Advanced search
Start date
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Comparison of rRNA-based reverse transcription PCR and rDNA-based PCR for the detection of streptococci in root canal infections

Full text
Prado, Lais Cunha [1] ; Gavini, Giulio [1] ; Silveira, Amanda da Costa [1] ; Nakamura, Vitor Cesar [1] ; Alves Mayer, Marcia Pinto [2] ; Pinheiro, Ericka Tavares [1]
Total Authors: 6
[1] Univ Sao Paulo, Fac Odontol, Sao Paulo, SP - Brazil
[2] Univ Sao Paulo, Fac Odontol, Inst Ciencias Biomed, Sao Paulo, SP - Brazil
Total Affiliations: 2
Document type: Journal article
Source: Journal of Applied Oral Science; v. 27, 2019.
Web of Science Citations: 0

Objective: The rDNA-based method is unable to distinguish between alive and dead cells. Alternatively, bacterial viability can be assessed by molecular methods based on ribosomal RNA (rRNA). Therefore, this study aimed to detect viable streptococci in root canal samples using rRNA-based reverse transcription polymerase chain reaction (RT-PCR), compared to an rDNA-based PCR assay. Methodology: Microbiological root canal samples were obtained from 32 teeth with primary endodontic infections before (S1) and after chemomechanical preparation (S2), and after removal of intracanal medication (S3). RNA and DNA were extracted, and complementary DNA (cDNA) was synthesized from RNA using RT reaction. cDNA and genomic DNA were subjected to PCR with primers complementary to the 16S rRNA sequences of Streptococcus spp. McNemar's test was used to compare the detection rate of both assays (P<0.05). Results: Streptococci were detected in 28.12% (9/32) and 37.5% (12/32) of S1 samples using rRNA- and rDNA-based PCR assays, respectively. In contrast, they were detected in only 6.25% (2/32) of S2 samples using rRNA-based RT-PCR, compared to 15.62% (5/32) using rDNA-based PCR. Finally, in S3 samples, streptococci were not detected by rRNA, whereas rDNA-based PCR still detected the bacteria in 12.5% (4/32) of the samples. The total number of PCR-positive reactions in the rDNA-based PCR was higher than in the rRNA-based assay (P<0.05). Conclusions: The rRNA-based RT-PCR showed a lower detection rate of streptococci when compared to the rDNA-based PCR, suggesting that the latter may have detected dead cells of streptococci in root canal samples. (AU)

FAPESP's process: 16/15473-0 - Analysis of Metabolically Active Bacterial Community of Persistent Endodontic Infections after Different Clinical Protocols of Disinfection: RNA- and DNA-based Molecular Study
Grantee:Ericka Tavares Pinheiro
Support Opportunities: Regular Research Grants
FAPESP's process: 13/02923-9 - Influence of passive ultrasonic irrigation on bacterial and endotoxin reduction from root canal system: a randomized clinical study
Grantee:Giulio Gavini
Support Opportunities: Regular Research Grants