CAMILA ALVES MAIA DA SILVA FONZAGHI

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LIM/46 - Laboratório de Parasitologia Médica, Hospital das Clínicas, Faculdade de Medicina

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  • article 9 Citação(ões) na Scopus
    Understanding Sabia virus infections (Brazilian mammarenavirus)
    (2022) NASTRI, Ana Catharina; DUARTE-NETO, Amaro Nunes; CASADIO, Luciana Vilas Boas; SOUZA, William Marciel de; CLARO, Ingra M.; MANULI, Erika R.; SELEGATTO, Gloria; SALOMA, Matias C.; FIALKOVITZ, Gabriel; TABORDA, Mariane; ALMEIDA, Bianca Leal de; MAGRI, Marcello C.; GUEDES, Ana Rubia; NETO, Laura Vieira Perdigao; SATAKI, Fatima Mitie; GUIMARAES, Thais; MENDES-CORREA, Maria Cassia; TOZETTO-MENDOZA, Tania R.; FUMAGALLI, Marcilio Jorge; HO, Yeh-Li; SILVA, Camila ALves Maia da; COLETTI, Thais M.; JESUS, Jacqueline Goes de; ROMANO, Camila M.; HILL, Sarah C.; PYBUS, Oliver; PINHO, Joao Renato Rebello; LEDESMA, Felipe Lourenco; CASAL, Yuri R.; KANAMURA, Cristina; ARAUJO, Leonardo Jose Tadeu de; FERREIRA, Camila Santos da Silva; GUERRA, Juliana Mariotti; FIGUEIREDO, Luiz Tadeu Moraes; DOLHNIKOFF, Marisa; FARIA, Nuno R.; SABINO, Ester C.; AVANCINI, Venacio; ALVES, Ferreira; LEVIN, Anna S.
    Background: Only two naturally occurring human Sabi ' a virus (SABV) infections have been reported, and those occurred over 20 years ago. Methods: We diagnosed two new cases of SABV infection using metagenomics in patients thought to have severe yellow fever and described new features of histopathological findings. Results: We characterized clinical manifestations, histopathology and analyzed possible nosocomial transmission. Patients presented with hepatitis, bleeding, neurological alterations and died. We traced twenty-nine hospital contacts and evaluated them clinically and by RT-PCR and neutralizing antibodies. Autopsies uncovered unique features on electron microscopy, such as hepatocyte ""pinewood knot"" lesions. Although previous reports with similar New-World arenavirus had nosocomial transmission, our data did not find any case in contact tracing. Conclusions: Although an apparent by rare, Brazilian mammarenavirus infection is an etiology for acute hemorrhagic fever syndrome. The two fatal cases had peculiar histopathological findings not previously described. The virological diagnosis was possible only by contemporary techniques such as metagenomic assays. We found no subsequent infections when we used serological and molecular tests to evaluate close contacts.
  • article 4 Citação(ões) na Scopus
    Identification of Leishmania species by next generation sequencing of hsp70 gene
    (2022) SOUZA, Regina Maia de; MARTINS, Roberta Cristina Ruedas; FRANCO, Lucas Augusto Moyses; TUON, Felipe Francisco; OLIVEIRA JUNIOR, Isael Gomes de; SILVA, Camila Alves Maia da; IMAMURA, Rui; AMATO, Valdir Sabbaga
    Leishmaniasis is a major public health problem worldwide. Although next generation sequencing technology has been widely used in the diagnosis of infectious diseases, it has been scarcely applied in identification of Leishmania species. The aim of this study was to compare the efficiency of MinIONTM nanopore sequencing and polymerase chain reaction restriction fragment length polymorphism in identifying Leishmania species. Our results showed that the MinIONTM sequencer was able to discriminate reference strains and clinical samples with high sensitivity in a cost and time effective manner without the prior need for culture.
  • article 16 Citação(ões) na Scopus
    First case report of monkeypox in Brazil: clinical manifestations and differential diagnosis with sexually transmitted infections
    (2022) LIMA, Evelyn Lepka de; BARRA, Luiz Alberto Costa; BORGES, Luciana Marques Sansao; MEDEIROS, Lucas Alberto; TOMISHIGE, Marcia Y. S.; SANTOS, Lucas de Souza Loureiro Abbud; SILVA, Anderson Jose Dias da; RODRIGUES, Camila Cristina Martini; AZEVEDO, Luiz Cesar Fernandes de; VILLAS-BOAS, Lucy Santos; SILVA, Camila Alves Maia da; COLETTI, Thais Moura; MANULI, Erika R.; CLARO, Ingra Morales; ROMANO, Camila Malta; RAMUNDO, Mariana Severo; MOUTINHO, Tomas; SABINO, Ester Cerdeira; LINDOSO, Jose Angelo Lauletta; FIGUEIREDO-MELLO, Claudia
    In 2022, an outbreak of monkeypox is being reported in non-endemic areas, with unusual clinical manifestations. The detailed clinical description of the first patient that received the diagnosis of monkeypox in Brazil is reported here, whose clinical manifestations can easily lead to misdiagnosis of sexually transmitted infections. A 41 years old male presented to an emergency room with a vesicular rash with eight days of evolution. He had traveled to Portugal and Spain and reported non-penetrative sexual involvement with three different male individuals. On the third day of symptoms, he sought medical care and received empirical treatment directed to sexually transmitted infections. As the symptoms did not improve, he sought medical attention at an infectious disease referral center presenting, on admission, an ulcerated penile lesion with central necrotic crusts, a disseminated pleomorphic skin rash and an oropharyngeal ulcer. The monkeypox diagnosis was suspected due to the characteristics of the lesions and the history of intimate contact with casual partners, and it was later confirmed by sequencing the almost complete monkeypox genome. The patient was hospitalized for pain control, which required opiate administration. He developed a secondary bacterial infection on the penile lesions, which were treated with oral antibiotics. He was discharged after 14 days, with lesions in process of re-epithelialization. Given the current outbreak, we must consider the possibility of monkeypox in patients with suggestive lesions, anywhere on the body (including the genitals), added to an epidemiological link or history of intimate contact with strangers or casual partners.
  • article 0 Citação(ões) na Scopus
    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.
  • article 43 Citação(ões) na Scopus
    Local Transmission of SARS-CoV-2 Lineage B.1.1.7, Brazil, December 2020
    (2021) CLARO, Ingra Morales; SALES, Flavia Cristina da Silva; RAMUNDO, Mariana Severo; CANDIDO, Darlan S.; SILVA, Camila A. M.; JESUS, Jaqueline Goes de; MANULI, Erika R.; OLIVEIRA, Cristina Mendes de; SCARPELLI, Luciano; CAMPANA, Gustavo; PYBUS, Oliver G.; SABINO, Ester Cerdeira; FARIA, Nuno Rodrigues; LEVI, Jose Eduardo
    In December 2020, research surveillance detected the B.1.1.7 lineage of severe acute respiratory syndrome coronavirus 2 in Sao Paulo, Brazil. Rapid genomic sequencing and phylogenetic analysis revealed 2 distinct introductions of the lineage. One patient reported no international travel. There may be more infections with this lineage in Brazil than reported.
  • article 879 Citação(ões) na Scopus
    Genomics and epidemiology of the P.1 SARS-CoV-2 lineage in Manaus, Brazil
    (2021) FARIA, Nuno R.; MELLAN, Thomas A.; WHITTAKER, Charles; CLARO, Ingra M.; CANDIDO, Darlan da S.; MISHRA, Swapnil; CRISPIM, Myuki A. E.; SALES, Flavia C.; HAWRYLUK, Iwona; MCCRONE, John T.; HULSWIT, Ruben J. G.; FRANCO, Lucas A. M.; RAMUNDO, Mariana S.; JESUS, Jaqueline G. de; ANDRADE, Pamela S.; COLETTI, Thais M.; FERREIRA, Giulia M.; SILVA, Camila A. M.; MANULI, Erika R.; PEREIRA, Rafael H. M.; PEIXOTO, Pedro S.; KRAEMER, Moritz U.; GABURO JR., Nelson; CAMILO, Cecilia da C.; HOELTGEBAUM, Henrique; SOUZA, William M.; ROCHA, Esmenia C.; SOUZA, Leandro M. de; PINHO, Mariana C. de; ARAUJO, Leonardo J. T.; V, Frederico S. Malta; LIMA, Aline B. de; SILVA, Joice do P.; ZAULI, Danielle A. G.; FERREIRA, Alessandro C. de S.; SCHNEKENBERG, Ricardo P.; LAYDON, Daniel J.; WALKER, Patrick G. T.; SCHLUETER, Hannah M.; SANTOS, Ana L. P. dos; VIDAL, Maria S.; CARO, Valentina S. Del; FILHO, Rosinaldo M. F.; SANTOS, Helem M. dos; AGUIAR, Renato S.; PROENCA-MODENA, Jose L. P.; NELSON, Bruce; HAY, James A.; MONOD, Melodie; MISCOURIDOU, Xenia; COUPLAND, Helen; SONABEND, Raphael; VOLLMER, Michaela; GANDY, Axel; PRETE JR., Carlos A.; NASCIMENTO, Vitor H.; SUCHARD, Marc A.; BOWDEN, Thomas A.; POND, Sergei L. K.; WU, Chieh-Hsi; RATMANN, Oliver; FERGUSON, Neil M.; DYE, Christopher; LOMAN, Nick J.; LEMEY, Philippe; RAMBAUT, Andrew; FRAIJI, Nelson A.; CARVALHO, Maria do P. S. S.; PYBUS, Oliver G.; FLAXMAN, Seth; BHATT, Samir; SABINO, Ester C.
    Cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Manaus, Brazil, resurged in late 2020 despite previously high levels of infection. Genome sequencing of viruses sampled in Manaus between November 2020 and January 2021 revealed the emergence and circulation of a novel SARS-CoV-2 variant of concern. Lineage P.1 acquired 17 mutations, including a trio in the spike protein (K417T, E484K, and N501Y) associated with increased binding to the human ACE2 (angiotensin-converting enzyme 2) receptor. Molecular clock analysis shows that P.1 emergence occurred around mid-November 2020 and was preceded by a period of faster molecular evolution. Using a two-category dynamical model that integrates genomic and mortality data, we estimate that P.1 may be 1.7- to 2.4-fold more transmissible and that previous (non-P.1) infection provides 54 to 79% of the protection against infection with P.1 that it provides against non-P.1 lineages. Enhanced global genomic surveillance of variants of concern, which may exhibit increased transmissibility and/or immune evasion, is critical to accelerate pandemic responsiveness.
  • article 5 Citação(ões) na Scopus
    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.
  • article 0 Citação(ões) na Scopus
    Case Report: Cutaneous Leishmaniasis in a Rheumatoid Arthritis Patient Receiving Methotrexate
    (2022) AMATO, Valdir Sabbaga; SOUZA, Regina Maia de; FRANCO, Lucas Augusto Moyses; MARTINS, Roberta Cristina Ruedas; SILVA, Camila Alves Maia da; EMORI, Christini Takemi; CELESTE, Beatriz Julieta; CASTANHEIRA, Gabriel Victor; TUON, Felipe Francisco
    The immunosuppressive effect of methotrexate has rarely been associated with reactivation of cutaneous leishmaniasis. Here we present a case of a cutaneous leishmaniasis patient with atypical clinical symptoms without splenomegaly but with cutaneous manifestations after treatment of rheumatoid arthritis with methotrexate and blood recovery of the parasite. Next-generation sequencing was used to identify Leishmania infantum chagasi in the patient's blood sample.
  • article 20 Citação(ões) na Scopus
    Shotgun metagenomic sequencing of the first case of monkeypox virus in Brazil, 2022
    (2022) CLARO, Ingra Morales; ROMANO, Camila Malta; CANDIDO, Darlan da Silva; LIMA, Evelyn Lepka de; LINDOSO, Jose Angelo Lauletta; RAMUNDO, Mariana Severo; MOREIRA, Filipe Romero Rebello; BARRA, Luiz Alberto Costa; BORGES, Luciana Marques Sansao; MEDEIROS, Lucas Alberto; TOMISHIGE, Marcia Y. S.; MOUTINHO, Tomas; SILVA, Anderson Jose Dias da; RODRIGUES, Camila Cristina Martini; AZEVEDO, Luiz Cesar Fernandes de; VILLAS-BOAS, Lucy Santos; SILVA, Camila Alves Maia da; COLETTI, Thais Moura; MANULI, Erika R.; O'TOOLE, Aine; QUICK, Joshua; LOMAN, Nicholas; RAMBAUT, Andrew; FARIA, Nuno R.; FIGUEIREDO-MELLO, Claudia; SABINO, Ester Cerdeira
    Monkeypox virus (MPXV), a zoonotic virus endemic to the African continent, has been reported in 33 non-endemic countries since May 2022. We report an almost complete genome of the first confirmed case of MPXV in Brazil. Shotgun metagenomic sequencing was completed in 18 hours, from DNA extraction to consensus sequence generation.
  • article 75 Citação(ões) na Scopus
    Neutralisation of SARS-CoV-2 lineage P.1 by antibodies elicited through natural SARS-CoV-2 infection or vaccination with an inactivated SARS-CoV-2 vaccine: an immunological study
    (2021) SOUZA, William M.; AMORIM, Mariene R.; SESTI-COSTA, Renata; COIMBRA, Lais D.; BRUNETTI, Natalia S.; TOLEDO-TEIXEIRA, Daniel A.; SOUZA, Gabriela F. de; MURARO, Stefanie P.; PARISE, Pierina L.; BARBOSA, Priscilla P.; BISPO-DOS-SANTOS, Karina; MOFATTO, Luciana S.; SIMEONI, Camila L.; CLARO, Ingra M.; DUARTE, Adriana S. S.; COLETTI, Thais M.; ZANGIROLAMI, Audrey B.; COSTA-LIMA, Carolina; GOMES, Arilson B. S. P.; I, Lucas Buscaratti; SALES, Flavia C.; COSTA, Vitor A.; FRANCO, Lucas A. M.; CANDIDO, Darlan S.; PYBUS, Oliver G.; JESUS, Jaqueline G. de; SILVA, Camila A. M.; RAMUNDO, Mariana S.; FERREIRA, Giulia M.; PINHO, Mariana C.; SOUZA, Leandro M.; ROCHA, Esmenia C.; ANDRADE, Pamela S.; CRISPIM, Myuki A. E.; MAKTURA, Grazielle C.; MANULI, Erika R.; SANTOS, Magnun N. N.; CAMILO, Cecilia C.; ANGERAMI, Rodrigo N.; MORETTI, Maria L.; SPILKI, Fernando R.; ARNS, Clarice W.; ADDAS-CARVALHO, Marcelo; BENITES, Bruno D.; VINOLO, Marco A. R.; MORI, Marcelo A. S.; GABURO, Nelson; DYE, Christopher; MARQUES-SOUZA, Henrique; MARQUES, Rafael E.; FARIAS, Alessandro S.; DIAMOND, Michael S.; FARIA, Nuno R.; SABINO, Ester C.; GRANJA, Fabiana; PROENCA-MODENA, Jose Luiz
    Background Mutations accrued by SARS-CoV-2 lineage P.1-first detected in Brazil in early January, 2021-include amino acid changes in the receptor-binding domain of the viral spike protein that also are reported in other variants of concern, including B.1.1.7 and B.1.351. We aimed to investigate whether isolates of wild-type P.1 lineage SARS-CoV-2 can escape from neutralising antibodies generated by a polyclonal immune response. Methods We did an immunological study to assess the neutralising effects of antibodies on lineage P.1 and lineage B isolates of SARS-CoV-2, using plasma samples from patients previously infected with or vaccinated against SARS-CoV-2. Two specimens (P.1/28 and P.1/30) containing SARS-CoV-2 lineage P.1 (as confirmed by viral genome sequencing) were obtained from nasopharyngeal and bronchoalveolar lavage samples collected from patients in Manaus, Brazil, and compared against an isolate of SARS-CoV-2 lineage B (SARS.CoV2/SP02.2020) recovered from a patient in Brazil in February, 2020. Isolates were incubated with plasma samples from 21 blood donors who had previously had COVID-19 and from a total of 53 recipients of the chemically inactivated SARS-CoV-2 vaccine CoronaVac: 18 individuals after receipt of a single dose and an additional 20 individuals (38 in total) after receipt of two doses (collected 17-38 days after the most recent dose); and 15 individuals who received two doses during the phase 3 trial of the vaccine (collected 134-230 days after the second dose). Antibody neutralisation of P.1/28, P.1/30, and B isolates by plasma samples were compared in terms of median virus neutralisation titre (VNT50, defined as the reciprocal value of the sample dilution that showed 50% protection against cytopathic effects). Findings In terms of VNT50, plasma from individuals previously infected with SARS-CoV-2 had an 8.6 times lower neutralising capacity against the P.1 isolates (median VNT50 30 [IQR <20-45] for P.1/28 and 30 [<20-40] for P.1/30) than against the lineage B isolate (260 [160-400]), with a binominal model showing significant reductions in lineage P.1 isolates compared with the lineage B isolate (p <= 0.0001). Efficient neutralisation of P.1 isolates was not seen with plasma samples collected from individuals vaccinated with a first dose of CoronaVac 20-23 days earlier (VNT(50)s below the limit of detection [<20] for most plasma samples), a second dose 17-38 days earlier (median VNT50 24 [IQR <20-25] for P.1/28 and 28 [<20-25] for P.1/30), or a second dose 134-260 days earlier (all VNT(50)s below limit of detection). Median VNT(50)s against the lineage B isolate were 20 (IQR 20-30) after a first dose of CoronaVac 20-23 days earlier, 75 (<20-263) after a second dose 17-38 days earlier, and 20 (<20-30) after a second dose 134-260 days earlier. In plasma collected 17-38 days after a second dose of CoronaVac, neutralising capacity against both P.1 isolates was significantly decreased (p=0.0051 for P.1/28 and p=0.0336 for P.1/30) compared with that against the lineage B isolate. All data were corroborated by results obtained through plaque reduction neutralisation tests. Interpretation SARS-CoV-2 lineage P.1 might escape neutralisation by antibodies generated in response to polyclonal stimulation against previously circulating variants of SARS-CoV-2. Continuous genomic surveillance of SARS-CoV-2 combined with antibody neutralisation assays could help to guide national immunisation programmes.