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LIM/46 - Laboratório de Parasitologia Médica, Hospital das Clínicas, Faculdade de Medicina
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- Phylogenetics, Epidemiology and Temporal Patterns of Dengue Virus in Araraquara, São Paulo State(2024) SOUZA, Caio Santos de; CALEIRO, Giovana Santos; CLARO, Ingra Morales; JESUS, Jaqueline Goes de; COLETTI, Thais Moura; SILVA, Camila Alves Maia da; COSTA, Angela Aparecida; INENAMI, Marta; RIBEIRO, Andreia C.; FELIX, Alvina Clara; PAULA, Anderson Vicente de; FIGUEIREDO, Walter M.; LUNA, Expedito Jose de Albuquerque; SABINO, Ester C.; ROMANO, Camila M.Dengue virus (DENV) is a prominent arbovirus with global spread, causing approximately 390 million infections each year. In Brazil, yearly epidemics follow a well-documented pattern of serotype replacement every three to four years on average. Araraquara, located in the state of Sao Paulo, has faced significant impacts from DENV epidemics since the emergence of DENV-1 in 2010. The municipality then transitioned from low to moderate endemicity in less than 10 years. Yet, there remains an insufficient understanding of virus circulation dynamics, particularly concerning DENV-1, in the region, as well as the genetic characteristics of the virus. To address this, we sequenced 37 complete or partial DENV-1 genomes sampled from 2015 to 2022 in Araraquara. Then, using also Brazilian and worldwide DENV-1 sequences we reconstructed the evolutionary history of DENV-1 in Araraquara and estimated the time to the most recent common ancestor (tMRCA) for serotype 1, for genotype V and its main lineages. Within the last ten years, there have been at least three introductions of genotype V in Araraquara, distributed in two main lineages (L Ia and L Ib, and L II). The tMRCA for the first sampled lineage (2015/2016 epidemics) was approximately 15 years ago (in 2008). Crucially, our analysis challenges existing assumptions regarding the emergence time of the DENV-1 genotypes, suggesting that genotype V might have diverged more recently than previously described. The presence of the two lineages of genotype V in the municipality might have contributed to the extended persistence of DENV-1 in the region.
- Characterization of insect-specific Culex flavivirus (Flaviviridae) nucleotide sequences in mosquitoes from urban parks in Sao Paulo, Brazil(2022) COLETTI, Thais de Moura; ROMANO, Camila Malta; URBINATTI, Paulo Roberto; TEIXEIRA, Renildo Souza; PEDROSA, Leila Weiss de Almeida; NARDI, Marcello Schiavo; NATAL, Delsio; COSTA, Antonio Charlys da; JESUS, Jaqueline Goes de; CLARO, Ingra Morales; SABINO, Ester Cerdeira; WITKIN, Steven S.; MARRELLI, Mauro Toledo; FERNANDES, Licia NatalBackground: Despite their worldwide occurrence, the distribution and role of insect-specific flaviviruses (ISFs) remain unclear. Methods: We evaluated the presence of ISFs in mosquitoes collected in Sao Paulo, Brazil, using reverse transcription and semi-nested polymerase chain reaction (PCR). Some of the positive samples were subjected to nanopore sequencing. Results: Twelve mosquito pools (2.8%) tested positive for flavivirus infection. Nanopore sequencing was successfully performed on six samples. Phylogenetic analysis grouped these sequences into genotype 2 of Culex flavivirus (CxFV). Conclusions: The identification of CxFV genotype 2 at new locations in Sao Paulo highlights the importance of understanding the role of ISFs in mosquito vector competence.
- First case of SARS-COV-2 sequencing in cerebrospinal fluid of a patient with suspected demyelinating disease(2020) DOMINGUES, Renan Barros; MENDES-CORREA, Maria Cassia; LEITE, Fernando Brunale Vilela de Moura; SABINO, Ester Cerdeira; SALARINI, Diego Zanotti; CLARO, Ingra; SANTOS, Daniel Wagner; JESUS, Jaqueline Goes de; FERREIRA, Noely Evangelista; ROMANO, Camila Malta; SOARES, Carlos Augusto SenneThe association between coronaviruses and central nervous system (CNS) demyelinating lesions has been previously shown. However, no case has been described of an association between the novel coronavirus (SARS-COV-2) and CNS demyelinating disease so far. SARS-COV-2 was previously detected in cerebrospinal fluid (CSF) sample of a patient with encephalitis. However, the virus identity was not confirmed by deep sequencing of SARS-COV-2 detected in the CSF. Here, we report a case of a patient with mild respiratory symptoms and neurological manifestations compatible with clinically isolated syndrome. The viral genome of SARS-COV-2 was detected and sequenced in CSF with 99.74-100% similarity between the patient virus and worldwide sequences. This report suggests a possible association of SARS-COV-2 infection with neurological symptoms of demyelinating disease, even in the absence of relevant upper respiratory tract infection signs.
- SARS-CoV-2 Detection and Culture in Different Biological Specimens from Immunocompetent and Immunosuppressed COVID-19 Patients Infected with Two Different Viral Strains(2023) MENDES-CORREA, Maria Cassia; SALOMAO, Matias Chiarastelli; GHILARDI, Fabio; TOZETTO-MENDOZA, Tania Regina; VILLAS-BOAS, Lucy Santos; PAULA, Anderson Vicente de; PAIAO, Heuder Gustavo Oliveira; COSTA, Antonio Charlys da; LEAL, Fabio E.; FERRAZ, Andrea de Barros Coscelli; SALES, Flavia C. S.; CLARO, Ingra M.; FERREIRA, Noely E.; PEREIRA, Geovana M.; JR, Almir Ribeiro da Silva; FREIRE, Wilton; ESPINOZA, Evelyn Patricia Sanchez; MANULI, Erika R.; ROMANO, Camila M.; JESUS, Jaqueline G. de; SABINO, Ester C.; WITKIN, Steven S.Introduction-The dynamics of SARS-CoV-2 shedding and replication in humans remain incompletely understood. Methods-We analyzed SARS-CoV-2 shedding from multiple sites in individuals with an acute COVID-19 infection by weekly sampling for five weeks in 98 immunocompetent and 25 immunosuppressed individuals. Samples and culture supernatants were tested via RT-PCR for SARS-CoV-2 to determine viral clearance rates and in vitro replication. Results-A total of 2447 clinical specimens were evaluated, including 557 nasopharyngeal swabs, 527 saliva samples, 464 urine specimens, 437 anal swabs and 462 blood samples. The SARS-CoV-2 genome sequences at each site were classified as belonging to the B.1.128 (ancestral strain) or Gamma lineage. SARS-CoV-2 detection was highest in nasopharyngeal swabs regardless of the virus strain involved or the immune status of infected individuals. The duration of viral shedding varied between clinical specimens and individual patients. Prolonged shedding of potentially infectious virus varied from 10 days up to 191 days, and primarily occurred in immunosuppressed individuals. Virus was isolated in culture from 18 nasal swab or saliva samples collected 10 or more days after onset of disease. Conclusions-Our findings indicate that persistent SARS-CoV-2 shedding may occur in both competent or immunosuppressed individuals, at multiple clinical sites and in a minority of subjects is capable of in vitro replication.
- SARS-CoV-2 reinfection caused by the P.1 lineage in Araraquara city, Sao Paulo State, Brazil(2021) ROMANO, Camila Malta; FELIX, Alvina Clara; PAULA, Anderson Vicente de; JESUS, Jaqueline Goes de; ANDRADE, Pamela S.; CANDIDO, Darlan; OLIVEIRA, Franciane M. de; RIBEIRO, Andreia C.; SILVA, Francini C. da; INEMAMI, Marta; COSTA, Angela Aparecida; LEAL, Cibele O. D.; FIGUEIREDO, Walter Manso; PANNUTI, Claudio Sergio; SOUZA, William M. de; FARIA, Nuno Rodrigues; SABINO, Ester CerdeiraReinfection by the severe acute respiratory syndrome coronavirus type 2 (SARS-COV-2) has been reported in many countries, suggesting that the virus may continue to circulate among humans despite the possibility of local herd immunity due to massive previous infections. The emergence of variants of concern (VOC) that are more transmissible than the previous circulating ones has raised particular concerns on the vaccines effectiveness and reinfection rates. The P.1 lineage was first identified in December 2020 in Manaus city and is now globally spread. We report the first case of reinfection of SARS-CoV-2 caused by the P.1 variant outside of Manaus. The potential of these new variants to escape naturally and vaccine-induced immunity highlights the need for a global vigilance.
- Prolonged presence of replication-competent SARS-CoV-2 in mildly symptomatic individuals: A report of two cases(2021) CORREA, Maria C. Mendes; LEAL, Fabio E.; BOAS, Lucy S. Villas; WITKIN, Steven S.; PAULA, Anderson de; MENDONZA, Tania R. Tozetto; FERREIRA, Noely E.; CURTY, Gislaine; CARVALHO, Pedro S. de; BUSS, Lewis F.; COSTA, Silvia F.; CARVALHO, Flavia M. da Cunha; KAWAKAMI, Joyce; TANIWAKI, Noemi N.; PAIAO, Heuder; BIZARIO, Joao C. da Silva; JESUS, Jaqueline G. de; SABINO, Ester C.; ROMANO, Camila M.; GREPAN, Regina M. Z.; SESSO, AntonioIt has been estimated that individuals with COVID-19 can shed replication-competent virus up to a maximum of 20 days after initiation of symptoms. The majority of studies that addressed this situation involved hospitalized individuals and those with severe disease. Studies to address the possible presence of SARS-CoV-2 during the different phases of COVID-19 disease in mildly infected individuals, and utilization of viral culture techniques to identify replication-competent viruses, have been limited. This report describes two patients with mild forms of the disease who shed replication-competent virus for 24 and 37 days, respectively, after symptom onset.
- Genomic detection of a virus lineage replacement event of dengue virus serotype 2 in Brazil, 2019(2020) JESUS, Jaqueline Goes de; DUTRA, Karina Rocha; SALES, Flavia Cristina da Silva; CLARO, Ingra Morales; TERZIAN, Ana Carolina; CANDIDO, Darlan da Silva; HILL, Sarah C.; THEZE, Julien; TORRES, Celeste; D'AGOSTINI, Tatiana Lang; FELIX, Alvina Clara; REIS, Andreia F. Negri; ALCANTAR, Luiz Carlos Junior; ABREU, Andre L. de; CRODA, Julio H. R.; OLIVEIRA, Wanderson K. de; FILIPIS, Ana Maria Bispo de; CAMIS, Maria do Carmo Rodrigues dos Santos; ROMANO, Camila Malta; LOMAN, Nick J.; PYBUS, Oliver G.; SABINO, Ester Cerdeira; NOGUEIRA, Mauricio L.; FARIA, Nuno RodriguesBACKGROUND Despite efforts to mitigate the impact of dengue virus (DENV) epidemics, the virus remains a public health problem in tropical and subtropical regions around the world. Most DENV cases in the Americas between January and July 2019 were reported in Brazil. Sao Paulo State in the southeast of Brazil has reported nearly half of all DENV infections in the country. OBJECTIVES To understand the origin and dynamics of the 2019 DENV outbreak. METHODS Here using portable nanopore sequencing we generated20 new DENV genome sequences from viremic patients with suspected dengue infection residing in two of the most-affected municipalities of Sao Paulo State, Araraquara and Sao Jose do Rio Preto. We conducted a comprehensive phylogenetic analysis with 1,630 global DENV strains to better understand the evolutionary history of the DENV lineages that currently circulate in the region. FINDINGS The new outbreak strains were classified as DENV2 genotype III (American/Asian genotype). Our analysis shows that the 2019 outbreak is the result of a novel DENV lineage that was recently introduced to Brazil from the Caribbean region. Dating phylogeographic analysis suggests that DENV2-III BR-4 was introduced to Brazil in or around early 2014, possibly from the Caribbean region. MAIN CONCLUSIONS Our study describes the early detection of a newly introduced and rapidly-expanding DENV2 virus lineage in Brazil.