Oral Research, v. 30, n. 1, 2016. (09/07760-5, 11/09047-4)

MENEZES NOBREGA, LETICIA MARIA; MONTAGNER, FRANCISCO; RIBEIRO, ADRIANA COSTA; PINTO MAYER, MARCIA ALVES; FIGUEIREDO DE ALMEIDA GOMES, BRENDA PAULA. Bacterial diversity of symptomatic primary endodontic infection by clonal analysis. BRAZILIAN ORAL RESEARCH, v. 30, n. 1, p. 9-pg., 2016-01-01. (09/07760-5, 11/09047-4)

LETÍCIA M. M. NÓBREGA; FRANCISCO MONTAGNER; ADRIANA C. RIBEIRO; MÁRCIA A. P. MAYER; BRENDA P. F. A. GOMES. Molecular Identification of Cultivable Bacteria From Infected Root Canals Associated With Acute Apical Abscess. Brazilian Dental Journal, v. 27, n. 3, p. 318-324, 2016-06-00. (11/09047-4, 09/07760-5)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

NÓBREGA, Letícia Maria Menezes. Study of bacterial diversity of infected root canal with acute apical abscess by culture, cloning and 16S rRNA sequencing. 2012. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba Piracicaba, SP.