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

Journal of Applied Oral Science : Revista FOB
L C PradoE T Pinheiro

Abstract

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. 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). 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...Continue Reading

References

Oct 31, 2006·Applied and Environmental Microbiology·Kazunori MatsudaKoji Nomoto
Jan 24, 2007·Oral Microbiology and Immunology·M SakamotoY Benno
May 15, 2007·Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontics·José F SiqueiraKenio C Lima
Jul 19, 2008·Journal of Endodontics·Isabela N RôçasJosé F Siqueira
Oct 22, 2008·Journal of Endodontics·José F Siqueira, Isabela N Rôças
Oct 16, 2009·Journal of Dental Research·J F Siqueira, I N Rôças
Nov 30, 2010·Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontics·Malin BrundinUlf Sjögren
Mar 10, 2012·Journal of Clinical Microbiology·Isabela N Rôças, José F Siqueira
Jan 17, 2013·Journal of Endodontics·Malin BrundinUlf Sjögren
Jul 9, 2013·International Endodontic Journal·M A S NevesJ F Siqueira
Jun 14, 2015·Journal of Endodontics·Ericka T PinheiroMárcia P A Mayer

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