JOSE EDUARDO LEVI

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

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Autor: LEVI, Jose E. ×

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  • 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 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 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 17 Citação(ões) na Scopus
    An autopsy-based study of Trypanosoma cruzi persistence in organs of chronic chagasic patients and its relevance for transplantation
    (2017) BENVENUTI, Luiz A.; ROGGERIO, Alessandra; CAVALCANTI, Marta M.; NISHIYA, Anna S.; LEVI, Jose E.
    BackgroundChagas' disease (CD) is caused by infection with the protozoan Trypanosoma cruzi. The disease can affect the heart and/or the gastrointestinal (GI) tract, but around 70% of infected individuals remain asymptomatic in the chronic form. Organ transplantation from T.cruzi-infected donors is often avoided because of the risk of disease transmission, previously reported after heart, kidney, or liver transplantation. MethodsWe investigated by histology, immunohistochemistry, and polymerase chain reaction (PCR) the persistence of T.cruzi in samples of the heart, lung, liver, kidney, pancreas, adrenal gland, esophagus, and GI tract of 21 chronic chagasic patients. ResultsParasite persistence was detected in 12/21 (57.1%) heart samples, mainly by PCR-based assays. T.cruzi parasites were detected by histology and immunohistochemistry in smooth muscle cells of the central vein from 1/21 (4.8%) adrenal gland samples. No samples of the lung, liver, kidney, pancreas, esophagus, or GI tract were found to have parasites by histology, immunohistochemistry, or PCR. ConclusionsWe concluded that, aside from the heart, the other solid organs of T.cruzi-infected donors can be used for transplantation with a lot of caution. Such organs are not safe in the view of previous reports of CD transmission, but seem to present a low T.cruzi load compared to the heart.
  • conferenceObject
    ZIKA VIRUS INFECTION IN A COHORT STUDY TO ASSESS THE INCIDENCE OF DENGUE, STATE OF SAO PAULO, BRAZIL, 2015, 2016
    (2017) FIGUEIREDO, Gerusa M.; LUNA, Expedito J.; CARDOSO, Maria Regina; LEVI, Jose E.; FELIX, Alvina C.; SOUZA, Nathalia C. C.; SOUZA, Ana C.; CAMPOS, Sergio R. Campos R.; FIGUEIREDO, Walter M.; COSTA, Angela A.; PANNUTI, Claudio S.
  • article 26 Citação(ões) na Scopus
    HCV Genotypes, Characterization of Mutations Conferring Drug Resistance to Protease Inhibitors, and Risk Factors among Blood Donors in Sao Paulo, Brazil
    (2014) NISHIYA, Anna S.; ALMEIDA-NETO, Cesar de; FERREIRA, Suzete C.; ALENCAR, Cecilia S.; DI-LORENZO-OLIVEIRA, Claudia; LEVI, Jose E.; SALLES, Nanci A.; MENDRONE JR., Alfredo; SABINO, Ester C.
    Background: Hepatitis C virus (HCV) infection is a global health problem estimated to affect almost 200 million people worldwide. The aim of this study is to analyze the subtypes and existence of variants resistant to protease inhibitors and their association with potential HCV risk factors among blood donors in Brazil. Methods: Repeat anti-HCV reactive blood donors are systematically asked to return for retest, notification, and counseling in which they are interviewed for risk factors for transfusion-transmitted diseases. We analyzed 202 donors who returned for counseling from 2007 to 2010 and presented enzyme immunoassay-and immunoblot-reactive results. The HCV genotypes and resistance mutation analyses were determined by the direct sequencing of the NS5b and NS3 regions, respectively. The HCV viral load was determined using an in-house real-time PCR assay targeting the 5'-NCR. Results: HCV subtypes 1b, 1a, and 3a were found in 45.5%, 32.0%, and 18.0% of the donors, respectively. The mean viral load of genotype 1 was significantly higher than that of the genotype 3 isolates. Subtype 1a was more frequent among young donors and 3a was more frequent among older donors. Protease inhibitor-resistant variants were detected in 12.8% of the sequenced samples belonging to genotype 1, and a higher frequency was observed among subtype 1a (20%) in comparison to 1b (8%). There was no difference in the prevalence of HCV risk factors among the genotypes or drug-resistant variants. Conclusions: We found a predominance of subtype 1b, with an increase in the frequency of subtype 1a, in young subjects. Mutations conferring resistance to NS3 inhibitors were frequent in treatment-naive blood donors, particularly those infected with subtype 1a. These variants were detected in the major viral population of HCV quasispecies, have replicative capacities comparable to nonresistant strains, and could be important for predicting the response to antiviral triple therapy.
  • article 9 Citação(ões) na Scopus
    Evaluation of the applicability and effectiveness of a molecular strategy for identifying weakD and DEL phenotype among D- blood donors of mixed origin exhibiting high frequency of RHD*
    (2018) DEZAN, Marcia Regina; GUARDALINI, Luis Giovani O.; PESSOA, Elaine; RIBEIRO, Ingrid Helena; OLIVEIRA, Valeria Brito; LUZ, Fabio; NOVAC, Denise Rossite; GALLUCCI, Antonio; BONIFACIO, Silvia; GOMES, Francisco; LEVI, Jose E.; PEREIRA, Alexandre C.; KRIEGER, Jose E.; MENDRONE-JUNIOR, Alfredo; ROCHA, Vanderson; DINARDO, Carla Luana
    BACKGROUNDMolecular tests designed to detect the presence of active RHD gene among D- donors have been successfully applied in people of European ancestry, but not in admixed populations with a considerable frequency of RHD*. Our goal was to evaluate the performance of a molecular screening tool for identifying active RHD alleles among Brazilian blood donors classified as D- C+ and/or E+. STUDY DESIGN AND METHODSPools of five DNA samples of serologically D- C+ and/or E+ donors were checked by a RHD polymerase chain reaction (PCR) assay specific for RHD Intron 4 and Exon 7. When a pool result was positive, samples were genotyped individually for RHD Intron 4 and Exon 7, RHD*, RHCE*Cc, and RHD zygosity. Donors suspected of active RHD gene were further evaluated by whole-coding region and flanking intron direct sequencing. RESULTSA total of 405 donors were included. Two percent exhibited active RHD gene, codifying D-weak (38 and 45) or DEL phenotype. The most prevalent DEL allele was RHD*DEL1 (c.1227G>A), which is proven to be immunogenic. A high frequency of RHD* was detected in the donors with nondeleted RHD alleles (31%), far superior to the frequency of RHD variant alleles (15.5%). The proposed approach presented sensitivity of 100% and specificity of 85.7% for identifying active RHD gene. CONCLUSIONThe strategy of checking D- donors with RHD PCR followed by exclusion of RHD* allele has proved efficient in identifying weak-D and DEL phenotype in the Brazilian population.
  • article 1 Citação(ões) na Scopus
    Prevalence of SMIM1 c.64_80del17 homozygotes in southeastern Brazil: the Vel-negative phenotype
    (2019) DEZAN, Marcia R.; DINARDO, Carla L.; ROCHA, Vanderson; MENDRONE-JUNIOR, Alfredo; LEVI, Jose E.
  • article 2 Citação(ões) na Scopus
    Chlamydia trachomatis infection in women living in remote areas in Amazonas, Brazil-a self-collection screening experience
    (2019) ROCHA, Danielle A. P.; MORAES, Cassia de Oliveira; ARAUJO, Adriene F.; BELTRAO, Emille dos Santos; SANTOS, Lainara Castelo dos; MATA, Ligia Menezes da; XAVIER, Antonio N. N.; REIS, Renato dos Santos; SOARES, Silvia C. C.; TORRES, Katia L.; LEVI, Jose E.; MARINO, Josiane M.
    Chlamydia trachomatis infection is the most common sexually transmitted bacterial infection among women. In Brazil, there is no organized screening program for C. trachomatis, and the actual prevalence of infection is unknown. This study aimed to determine the prevalence of C. trachomatis infection in women living in riverside communities in the Amazon, using self-collection employing the Evalyn (R) Brush and polymerase chain reaction. A total of 299 riverine women aged 18-81 years, mean age 35.7 (+/- 12.8) years, predominantly agricultural workers, with low schooling and living with a partner, participated in this study. The prevalence of C. trachomatis infection was found to be 3.7% (95% CI 1.8-6.5), most of them being symptomatic. The mean age of the first sexual intercourse reported by women was 15.2 (+/- 2.3) years, and the majority reported having had none or only one partner in the last 12 months, with very low adherence to consistent condom use (15.4%). Most women (98.3%) reported having approved using the vaginal self-collecting brush, and only 4.7% reported having difficulty in handling the brush. We consider that a vaginal self-collecting device is adequate for diagnosing C. trachomatis infection in women living in remote, hard-to-reach areas.
  • article 31 Citação(ões) na Scopus
    Malaria diagnosis from pooled blood samples: comparative analysis of real-time PCR, nested PCR and immunoassay as a platform for the molecular and serological diagnosis of malaria on a large-scale
    (2011) LIMA, Giselle F. M. C.; LEVI, Jose E.; GERALDI, Marcelo P.; SANCHEZ, Maria Carmen A.; SEGURADO, Aluisio A. C.; HRISTOV, Angelica D.; INOUE, Juliana; COSTA-NASCIMENTO, Maria de Jesus; SANTI, Silvia M. Di
    Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis.