Performance of Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Targeting the RNA Polymerase Gene for the Direct Detection of SARS-CoV2 in Nasopharyngeal Swabs.

In 2020, a global pandemic caused by SARS-CoV-2 was declared. Different institutes proposed diagnostic molecular methods to detect the virus in clinical samples. This study aims to validate and standardize the use of a loop-mediated isothermal amplification (LAMP)-based methodology targeting the viral RP gene, as a faster and low-cost diagnostic method for SARS-CoV-2 infections. The results obtained with RT-LAMP (Reverse Transcriptase) were compared to the results of real-time polymerase chain reaction (RT-PCR) to assess its sensitivity and specificity. In total, 115 samples (nasopharyngeal samples) were used for detecting SARS-CoV-2 by RT-LAMP, with 43 positives and 72 negatives. The study showed a positive predictive value (PPV) of 90.7% and a negative predictive value (VPN) of 100%. The LAMP assay also demonstrated a high sensitivity of 90.7% and a specificity of 100% (confidence interval 77.9-97.4%) when using the lower detection limit of 40 copies/µL. The RT-LAMP described here has the potential to detect even the new variants of SARS-CoV-2, suggesting that it may not be significantly affected by gene mutations. The RT-LAMP targeting the RP viral region is faster and less expensive than other molecular approaches, making it an alternative for developing countries.

Influência da adesão bacteriana em resina composta Bulk fill submetida a diferentes protocolos de acabamento e polimento: estudo in vitro / Influence of bacterial adhesion on Bulk fill composite resin submitted to different finishing and polishing protocols: in vitro study

Objetivo: avaliar a adesão bacteriana do Streptococcus mutans na superfície de uma resina composta do tipo incremento único submetida a diferentes protocolos de polimento. Materiais e métodos: foram realizadas 60 amostras nas quais foram divididas aleatoriamente em 6 grupos (n=10) de diferentes tratamentos de superfície. Cinco amostras de cada grupo foram separadas e submetidas ao estudo de adesão bacteriana, das quais duas foram analisadas pela microscopia eletrônica de varredura. Foram contabilizadas as unidades formadoras de colônias UFC/ml de modo manual e realizada a média e desvio padrão de cada grupo. De acordo com os resultados analisados através do teste de One Way ANOVA e comparações múltiplas de Tukey observou-se uma diferença estatisticamente significativa entre os grupos. Resultados: os valores de UFC/ mL variaram de 0 para o grupo American Burrs a 8,64 para o grupo Dhpro. Os grupos Jota e Dhpro não diferiram estatisticamente entre si (p=0,71), porém diferiram dos demais grupos avaliados (p=0,45). Os grupos American Burrs e o controle negativo não diferiram estatisticamente entre si (p>0.999) e diferiram dos demais grupos testados (p=0,20). O grupo de controle positivo diferiu estatisticamente dos outros grupos (p=0,02) assim como o grupo KG (p=0,01). Conclusão: Diante dos resultados obtidos, pode-se concluir que, a superfície da resina Bulk Fill One submetida a diferentes protocolos de polimento e mesmo sem ter passado por nenhum tratamento de superfície é passível de adesão bacteriana seja por contagem manual ou microscopia eletrônica de varredura(AU)

Objective: To evaluate the bacterial adhesion of Streptococcus mutans on the surface of a single increment composite resin submitted to different polishing protocols. Materials and methods: 60 samples were randomly divided into 6 groups (n = 10) of different surface treatments. Five samples from each group were separated and submitted to the bacterial adhesion study, two of which were analyzed by scanning electron microscopy. The UFC/ml colony forming units were accounted for manually and the mean and standard deviation of each group were performed. According to the results analyzed by the One Way ANOVA test and Tukey's multiple comparisons, a statistically significant difference was observed between the groups. Results: The values ranged from 0 for the American Burrs group to 8.64 for the Dhpro group. The Jota and Dhpro groups did not differ statistically (p = 0.71), but differed from the other groups evaluated (p = 0.45). The American Burrs and negative control groups did not differ statistically (p> 0.999) and differed from the other groups tested (p = 0.20). The positive control group differed statistically from the other groups (p = 0.02) as did the KG group (p = 0.01). Conclusion: It can be concluded that the surface of the Bulk Fill One resin submitted to different polishing protocols and even without any surface treatment is susceptible to bacterial adhesion either by manual counting or scanning electron microscopy(AU)