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LIM/52 - Laboratório de Virologia, Hospital das Clínicas, Faculdade de Medicina

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Agora exibindo 1 - 10 de 136
  • article 9 Citação(ões) na Scopus
    Critical Analyses of the Introduction of Liquid-Based Cytology in a Public Health Service of the State of Sao Paulo, Brazil
    (2015) LONGATTO-FILHO, Adhemar; LEVI, Jose Eduardo; MARTINS, Toni Ricardo; COHEN, Diane; CURY, Lise; VILLA, Luisa Lina; ELUF-NETO, Jose
    Objective: The aim of this study was to compare the performance of the current conventional Pap smear with liquidbased cytology (LBC) preparations. Study Design: Women routinely undergoing their cytopathological and histopathological examinations at Fundacao Oncocentro de Sao Paulo (FOSP) were recruited for LBC. Conventional smears were analyzed from women from other areas of the State of Sao Paulo with similar sociodemographic characteristics. Results: A total of 218,594 cases were analyzed, consisting of 206,999 conventional smears and 11,595 LBC. Among the conventional smears, 3.0% were of unsatisfactory preparation; conversely, unsatisfactory LBC preparations accounted for 0.3%. The ASC-H (atypical squamous cells -cannot exclude high-grade squamous intraepithelial lesion) frequency did not demonstrate any differences between the twomethods. In contrast, the incidence of ASC-US (atypical squamous cells of undetermined significance) was almost twice as frequent between LBC and conventional smears, at 2.9 versus 1.6%, respectively. An equal percentage of highgrade squamous intraepithelial lesions were observed for the two methods, but not for low-grade squamous intraepithelial lesions, which were more significantly observed in LBC preparations than in conventional smears (2.2 vs. 0.7%). The index of positivity was importantly enhanced from 3.0% (conventional smears) to 5.7% (LBC). Conclusions : LBC performed better than conventional smears, and we are truly confident that LBC can improve public health strategies aimed at reducing cervical lesions through prevention programs. (C) 2015 S. Karger AG, Basel
  • article 0 Citação(ões) na Scopus
    Detection and analysis of blood donors seropositive for syphilis
    (2021) ATTIE, Adriana; ALMEIDA-NETO, Cesar de; WITKIN, Steven S.; DERRIGA, Juliana; NISHIYA, Anna S.; FERREIRA, Jerenice E.; COSTA, Natalia de Souza Xavier; SALLES, Nanci Alves; FACINCANI, Tila; LEVI, Jose E.; SABINO, Ester C.; ROCHA, Vanderson; MENDRONE-JR, Alfredo; FERREIRA, Suzete C.
    Background The increasing incidence of syphilis worldwide has called attention to the risk of transmission by transfusion. Aims To determine the prevalence of active syphilis in blood donors and characterise the serological profile of syphilis-positive donors. Methods Samples positive for Treponema pallidum using the chemiluminescent microparticle immunoassay (CMIA) during blood donor screening from 2017 to 2018 were tested by the Venereal Disease Research Laboratory (VDRL) non-treponemal test and for anti-T. pallidum IgM by ELISA (Immunoassay Enzyme test for detection of IgM antibodies). The INNO-LIA Syphilis test (Line Immuno Assay solid test for confirmation antibodies to Treponema pallidum) was performed as a confirmatory test on samples that were positive on ELISA-IgM but negative on VDRL. ELISA-IgM (+) samples were also tested for T. pallidum DNA in sera by real-time polymerase chain reaction (PCR). Results Of 248 542 samples screened, 1679 (0.67%) were positive for syphilis by CMIA. Further analysis was performed on 1144 (68.1%) of these samples. Of those tested, 16% were ELISA IgM(+)/VDRL(+), 16.5% were ELISA IgM(-)/VDRL(+), 4.1% were ELISA IgM(+)/VDRL(-), and 63.4% were ELISA IgM (-)/VDRL(-). The INNO-LIA Syphilis test results were 33 (3%) positive, 2 (0.2%) undetermined and 12 (1%) negative. Of the 230 EIA-IgM(+) samples (20.1%), 5 (2.2%) were PCR positive. The prevalence of active syphilis in 2017 and 2018 was 0.1% and 0.07%, respectively, and overall prevalence of serologic markers for syphilis was highest among male, unmarried, 25-34-year-olds with a high school education and who were first-time donors. Conclusion There is a risk of transfusion-transmitted syphilis in blood banks that exclusively use the VDRL test for donor screening, as is currently the situation in some Brazilian blood centres, as well as in other blood centres around the world.
  • article 8 Citação(ões) na Scopus
    Clinical characteristics of women diagnosed with carcinoma who tested positive for cervical and anal high-risk human papillomavirus DNA and E6 RNA
    (2015) VEO, Carlos A. R.; SAAD, Sarhan S.; FREGNANI, Jose Humberto T. G.; SCAPULATEMPO-NETO, Cristovam; TSUNODA, Audrey Tieko; RESENDE, Julio Cesar Possati; LORENZI, Adriana Tarla; MAFRA, Allini; CINTI, Claudia; COTRIM, Ismael Dale; ROSA, Luciana Albina Reis; OLIVEIRA, Cristina Mendes de; MARTINS, Toni Ricardo; CENTRONE, Cristiane; LEVI, Jose Eduardo; LONGATTO-FILHO, Adhemar
    High-risk human papillomavirus (hrHPV) is an essential cause of cervical carcinoma and is also strongly related to anal cancer development. The hrHPV E6 oncoprotein plays a major role in carcinogenesis. We aimed to evaluate the frequency of hrHPV DNA and E6 oncoprotein in the anuses of women with cervical carcinoma. We analyzed 117 women with cervical cancer and 103 controls for hrHPV and the E6 oncogene. Positive test results for a cervical carcinoma included 66.7 % with hrHPV-16 and 7.7 % with hrHPV-18. One case tested positive for both HPV variants (0.9 %). The samples from the anal canal were positive for HPV-16 in 59.8 % of the cases. Simultaneous presence of HPV in the cervix and anal canal was found in 53.8 % of the cases. Regarding expression of E6 RNA, positivity for HPV-16 in the anal canal was found in 21.2 % of the cases, positivity for HPV-16 in the cervix was found in 75.0 %, and positivity for HPV-18 in the cervix was found in 1.9 %. E6 expression in both the cervix and anal canal was found in 19.2 % of the cases. In the controls, 1 % tested positive for HPV-16 and 0 % for HPV-18. Anal samples from the controls showed a hrHPV frequency of 4.9 % (only HPV16). The presence of hrHPV in the anal canal of women with cervical cancer was detected at a high frequency. We also detected E6 RNA expression in the anal canal of women with cervical cancer, suggesting that these women are at risk for anal hrHPV infection.
  • conferenceObject
    (2018) LUNA, Expedito; FIGUEIREDO, Gerusa; LEVI, Jose; CAMPOS, Sergio; FIGUEIREDO, Walter; COSTA, Angela; FELIX, Alvina; SOUZA, Nathalia; PANNUTI, Claudio
  • article 15 Citação(ões) na Scopus
    Genomic analysis of head and neck cancer cases from two high incidence regions
    (2018) PERDOMO, Sandra; ANANTHARAMAN, Devasena; FOLL, Matthieu; ABEDI-ARDEKANI, Behnoush; DURAND, Geoffroy; ROSA, Luciana Albina Reis; HOLMILA, Reetta; CALVEZ-KELM, Florence Le; TAJARA, Eloiza H.; WUNSCH-FILHO, Victor; LEVI, Jose Eduardo; VILENSKY, Marta; POLESEL, Jerry; HOLCATOVA, Ivana; SIMONATO, Lorenzo; CANOVA, Cristina; LAGIOU, Pagona; MCKAY, James D.; BRENNAN, Paul
    We investigated how somatic changes in HNSCC interact with environmental and host risk factors and whether they influence the risk of HNSCC occurrence and outcome. 180-paired samples diagnosed as HNSCC in two high incidence regions of Europe and South America underwent targeted sequencing (14 genes) and evaluation of copy number alterations (SCNAs). TP53, PIK3CA, NOTCH1, TP63 and CDKN2A were the most frequently mutated genes. Cases were characterized by a low copy number burden with recurrent focal amplification in 11q13.3 and deletion in 15q22. Cases with low SCNAs showed an improved overall survival. We found significant correlations with decreased overall survival between focal amplified regions 4p16, 10q22 and 22q11, and losses in 12p12, 15814 and 15q22. The mutational landscape in our cases showed an association to both environmental exposures and clinical characteristics. We confirmed that somatic copy number alterations are an important predictor of HNSCC overall survival.
  • article 1 Citação(ões) na Scopus
    Low mutation percentage of KRAS and BRAF genes in Brazilian anal tumors
    (2016) BIDINOTTO, Lucas Tadeu; VEO, Carlos A. R.; LOAIZA, Edgar Aleman; FRANCA, Alessandra Paulino Santos De; LORENZI, Adriana Tarla; ROSA, Luciana Albina Reis; OLIVEIRA, Cristina Mendes De; LEVI, Jose Eduardo; SCAPULATEMPO-NETO, Cristovam; LONGATTO-FILHO, Adhemar; REIS, Rui Manuel
    Anal cancer is a rare type of digestive tract disease, which has had a crescent incidence in a number of regions. Carcinomas are most frequently found, with squamous cell carcinoma (SCC) comprising similar to 95% of all anal tumors. The major risk factor for development of this type of tumor is human papillomavirus (HPV) infection. However, previous studies have identified patients with anal cancer that are HPV-/p16-and observed that they have a poorer outcome compared with HPV+/p16+ patients. This suggests that molecular profile may drive anal cancer progression. The aim of the present study was to evaluate the mutational status of two important oncogenes, KRAS and BRAF, in a series of anal cancer lesions. Resected tumors of the anal canal (n=43) were evaluated, nine of these were high-grade squamous intra-epithelial lesion cases (HSIL), 11 were adenocarcinomas, and 23 SCCs. Direct sequencing of KRAS proto-oncogene, GTPase (KRAS; codons 12 and 13) and B-Raf proto-oncogene, serine/threonine kinase (BRAF; codon 600) was performed and associated with patient clinicopathological and molecular features. There was a trend of poorer prognosis of adenocarcinoma compared with HSIL and SCC. Analysis indicated one SCC patient (2.3%) exhibited a KRAS p.G13D mutation, and one adenocarcinoma patient (2.3%) exhibited a BRAF p.V600E mutation. It was observed that, these mutations are rare in anal tumors, and certain patients may be at a disadvantage using targeted therapies based on KRAS and BRAF mutational status. As there is a low mutation percentage in SCCs, adenocarcinomas and HSIL, there may exist other underlying molecular alterations that result in anal cancer development, which require further elucidation.
  • article 64 Citação(ões) na Scopus
    Microbial Translocation Is Associated with Extensive Immune Activation in Dengue Virus Infected Patients with Severe Disease
    (2013) WEG, Cornelia A. M. van de; PANNUTI, Claudio S.; ARAUJO, Evaldo S. A. de; HAM, Henk-Jan van den; ANDEWEG, Arno C.; BOAS, Lucy S. V.; FELIX, Alvina C.; CARVALHO, Karina I.; MATOS, Andreia M. de; LEVI, Jose E.; ROMANO, Camila M.; CENTRONE, Cristiane C.; RODRIGUES, Celia L. de Lima; LUNA, Expedito; GORP, Eric C. M. van; OSTERHAUS, Albert D. M. E.; MARTINA, Byron E. E.; KALLAS, Esper G.
    Background: Severe dengue virus (DENV) disease is associated with extensive immune activation, characterized by a cytokine storm. Previously, elevated lipopolysaccharide (LPS) levels in dengue were found to correlate with clinical disease severity. In the present cross-sectional study we identified markers of microbial translocation and immune activation, which are associated with severe manifestations of DENV infection. Methods: Serum samples from DENV-infected patients were collected during the outbreak in 2010 in the State of Sao Paulo, Brazil. Levels of LPS, lipopolysaccharide binding protein (LBP), soluble CD14 (sCD14) and IgM and IgG endotoxin core antibodies were determined by ELISA. Thirty cytokines were quantified using a multiplex luminex system. Patients were classified according to the 2009 WHO classification and the occurrence of plasma leakage/shock and hemorrhage. Moreover, a (non-supervised) cluster analysis based on the expression of the quantified cytokines was applied to identify groups of patients with similar cytokine profiles. Markers of microbial translocation were linked to groups with similar clinical disease severity and clusters with similar cytokine profiles. Results: Cluster analysis indicated that LPS levels were significantly increased in patients with a profound pro-inflammatory cytokine profile. LBP and sCD14 showed significantly increased levels in patients with severe disease in the clinical classification and in patients with severe inflammation in the cluster analysis. With both the clinical classification and the cluster analysis, levels of IL-6, IL-8, sIL-2R, MCP-1, RANTES, HGF, G-CSF and EGF were associated with severe disease. Conclusions: The present study provides evidence that both microbial translocation and extensive immune activation occur during severe DENV infection and may play an important role in the pathogenesis.
  • article 9 Citação(ões) na Scopus
    PCR-RFLP assay as an option for primary HPV test
    Persistent human papillomavirus (HPV) infection is an essential factor of cervical cancer. This study evaluated the analytical performance of restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) assay compared to PapilloChecks (R) microarray to identify human papilloma virus (HPV) in cervical cells. Three hundred and twenty-five women were analyzed. One sample was used for conventional cytology and another sample was collected using BD SurePath (TM) kit for HPV tests. Eighty samples (24.6%) were positive for HPV gene by PCR-Multiplex and were then submitted to PCR-RFLP and PapilloChecks (R) microarray. There was a genotyping agreement in 71.25% (57/80) on at least one HPV type between PCRRFLP and PapilloChecks (R) microarray. In 22 samples (27.5%), the results were discordant and those samples were additionally analyzed by DNA sequencing. HPV 16 was the most prevalent HPV type found in both methods, followed by HPVs 53, 68, 18, 39, and 66 using PCR-RFLP analysis, and HPVs 39, 53, 68, 56, 31, and 66 using PapilloChecks (R) microarray. In the present study, a perfect agreement using Cohen's kappa (kappa) was found in HPV 33 and 58 (kappa=1), very good for HPV 51, and good for types 16, 18, 53, 59, 66, 68, 70, and 73. PCR-RFLP analysis identified only 25% (20/80) HPV coinfection, and PapilloChecks (R) microarray found 62.5% (50/80). Our Cohen's kappa results indicate that our in-house HPV genotyping testing (PCR-RFLP analysis) could be applied as a primary HPV test screening, especially in low income countries. If multiple HPV types are found in this primary test, a more descriptive test, such as PapilloChecks (R) microarray, could be performed.
  • article 13 Citação(ões) na Scopus
    Asymptomatic infections in blood donors harbouring Plasmodium: an invisible risk detected by molecular and serological tools
    (2018) LIMA, Giselle F. M. C.; SANCHEZ, Maria C. Arroyo; LEVI, Jose E.; FUJIMORI, Mahyumi; CARAMELO, Luiza da Cruz; SANCHEZ, Arianni Rondelli; RAMOS-SANCHEZ, Eduardo M.; INOUE, Juliana; COSTA-NASCIMENTO, Maria de Jesus; MENDRONE JUNIOR, Alfredo; SANTI, Silvia M. Di
    Background. Transfusion-transmitted malaria due to asymptomatic Plasmodium infections is a challenge for blood banks. There is a lack of data on the prevalence of asymptomatic infected blood donors and the incidence of transfusion-transmitted malaria in low endemicity areas worldwide. We estimated the frequency of blood donors harbouring Plasmodium in an area in which asymptomatic infections have been reported. Material and methods. To estimate the frequency of blood donors harbouring Plasmodium we used microscopy and molecular tools. Serological tests were applied to measure the exposure of candidates to Plasmodium antigens. Venous blood was collected from 91 candidates attending the ""Pro-Sangue"" Blood Centre Foundation in Sao Paulo, who lived in the municipality of Juquitiba, Sao Paulo, Brazil, where sporadic autochthonous cases of malaria have been described. Blood samples were used for parasitological, molecular and serological studies. Results. Among the 91 samples examined, rare Plasmodium forms were observed in two donors. Genus real-time polymerase chain reaction analysis demonstrated Plasmodium amplification in three candidates and species-specific nested polymerase chain reaction identified P. malariae in two. ELISA-IgG was reactive in 42.9% of samples for P. vivax (Pv-MSP1 19) and in 6.6% for P. falciparum (Pf-Zw). ELISA-IgM was reactive in 2.2% of samples for P. vivax and in 4.4% for P. falciparum. An indirect immunofluorescence assay was reactive for P. malariae in 15.4% of cases. Discussion. Reservoirs of Plasmodium represent a challenge for blood banks, since studies have shown that high levels of submicroscopic infections can occur in low transmission areas. The risk of transfusion-transmitted malaria presented here points to the need to conduct molecular investigations of candidate donors with any positive malarial antibody test.
  • article 42 Citação(ões) na Scopus
    West Nile virus surveillance, Brazil, 2008-2010
    (2013) OMETTO, Tatiana; DURIGON, Edison Luiz; ARAUJO, Jansen de; APRELON, Rosalie; AGUIAR, Daniel Moura de; CAVALCANTE, Guacyara Tenorio; MELO, Rosane Marini; LEVI, Jose Eduardo; AZEVEDO JUNIOR, Severino Mendes de; PETRY, Maria Virginia; NETO, Isaac Simao; SERAFINI, Patricia; VILLALOBOS, Eliana; CUNHA, Elenice Maria Sequetin; LARA, Maria do Carmo Custodio S. H.; NAVA, Alessandra Ferreira Dales; NARDI, Marcello Schiavo; HURTADO, Renata; RODRIGUES, Roberta; SHERER, Angelo Luis; SHERER, Janete de Fatima Martins; GERALDI, Marcelo Plaisant; SEIXAS, Marina Maria Moraes de; PETERKA, Cassio; BANDEIRA, Debora de Souza; PRADEL, Jennifer; VACHIERY, Nathalie; LABRUNA, Marcelo Bahia; CAMARGO, Luiz Marcelo Aranha de; LANCIOTTI, Robert; LEFRANCOIS, Thierry
    Background: West Nile virus (WNV) is an emergent pathogen that is widely distributed in North and Central America. The recent introduction in South America has focused attention on the spread of WNV across Southern American countries. The transmission network involves mosquitoes, birds, horses and humans. Methods: The serological evaluation of sera from 678 equids and 478 birds was performed using a WNV-specific blocking ELISA, and only the positive results were confirmed by plaque reduction neutralisation tests (PRNTs). Molecular analysis was performed on sera from 992 healthy equids and on 63 macerates of brains from equids that died of encephalitis and had previously tested negative for other pathogens. We also tested swabs from 928 birds. The samples analysed were collected in different biomes of Brazil. Results: We identified WNV antibodies by ELISA in thirteen equids and five birds, and PRNT90 confirmed WNV positivity in four equid samples collected in 2009 in an area between the Amazon and the Pantanal. None of the ELISA positive bird samples were confirmed by PRNT90, and all samples tested by RT-PCR were negative. Conclusion: WNV circulation is confirmed by this large scale survey even in the absence of detection of clinical cases.