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

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.

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

Abstract 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.

Lineage variability in surface components expression within Porphyromonas gingivalis.

The periodontopathogen Porphyromonas gingivalis is represented by a spectrum of phenotypes ranging from commensals to pathogenic lineages. Capsule and fimbriae are considered key virulence factors in this specie, involved in colonization and host defenses evasion. Since these virulence traits may not be expressed by certain strains, we aimed to test the hypothesis that certain clusters or genotypes of P. gingivalis correlate with the production of capsule and fimbriae. Sixteen P. gingivalis isolates were evaluated. Capsule (K) was detected by optical microscopy of negatively stained cells. The presence of fimbriae (F) was determined by TEM. Genotypes were determined by NotI macrorestriction fragments analysis through Pulsed-Field Gel Electrophoresis (PFGE) and Multi-locus sequence typing (MLST) based on seven house-keeping genes. The phenotypes included F(+)K(+) (n = 4), F(-)K(+) (n = 5), F(+)K(-) (n = 5) and F(-)K(-) (n = 2). The analysis of whole genome macrorestriction fragments revealed 14 different clusters. MLST data also revealed extensive genetic diversity; however, PFGE and MLST profiles showed evident differences. There was no association between P. gingivalis clusters and encapsulated and/or fimbriated phenotypes. Genotyping methods were not able to discriminate isolates according to the production of virulence factors such as capsule and major fimbriae, indicating that recombination played a key role in the expression of capsule and fimbriae in P. gingivalis.

Prevalence of traumatic dental injuries and associated factors among Brazilian schoolchildren.

PURPOSE: To assess the prevalence of traumatic dental injuries to permanent anterior teeth in 9- to 14-year-old schoolchildren attending public schools in Anápolis, Brazil, and to investigate the association between the occurrence of these injuries and the size of incisal overjet and type of lip coverage. MATERIALS AND METHODS: A cross-sectional survey and a two-stage cluster sampling technique were used. The sample size included 765 9- to 14-year-old schoolchildren. Data were collected through clinical examinations and interviews carried out by a trained, calibrated dentist. Oral examinations dealt with the type of traumatic dental injury (TDI), the treatment received, the size of incisal overjet and the type of lip coverage. The teeth examined were maxillary and mandibular incisors. RESULTS: A 16.5% prevalence of dental trauma was found. Boys experienced double the number of girls' injuries. The maxillary central incisors were the teeth most affected, totaling 84.8%. The most frequent type of injury found was enamel fracture (66%), followed by enamel-dentin fracture (27%) and enamel cracks (5%). Only 26% of traumatised teeth were restored. Children with an overjet size > 3 mm were 1.78 times (CI= 1.18 - 2.69) more likely to have a dental injury than children with an overjet size <= 3 mm. Children with inadequate lip coverage were 2.18 times (CI= 1.27 - 3.76) more likely to experience dental trauma than children whose lip coverage was adequate. CONCLUSION: This study shows that the prevalence of traumatic dental injuries among schoolchildren in Anápolis, Brazil is similar to that of other regions in Brazil. The teeth most affected by dental trauma are the maxillary central incisors. Boys run a 2.03-times higher risk of crown fracture than girls, and children with an overjet size > 3 mm are 1.78 times more likely to have dental injuries. In addition, children with inadequate lip coverage are 2.18 times more likely to present traumatic dental injuries than children with adequate lip coverage.

Gemella morbillorum in primary and secondary/persistent endodontic infections.

OBJECTIVE: To investigate the presence of Gemella morbillorum by culture or nested PCR in primary and secondary/persistent endodontic infections. STUDY DESIGN: Microbial samples were taken from 50 cases with primary and 50 cases with secondary/persistent endodontic infections. Microbiologic techniques were used for culture and identification. The DNA extracted from the samples was analyzed for the presence of the target species using species-specific primers. RESULTS: Culture and polymerase chain reaction (PCR) identified the species in 23 and 77, respectively, of 100 root canals. Culture yielded the test organism in 19 of 50 (38%) of root canal samples from primary and in 4 of 50 (8%) from secondary/persistent infections. PCR yielded the test organisms in 41 of 50 (82%) and 36 of 50 (72%) of the, respectively, primary and secondary/persistent root canal infections studied. CONCLUSION: Gemella morbillorum was identified more frequently in primary endodontic infections than in secondary/persistent ones. A higher frequency of the target species was detected by PCR than by culture.

Root canal microbiota of dogs' teeth with periapical lesions induced by two different methods.

OBJECTIVE: The microbial composition was investigated in root canals of dogs' teeth with periapical lesions induced by 2 different methods: open versus sealed canals. STUDY DESIGN: Teeth from Group I (n = 16) were left open for a week, then sealed with composite resin for 120 days. The teeth from Group II (n = 16) were left open for the same period. Microbiological samples from the root canals were collected and processed by the anaerobic technique for identification and counting of microorganisms after establishment of periapical reactions. RESULTS: Seventy-four cultivable isolates were recovered in sealed canals (Group I). Strict anaerobes accounted for 64.9% of all species isolated, and gram-negative microorganisms accounted for 55.4%. Microbial genera most frequently isolated were Prevotella, Fusobacterium, Peptostreptococcus, Streptococcus, Enterococcus, Clostridium, and Porphyromonas. Statistical analysis by Pearson chi-square or Fisher's test revealed positive association between sealed teeth and strict anaerobes (P < .05). In open canals (Group II), from a total of 58 cultivable isolates, 19% were strict anaerobes and 81% facultative anaerobes, with predominance of gram-positive species (75.8%). Genera most frequently isolated were Streptococcus, Propionibacterium, Staphylococcus, Neisseria, and Prevotella. CONCLUSION: Strict anaerobes were most frequently found in sealed teeth rather than in the teeth with canals left exposed to the oral cavity for 4 months. Therefore, the method that induced periapical inflammatory lesions by intentional oral exposure, followed by tooth sealing, produced root canal microbiota similar to the same found in humans.

Bacteriological study of root canals associated with periapical abscesses.

OBJECTIVE: The aim of this study was to identify microorganisms from root canals with periapical abscesses and to ascertain the susceptibility of Peptostreptococcus prevotii and Fusobacterium necrophorum to antimicrobials. Study design Thirty root canals were microbiologically sampled by using sterile paper points. The concomitant microorganisms were identified through the use of established methods. The susceptibility of P prevotii and F necrophorum to antimicrobials was evaluated by using the E test method. RESULTS: A total of 117 different bacterial strains were recovered, including 75 strict anaerobes or microphilic species. The most frequently isolated strict anaerobes were P prevotii, Peptostreptococcus micros, and F necrophorum. Facultative bacteria such as Gemella morbillorum and Streptococcus mitis were also found, albeit less frequently. The data revealed that P prevotii and F necrophorum were susceptible to the tested antibiotics. CONCLUSIONS: Gram-positive anaerobic bacteria predominate in the mixed microbiota of root canals with periapical abscesses. Moreover, P prevotii and F necrophorum are susceptible to the tested antibiotics.

Cistos Peri-radiculares: uma proposta de tratamento / Periradicular cysts: a treatment proposal

O objetivo deste trabalho foi de realizar uma revisäo de literatura sobre cisto radicular, procurando elucidar a etiologia e formaçäo, o diagnóstico diferencial com o granuloma, a classificaçäo e as opçöes de tratamento. Baseada nesa revisäo, como clínica e radiograficamente o diagnóstico diferencial näo é possível em virtude das características serem muito semelhantes entre si, o tratamento inicial proposto é o endodôntico. Se a terapia endodôntica for bem conduzida, tanto o granuloma como o cisto baia deveräo ser reparados, enquanto o cisto verdadeiro necessitará ser removido cirurgicamente, pois é pouco provável que haja reparo somente com o tratamento endodôntico