SARS-COV-2 detection in saliva and nasopharyngeal swabs using RT-PCR was similar.

The World Health Organization has declared the widespread spread of SARS-CoV-2 and its associated disease (COVID-19) a public health emergency. The standard gold test for detecting the virus is the RT-PCR, performed from nasopharyngeal swab (NPS) samples. However, this test may be uncomfortable for the patient and requires specific training and attire from the health professional responsible for collecting the sample. Therefore, the search for alternative ways to collect samples that may be used in the diagnosis of COVID-19 is relevant. This study aimed to compare the results obtained from NPS and saliva samples. NPS and saliva samples were collected from 189 symptomatic outpatients suspected of COVID-19, who came to Piquet Carneiro Polyclinic. RNA extraction was performed using the Bio-Gene DNA/RNA Viral Extraction kit (Bioclin®). Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) reactions used the Molecular SARS-CoV-2 (E / RP) kit (Bio-Manguinhos). The results indicated that 142 showed a non-detectable result (ND), while 47 showed a detectable result (D). Among the 142 "ND", 137 (94.4%) saliva samples obtained the same result, while 5 samples (3.4%) were "D". Among the 47 "D" swab samples, 35 (74.4%) showed the same result in the saliva samples. The sensitivity of the saliva test was 0.74 and the specificity was 0.97. The positive predictive value was 0.88 while the negative predictive value was 0.92. The results showed that detection of Sars-CoV-2 using saliva samples showed high sensitivity and specificity compared to nasopharyngeal swabs.

SARS-COV-2 detection in saliva and nasopharyngeal swabs using RT-PCR was similar

Abstract The World Health Organization has declared the widespread spread of SARS-CoV-2 and its associated disease (COVID-19) a public health emergency. The standard gold test for detecting the virus is the RT-PCR, performed from nasopharyngeal swab (NPS) samples. However, this test may be uncomfortable for the patient and requires specific training and attire from the health professional responsible for collecting the sample. Therefore, the search for alternative ways to collect samples that may be used in the diagnosis of COVID-19 is relevant. This study aimed to compare the results obtained from NPS and saliva samples. NPS and saliva samples were collected from 189 symptomatic outpatients suspected of COVID-19, who came to Piquet Carneiro Polyclinic. RNA extraction was performed using the Bio-Gene DNA/RNA Viral Extraction kit (Bioclin®). Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) reactions used the Molecular SARS-CoV-2 (E / RP) kit (Bio-Manguinhos). The results indicated that 142 showed a non-detectable result (ND), while 47 showed a detectable result (D). Among the 142 "ND", 137 (94.4%) saliva samples obtained the same result, while 5 samples (3.4%) were "D". Among the 47 "D" swab samples, 35 (74.4%) showed the same result in the saliva samples. The sensitivity of the saliva test was 0.74 and the specificity was 0.97. The positive predictive value was 0.88 while the negative predictive value was 0.92. The results showed that detection of Sars-CoV-2 using saliva samples showed high sensitivity and specificity compared to nasopharyngeal swabs.

Resumo A Organização Mundial da Saúde declarou a disseminação generalizada do SARS-CoV-2 e sua doença associada (COVID-19) uma emergência de saúde pública. O teste padrão ouro para detecção do vírus é o RT-PCR, realizado a partir de amostras de swab nasofaríngeo (NPS). No entanto, esse exame pode ser desconfortável para o paciente e requer treinamento específico e vestimenta do profissional de saúde responsável pela coleta da amostra. Portanto, a busca por formas alternativas de coleta de amostras que possam ser utilizadas no diagnóstico de COVID-19 é relevante. O objetivo deste estudo foi comparar os resultados obtidos em amostras de NPS e saliva. Amostras de NPS e saliva foram coletadas de 189 pacientes ambulatoriais sintomáticos com suspeita de COVID-19, que procuraram a Policlínica Piquet Carneiro. A extração de RNA foi realizada com o kit Bio-Gene DNA / RNA Viral Extraction (Bioclin®) e as reações em tempo real da reação em cadeia da polimerase-transcriptase reversa (RT-PCR) usaram o kit Molecular SARS-CoV-2 (E / RP) (Bio-Manguinhos). Os resultados indicaram que 142 apresentaram resultado não detectável (ND), enquanto 47 apresentaram resultado detectável (D). Entre os 142 "ND", 137 (94,4%) amostras de saliva obtiveram o mesmo resultado, enquanto 5 amostras (3,4%) foram "D". Dentre as 47 amostras de swab "D", 35 (74,4%) apresentaram o mesmo resultado nas amostras de saliva. A sensibilidade do teste de saliva foi de 0,74 e a especificidade foi de 0,97. O valor preditivo positivo foi de 0,88, enquanto o valor preditivo negativo foi de 0,92. Os resultados mostraram que a detecção de Sars-CoV-2 em amostras de saliva apresentou alta sensibilidade e especificidade quando comparada com swabs nasofaríngeos.

Surface roughness evaluation and whitening efficiency on tooth enamel after using whitening toothpaste: a randomized double-blinded study

The aim of this randomized double-blinded study was to evaluate the enamel surface roughness and color change after one month of whitening toothpaste use and the color stability obtained 1 month after its interruption. 30 volunteers were divided into 3 groups (n = 10) corresponding to the dentifrices: 1) Colgate Total 12 Clean Mint (TD) (Control), 2) Colgate Luminous White (LW) and 3) Sensodyne Whitening Extra Fresh (SB). The volunteers were impression with addition silicone to obtain an epoxy resin replica of the upper central incisor for the initial surface roughness evaluation using a profilometer and the initial color of the incisors and canines was evaluated with a spectrophotometer after one week of wash-out. After 1 month, the color of the central incisors and canines was measured again, and the volunteers were molded to obtain a second replica to the final roughness analysis. Data were submitted to ANOVA-one way (p≤0.05). The results showed that there was no statistical difference between the dentifrices for color difference and surface roughness for all the studied conditions. It was possible to conclude that the whitening dentifrices used in this study were not able to alter the initial color of the teeth and did not cause changes in the surface roughness of enamel.

Ellagic acid on the quality of the adhesive interface of class I composite resin restorations after aging

To evaluate the effect of ellagic acid on the inhibition of matrix metalloproteinase by analyzing the quality of the adhesive interface with bond strength measures in periods of 24 hours and six months of storage. Method: 40 healthy human third molars were prepared with class I cavities (5x4x3mm). The teeth were divided into four experimental groups: Group 1- without application of ellagic acid and storage time of 24 hours; Group 2- with ellagic acid/24 hours; G3- without ellagic acid/six months; Group 4- with ellagic acid/six months. Then, the cavities were restored with Single Bond Universal adhesive and Z350 composite resin, with and without the previous application of ellagic acid. Subsequently, hourglass-shaped specimens were obtained and subjected to the bond strength (BS) test (n = 10) in a universal testing machine. The bond test was performed after 24 hours and six months of storage. For the standard evaluation (n = 3) the samples were infiltrated with silver nitrate and placed in a developing solution for analysis in a Scanning Electron Microscope (SEM). The data obtained were analyzed with the Kruskal-Wallis non-parametric test, showing a statistically significant difference. Results: The highest bond strength values were found for the 24-hour groups followed by the groups with six months of storage. For nano-infiltration, groups G1 and G2 showed lower infiltration than groups G3 and G4. Conclusion: The previous application of ellagic acid did not affect the BS of the adhesive interface of the adhesive system analyzed, regardless of storage time.

Reducing the viral load of SARS-CoV-2 in the saliva of patients with COVID-19.

OBJECTIVE: To evaluate the reduction in the salivary viral load using oral antiseptic mouthwashes in patients testing positive for COVID-19. METHODS: Sixty-three individuals were recruited after testing positive for COVID-19 by real-time RT-PCR assay and divided into 5 groups. Group 1 received sterile water, group 2 received 1.5% hydrogen peroxide solution (HP), group 3 received 0.12% chlorhexidine (CHX), group 4 received 0.1% sodium hypochlorite solution (NaClO), and group 5 received sequential rinses using CHX and HP. After collecting the initial saliva sample, individuals were asked to use the designated mouthwash for 1 min. Additional saliva samples were collected immediately after rinsing, 15, and 30 min after rinsing. Real-time RT-PCR assays for RNA detection of SARS-CoV-2 were performed on the saliva samples. RESULTS: There were no significant differences among the experimental groups and the control group in any period. Compared to the baseline values, there was a significant reduction in the number of copies of SARS-CoV-2 after 30 min in group 2 and immediately after the initial mouthwash in group 4. CONCLUSIONS: No experimental group demonstrated a significant reduction in the viral load compared to the control group.

Morphological alterations in tongue epithelial cells infected by SARS-CoV-2: A case-control study.

OBJECTIVE: The aim of the present case-control study was to evaluate the morphological aspects of the epithelial cells from the dorsum of the tongue and the expression of the SARS-CoV-2 Spike protein in these cells, in patients with and without COVID-19 infection. METHODS: 24 individuals with at least one symptom of COVID-19 were recruited among inpatients from Hospital Universitário Pedro Ernesto (Rio de Janeiro, Brazil). 14 patients who tested positive for COVID-19 by RT-PCR were included in the case group, and 10 patients who tested negative were included in the control group. Cytological smears from the dorsum of the tongue were obtained from all patients and analyzed using immunohistochemistry directed against SARS-CoV-2-Spike protein. Morphological changes in epithelial cells were analyzed using light microscopy. RESULTS: Immunohistochemistry showed that 71% of the COVID-19 patients presented epithelial cells positive for the presence of the SARS-CoV-2 Spike protein, and all cells coming from patients in the control group were negative. Cytological analysis showed significant differences when comparing epithelial cells from COVID-19-positive and COVID-19-negative patients. CONCLUSION: COVID-19 may generate dimensional changes in tongue epithelial cells; however, further studies are necessary to understand how this happens.

Mouthguard use and attitudes regarding dental trauma among elite cross-country mountain biking and field hockey athletes.

BACKGROUND/AIMS: Cross-country mountain biking and field hockey are two Olympic sports that pose a potential risk for dentofacial trauma. However, mouthguard use is not mandatory in either of these sports and knowledge about tooth rescue among athletes is often neglected. The aim of this cross-sectional epidemiological survey was to evaluate the prevalence of sports-related dentofacial injuries, mouthguard use, and attitudes regarding tooth rescue among cross-country mountain biking and field hockey athletes participating at pre-Olympic competitions held in Rio de Janeiro, Brazil. MATERIALS AND METHODS: A convenience sample of 217 athletes from 33 countries participating in cross-country mountain biking (n = 82; mean age = 30.96 ± 8.52 years) and field hockey (n = 135; mean age = 19.72 ± 2.46 years) pre-Olympic competitions were examined clinically, and they answered a questionnaire regarding previous history of sports-related dentofacial injuries, attitudes toward mouthguard use, and tooth rescue. RESULTS: There were 120 (55.30%) males and 97 (44.70%) females who participated in the study. The prevalence of facial trauma was higher in mountain biking (54.88%) than in field hockey (26.66%, P < .001). Fracture was the most prevalent facial injury in mountain biking (80%), while laceration was the most prevalent in field hockey (30.55%). Extensive dental injuries occurred more often in mountain biking (47.37%) than in hockey (12.50%). The teeth most often affected were the maxillary central incisors in both mountain biking (57.89%) and field hockey (93.75%). Mouthguard use was more frequent among field hockey athletes (41.48%) than in mountain biking (1.22%, P < .001). The overall knowledge about tooth rescue was low, and most of the athletes in both sports did not know what to do in case of an avulsed tooth. CONCLUSIONS: Prevalence of dentofacial injuries among XCO-MTB and field hockey athletes participating at this pre-Olympic event was high. The majority of the athletes in this study did not use mouthguards and were unaware of recommendations in the case of an avulsed tooth.

Efeito antimicrobiano do bacteriófago geneticamente modificado φEf11/φFL1C(Δ36)PnisA sobre cepas de E. faecalis em biofilme estático e em canais radiculares infectados / Antibacterial effect of a Genetically-Engineered Bacteriophage φEf11/φFL1C(Δ36)PnisA on static biofilms and on Dentin Infected with E. faecalis

A presença de micro-organismos no sistema de canais radiculares (SCR) tem sido apontada como uma das principais causas de insucesso da terapia endodôntica. A capacidade de formar biofilme e penetrar nos túbulos dentinários são fatores de sobrevivência que favorecem a perpetuação de micro-organismos no interior do SCR. Terapias que promovam a desorganizazão de biofilmes e eliminação de bactérias dentro dos túbulos dentinários são fundamentais para a desinfecção endodôntica. Uma terapia descoberta há um século, denominada de Bacteriofagoterapia, baseia-se na utilização de vírus capazes de infectar e matar bactérias. Esta abordagem antimicrobiana tem recebido bastante atenção atualmente por representar uma alternativa para o tratamento de doenças causadas por bactérias multirresistentes aos antibióticos. O objetivo deste estudo foi avaliar a eficácia de um bacteriófago modificado geneticamente, φEf11/φFL1C(Δ36)PnisA, para eliminar biofilmes de duas cepas de E. faecalis: JH2-2 (sensível à vancomicina e resistente ao ácido fusídico e à rifampicina) e V583 (resistente à vancomicina). Este estudo foi dividido em dois experimentos distintos. No primeiro experimento, biofilmes estáticos de 48 horas de cepas de E. faecalis JH2-2 (pMSP3535 nisR / K) ou V583 (pMSP3535 nisR / K) formados em lâminulas de vidro (coverslips) foram inoculados por suspensão do bacteriófago φEf11/φFL1C(Δ36)PnisA. Após 48 horas de incubação, a biomassa bacteriana foi fotografada por microscopia confocal e as células viáveis foram quantificadas por medição de unidades formadoras de colônias (UFC). No segundo experimento, segmentos radiculares de dentes humanos extraídos foram cimentados em dispositivos vedáveis de duas câmaras para formar modelos ex vivo com dentina infectada, contendo solução tampão na câmara inferior. Os modelos foram inoculados com uma suspensão de E. faecalis V583 ou E. faecalis JH2-2. Após sete dias de incubação a 37°C, adicionou-se ao canal de cada segmento de dentina infectada dos grupos 2 e 5 uma suspensão do fago geneticamente modificado, φEf11/φFL1C(Δ36)PnisA e manteve-se a incubação por mais 72 horas. Os segmentos de dentina foram instrumentados com Gates Glidden e a solução tampão foi aliquotada para semeadura e contagem de UFC e aferição do título residual de células de E. faecalis. Os resultados do primeiro experimento mostraram uma diminuição de 10-100 vezes (p≤ 0,05) das células viáveis (UFC / biofilme) após tratamento com bacteriófago, o que foi consistente com a comparação das imagens de biofilme tratado e não tratado visualizadas com projeções máximas da série Z. No segundo experimento a titulação de E. faecalis verificada após tratamento com o bacteriófago foi reduzida em 18% para os modelos infectados com JH2-2 e em 99% (p≤ 0,05) nos modelos infectados com V583. Com base nesses resultados, pode-se concluir que a biomassa dos biofilmes de E. faecalis, tanto sensíveis quanto resistentes à vancomicina, foi significantemente reduzida após a infecção pelo bacteriófago φEf11/φFL1C(Δ36)PnisA. Além disso, o tratamento da dentina infectada por E. faecalis com bacteriófago φEf11/φFL1C(Δ36)PnisA resultou em diminuição da população bacteriana residual de cepas sensíveis e resistentes à vancomicina, alcançando significância estatística no grupo que utilizou a cepa V583.

Residual microorganisms in the root canal system (RCS) have been identified as the main cause of endodontic therapy failure. The ability to form a biofilm and penetrate the dentin tubules are survival factors that favor the perpetuation of microorganisms within the RCS. Therapies that promote the disorganization of biofilms and elimination of bacteria within the dentinal tubules are essential for endodontic disinfection. A therapy discovered a century ago called Bacteriophage Therapy is based on the use of viruses capable of infecting and killing bacteria. Recently, this antimicrobial approach has been receiving considerable attention since it represents an alternative for the treatment of diseases caused by multiresistant antibiotic bacteria. The aim of this study was to evaluate the efficacy of a genetically engineered bacteriophage, φEf11/φFL1C(Δ36)PnisA, to disrupt biofilms of two Enterococcus faecalis strains: JH2-2 (vancomycin sensitive) and V583 (vancomycin resistant). This study was divided into two separate experiments. In the first experiment, 24 hour static biofilms of E. faecalis strains JH2-2(pMSP3535 nisR/K) and V583(pMSP3535 nisR/K) formed on cover slips were inoculated with bacteriophage φEf11/φFL1C(Δ36)PnisA. After 24 and 48 hours incubation, the bacterial biomass was imaged by confocal microscopy and viable cells were quantified by colony forming unit (CFU) measurement. In the second experiment, extracted human dentin root segments were cemented into sealable two-chamber devices, fabricated from syringe needle caps to form in vitro infected-dentin models. The models were inoculated with an overnight suspension of either E. faecalis V583 (vancomycin resistant strain) or E. faecalis JH2-2 (fusidic acid and rifampin resistant, vancomycin sensitive strain). After 7 days of incubation at 37°C, a suspension of a genetically engineered phage, φEf11/φFL1C(Δ36)PnisA, was added to the root canal of each infected dentin segment, and the incubation was continued for an additional 72-hours. Dentin was harvested from the walls of each root canal and assayed for the residual titer of E. faecalis cells. The results from the first experiment showed a 10-100-fold fewer decrease in viable cells (CFU/biofilm) after bacteriophage treatment, which was consistent with comparisons of treated and untreated biofilm images visualized as max projections of the Z-series. On the second experiment, the recovered E. faecalis titer was reduced by 18% for the JH2-2 infected models, and by 99% for the V583 infected models. These results suggest that the biomass of E. faecalis biofilms, both sensitive and resistant to vancomycin, was significantly reduced after infection by bacteriophage φEf11/φFL1C(Δ36)PnisA. In addition, treatment of E. faecalis-infected dentin with the phage resulted in the decrease of the residual bacterial population for both susceptible and vancomycin resistant strains, reaching statistical significance in strain V583 group.

Epidemiological evaluation of apical periodontitis prevalence in an urban Brazilian population.

The present study aimed to assess the prevalence of apical periodontitis (AP) in an urban Brazilian population according to gender, age group and tooth type. Data were collected from clinical files containing the medical and dental histories and periapical radiographs of 1,126 patients treated at the School of Dentistry at Universidade do Estado do Rio de Janeiro between March 2000 and December 2010. A total of 15,724 periapical radiographs were evaluated. All the radiographs were evaluated by two independent, previously calibrated endodontists (kappa = 0.88). Periapical areas on the radiographs were classified as N (normal) or AR (apical radiolucency). The frequency of AP and the 95% Confidence Interval (95%CI) were calculated according to gender, age group and tooth type. Differences between groups were calculated using the Z-test at a significance level of 5% (p < 0.05). AP was present in 7.87% of the samples, with 16.70% occurring on previously endodontically treated teeth and 44.65% occurring on teeth referred for endodontic treatment (TR-RCT). The frequency of AP was higher among females (64%) than among males (35%). The central and lateral maxillary incisors were the most frequently affected teeth. The frequency of AP was higher among individuals between 30 and 49 years of age. In this population, AP was more prevalent among females and among individuals between 30 and 49 years of age, and the central and lateral maxillary incisors were the most frequently affected teeth.

Epidemiological evaluation of apical periodontitis prevalence in an urban Brazilian population

The present study aimed to assess the prevalence of apical periodontitis (AP) in an urban Brazilian population according to gender, age group and tooth type. Data were collected from clinical files containing the medical and dental histories and periapical radiographs of 1,126 patients treated at the School of Dentistry at Universidade do Estado do Rio de Janeiro between March 2000 and December 2010. A total of 15,724 periapical radiographs were evaluated. All the radiographs were evaluated by two independent, previously calibrated endodontists (kappa = 0.88). Periapical areas on the radiographs were classified as N (normal) or AR (apical radiolucency). The frequency of AP and the 95% Confidence Interval (95%CI) were calculated according to gender, age group and tooth type. Differences between groups were calculated using the Z-test at a significance level of 5% (p < 0.05). AP was present in 7.87% of the samples, with 16.70% occurring on previously endodontically treated teeth and 44.65% occurring on teeth referred for endodontic treatment (TR-RCT). The frequency of AP was higher among females (64%) than among males (35%). The central and lateral maxillary incisors were the most frequently affected teeth. The frequency of AP was higher among individuals between 30 and 49 years of age. In this population, AP was more prevalent among females and among individuals between 30 and 49 years of age, and the central and lateral maxillary incisors were the most frequently affected teeth.

Avaliação da extrusão bacteriana apical após instrumentação mecanizada com sistemas reciprocantes de instrumento único comparados a um sistema rotatório multi-instrumento / Apical extrusion of bacteria by reciprocating single file systems and rotary multi-file system

O objetivo deste estudo foi avaliar ex vivo a extrusão bacteriana apical após instrumentação mecanizada com sistemas reciprocantes de instrumento único e movimento reciprocante (WaveOne® and Reciproc®) comparados a um sistema multi-instrumentos (BioRaCe®). Quarenta e cinco incisivos inferiores humanos unirradiculares, ovais e de anatomia semelhante foram utilizados. Os dentes foram acessados e seus canais radiculares foram contaminados com uma suspensão de Enterococcus faecalis e incubados por 30 dias possibilitando crescimento bacteriano em biofilme. Os dentes contaminados foram divididos em três grupos com 15 espécimes cada (RE - Reciproc®, WO -WaveOne® e BR - BioRaCe®). Foram utilizados oito dentes para grupos controle de crescimento bacteriano positivo e negativo. As bactérias extruídas apicalmente durante a instrumentação foram coletadas em frascos de vidro contendo 0,9% de NaCl. As amostras microbiológicas foram retiradas dos frascos e incubadas em meio BHI ágar, durante 24 horas. O crescimento bacteriano foi contado e os resultados foram expressos em unidades formadoras de colônia (UFC). Os dados foram analisados pelos testes estatísticos de Wilcoxon e Kruskal-Wallis. Não houve diferença estatisticamente significante no número de UFC entre os dois sistemas reciprocantes (p>0,05). Em contrapartida, o sistema de instrumentos rotatórios mostrou uma quantidade de UFC significativamente maior do que os dois outros grupos (p <0,05). A partir da análise dos resultados e dentro das limitações deste estudo foi possível concluir que todos os sistemas de instrumentação testados extruem bactérias apicalmente. No entanto, ambos os sistemas de instrumento único e movimento reciprocante extruem menos bactérias apicalmente do que o sistema rotatório multi-instrumentos de referência

The aim of this work was to evaluate ex vivo the apical bacterial extrusion of reciprocating single-file systems (Wave One® and Reciproc®) compared to a multi-file rotary system (BioRaCe®). This was tested in the current study using extracted, single-rooted teeth with oval canals (n = 45) with similar anatomy. Endodontic access cavities were prepared and root canals were contaminated with an Enterococcus faecalis suspension and incubated for thirty days to promote bacterial biofilm growth. The contaminated teeth were divided into three groups with 15 specimens each (RE - Reciproc®, WO- Wave One® and BR - BioRaCe®). Positive and negative bacterial growth control groups were done. Bacterial extruded from the apical foramen during instrumentation were collected into vials containing 0.9% NaCl. The microbiological samples were taken from the vials and incubated in BHI agar medium during 24h. Bacterial growth was counted and the results were given by colony-forming units (CFU). These data were analyzed by Wilcoxon matched-pairs signed rank test and Kruskal–Wallis H-test. After data analysis, no significant difference was found in the number of CFU between the two reciprocating systems (p > 0.05). Conventional multi-file rotary system group showed significantly higher CFU than both of the other two reciprocating groups (p<0.05). Based on the results evaluation and within the limitation of this study, it was possible to conclude that all tested instrumentation systems extruded bacteria beyond the apex. However, both reciprocation single-file systems apically extruded fewer bacteria than the conventional multi-file rotary system