PAULO HENRIQUE BRAZ DA SILVA
Projetos de Pesquisa
Unidades Organizacionais
ODE, FO - Docente
LIM/52 - Laboratório de Virologia, Hospital das Clínicas, Faculdade de Medicina
LIM/52 - Laboratório de Virologia, Hospital das Clínicas, Faculdade de Medicina
10 resultados
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- Rapid diagnosis of Zika virus through saliva and urine by Loop-mediated isothermal amplification (LAMP)(2018) CASTRO, Talita; SABALZA, Maite; BARBER, Cheryl; ABRAMS, William; COSTA, Antonio Charlys Da; MILAGRES, Flavio Augusto De Padua; BRAZ-SILVA, Paulo Henrique; MALAMUD, Daniel; GALLOTTINI, MarinaBackground: Zika virus (ZIKV) is a single-stranded RNA virus and member of the Flaviviridae family. Recent studies have reported that saliva can be an important alternative to detect ZIKV. Saliva requires less processing than blood greatly simplifying the assay. Loop-mediated Isothermal Amplification (LAMP) is a rapid assay that detects nucleic acids, including ZIKV RNA. Aim: The aim of this study was to evaluate the efficacy of saliva and urine to diagnose ZIKV infection in subjects during the acute phase, through ZIKV RNA detection by LAMP. Method: A total of 131 samples (68 saliva and 63 urine samples) from 69 subjects in the acute phase of ZIKV infection, and confirmed positive for ZIKV by blood analysis through real time-PCR, were collected and analyzed by Reverse Transcriptase Loop-mediated Isothermal Amplification (RT-LAMP). Results: From the 68 saliva samples, 45 (66.2%) were positive for ZIKV with an average time to positivity (Tp) of 13.5min, and from the 63 urine samples, 25 (39.7%) were positive with the average Tp of 15.8min. Saliva detected more samples (p=0.0042) and had faster Tp (p=0.0176) as compared with urine. Conclusion: Saliva proved to be a feasible alternative to diagnose ZIKV infection during the acute phase by LAMP.
- Longitudinal study on oral shedding of human betaherpesviruses 6 and 7 in renal transplant recipients reveals active replication(2020) RAPOSO, Jessica Vasques; SARMENTO, Dmitry Jose De Santana; PINTO, Rafaela Barbosa Da Silva; LOPES, Amanda Oliveira; GALLOTTINI, Marina; TOZETTO-MENDOZA, Tania Regina; BRAZ-SILVA, Paulo Henrique; PAULA, Vanessa Salete deBackgroung Roseolovirus latency and persistence in salivary glands that are frequently reactivated after renal transplantation to cause infection have been reported. However, limited information is available on the persistence and excretion of HHV-6 and HHV-7 during and after transplant. Methods 32 renal transplant recipients were followed up before (T1) and after transplant (T2 and T3) and viral replication (via assessment of mRNA) in oral fluid samples investigated. Roseolovirus DNA was detected and quantified via multiplex qPCR. For evaluation of mRNA replication, positive samples were subjected to nested RT-PCR. Results Viral replication of HHV-7 was significantly increased during T3 (72.9%), compared to the pre-transplant period T1 (25%; McNemar Test, p= 0.001). Analysis of the viral replicative to quantitative ratio disclosed ahigher number of DNA copies (>10(6)) in positive cases of replication (p < 0.001). Astrong positive correlation (Spearman correlation coefficient = 0.781; p< 0.001) was evident between viral quantities of Roseoloviruses. Conclusion Our findings consistently suggest that the salivary gland is an important site of active and persistent infection by roseoloviruses. In view of the increasing problem of Roseoloviruses, pre- and post-transplantation, viral surveillance and monitoring of active replication are pivotal steps for effective screening and treatment of renal transplant patients.
- Prospective study of human herpesvirus 8 oral shedding, viremia, and serological status among human immunodeficiency virus seropositive and seronegative individuals in Sao Paulo, Brazil(2017) BRAZ-SILVA, Paulo H.; TOZETTO-MENDOZA, Tania R.; SUMITA, Laura M.; FREIRE, Wilton; PALMIERI, Michelle; CANTO, Alan M. do; AVELINO-SILVA, Vivian I.; GALLOTTINI, Marina; MAYAUD, Philippe; PANNUTI, Claudio S.Human herpesvirus 8 (HHV-8) is a gamma-herpesvirus and etiological agent of all forms of Kaposi sarcoma (KS). Saliva may play an important role in HHV-8 transmission in specific populations. Little is known about HHV-8 oral shedding pattern and the possible correlation with the HHV-8 serological profile and viremia. A prospective study was conducted of HHV-8 salivary excretion among human immunodeficiency virus HIV-seronegative (n = 47) and -seropositive (n = 44) homosexual men and HIV-seropositive women (n = 32) over a 6-month period with monthly HHV-8 serologies (immunofluorescence assays to identify antibodies to latent and lytic HHV-8 viral proteins, and a whole-virus HHV-8 enzyme-linked immunosorbent assay [ELISA]), monthly HHV-8 DNA serum/plasma detection, and daily self-collected oral rinses for HHV-8-DNA detection using real-time polymerase chain reaction. HHV-8 seropositivity was 51.1%, 63.6%, and 37.5%, in the three studied groups. There was no case of HHV-8 DNA detection in serum/plasma. Intermittent detection of oral HHV-8 DNA was observed during 5.1% (110/2,160) of visits among 28% (18/64) of HHV-8-seropositive individuals, all of whom were males and HHV-8 ELISA seropositive. In immunologically controlled populations of Brazil, HHV-8 oral shedding was limited to HHV-8-seropositive men, occurred infrequently and intermittently, and was not linked to HHV-8 viremia, suggesting a limited potential for oral or blood transmission.
- Lack of direct association between oral mucosal lesions and SARS-CoV-2 in a cohort of patients hospitalised with COVID-19(2022) SCHWAB, Gabriela; PALMIERI, Michelle; ZERBINATI, Rodrigo M.; SARMENTO, Dmitry J. S.; REIS, Thais; ORTEGA, Karem L.; KANO, Italo T.; V, Rafael A. Caixeta; HASSEUS, Bengt; SAPKOTA, Dipak; JUNGES, Roger; GIANNECCHINI, Simone; COSTA, Andre L. F.; JALES, Sumatra M. C. P.; LINDOSO, Jose A. L.; GALLO, Camila Barros; BRAZ-SILVA, Paulo H.Background COVID-19 is a disease affecting various human organs and systems, in which the virus seeks to interact with angiotensin-converting enzyme 2 receptors. These receptors are present in the oral cavity, but the direct relationship between such an interaction and possible oral manifestations of COVID-19 is still unclear. Aim The present study evaluated oral manifestations in a cohort of COVID-19 patients during the period of hospitalisation. Methods In total, 154 patients presenting moderate-to-severe forms of COVID-19 had their oral mucosa examined twice a week until the final outcome, either discharge or death. The oral alterations observed in the patients were grouped into Group 1 (pre-existing conditions and opportunistic oral lesions) and Group 2 (oral mucosal changes related to hospitalization). Results Oral lesions found in the patients of Group 1 are not suggestive of SARS-CoV-2 infection as they are mainly caused by opportunistic infections. On the other hand, oral alterations found in the patients of Group 2 were statistically (P < 0.001) related to intubation and longer period of hospitalisation. Conclusion It is unlikely that ulcerative lesions in the oral cavity are a direct manifestation of SARS-CoV-2 or a marker of COVID-19 progression.
- Periodontal tissues are targets for Sars-Cov-2: a post-mortem study(2021) MATUCK, Bruno Fernandes; DOLHNIKOFF, Marisa; MAIA, Gilvan V. A.; SENDYK, Daniel Isaac; ZARPELLON, Amanda; GOMES, Sara Costa; DUARTE-NETO, Amaro Nunes; PINHO, Joao Renato Rebello; GOMES-GOUVEA, Michele Soares; SOUSA, Suzana C. O. M.; MAUAD, Thais; SALDIVA, Paulo Hilario do Nascimento; BRAZ-SILVA, Paulo H.; SILVA, Luiz Fernando Ferraz daBackground: The ability of coronavirus SARS-CoV-2 to spread is one of the determinants of the COVID-19 pandemic status. Until June 2020, global COVID-19 cases surpassed 10 million. Asymptomatic patients, with no respiratory impairment, are believed to be responsible for more than 80% of the transmission. Other viruses have been consistently detected in periodontal tissues. Objective: The aim of this study was to investigate the presence of SARS-CoV-2 in periodontal tissue. Methods: We conducted video-endoscope minimally invasive post-mortem biopsy in seven fatal cases of COVID-19, using a regular endoscope video system associated with a smartphone to locate periodontal tissue. We analyzed the samples using RT-PCR, to identify the SARS-CoV-2 RNA and histopathological analysis. Results: The seven studied autopsies with positive laboratory tests for COVID-19 included 57.14% of female patients at the average age of 47.4 (range 8 to 74). In five cases, periodontal tissue was positive for SARS-CoV-2 (RT-PCR). Histopathologic analyses showed morphologic alterations in the keratinocytes of the junctional epithelium, a vacuolization of the cytoplasm and nucleus and nuclear pleomorphism. Conclusion: We presented a biomolecular analysis obtained from minimally invasive autopsies. This is the first study to demonstrate the presence of SARS-CoV-2 in periodontal tissue in COVID-19 positive patients.
- MALDI-TOF mass spectrometry of saliva samples as a prognostic tool for COVID-19(2022) LAZARI, Lucas C.; ZERBINATI, Rodrigo M.; ROSA-FERNANDES, Livia; SANTIAGO, Veronica Feijoli; ROSA, Klaise F.; ANGELI, Claudia B.; SCHWAB, Gabriela; PALMIERI, Michelle; SARMENTO, Dmity J. S.; MARINHO, Claudio R. F.; ALMEIDA, Janete Dias; TO, Kelvin; GIANNECCHINI, Simone; WRENGER, Carsten; SABINO, Ester C.; MARTINHO, Herculano; LINDOSO, Jose A. L.; DURIGON, Edison L.; BRAZ-SILVA, Paulo H.; PALMISANO, GiuseppeBackground The SARS-CoV-2 infections are still imposing a great public health challenge despite the recent developments in vaccines and therapy. Searching for diagnostic and prognostic methods that are fast, low-cost and accurate are essential for disease control and patient recovery. The MALDI-TOF mass spectrometry technique is rapid, low cost and accurate when compared to other MS methods, thus its use is already reported in the literature for various applications, including microorganism identification, diagnosis and prognosis of diseases. Methods Here we developed a prognostic method for COVID-19 using the proteomic profile of saliva samples submitted to MALDI-TOF and machine learning algorithms to train models for COVID-19 severity assessment. Results We achieved an accuracy of 88.5%, specificity of 85% and sensitivity of 91.5% for classification between mild/moderate and severe conditions. When we tested the model performance in an independent dataset, we achieved an accuracy, sensitivity and specificity of 67.18, 52.17 and 75.60% respectively. Conclusion Saliva is already reported to have high inter-sample variation; however, our results demonstrates that this approach has the potential to be a prognostic method for COVID-19. Additionally, the technology used is already available in several clinics, facilitating the implementation of the method. Further investigation using a larger dataset is necessary to consolidate the technique.
- Performance of at-home self-collected saliva and nasal-oropharyngeal swabs in the surveillance of COVID-19(2021) BRAZ-SILVA, Paulo H.; MAMANA, Ana C.; ROMANO, Camila M.; FELIX, Alvina C.; V, Anderson de Paula; FEREIRA, Noeli E.; BUSS, Lewis F.; TOZETTO-MENDOZA, Tania R.; V, Rafael A. Caixeta; LEAL, Fabio E.; GRESPAN, Regina M. Z.; BIZARIO, Joao C. S.; FERRAZ, Andrea B. C.; SAPKOTA, Dipak; GIANNECCHINI, Simone; TO, Kelvin K.; DOGLIO, Alain; MENDES-CORREA, Maria C.Background: SARS-CoV-2 quickly spreads in the worldwide population, imposing social restrictions to control the infection, being the massive testing another essential strategy to break the chain of transmission. Aim: To compare the performance of at-home self-collected samples - saliva and combined nasal-oropharyngeal swabs (NOP) - for SARS-CoV-2 detection in a telemedicine platform for COVID-19 surveillance. Material and methods: We analyzed 201 patients who met the criteria of suspected COVID-19. NOP sampling was combined (nostrils and oropharynx) and saliva collected using a cotton pad device. Detection of SARS-COV-2 was performed by using the Altona RealStar (R) SARS-CoV-2 RT-PCR Kit 1.0. Results: There was an overall significant agreement (kappa coefficient value of 0.58) between saliva and NOP. Considering results in either sample, 70 patients positive for SARS-CoV-2 were identified, with 52/70 being positive in NOP and 55/70 in saliva. This corresponds to sensitivities of 74.2% (95% CI; 63.7% to 83.1%) for NOP and 78.6% (95% CI; 67.6% to 86.6%) for saliva. Conclusion: Our data show the feasibility of using at-home self-collected samples (especially saliva), as an adequate alternative for SARS-CoV-2 detection. This new approach of testing can be useful to develop strategies for COVID-19 surveillance and for guiding public health decisions.
- Detection of polyomavirus microRNA-5p expression in saliva shortly after kidney transplantation(2021) MAMANA, Ana Carolina; STINCARELLI, Maria; SARMENTO, Dmitry Jose De Santana; BATISTA, Alexandre Mendes; TOZETTO-MENDOZA, Tania Regina; GALLOTTINI, Marina; PESTANA, Jose Osmar Medina De Abreu; BRAZ-SILVA, Paulo Henrique; GIANNECCHINI, SimoneBackground: MicroRNAs (miRNAs) of polyomavirus (PyV) are present in several biological fluids and are suggested to be relevant viral factors for monitoring its persistence. Aim: To evaluate the effect of an immunosuppressive regimen on the status of PyV-miRNA-5p in the oral cavity. Materials and Methods: The JCPyV, BKPyV, MCPyV miRNA-5p were investigated in paired saliva and plasma samples obtained from 23 patients before and shortly after renal-transplantation by using real-time RT-PCR. Results: Overall, within a short-time after transplantation, patients exhibited decreased numbers of leukocyte and lymphocyte as well as low levels of creatinine. During the clinical management of the patients, a significant amount of saliva samples were positive for JCPyV and BKPyV miRNA-5p (range: 26%-91%) compared to paired plasma samples (range: 9%-35%). Among the two polyomaviruses showing positive expression of miRNA-5p, BKPyV presented the highest positivity in saliva (91%) and MCPyV-miRNA-5p was constantly negative in both saliva and plasma samples. Compared to the time before transplantation, a significant reduction in the expression of JCPyV-miRNA-5p was observed in saliva samples obtained after transplantation. Conclusions: Altogether, these data suggest that additional investigations of polyomavirus miRNA-5p in saliva should be performed shortly after renal-transplantation to evaluate the potential role in early viral reactivation.
- Oral shedding of human herpesviruses in patients undergoing radiotherapy/chemotherapy for head and neck squamous cell carcinoma is not affected by xerostomia (vol 10, pg 1, 2018)(2020) PALMIERI, M.; ORNAGHI, M.; MARTINS, V. A. O.; CORREA, L.; BRANDAO, T. B.; RIBEIRO, A. C. P.; SUMITA, L. M.; TOZETTO-MENDOZA, T. R.; PANNUTI, C. S.; BRAZ-SILVA, P.
- Oral shedding at human herpesviruses in patients undergoing radiotherapy/chemotherapy for head and neck squamous cell carcinoma is not affected by xerostomia(2018) PALMIERI, Michelle; ORNAGHI, Mariana; MARTINS, Victor Adriano de Oliveira; CORREA, Luciana; BRANDAO, Thais Bianca; RIBEIRO, Ana Carolina do Prado; SUMITA, Laura Masami; TOZETTO-MENDOZA, Tania Regina; PANNUTI, Claudio Sergio; BRAZ-SILVA, Paulo HenriqueBackground: Xerostomia is a very relevant and frequent complication of radiotherapy, causing the irradiated oral mucosa to be affected by bacterial, fungal and viral infections. Objective: The objective of this study was to evaluate a possible relationship between oral shedding of human herpesviruses and xerostomia in patients with squamous cell carcinoma of head and neck submitted to radio/chemotherapy. Methods: In this study, oral rinse samples were collected weekly from 20 patients during radiotherapy. The samples were submitted to PCR and enzymatic digestion for detection of human herpesviruses. Xerostomia was evaluated according to the Seminars in Radiation Oncology criteria. Results: There was a higher frequency of grade 1 xerostomia (51.4%), observed first in the 1st week of radiotherapy. In the 4th week of radiotherapy, all patients presented some degree of xerostomia. Analysis of herpesviruses showed oral shedding of EBV, HHV-6 and HHV-7 in all weeks. Considering all the periods, the highest frequency was in patients with EBV excretion (55.0%), which was significantly higher than that of other viruses. Conclusion: We observed that oral shedding of herpesviruses was not affected by xerostomia as there was a progression in their excretion, even with the evolution of xerostomia. This suggested that there is a local replication in the oral cavity that is not completely dependent of salivary excretion.