JOSE EDUARDO LEVI
Projetos de Pesquisa
Unidades Organizacionais
LIM/52 - Laboratório de Virologia, Hospital das Clínicas, Faculdade de Medicina
20 resultados
Resultados de Busca
Agora exibindo 1 - 10 de 20
- PCR-RFLP assay as an option for primary HPV test(2018) GOLFETTO, L.; ALVES, E. V.; MARTINS, T. R.; SINCERO, T. C. M.; CASTRO, J. B. S.; DANNEBROCK, C.; OLIVEIRA, J. G.; LEVI, J. E.; ONOFRE, A. S. C.; BAZZO, M. L.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.
- Occult hepatitis B virus infection among blood donors from the Brazilian Amazon: implications for transfusion policy(2014) MORESCO, M. N. dos S.; VIRGOLINO, H. de A.; MORAIS, M. P. E. de; MOTTA-PASSOS, I. da; GOMES-GOUVEA, M. S.; ASSIS, L. M. S. de; AGUIAR, K. R. de L.; LOMBARDI, S. C. F.; MALHEIRO, A.; CAVALHEIRO, N. de P.; LEVI, J. E.; TORRES, K. L.Background Brazil requires the performance of both a test for hepatitis B surface antigen (HBsAg) and a test for antibodies to the core of hepatitis B for blood donor screening. Blood centres in regions of high HBV endemicity struggle to maintain adequate stocks in face of the high discard rates due to anti-HBc reactivity. We evaluated the potential infectivity of donations positive for anti-HBc in search of a rational approach for the handling of these collections. Study Design and Methods We tested anti-HBc reactive blood donations from the state of Amazonas for the presence of HBV DNA and for titres of anti-HBs. The study population consists of village-based donors from the interior of Amazonas state. Results Among 3600 donations, 799 were anti-HBc reactive (22.2%). We were able to perform real-time PCR for the HBV S gene on specimens from 291 of these donors. Eight of these samples were negative for HBsAg and positive for HBV DNA and were defined as occult B virus infections (2.7%). Six of those eight specimens had anti-HBs titres above 100 mIU/ml, indicating the concomitant presence of the virus with high antibody titres. Conclusion A small proportion of anti-HBc reactive donors carry HBV DNA and anti-HBs testing is not useful for predicting viremia on them. This finding indicates the possibility of HBV transmission from asymptomatic donors, especially in areas of high HBV prevalence. Sensitive HBV DNA nucleic acid testing may provide another level of safety, allowing eventual use of anti-HBc reactive units in critical situations.
- HPV infection-associated anogenital cyto-colpo-histological findings and molecular typing in HIV-positive women(2015) TSO, F. K.; RODRIGUES, C. L. L.; LEVI, J. E.; FERRAZ, M. G. Mattosinho de Castro; SPECK, N. M. G.; RIBALTA, J. C. L.HIV and human papillomavirus (HPV) coinfection is increasing, especially in the anal canal (AC) and cervico-vaginal regions. We identified anal epithelium abnormalities related to high-risk HPV (HR-HPV) lesions in the lower genital tracts (LGTs) of HIV-positive women, described the HPV genotypes identified, and assessed the expression of E6/E7 oncogenes in coinfected patients. Ninety-eight women were enrolled in groups combining HIV status and presence or absence of HPV in the LGT. Anal and cervical smears were collected for cytology and HR-HPV assays using Cobas(R) and/or PapilloCheck(R). Samples with highly oncogenic HPV genotypes were confirmed by NucliSENS EasyQ(R). Forty-two HIV-positive (25-52; mean age 39.5) and 56 HIV-negative (18-58; mean age 35.7) patients were included. E2 and C1 groups presented AC alterations (P = 0.002); altered images for high-resolution anoscopy were higher in E1 and C2 (P < 0.001). Of the 29 women with alterations, 41.38% were HIV-negative and 58.62% were HIV-positive (P < 0.001). HIV-positive patients accounted for 29% of the anal high-grade squamous intraepithelial lesions (P = 0.015). The Cobas(R) positive result frequency was higher in three AC groups than in the other groups. There was variation in the number of HPV types in the cervicovaginal samples among the study groups (P < 0.001). Anal cytology and anoscopy showed more altered findings in HIV-positive patients with HPV in the LGT. HR-HPV anal infections by various genotypes are common and are associated with cervical infections in HIV-positive patients. E6/E7 expression is apparently more common in the AC of HIV-positive women.
conferenceObject An autopsy-based study of Trypanosoma cruzi persistence in chronic chagasic patients(2016) BENVENUTI, L. A.; ROGGERIO, A.; CAVALCANTI, M. M.; NISHYIA, A.; LEVI, J. E.article 24 Citação(ões) na Scopus Education, tobacco smoking, alcohol consumption, and IL-2 and IL-6 gene polymorphisms in the survival of head and neck cancer(2011) LOPEZ, R. V. M.; ZAGO, M. A.; ELUF-NETO, J.; CURADO, M. P.; DAUDT, A. W.; SILVA-JUNIOR, W. A. da; ZANETTE, D. L.; LEVI, J. E.; CARVALHO, M. B. de; KOWALSKI, L. P.; ABRAHAO, M.; GOIS-FILHO, J. F. de; BOFFETTA, P.; WUENSCH-FILHO, V.The association of education, tobacco smoking, alcohol consumption, and interleukin-2 (IL-2 +114 and -384) and -6 (IL-6 -174) DNA polymorphisms with head and neck squamous cell carcinoma (HNSCC) was investigated in a cohort study of 445 subjects. IL-2 and IL-6 genotypes were determined by real-time PCR. Cox regression was used to estimate hazard ratios (HR) and 95% confidence intervals (95% CI) of disease-specific survival according to anatomical sites of the head and neck. Mean age was 56 years and most patients were males (87.6%). Subjects with 5 or more years of schooling had better survival in larynx cancer. Smoking had no effect on HNSCC survival, but alcohol consumption had a statistically significant effect on larynx cancer. IL-2 gene +114 G/T (HR = 0.52; 95% CI = 0.15-1.81) and T/T (HR = 0.22; 95% CI = 0.02-3.19) genotypes were associated with better survival in hypopharynx cancer. IL-2 + 114 G/T was a predictor of poor survival in oral cavity/oropharynx cancer and larynx cancer (HR = 1.32; 95% CI = 0.61-2.85). IL-2 -384 G/T was associated with better survival in oral cavity/oropharynx cancer (HR = 0.80; 95% CI = 0.45-1.42) and hypopharynx cancer (HR = 0.68; 95% CI = 0.21-2.20), but an inverse relationship was observed for larynx cancer. IL-6 -174 G/C was associated with better survival in hypopharynx cancer (HR = 0.68; 95% CI = 0.26-1.78) and larynx cancer (HR = 0.93; 95% CI = 0.42-2.07), and C/C reduced mortality in larynx cancer. In general, our results are similar to previous reports on the value of education, smoking, alcohol consumption, and IL-2 and IL-6 genetic polymorphisms for the prognosis of HNSCC, but the risks due to these variables are small and estimates imprecise.- HPV genotyping and p16 expression in Xingu Indigenous Park, Brazil(2016) FREITAS, V. G.; FOCCHI, G. R.; PEREIRA, E. R.; LEVI, J. E.; SPECK, N. M. G.; RIBALTA, J. C.The association between high-risk human papillomavirus (HPV) genotypes and p16 expression in indigenous women from the Xingu Indigenous Park, Brazil, was unknown. This study evaluated p16 expression in women with a histological diagnosis of cervical intraepithelial neoplasia (CIN) 3 or higher and correlated this expression with HPV genotypes to determine possible discrepancies in the expression of this marker. We evaluated 37 previously collected samples with different HPV genotypes and high-grade lesions diagnosed based on cytology, histology, and colposcopy. Immunohistochemical analysis was performed using paraffin-embedded tissue sections and the CINtec (R) Histology Kit. p16 protein expression was investigated by immunostaining with an anti-p16 antibody. HPV genotyping was performed by reverse hybridization. The age of the study population ranged from 22-75 years (43.81 +/- 15.89 years) and parity ranged from 1-11 (5.92 +/- 2.58). Thirteen different HPV genotypes were found using the INNO-LiPA kit. Single and multiple infections by HPV were found with prevalence of single infections (P = 0.029). Comparison between HPV genotype and simple or multiple infections was highly significant; it was observed more HPV 52 followed by HPV 16 in single infections (P < 0.001). p16 expression was predominantly diffuse, which was observed in 91.7% of lesions, whereas 8.3% were focal (P < 0.001). HPV 52, HPV 16 and 31 were the most prevalent HPV types in high-grade CIN in these indigenous women. Diffuse p16 expression in high-grade CIN was not influenced by the viral genotype; however, more studies are necessary to further our understanding of this restricted group.
conferenceObject Increase of Trypanosoma cruzi parasitic load in endomyocardial biopsies anticipates Chagas disease reactivation after heart transplantation(2018) BENVENUTI, L. A.; ROGGERIO, A.; NISHIYA, A. S.; LEVI, J. E.- Rapid viral metagenomics using SMART-9N amplification and nanopore sequencing(2023) CLARO, I. M.; RAMUNDO, M. S.; COLETTI, T. M.; SILVA, C. A. M. da; VALENCA, I. N.; CANDIDO, D. S.; SALES, F. C. S.; MANULI, E. R.; JESUS, J. G. de; PAULA, A. de; FELIX, A. C.; ANDRADE, P. D. S.; PINHO, M. C.; SOUZA, W. M.; AMORIM, M. R.; PROENCA-MODENA, J. L.; KALLAS, E. G.; LEVI, J. E.; FARIA, N. R.; SABINO, E. C.; LOMAN, N. J.; QUICK, J.Emerging and re-emerging viruses are a global health concern. Genome sequencing as an approach for monitoring circulating viruses is currently hampered by complex and expensive methods. Untargeted, metagenomic nanopore sequencing can provide genomic information to identify pathogens, prepare for or even prevent outbreaks. SMART (Switching Mechanism at the 5′ end of RNA Template) is a popular approach for RNA-Seq but most current methods rely on oligo-dT priming to target polyadenylated mRNA molecules. We have developed two random primed SMART-Seq approaches, a sequencing agnostic approach ‘SMART-9N’ and a version compatible rapid adapters available from Oxford Nanopore Technologies ‘Rapid SMART-9N’. The methods were developed using viral isolates, clinical samples, and compared to a gold-standard amplicon-based method. From a Zika virus isolate the SMART-9N approach recovered 10kb of the 10.8kb RNA genome in a single nanopore read. We also obtained full genome coverage at a high depth coverage using the Rapid SMART-9N, which takes only 10 minutes and costs up to 45% less than other methods. We found the limits of detection of these methods to be 6 focus forming units (FFU)/mL with 99.02% and 87.58% genome coverage for SMART-9N and Rapid SMART-9N respectively. Yellow fever virus plasma samples and SARS-CoV-2 nasopharyngeal samples previously confirmed by RT-qPCR with a broad range of Ct-values were selected for validation. Both methods produced greater genome coverage when compared to the multiplex PCR approach and we obtained the longest single read of this study (18.5 kb) with a SARS-CoV-2 clinical sample, 60% of the virus genome using the Rapid SMART-9N method. This work demonstrates that SMART-9N and Rapid SMART-9N are sensitive, low input, and long-read compatible alternatives for RNA virus detection and genome sequencing and Rapid SMART-9N improves the cost, time, and complexity of laboratory work.
- Absence of nonprimate hepacivirus-related genomes in blood donors seroreactive for hepatitis C virus displaying indeterminate blot patterns(2014) LEVI, J. E.; CABRAL, S. P. N.; NISHIYA, A.; FERREIRA, S.; ROMANO, C. M.; POLITE, M. B. C.; PEREIRA, R. A. A.; MOTA, M. A.; KUTNER, J. M.Despite intensive search, no primate homologue to the Hepatitis C Virus (HCV) has ever been found. The search for a zoonotic origin for HCV has been renewed recently when a virus, now known as non-primate hepacivirus (NPHV), with a high homology to HCV was found in dogs. A variable proportion of anti-HCV reactive blood donors submitted to the immunoblot (IB) to confirm their HCV status, present indeterminate results. The degree of homology between HCV and NPHV suggests that humans may be infected by NPHV or NPHV-like viruses. Maximum similarity between NHPV and HCV is observed in the nonstructural regions 3 and 5. Peptides representing both domains are present in IB assays, so it is reasonable to suppose that blood donors harboring such viruses may display cross-reactivity to the HCV antigenic fractions. Fifty-nine plasma samples from blood donors found reactive for anti-HCV and presenting IB indeterminate results were submitted to five distinct PCR reactions under low-stringency conditions, employing primers targeting GBV-C 5'UTR and NS3, Flavivirus-genus NS5 and NPHV 5'UTR and NS3. No amplification was obtained with all primer pairs tested except for five samples that amplified both 5'UTR and NS3 fragments from GBV-C. Unbiased next-generation sequencing may prove or rule out the existence of HCV-related viruses in IB indeterminate samples.
- Low prevalence after the first Zika virus epidemic wave in Southeastern Brazil(2018) LUNA, E.; ROMANO, C.; ARAUJO, E.; FELIX, A. C.; NAKASONE, O.; CAMPOS, S.; FERNANDES, L.; LEVI, J. E.; SANTIAGO, N.; FERNANDES, J.; FRAGOSO, D.; KALLAS, E.; PANNUTI, C.