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Thalita Souza Arantes; Rodrigo Araújo Lima Rodrigues; Ludmila Karen Dos Santos Silva; Graziele Pereira Oliveira; Helton Luís de Souza; Jacques Y. B. Khalil; Danilo Bretas de Oliveira; Alice Abreu Torres; Luis Lamberti da Silva; Philippe Colson; Erna Geessien Kroon; Flávio Guimarães da Fonseca; Cláudio Antônio Bonjardim; Bernard La Scola; Jônatas Santos Abrahão. Correction for Arantes et al., “The Large Marseillevirus Explores Different Entry Pathways by Forming Giant Infectious Vesicles”. Journal of Virology 2021, 95, 1 .
AMA StyleThalita Souza Arantes, Rodrigo Araújo Lima Rodrigues, Ludmila Karen Dos Santos Silva, Graziele Pereira Oliveira, Helton Luís de Souza, Jacques Y. B. Khalil, Danilo Bretas de Oliveira, Alice Abreu Torres, Luis Lamberti da Silva, Philippe Colson, Erna Geessien Kroon, Flávio Guimarães da Fonseca, Cláudio Antônio Bonjardim, Bernard La Scola, Jônatas Santos Abrahão. Correction for Arantes et al., “The Large Marseillevirus Explores Different Entry Pathways by Forming Giant Infectious Vesicles”. Journal of Virology. 2021; 95 (18):1.
Chicago/Turabian StyleThalita Souza Arantes; Rodrigo Araújo Lima Rodrigues; Ludmila Karen Dos Santos Silva; Graziele Pereira Oliveira; Helton Luís de Souza; Jacques Y. B. Khalil; Danilo Bretas de Oliveira; Alice Abreu Torres; Luis Lamberti da Silva; Philippe Colson; Erna Geessien Kroon; Flávio Guimarães da Fonseca; Cláudio Antônio Bonjardim; Bernard La Scola; Jônatas Santos Abrahão. 2021. "Correction for Arantes et al., “The Large Marseillevirus Explores Different Entry Pathways by Forming Giant Infectious Vesicles”." Journal of Virology 95, no. 18: 1.
Genetic variants of SARS-CoV-2 have been emerging and circulating in many places across the world. Rapid detection of these variants is essential since their dissemination can impact transmission rates, diagnostic procedures, disease severity, response to vaccines or patient management. Sanger sequencing has been used as the preferred approach for variant detection among circulating human immunodeficiency and measles virus genotypes. Using primers to amplify a fragment of the SARS-CoV-2 genome encoding part of the Spike protein, we showed that Sanger sequencing allowed us to rapidly detect the introduction and spread of three distinct SARS-CoV-2 variants in two major Brazilian cities. In both cities, after the predominance of variants closely related to the virus first identified in China, the emergence of the P.2 variant was quickly followed by the identification of the P1 variant, which became dominant in less than one month after it was first detected.
Erick Gustavo Dorlass; Karine Lima Lourenco; Rubens Daniel Miserani Magalhaes; Hugo Sato; Alex Fiorini; Renata Peixoto; Helena Perez Coelho; Bruna Larotonda Telezynski; Guilherme Pereira Scagion; Tatiana Ometto; Luciano Matsumiya Thomazelli; Danielle Bruna Leal Oliveira; Ana Paula Fernandes; Edison Luiz Durigon; Flavio Guimaraes da Fonseca; Santuza Maria Ribeiro Teixeira. Survey of SARS-CoV-2 genetic diversity in two major Brazilian cities using a fast and affordable Sanger sequencing strategy. 2021, 1 .
AMA StyleErick Gustavo Dorlass, Karine Lima Lourenco, Rubens Daniel Miserani Magalhaes, Hugo Sato, Alex Fiorini, Renata Peixoto, Helena Perez Coelho, Bruna Larotonda Telezynski, Guilherme Pereira Scagion, Tatiana Ometto, Luciano Matsumiya Thomazelli, Danielle Bruna Leal Oliveira, Ana Paula Fernandes, Edison Luiz Durigon, Flavio Guimaraes da Fonseca, Santuza Maria Ribeiro Teixeira. Survey of SARS-CoV-2 genetic diversity in two major Brazilian cities using a fast and affordable Sanger sequencing strategy. . 2021; ():1.
Chicago/Turabian StyleErick Gustavo Dorlass; Karine Lima Lourenco; Rubens Daniel Miserani Magalhaes; Hugo Sato; Alex Fiorini; Renata Peixoto; Helena Perez Coelho; Bruna Larotonda Telezynski; Guilherme Pereira Scagion; Tatiana Ometto; Luciano Matsumiya Thomazelli; Danielle Bruna Leal Oliveira; Ana Paula Fernandes; Edison Luiz Durigon; Flavio Guimaraes da Fonseca; Santuza Maria Ribeiro Teixeira. 2021. "Survey of SARS-CoV-2 genetic diversity in two major Brazilian cities using a fast and affordable Sanger sequencing strategy." , no. : 1.
There is a massive demand to identify alternative methods to detect new cases of COVID-19 as well as to investigate the epidemiology of the disease. In many countries, importation of commercial kits poses a significant impact on their testing capacity and increases the costs for the public health system. We have developed an ELISA to detect IgG antibodies against SARS-CoV-2 using a recombinant viral nucleocapsid (rN) protein expressed in E. coli. Using a total of 894 clinical samples we showed that the rN-ELISA was able to detect IgG antibodies against SARS-CoV-2 with high sen-sitivity (97.5%) and specificity (96.3%) when compared to a commercial antibody test. After three external validation studies, we showed that the test accuracy was higher than 90%. The rN-ELISA IgG kit constitutes a convenient and specific method for the large-scale determination of SARS-Cov-2 antibodies in human sera with high reliability.
Flávia F. Bagno; Sarah A. R. Sergio; Maria Marta Figueiredo; Lara C. Godoi; Luis Adan F Andrade; Natália C. Salazar; Camila P. Soares; Andressa Aguiar; Flavia Jaqueline Almeida; Edimilson D. da Silva; Antonio G. P. Ferreira; Edison Luiz Durigon; Ricardo T. Gazzinelli; Santuza M. R. Teixeira; Ana Paula S. M. Fernandes; Flavio G. da Fonseca. DEVELOPMENT AND VALIDATION OF AN ENZYME-LINKED IMMUNOASSAY KIT FOR DIAGNOSIS AND SURVEILLANCE OF COVID-19. 2021, 1 .
AMA StyleFlávia F. Bagno, Sarah A. R. Sergio, Maria Marta Figueiredo, Lara C. Godoi, Luis Adan F Andrade, Natália C. Salazar, Camila P. Soares, Andressa Aguiar, Flavia Jaqueline Almeida, Edimilson D. da Silva, Antonio G. P. Ferreira, Edison Luiz Durigon, Ricardo T. Gazzinelli, Santuza M. R. Teixeira, Ana Paula S. M. Fernandes, Flavio G. da Fonseca. DEVELOPMENT AND VALIDATION OF AN ENZYME-LINKED IMMUNOASSAY KIT FOR DIAGNOSIS AND SURVEILLANCE OF COVID-19. . 2021; ():1.
Chicago/Turabian StyleFlávia F. Bagno; Sarah A. R. Sergio; Maria Marta Figueiredo; Lara C. Godoi; Luis Adan F Andrade; Natália C. Salazar; Camila P. Soares; Andressa Aguiar; Flavia Jaqueline Almeida; Edimilson D. da Silva; Antonio G. P. Ferreira; Edison Luiz Durigon; Ricardo T. Gazzinelli; Santuza M. R. Teixeira; Ana Paula S. M. Fernandes; Flavio G. da Fonseca. 2021. "DEVELOPMENT AND VALIDATION OF AN ENZYME-LINKED IMMUNOASSAY KIT FOR DIAGNOSIS AND SURVEILLANCE OF COVID-19." , no. : 1.
Background The vaccinia virus (VACV) isolates, Guarani P1 virus (GP1V) and Passatempo virus (PSTV), were isolated during zoonotic outbreaks in Brazil. Each one of them belongs to two different VACV clades, defined by biological aspects that include virulence in mice and phylogenetic analysis. Considering that information about how vaccinia viruses from different groups elicit immune responses in animals is scarce, we investigated such responses in mice infected either by GP1V (group 2) or PSTV (group 1), using VACV Western Reserve strain (VACV-WR) as control. Methods The severity of the infections was evaluated in BALB/c mice considering diverse clinical signs and defined scores, and the immune responses triggered by GP1V and PSTV infections were analysed by immune cell phenotyping and intra-cytoplasmic cytokines detection. Results We detected a reduction in total lymphocytes (CD3 +), macrophages (CD14 +), and NK cells (CD3-CD49 +) in animals infected with VACV-WR or GP1V. The VACV-WR and GP1V viruses, belonging to the most virulent group in a murine model, were able to down-modulate the cell immune responses upon mice infection. In contrast, PSTV, a virus considered less virulent in a murine model, showed little ability to down-modulate the mice immune responses. Mice infected with VACV-WR and GP1V viruses presented significant weight loss and developed lesions in their spleens, as well as damage to liver and lungs whereas mice infected with PSTV developed only moderate clinical signs. Conclusions Our results suggest that VACV immunomodulation in vivo is clade-related and is proportional to the strain’s virulence upon infection. Our data corroborate the classification of the different Brazilian VACV isolates into clades 1 and 2, taking into account not only phylogenetic criteria, but also clinical and immunological data.
Karine Lima Lourenço; Leandro Andrade Chinália; Lethícia Ribeiro Henriques; Rodrigo Araújo Lima Rodrigues; Flávio Guimarães da Fonseca. Zoonotic vaccinia virus strains belonging to different genetic clades exhibit immunomodulation abilities that are proportional to their virulence. Virology Journal 2021, 18, 1 -12.
AMA StyleKarine Lima Lourenço, Leandro Andrade Chinália, Lethícia Ribeiro Henriques, Rodrigo Araújo Lima Rodrigues, Flávio Guimarães da Fonseca. Zoonotic vaccinia virus strains belonging to different genetic clades exhibit immunomodulation abilities that are proportional to their virulence. Virology Journal. 2021; 18 (1):1-12.
Chicago/Turabian StyleKarine Lima Lourenço; Leandro Andrade Chinália; Lethícia Ribeiro Henriques; Rodrigo Araújo Lima Rodrigues; Flávio Guimarães da Fonseca. 2021. "Zoonotic vaccinia virus strains belonging to different genetic clades exhibit immunomodulation abilities that are proportional to their virulence." Virology Journal 18, no. 1: 1-12.
An alarming disease caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) named COVID-19 has emerged as an unprecedented public health problem and ignited a world health crisis. As opposed to what was believed at the beginning of the pandemic, the virus has not only spread but persevere causing secondary waves and challenging the concept of herd immunity against viral infections. While the majority of SARS-CoV-2-infected individuals may remain asymptomatic, a fraction of individuals may develop low to high-grade severity signs and symptoms of COVID-19. The disease is multifactorial and can progress quickly, leading to severe complications and even death in a few days. Therefore, understanding the pre-existing factors for disease development has never been so pressing. In this scenario, the insights on the mechanisms underlying disease allied to the immune response developed during the viral invasion could shed light on novel predictive factors and prognostic tools for COVID-19 management and interventions. A recent genome-wide association study (GWAS) revealed several molecules that significantly impacted critically ill COVID-19 patients, leading to the core mechanisms of COVID-19 pathogenesis. Considering these findings and the fact that ACE-2 polymorphisms alone cannot explain disease progress and severity, this review aims at summarizing the most important and recent findings of the research and expert consensus of possible cytokine-related polymorphisms existing in the differential expression of paramount immune molecules that could be crucial for providing guidelines for decision-making and appropriate clinical management of COVID-19.
Adriana Alves Oliveira Paim; Ágata Lopes-Ribeiro; Daniele S.O. Daian e Silva; Luis Adan F. Andrade; Thais F.S. Moraes; Edel F. Barbosa-Stancioli; Flávio Guimarães da Fonseca; Jordana G. Coelho-Dos-Reis. Will a little change do you good? A putative role of polymorphisms in COVID-19. Immunology Letters 2021, 235, 9 -14.
AMA StyleAdriana Alves Oliveira Paim, Ágata Lopes-Ribeiro, Daniele S.O. Daian e Silva, Luis Adan F. Andrade, Thais F.S. Moraes, Edel F. Barbosa-Stancioli, Flávio Guimarães da Fonseca, Jordana G. Coelho-Dos-Reis. Will a little change do you good? A putative role of polymorphisms in COVID-19. Immunology Letters. 2021; 235 ():9-14.
Chicago/Turabian StyleAdriana Alves Oliveira Paim; Ágata Lopes-Ribeiro; Daniele S.O. Daian e Silva; Luis Adan F. Andrade; Thais F.S. Moraes; Edel F. Barbosa-Stancioli; Flávio Guimarães da Fonseca; Jordana G. Coelho-Dos-Reis. 2021. "Will a little change do you good? A putative role of polymorphisms in COVID-19." Immunology Letters 235, no. : 9-14.
Accurate testing to detect SARS-CoV-2 RNA is key to counteract the virus spread. Nonetheless, the number of diagnostic laboratories able to perform qPCR tests is limited, particularly in developing countries. We describe the use of a virus-inactivating, denaturing solution (DS) to decrease virus infectivity in clinical specimens without affecting RNA integrity. Swab samples were collected from infected patients and from laboratory personnel using a commercially available viral transport solution and the in-house DS. Samples were tested by RT-qPCR, and exposure to infective viruses was also accessed by ELISA. The DS used did not interfere with viral genome detection and was able to maintain RNA integrity for up to 16 days at room temperature. Furthermore, virus loaded onto DS were inactivated, as attested by attempts to grow SARS-CoV-2 in cell monolayers after DS desalt filtration to remove toxic residues. The DS described here provides a strategy to maintain diagnostic accuracy and protects diagnostic laboratory personnel from accidental infection, as it has helped to protect our lab crew.
Alex F. Carvalho; Raissa P. Rocha; Andreza P. Gonçalves; Thaís B. S. Silva; Hugo I. Sato; Larissa Vuitika; Flavia F. Bagno; Sarah A. R. Sérgio; Maria M. Figueiredo; Ronaldo B. Martins; Juliano P. Souza; Eurico Arruda; Ana P. S. M. Fernandes; Pedro A. Alves; Santuza M. R. Teixeira; Flavio G. da Fonseca. The use of denaturing solution as collection and transport media to improve SARS-CoV-2 RNA detection and reduce infection of laboratory personnel. Brazilian Journal of Microbiology 2021, 52, 531 -539.
AMA StyleAlex F. Carvalho, Raissa P. Rocha, Andreza P. Gonçalves, Thaís B. S. Silva, Hugo I. Sato, Larissa Vuitika, Flavia F. Bagno, Sarah A. R. Sérgio, Maria M. Figueiredo, Ronaldo B. Martins, Juliano P. Souza, Eurico Arruda, Ana P. S. M. Fernandes, Pedro A. Alves, Santuza M. R. Teixeira, Flavio G. da Fonseca. The use of denaturing solution as collection and transport media to improve SARS-CoV-2 RNA detection and reduce infection of laboratory personnel. Brazilian Journal of Microbiology. 2021; 52 (2):531-539.
Chicago/Turabian StyleAlex F. Carvalho; Raissa P. Rocha; Andreza P. Gonçalves; Thaís B. S. Silva; Hugo I. Sato; Larissa Vuitika; Flavia F. Bagno; Sarah A. R. Sérgio; Maria M. Figueiredo; Ronaldo B. Martins; Juliano P. Souza; Eurico Arruda; Ana P. S. M. Fernandes; Pedro A. Alves; Santuza M. R. Teixeira; Flavio G. da Fonseca. 2021. "The use of denaturing solution as collection and transport media to improve SARS-CoV-2 RNA detection and reduce infection of laboratory personnel." Brazilian Journal of Microbiology 52, no. 2: 531-539.
Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) to detect SARS-CoV-2 RNA is an essential test to monitor the occurrence of COVID-19. A methodology is proposed for the determination of maximum pool size and adjustments of cut-off values of cycle threshold (
Murilo Costa; Hugo Sato; Raissa Rocha; Alex Carvalho; Nathalia Guimarães; Elaine Machado; Claudia Alves; Santuza Teixeira; Ricardo Takahashi; Unaí Tupinambás; Flávio da Fonseca. Adjusting the Cut-Off and Maximum Pool Size in RT-qPCR Pool Testing for SARS-CoV-2. Viruses 2021, 13, 557 .
AMA StyleMurilo Costa, Hugo Sato, Raissa Rocha, Alex Carvalho, Nathalia Guimarães, Elaine Machado, Claudia Alves, Santuza Teixeira, Ricardo Takahashi, Unaí Tupinambás, Flávio da Fonseca. Adjusting the Cut-Off and Maximum Pool Size in RT-qPCR Pool Testing for SARS-CoV-2. Viruses. 2021; 13 (4):557.
Chicago/Turabian StyleMurilo Costa; Hugo Sato; Raissa Rocha; Alex Carvalho; Nathalia Guimarães; Elaine Machado; Claudia Alves; Santuza Teixeira; Ricardo Takahashi; Unaí Tupinambás; Flávio da Fonseca. 2021. "Adjusting the Cut-Off and Maximum Pool Size in RT-qPCR Pool Testing for SARS-CoV-2." Viruses 13, no. 4: 557.
The year 2020 was profoundly marked by the emergence and spread of SARS-CoV-2, causing COVID-19, which represents the greatest pandemic of the 21st century until now, and a major challenge for virologists in the scientific and medical communities. Increased numbers of SARS-CoV-2 infection all over the world imposed social and travel restrictions, including avoidance of face-to-face scientific meetings. Therefore, for the first time in history, the 2020 edition of the Brazilian Society of Virology (SBV) congress was totally online. Despite the challenge of the new format, the Brazilian society board and collaborators were successful in virtually congregating more than 921 attendees, which was the greatest SBV participant number ever reached. Seminal talks from prominent national and international researchers were presented every night, during a week, and included discussions about environmental, basic, animal, human, plant and invertebrate virology. A special roundtable debated exclusively new data and perspectives regarding COVID-19 by some of the greatest Brazilian virologists. Women scientists were very well represented in another special roundtable called “Young Women Inspiring Research”, which was one of the most viewed and commented section during the meeting, given the extraordinary quality of the presented work. Finally, SBV offered the Helio Gelli Pereira award for one graduate and one undergraduate student, which has also been a fruitful collaboration between the society and Viruses journal. The annual SBV meeting has, therefore, reached its goals to inspire young scientists, stimulate high-quality scientific discussion and to encourage global collaboration between virologists.
Luciana Arruda; Fabrício Campos; Jônatas Abrahão; Flávio da Fonseca; João Araújo Junior; Fernando Rosado Spilki. 31st Brazilian Online Society for Virology (SBV) 2020 Annual Meeting. Viruses 2021, 13, 414 .
AMA StyleLuciana Arruda, Fabrício Campos, Jônatas Abrahão, Flávio da Fonseca, João Araújo Junior, Fernando Rosado Spilki. 31st Brazilian Online Society for Virology (SBV) 2020 Annual Meeting. Viruses. 2021; 13 (3):414.
Chicago/Turabian StyleLuciana Arruda; Fabrício Campos; Jônatas Abrahão; Flávio da Fonseca; João Araújo Junior; Fernando Rosado Spilki. 2021. "31st Brazilian Online Society for Virology (SBV) 2020 Annual Meeting." Viruses 13, no. 3: 414.
BackgroundThe Vaccinia virus (VACV) isolates, Guarani P1 virus (GP1V) and Passatempo virus (PSTV), were isolated from zoonotic outbreaks in Brazil and belong to two different VACV clades, as defined by biological aspects that include virulence in mice and phylogenetic analysis. Considering that information about how vaccinia viruses from different groups elicit immune responses in animals is scarce, we investigated such responses in mice infected by GP1V (group 2) or PSTV (group 1) using VACV Western Reserve strain (WR) as control. MethodsThe severity of the infections was evaluated in BALB/c mice considering diverse clinical signs and defined scores, and the immune responses triggered by GP1V and PSTV infections were analysed by immune cell phenotyping and intra-cytoplasmic cytokines detection. ResultsInfected mice showed significant weight loss and developed spleen lesions as well as liver and lung damage. Mice infected with PSTV, however, developed only moderate clinical signs. We detected a reduction of total lymphocytes (CD3+), macrophages (CD14+) and NK cells (CD3-CD49+) in animals infected with VACV-WR or GP1V. VACV-WR was able to significantly downmodulate cell immune responses upon mice infection, and GP1V-infected animals also showed intense downmodulation in cell responses. Contrarily, PSTV presented little ability to downmodulate mice immune responses. ConclusionsOur results suggest that VACV immunomodulation in vivo is clade-related and is proportional to the strain virulence upon infection. Our data corroborate the classification of the different Brazilian VACV isolates in clades 1 and 2, taking into account not only phylogenetic criteria, but also clinical and immunological data.
Karine Lima Lourenço; Leandro Andrade Chinália; Lethícia Rodrigues Henriques; Rodrigo Araújo Lima Rodrigues; Flávio Guimarães da Fonseca. Zoonotic Vaccinia Viruses Belonging to Different Genetic Clades Exhibit Immunomodulation Abilities That Are Proportional to Their Pathogenic Potential. 2021, 1 .
AMA StyleKarine Lima Lourenço, Leandro Andrade Chinália, Lethícia Rodrigues Henriques, Rodrigo Araújo Lima Rodrigues, Flávio Guimarães da Fonseca. Zoonotic Vaccinia Viruses Belonging to Different Genetic Clades Exhibit Immunomodulation Abilities That Are Proportional to Their Pathogenic Potential. . 2021; ():1.
Chicago/Turabian StyleKarine Lima Lourenço; Leandro Andrade Chinália; Lethícia Rodrigues Henriques; Rodrigo Araújo Lima Rodrigues; Flávio Guimarães da Fonseca. 2021. "Zoonotic Vaccinia Viruses Belonging to Different Genetic Clades Exhibit Immunomodulation Abilities That Are Proportional to Their Pathogenic Potential." , no. : 1.
This Special Issue of Viruses is a collection of the current knowledge on a broad range of emerging human, animal, and plant viral diseases
Fabrício Campos; Luciana de Arruda; Flávio da Fonseca. Special Issue “Emerging Viruses 2020: Surveillance, Prevention, Evolution and Control”. Viruses 2021, 13, 251 .
AMA StyleFabrício Campos, Luciana de Arruda, Flávio da Fonseca. Special Issue “Emerging Viruses 2020: Surveillance, Prevention, Evolution and Control”. Viruses. 2021; 13 (2):251.
Chicago/Turabian StyleFabrício Campos; Luciana de Arruda; Flávio da Fonseca. 2021. "Special Issue “Emerging Viruses 2020: Surveillance, Prevention, Evolution and Control”." Viruses 13, no. 2: 251.
By analysing the evolution of the COVID‐19 epidemic in the state of Minas Gerais, Brazil, we showed the importance of considering the sub‐notification not only of deaths but also of infected cases. It was shown that the largely used criteria of a historical all‐deaths baseline is not approachable in this case, where most of the deaths are associated with causes that should decrease due to social distancing and reduction of economic activities. A quite simple and intuitive model based on the Gompertz function was applied to estimate excess deaths and excess of infected cases. It fits well the data and predicts the evolution of the epidemic adequately. Based on these analyses, an excess of 21.638 deaths and 557.216 infected cases is predicted until the end of 2020, with an upper bound of the case fatality rate of around 2.4 % and a prevalence of 2.6 %. The geographical distribution of cases and deaths and its ethnic correlation are also presented. This study points out the necessity of governmental and private organizations working together to improve public awareness and stimulate social distancing to curb the viral infection, especially in critical places with high poverty.
Paulo H. R. Amaral; Lidia M. Andrade; Flavio G. Da Fonseca; Juan Carlos González Perez. Impact of COVID‐19 in Minas Gerais, Brazil: Excess deaths, sub‐notified cases, geographic and ethnic distribution. Transboundary and Emerging Diseases 2020, 1 .
AMA StylePaulo H. R. Amaral, Lidia M. Andrade, Flavio G. Da Fonseca, Juan Carlos González Perez. Impact of COVID‐19 in Minas Gerais, Brazil: Excess deaths, sub‐notified cases, geographic and ethnic distribution. Transboundary and Emerging Diseases. 2020; ():1.
Chicago/Turabian StylePaulo H. R. Amaral; Lidia M. Andrade; Flavio G. Da Fonseca; Juan Carlos González Perez. 2020. "Impact of COVID‐19 in Minas Gerais, Brazil: Excess deaths, sub‐notified cases, geographic and ethnic distribution." Transboundary and Emerging Diseases , no. : 1.
Chikungunya virus (CHIKV) is a mosquito-borne pathogen that causes a disease characterized by the acute onset of fever accompanied by arthralgia and intense joint pain. Clinical similarities and cocirculation of this and other arboviruses in many tropical countries highlight the necessity for efficient and accessible diagnostic tools. CHIKV envelope proteins are highly conserved among alphaviruses and, particularly, the envelope 2 glycoprotein (CHIKV-E2) appears to be immunodominant and has a considerable serodiagnosis potential. Here, we investigate how glycosylation of CHIKV-E2 affects antigen/antibody interaction and how this affects the performance of CHIKV-E2-based Indirect ELISA tests. We compare two CHIKV-E2 recombinant antigens produced in different expression systems: prokaryotic-versus eukaryotic-made recombinant proteins. CHIKV-E2 antigens are expressed either in E. coli BL21(DE3)—a prokaryotic system unable to produce post-translational modifications—or in HEK-293T mammalian cells—a eukaryotic system able to add post-translational modifications, including glycosylation sites. Both prokaryotic and eukaryotic recombinant CHIKV-E2 react strongly to anti-CHIKV IgG antibodies, showing accuracy levels that are higher than 90%. However, the glycan-added viral antigen presents better sensitivity and specificity (85 and 98%) than the non-glycosylated antigen (81 and 71%, respectively) in anti-CHIKV IgM ELISA assays.
Flávia Fonseca Bagno; Lara Carvalho Godói; Maria Marta Figueiredo; Sarah Aparecida Rodrigues Sérgio; Thaís De Fátima Silva Moraes; Natália De Castro Salazar; Young Chan Kim; Arturo Reyes-Sandoval; Flávio Guimarães Da Fonseca. Chikungunya E2 Protein Produced in E. coli and HEK293-T Cells—Comparison of Their Performances in ELISA. Viruses 2020, 12, 939 .
AMA StyleFlávia Fonseca Bagno, Lara Carvalho Godói, Maria Marta Figueiredo, Sarah Aparecida Rodrigues Sérgio, Thaís De Fátima Silva Moraes, Natália De Castro Salazar, Young Chan Kim, Arturo Reyes-Sandoval, Flávio Guimarães Da Fonseca. Chikungunya E2 Protein Produced in E. coli and HEK293-T Cells—Comparison of Their Performances in ELISA. Viruses. 2020; 12 (9):939.
Chicago/Turabian StyleFlávia Fonseca Bagno; Lara Carvalho Godói; Maria Marta Figueiredo; Sarah Aparecida Rodrigues Sérgio; Thaís De Fátima Silva Moraes; Natália De Castro Salazar; Young Chan Kim; Arturo Reyes-Sandoval; Flávio Guimarães Da Fonseca. 2020. "Chikungunya E2 Protein Produced in E. coli and HEK293-T Cells—Comparison of Their Performances in ELISA." Viruses 12, no. 9: 939.
The Flaviviridae virus family was named after the Yellow-fever virus, and the latin term flavi means “of golden color”. Dengue, caused by Dengue virus (DENV), is one of the most important infectious diseases worldwide. A sensitive and differential diagnosis is crucial for patient management, especially due to the occurrence of serological cross-reactivity to other co-circulating flaviviruses. This became particularly important with the emergence of Zika virus (ZIKV) in areas were DENV seroprevalence was already high. We developed a sensitive and specific diagnostic test based on gold nanorods (GNR) functionalized with DENV proteins as nanosensors. These were able to detect as little as one picogram of anti-DENV monoclonal antibodies and highly diluted DENV-positive human sera. The nanosensors could differentiate DENV-positive sera from other flavivirus-infected patients, including ZIKV, and were even able to distinguish which DENV serotype infected individual patients. Readouts are obtained in ELISA-plate spectrophotometers without the need of specific devices.
Alice Versiani; Estefânia M. N. Martins; Lidia M. Andrade; Laura Cox; Glauco C. Pereira; Edel F. Barbosa-Stancioli; Mauricio L. Nogueira; Luiz O. Ladeira; Flávio G. Da Fonseca. Nanosensors based on LSPR are able to serologically differentiate dengue from Zika infections. Scientific Reports 2020, 10, 1 -17.
AMA StyleAlice Versiani, Estefânia M. N. Martins, Lidia M. Andrade, Laura Cox, Glauco C. Pereira, Edel F. Barbosa-Stancioli, Mauricio L. Nogueira, Luiz O. Ladeira, Flávio G. Da Fonseca. Nanosensors based on LSPR are able to serologically differentiate dengue from Zika infections. Scientific Reports. 2020; 10 (1):1-17.
Chicago/Turabian StyleAlice Versiani; Estefânia M. N. Martins; Lidia M. Andrade; Laura Cox; Glauco C. Pereira; Edel F. Barbosa-Stancioli; Mauricio L. Nogueira; Luiz O. Ladeira; Flávio G. Da Fonseca. 2020. "Nanosensors based on LSPR are able to serologically differentiate dengue from Zika infections." Scientific Reports 10, no. 1: 1-17.
Background Since the emergence of the COVID-19, health officials have struggled to devise strategies to counteract the speed of the pandemic’s spread across the globe. It became imperative to implement accurate diagnostic tests for the detection of SARS-CoV-2 RNA on respiratory samples. In many places, however, besides the limited availability of test reagents, laboratory personnel face the challenge of adapting their working routines to manipulate highly infective clinical samples. Here, we proposed the use of a virus-inactivating solution as part of a sample collection kit to decrease the infectious potential of the collected material without affecting the integrity of RNA samples used in diagnostic tests based on RT-qPCR. Methods Nasopharyngeal and oropharyngeal swab samples were collected from SARS-CoV-2-infected patients and from laboratory personnel using a commercially available viral transport solution (VTM) and the denaturing solution (DS) described here. RNA extracted from all samples was tested by RT-qPCR using probes for viral and human genes. Exposure of laboratory personnel to infective viruses was also accessed using ELISA tests. Findings The use of the DS did not interfere with the detection of viral genome or the endogenous human mRNA, since similar results were obtained from samples collected with VTM or DS. In addition, all tests of laboratory personnel for the presence of viral RNA and IgG antibodies against SARS-CoV-2 were negative. Interpretation The methodology described here provides a strategy that allow high diagnostic accuracy as well as safe manipulation of clinical samples by those involved with diagnostic procedures. Funding CAPES, FAPEMIG, CNPq, MCTIC, FIOCRUZ and the UK Global Challenges Research Fund (GCRF).
Alex Fiorini de Carvalho; Andreza Parreiras Gonçalves; Thaís Bárbara De Souza Silva; Hugo Itaru Sato; Larissa Vuitika; Flavia Fonseca Bagno; Sarah Aparecida Rodrigues Sérgio; Maria Marta Figueiredo; Raissa Prado Rocha; Ana Paula Salles Moura Fernandes; Pedro Augusto Alves; Santuza Maria Ribeiro Teixeira; Flavio Guimarães da Fonseca. THE USE OF DENATURING SOLUTION AS COLLECTION AND TRANSPORT MEDIA TO IMPROVE SARS-COV-2 RNA DETECTION AND REDUCE INFECTION OF LABORATORY PERSONNEL. 2020, 1 .
AMA StyleAlex Fiorini de Carvalho, Andreza Parreiras Gonçalves, Thaís Bárbara De Souza Silva, Hugo Itaru Sato, Larissa Vuitika, Flavia Fonseca Bagno, Sarah Aparecida Rodrigues Sérgio, Maria Marta Figueiredo, Raissa Prado Rocha, Ana Paula Salles Moura Fernandes, Pedro Augusto Alves, Santuza Maria Ribeiro Teixeira, Flavio Guimarães da Fonseca. THE USE OF DENATURING SOLUTION AS COLLECTION AND TRANSPORT MEDIA TO IMPROVE SARS-COV-2 RNA DETECTION AND REDUCE INFECTION OF LABORATORY PERSONNEL. . 2020; ():1.
Chicago/Turabian StyleAlex Fiorini de Carvalho; Andreza Parreiras Gonçalves; Thaís Bárbara De Souza Silva; Hugo Itaru Sato; Larissa Vuitika; Flavia Fonseca Bagno; Sarah Aparecida Rodrigues Sérgio; Maria Marta Figueiredo; Raissa Prado Rocha; Ana Paula Salles Moura Fernandes; Pedro Augusto Alves; Santuza Maria Ribeiro Teixeira; Flavio Guimarães da Fonseca. 2020. "THE USE OF DENATURING SOLUTION AS COLLECTION AND TRANSPORT MEDIA TO IMPROVE SARS-COV-2 RNA DETECTION AND REDUCE INFECTION OF LABORATORY PERSONNEL." , no. : 1.
The 30th meeting of the Brazilian Society for Virology (SBV) was held, for the first time in its 30 years of existence, in Cuiabá, the capital of Mato Grosso State, Central Western Brazil, a tropical region between the three richest biomes in the world: Amazon Florest, Cerrado and Pantanal. In recent years, the field of virology has been built in the State. The aim of this report is to support participants and virologists to receive the most up-to-date information about the meeting, which occurred from 16 to 19 October 2019. National and international speakers gave SBV the opportunity to learn about their experience on their virology fields, sharing recent scientific findings, compiling conferences, round table presentations and work presentations in oral and poster sessions. The meeting held over 300 attendants, who were also involved on oral and poster presentations, showing a great variety of recent unpublished studies on environmental, basic, animal, human, plant and invertebrate virology. In addition, SBV offered the Helio Gelli Pereira award for the best research studies in each field presented during the meeting. The 30th meeting of SBV was very productive and has also encouraged scientific partnership and collaboration among virologists worldwide.
Renata Dezengrini Slhessarenko; Marcelo Adriano Mendes Dos Santos; Michele Lunardi; Bruno Moreira Carneiro; Juliana Helena Chavez-Pavoni; Daniel Moura De Aguiar; Ana Claudia Pereira Terças Trettel; Carla Regina Andrighetti; Flávio Guimarães Da Fonseca; João Pessoa Araújo Junior; Fabrício Souza Campos; Luciana Barros De Arruda; Jônatas Santos Abrahão; Fernando Rosado Spilki. 30th Brazilian Society for Virology 2019 Annual Meeting—Cuiabá, Mato Grosso, Brazil. Viruses 2020, 12, 494 .
AMA StyleRenata Dezengrini Slhessarenko, Marcelo Adriano Mendes Dos Santos, Michele Lunardi, Bruno Moreira Carneiro, Juliana Helena Chavez-Pavoni, Daniel Moura De Aguiar, Ana Claudia Pereira Terças Trettel, Carla Regina Andrighetti, Flávio Guimarães Da Fonseca, João Pessoa Araújo Junior, Fabrício Souza Campos, Luciana Barros De Arruda, Jônatas Santos Abrahão, Fernando Rosado Spilki. 30th Brazilian Society for Virology 2019 Annual Meeting—Cuiabá, Mato Grosso, Brazil. Viruses. 2020; 12 (5):494.
Chicago/Turabian StyleRenata Dezengrini Slhessarenko; Marcelo Adriano Mendes Dos Santos; Michele Lunardi; Bruno Moreira Carneiro; Juliana Helena Chavez-Pavoni; Daniel Moura De Aguiar; Ana Claudia Pereira Terças Trettel; Carla Regina Andrighetti; Flávio Guimarães Da Fonseca; João Pessoa Araújo Junior; Fabrício Souza Campos; Luciana Barros De Arruda; Jônatas Santos Abrahão; Fernando Rosado Spilki. 2020. "30th Brazilian Society for Virology 2019 Annual Meeting—Cuiabá, Mato Grosso, Brazil." Viruses 12, no. 5: 494.
Porcine circovirus 2 (PCV2) infections are related to a number of syndromes and clinical manifestations, generally known as Porcine circovirus-associated diseases, which are related to losses in the swine industry. There are commercially available vaccines and new vaccines being tested, however, persistency of the PCV2 as an important pig pathogen, and the growing number of affected farms in different countries have suggested that there is room for vaccine improvement. In this study, we describe the construction and testing of a recombinant live vaccine based on a modified Vaccinia virus Ankara (MVA) vector expressing the PCV2b capsid protein (CAP). Using a two-dose homologous vaccination regimen, in mice, we demonstrated that the vaccine induced high titers of anti-PCV2 antibodies. The vaccine is stable upon lyophilization, and, together with the good immunogenicity potential observed, the results support further evaluation of the MVA-CAP vaccine in the target species.
Danielle Soares De Oliveira Daian E Silva; Edel Barbosa-Stancioli; Jordana Graziela Alves Coelho-Dos-Reis; Flávio Guimarães Da Fonseca. Short communication: a modified Vaccinia virus Ankara-based Porcine circovirus 2 vaccine elicits strong antibody response upon prime-boost homologous immunization in a preclinical model. Brazilian Journal of Microbiology 2020, 51, 1439 -1445.
AMA StyleDanielle Soares De Oliveira Daian E Silva, Edel Barbosa-Stancioli, Jordana Graziela Alves Coelho-Dos-Reis, Flávio Guimarães Da Fonseca. Short communication: a modified Vaccinia virus Ankara-based Porcine circovirus 2 vaccine elicits strong antibody response upon prime-boost homologous immunization in a preclinical model. Brazilian Journal of Microbiology. 2020; 51 (3):1439-1445.
Chicago/Turabian StyleDanielle Soares De Oliveira Daian E Silva; Edel Barbosa-Stancioli; Jordana Graziela Alves Coelho-Dos-Reis; Flávio Guimarães Da Fonseca. 2020. "Short communication: a modified Vaccinia virus Ankara-based Porcine circovirus 2 vaccine elicits strong antibody response upon prime-boost homologous immunization in a preclinical model." Brazilian Journal of Microbiology 51, no. 3: 1439-1445.
Dengue is currently one of the most important arbovirus infections worldwide. Early diagnosis is important for disease outcome, particularly for those afflicted with the severe forms of infection. The goal of this work was to identify conserved and polymorphic linear B-cell Dengue virus (DENV) epitopes that could be used for diagnostic purposes. To this end, we aligned the predicted viral proteome of the four DENV serotype and performed in silico B-cell epitope mapping. We developed a script in Perl integrating alignment and prediction information to identify potential serotype-specific epitopes. We excluded epitopes that were similarly present in the yellow fever and zika viruses’ proteomes. A total of 15 polymorphic and nine conserved peptides among DENV serotypes were selected. Peptides were spotted on cellulose membranes and tested against sera from rabbits that were monoinfected with each DENV serotype. Although serotype-specific peptides failed to recognize any sera, three conserved peptides were recognized by all anti-dengue sera and were included on an ELISA test employing a well-characterized human sera bank. Of the three peptides, one was able to efficiently identify sera from all four DENV serotypes and to discriminate them from Zika virus positive sera.
Alice Versiani; Raissa Prado Rocha; Tiago A. O. Mendes; Glauco C. Pereira; Jordana Graziella A. Coelho Dos Reis; Daniella C. Bartholomeu; Flávio G. Da Fonseca; Jordana Coelho Dos Reis. Identification of B-Cell Epitopes with Potential to Serologicaly Discrimnate Dengue from Zika Infections. Viruses 2019, 11, 1079 .
AMA StyleAlice Versiani, Raissa Prado Rocha, Tiago A. O. Mendes, Glauco C. Pereira, Jordana Graziella A. Coelho Dos Reis, Daniella C. Bartholomeu, Flávio G. Da Fonseca, Jordana Coelho Dos Reis. Identification of B-Cell Epitopes with Potential to Serologicaly Discrimnate Dengue from Zika Infections. Viruses. 2019; 11 (11):1079.
Chicago/Turabian StyleAlice Versiani; Raissa Prado Rocha; Tiago A. O. Mendes; Glauco C. Pereira; Jordana Graziella A. Coelho Dos Reis; Daniella C. Bartholomeu; Flávio G. Da Fonseca; Jordana Coelho Dos Reis. 2019. "Identification of B-Cell Epitopes with Potential to Serologicaly Discrimnate Dengue from Zika Infections." Viruses 11, no. 11: 1079.
Vaccinia virus was used as vaccine against smallpox and was instrumental in the successful eradication of that disease. Although smallpox vaccination is no longer in place in the overall population, the use of vaccinia virus in the development of viral vector-based vaccines has become popular. Nonetheless, different vaccinia virus strains are known and induce different immune responses. To look into this, we compared immune responses triggered by mouse infections with the nonreplicative MVA strain, the attenuated Lister strain, or the virulent WR strain. We observed that the WR strain was capable of downmodulating mouse cell responses, whereas the highly attenuated MVA strain induced high levels of cell-mediated immunity. Infections by the intermediately attenuated Lister strain induced cell responses that were intermediary between those induced by WR and MVA. We propose that the virulence of a vaccinia virus strain is directly proportional to its ability to downmodulate specific compartments of antiviral cell responses.
Lorena F. D. de Freitas; Rafael P. Oliveira; Mariana C. G. Miranda; Raíssa P. Rocha; Edel F. Barbosa-Stancioli; Ana Maria Faria; Flávio G. da Fonseca. The Virulence of Different Vaccinia Virus Strains Is Directly Proportional to Their Ability To Downmodulate Specific Cell-Mediated Immune Compartments In Vivo. Journal of Virology 2019, 93, 1 .
AMA StyleLorena F. D. de Freitas, Rafael P. Oliveira, Mariana C. G. Miranda, Raíssa P. Rocha, Edel F. Barbosa-Stancioli, Ana Maria Faria, Flávio G. da Fonseca. The Virulence of Different Vaccinia Virus Strains Is Directly Proportional to Their Ability To Downmodulate Specific Cell-Mediated Immune Compartments In Vivo. Journal of Virology. 2019; 93 (6):1.
Chicago/Turabian StyleLorena F. D. de Freitas; Rafael P. Oliveira; Mariana C. G. Miranda; Raíssa P. Rocha; Edel F. Barbosa-Stancioli; Ana Maria Faria; Flávio G. da Fonseca. 2019. "The Virulence of Different Vaccinia Virus Strains Is Directly Proportional to Their Ability To Downmodulate Specific Cell-Mediated Immune Compartments In Vivo." Journal of Virology 93, no. 6: 1.
Recombinant virus vectors represent a promising strategy for vaccine research. Among available viral vectors, members of the Poxviridae family—especially the modified Vaccinia virus Ankara (MVA)—stand out as immunogenic and safe vaccine platforms. Because MVA usually does not produce plaques in cell culture, visible selection markers such as the green fluorescent protein (GFP) are frequently incorporated into the constructions in order to facilitate the recognition of recombinants. However, these genetic markers have to be removed before any clinical trial. Here, we evaluated the acute responses generated in mice immunized with a MVA vector in which the GFP marker was not removed. We observed no differences in neutrophil, monocyte, or total leucocyte recruitment among animals inoculated with MVA or MVA-GFP. Likewise, there were no differences in neutrophil activation between mice groups. Hepatic functions were not altered in either MVA or MVA-GFP-inoculated mice, and we observed no histopathological alterations in different tissues from virus-inoculated animals. In conclusion, the presence of GFP is innocuous to immunized animals and do not alter acute physiopathological responses to the MVA vector. We suggest that keeping the GFP marker may be a good strategy for vaccine development, production, and evaluation.
D. S. O. Daian e Silva; T. M. G. Pinho; M. A. Rachid; D. F. Barbosa-Stancioli; F. G. Da Fonseca. The Perennial Use of the Green Fluorescent Protein Marker in a Live Vaccinia Virus Ankara Recombinant Platform Shows No Acute Adverse Effects in Mice. Brazilian Journal of Microbiology 2019, 50, 347 -355.
AMA StyleD. S. O. Daian e Silva, T. M. G. Pinho, M. A. Rachid, D. F. Barbosa-Stancioli, F. G. Da Fonseca. The Perennial Use of the Green Fluorescent Protein Marker in a Live Vaccinia Virus Ankara Recombinant Platform Shows No Acute Adverse Effects in Mice. Brazilian Journal of Microbiology. 2019; 50 (2):347-355.
Chicago/Turabian StyleD. S. O. Daian e Silva; T. M. G. Pinho; M. A. Rachid; D. F. Barbosa-Stancioli; F. G. Da Fonseca. 2019. "The Perennial Use of the Green Fluorescent Protein Marker in a Live Vaccinia Virus Ankara Recombinant Platform Shows No Acute Adverse Effects in Mice." Brazilian Journal of Microbiology 50, no. 2: 347-355.
Chikungunya virus (CHIKV) causes a disease characterized by acute onset of fever accompanied by arthralgia. Clinical similarities and co-circulation of other arboviruses such as Dengue virus (DENV) and Zika virus (ZIKV), have complicated their differentiation, making their diagnoses a challenge for the health authorities. Misdiagnosis is a serious issue to the management of patients and development of public health measures. We carried out further screening of CHIKV, DENV and ZIKV cases in Minas Gerais, Brazil, after diagnostics were already issued by a state laboratory and according to the Brazilian Ministry of Health (BMH) policy. Our aim was to look for possible co-infections or previous arboviruses’ exposure. Sera from 193 patients with symptoms of arboviral infections were tested for DEV, ZKV and/or CHIKV by the State laboratory, according to clinical suspicion and following standard BMH guidelines. After an official diagnosis was issued for each patient, we retested samples applying a broader panel of ELISA-based serological tests. We identified 13 patients with concurrent or consecutive infections (IgM positive for more than one arbovirus), including 11 individuals that were positive for CHIKV and other previously confirmed arbovirus infection. Guidelines established in many arbovirus-endemic countries prioritizes the diagnosis of Zika and Dengue and no further analyzes are done when samples are positive for those viruses. As a result, possible cases of co-infections with chikungunya are neglected, which affects the epidemiological assessments of virus circulation, patient management, and the development of public health policies.
Flávia Fonseca Bagno; Maria Marta Figueiredo; Jannely Villarreal; Glauco De Carvalho Pereira; Lara Carvalho Godoi; Flávio Guimarães da Fonseca. Undetected Chikungunya virus co-infections in a Brazilian region presenting hyper-endemic circulation of Dengue and Zika. Journal of Clinical Virology 2019, 113, 27 -30.
AMA StyleFlávia Fonseca Bagno, Maria Marta Figueiredo, Jannely Villarreal, Glauco De Carvalho Pereira, Lara Carvalho Godoi, Flávio Guimarães da Fonseca. Undetected Chikungunya virus co-infections in a Brazilian region presenting hyper-endemic circulation of Dengue and Zika. Journal of Clinical Virology. 2019; 113 ():27-30.
Chicago/Turabian StyleFlávia Fonseca Bagno; Maria Marta Figueiredo; Jannely Villarreal; Glauco De Carvalho Pereira; Lara Carvalho Godoi; Flávio Guimarães da Fonseca. 2019. "Undetected Chikungunya virus co-infections in a Brazilian region presenting hyper-endemic circulation of Dengue and Zika." Journal of Clinical Virology 113, no. : 27-30.