JOSE EDUARDO LEVI
Projetos de Pesquisa
Unidades Organizacionais
LIM/52 - Laboratório de Virologia, Hospital das Clínicas, Faculdade de Medicina
5 resultados
Resultados de Busca
Agora exibindo 1 - 5 de 5
- Comparison of four different human papillomavirus genotyping methods in cervical samples: Addressing method-specific advantages and limitations(2024) SIQUEIRA, Juliana D.; ALVES, Brunna M.; BRANCO, Adriana B. C. Castelo; DUQUE, Kristiane C. D.; BUSTAMANTE-TEIXEIRA, Maria Teresa; SOARES, Esmeralda A.; LEVI, Jose Eduardo; SILVA, Gulnar Azevedo e; SOARES, Marcelo A.Since human papillomavirus (HPV) is recognized as the causative agent of cervical cancer and associated with anogenital non-cervical and oropharyngeal cancers, the characterization of the HPV types circulating in different geographic regions is an important tool in screening and prevention. In this context, this study compared four methodologies for HPV detection and genotyping: real-time PCR (Cobas (R) HPV test), nested PCR followed by conventional Sanger sequencing, reverse hybridization (High + Low PapillomaStrip (R) kit) and next-generation sequencing (NGS) at an Illumina HiSeq2500 platform. Cervical samples from patients followed at the Family Health Strategy from Juiz de Fora, Minas Gerais, Brazil, were collected and subjected to the real-time PCR. Of those, 114 were included in this study according to the results obtained with the real-time PCR, considered herein as the gold standard method. For the 110 samples tested by at least one methodology in addition to real-time PCR, NGS showed the lowest concordance rates of HPV and high-risk HPV identification compared to the other three methods (67-75 %). Real-time PCR and Sanger sequencing showed the highest rates of concordance (97-100 %). All methods differed in their sensitivity and specificity. HPV genotyping contributes to individual risk stratification, therapeutic decisions, epidemiological studies and vaccine development, supporting approaches in prevention, healthcare and management of HPV infection.
- Prevention of multiple whole blood donations by an individual at the same month through the creation of a national Deferred Donor Registry (DDR)(2023) BERMUDEZ-FORERO, Maria-Isabel; ANZOLA-SAMUDIO, Diego-Alexander; LEVI, Jose-Eduardo; GARCIA-OTALORA, Michel-AndresIntroduction: The Colombian National Institute of Health administers the National Information System of Haemovigilance (SIHEVI-INS). Today, SIHEVI-INS constitutes a national blood donor and recipient database, which contains a national deferred donor registry (DDR), allowing blood banks to take acceptance or rejection decisions of a potential donor in real time. The study aimed to determine the rate of people who have made more than one whole blood donation monthly in Colombia, violating the national guideline of intervals between donations (three months for men and four for women), since DDR implementation.Methods: We detected the unique personal identification number of people who, in 30 calendar days, made more than one whole blood donation at any of the 83 blood banks set up in Colombia. There were three comparison periods: 01/01/2018-08/31/2019 (launch of SIHEVI-INS and first national feedback); 09/01/2019-12/31/2020 (second feedback) and 01/01/2021-09/30/2022 (massive incorporation of web services).Results: For the first period, blood banks accepted 18.0 donations per 1000 people. There was a rate of 28.8 people/10,000 donations who had donated whole blood twice within 30 days. In the second period, there were 17.0 donations/1000 people and a rate of 2.1 people/10,000 donations (OR:14.0 CI95 %:12.2-16.0). For the last period, there were 18.2 donations/1000 people and a rate of 0.9 individuals/10,000 donations (OR:31.3 CI95 %:26.6-36.9, p < 0.001).Conclusion: DDR reduced by 31 times the acceptance of blood donors who made more than one whole blood donation in the same month. It was necessary to provide periodic feedback and promote web service implementation to reduce this risky behavior.
- 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.
- Early Emergence and Dispersal of Delta SARS-CoV-2 Lineage AY.99.2 in Brazil(2022) ROMANO, Camila Malta; OLIVEIRA, Cristina Mendes de; SILVA, Luciane Sussuchi da; LEVI, Jose EduardoThe year of 2021 was marked by the emergence and dispersal of a number of SARS-CoV-2 lineages, resulting in the ""third wave"" of COVID-19 in several countries despite the level of vaccine coverage. Soon after the first confirmed cases of COVID-19 by the Delta variant in Brazil, at least seven Delta sub-lineages emerged, including the globally spread AY.101 and AY.99.2. In this study we performed a detailed analysis of the COVID-19 scenario in Brazil from April to December 2021 by using data collected by the largest private medical diagnostic company in Latin America (Dasa), and SARS-CoV-2 genomic sequences generated by its SARS-CoV-2 genomic surveillance project (GENOV). For phylogenetic and Bayesian analysis, SARS-CoV-2 genomes available at GISAID public database were also retrieved. We confirmed that the Brazilian AY.99.2 and AY.101 were the most prevalent lineages during this period, overpassing the Gamma variant in July/August. We also estimated that AY.99.2 likely emerged a few weeks after the entry of the B.1.617.2 in the country, at some point between late April and May and rapidly spread to other countries. Despite no increased fitness described for the AY.99.2 lineage, a rapid shift in the composition of Delta SARS-CoV-2 lineages prevalence in Brazil took place. Understanding the reasons leading the AY.99.2 to become the dominant lineage in the country is important to understand the process of lineage competitions that may inform future control measures.
- Characterization of hepatitis C virus in chronic hepatitis patients: genotypes in the State of Amazonas, Brazil(2011) ARAUJO, Ana Ruth; ALMEIDA, Carlos Maurico de; FRAPORTI, Liziara; GARCIA, Nadja; LIMA, Tatiane Amabili de; MAIA, Laura Patricia Viana; TORRES, Katia Luz; TARRAGO, Andrea Monteiro; VICTORIA, Flamir; VICTORIA, Marilu; TATENO, Adriana; LEVI, Jose Eduardo; TALHARI, Sinesio; MALHEIRO, AdrianaIntroduction: In the State of Amazonas, data regarding the prevalence of different genotypes of hepatitis C virus remains scarce. Methods: The genotype of 69 HCV positive patients was determined. An in-house standardized nested-PCR was used to detect HCV RNA. Genotype assignment was based on type-specific motifs on the sequenced amplicons delimited by primers HC11/HC18 from the 5' untranslated region. Results: Of the 69 patients studied, 65.2% were male and 34.8% were female. Genotype 1 showed the greatest prevalence, followed by 3 and 2. Conclusions: These data suggesting that Manaus is the point of arrival of HCV in the State of Amazonas.