Investigation of microbial diversity from primary endodontic infection by clonal analysis and 16S rRNA sequencing

• Grant number: 11/09047-4
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2012
• End date: July 31, 2014
• Field of knowledge: Health Sciences - Dentistry - Endodontics
• Principal Investigator: Brenda Paula Figueiredo de Almeida Gomes
• Grantee: Brenda Paula Figueiredo de Almeida Gomes
• Host Institution: Faculdade de Odontologia de Piracicaba (FOP). Universidade Estadual de Campinas (UNICAMP). Piracicaba , SP, Brazil

Abstract

Microorganisms are important for establishing of endodontic diseases. Different from the phenotypic methods, by means of culture and biochemical tests, molecular analysis of 16S rRNA genes allows a better identification of microorganisms, providing a wider view of the microbiota. This makes it possible to identify species that was believed not being cultivated, or find species that have never been found. Cloning and sequencing of the 16S is independent of culture and captures up to non-viable microorganisms. The objective of this research is to analyze bacterial diversity in primary infected root canal by means of clonal analysis and sequencing of 16S RNA. Microbial samples will be taken from 20 symptomatic teeth with periapical lesions using sterile paper points which wiil de immediately placed into the VMGA III transport medium. Microbial colonies isolated by culture and identified by biochemical methods will be subjected to PCR to amplify the 16S rRNA and then will be carried out the genetic sequencing of these samples. In addition, 10 out of the 20 samples collected will also be cloned and sequenced. The 16S rRNA gene sequences will be edited using the BioEdit software. BLAST will compare nucleic acid sequences against nucleotide sequence database from NCBI. The data will be tabulated and statistically analyzed by SPSS for Windows. (AU)

Scientific publications

(10)

(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)

NAGATA, JULIANA Y.; SOARES, ADRIANA J.; SOUZA-FILHO, FRANCISCO J.; ZAIA, ALEXANDRE A.; FERRAZ, CAIO C. R.; ALMEIDA, JOSE F. A.; GOMES, BRENDA P. F. A.. Microbial Evaluation of Traumatized Teeth Treated with Triple Antibiotic Paste or Calcium Hydroxide with 2% Chlorhexidine Gel in Pulp Revascularization. JOURNAL OF ENDODONTICS, v. 40, n. 6, p. 778-783, JUN 2014. (11/09047-4)

SIGNORETTI, FERNANDA G. C.; GOMES, BRENDA P. F. A.; MONTAGNER, FRANCISCO; JACINTO, ROGERIO C.. Investigation of Cultivable Bacteria Isolated from Longstanding Retreatment-resistant Lesions of Teeth with Apical Periodontitis. JOURNAL OF ENDODONTICS, v. 39, n. 10, p. 1240-1244, OCT 2013. (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)

PRADO, MAIRA; NOGUEIRA LEAL DA SILVA, EMMANUEL JOAO; DUQUE, THAIS MAGESTE; ZAIA, ALEXANDRE AUGUSTO; RANDI FERRAZ, CAIO CEZAR; AFFONSO DE ALMEIDA, JOSE FLAVIO; FIGUEIREDO DE ALMEIDA GOMES, BRENDA PAULA. Antimicrobial and cytotoxic effects of phosphoric acid solution compared to other root canal irrigants. Journal of Applied Oral Science, v. 23, n. 2, p. 158-163, MAR-APR 2015. (11/09047-4)

SOUSA, EZILMARA L. R.; MARTINHO, FREDERICO C.; LEITE, FABIO R. M.; NASCIMENTO, GUSTAVO G.; GOMES, BRENDA P. F. A.. Macrophage Cell Activation with Acute Apical Abscess Contents Determined by Interleukin-1 Beta and Tumor Necrosis Factor Alpha Production. JOURNAL OF ENDODONTICS, v. 40, n. 11, p. 1752-1757, NOV 2014. (10/19136-1, 10/17877-4, 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, 2016. (09/07760-5, 11/09047-4)

FREDERICO C. MARTINHO; FÁBIO R. M. LEITE; LETÍCIA M.M. NÓBREGA; MARCOS S. ENDO; GUSTAVO G. NASCIMENTO; RICHARD P. DARVEAU; BRENDA P. F. A. GOMES. Comparison of Fusobacterium nucleatum and Porphyromonas gingivalis Lipopolysaccharides Clinically Isolated from Root Canal Infection in the Induction of Pro-Inflammatory Cytokines Secretion. Brazilian Dental Journal, v. 27, n. 2, p. 202-207, 2016-04-00. (10/17877-4, 10/19136-1, 11/09047-4)

SALUSTIANO MARINHO, ARIANE CASSIA; MARTINHO, FREDERICO CANATO; ZAIA, ALEXANDRE AUGUSTO; RANDI FERRAZ, CAIO CEZAR; FIGUEIREDO DE ALMEIDA GOMES, BRENDA PAULA. Monitoring the effectiveness of root canal procedures on endotoxin levels found in teeth with chronic apical periodontitis. Journal of Applied Oral Science, v. 22, n. 6, p. 490-495, NOV-DEC 2014. (13/02402-9, 10/17877-4, 10/19136-1, 11/09047-4, 10/13743-3)

MARINHO, A. C. S.; MARTINHO, F. C.; GONCALVES, L. M.; RABANG, H. R. C.; GOMES, B. P. F. A.. Does the Reciproc file remove root canal bacteria and endotoxins as effectively as multifile rotary systems?. International Endodontic Journal, v. 48, n. 6, p. 542-548, JUN 2015. (13/02402-9, 10/13498-9, 10/17877-4, 13/17928-6, 10/19136-1, 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)