Flávio Lauretti

Universidade de São Paulo

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Carlos Nozawa

Universidade Federal do Ceará

Rosa Elisa Carvalho Linhares

Universidade Federal do Ceará

Fabrı́cio José Benati

Universidade Federal do Rio de Janeiro

Norma Santos

Universidade Federal de Juiz de Fora

Veronica Lippi

Universidade de São Paulo

Luíz Tadeu Moraes Figueiredo

Universidade de São Paulo

Mário Luis Garcia de Figueiredo

Universidade de São Paulo

Helda Liz Alfonso

Universidade de São Paulo

Fernando L. Melo

Universidade de Brasília

Víctor Hugo Aquino

Universidad Nacional de Asunción

Cooperative Institutions

Universidade de São Paulo

Universidade de Ribeirão Preto

Universidade Estadual de Londrina

Universidade Brasil

Clinics Hospital of Ribeirão Preto

Fundação Oswaldo Cruz

Universidade Federal do Rio de Janeiro

University of Queensland

Australian e-Health Research Centre

Fundação Faculdade de Medicina

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Use of acridine orange staining for the detection of rotavirus RNA in polyacrylamide gels

Journal of Virological Methods (2003)

Flávio Lauretti Fernando L. Melo Fabrı́cio José Benati Eduardo de Mello Volotão Norma Santos

Acridine orange

Ethidium bromide

Metachromasia

Acridine

10.1016/j.jviromet.2003.08.005

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Citations (32)

SEVERE DIE- BACK OF PLUM IN CENTRAL ITALY IS ASSOCIATED WITH MLOS AND VIRUS INFECTIONS

Acta Horticulturae (1995)

Massimo Pilotti F. Faggioli Flávio Lauretti M. Barba

10.17660/actahortic.1995.386.15

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Citations (4)

BIOLOGICAL, MOLECULAR AND IMMUNOCYTOCHEMICAL CHARACTERIZATION OF CELL CULTURE ADAPTED HUMAN ROTAVIRUS STRAINS DETECTED IN THE CITY OF PONTA GROSSA, PARANA, BRAZIL

VIRUS Reviews & Research (2014)

Alessandra Cristina Santos Fabrı́cio José Benati Flávio Lauretti Rosa Elisa Carvalho Linhares Carlos Nozawa

Rotaviruses (RV) are the single most important cause of severe diarrheal illness in infants and young children in both developed and developing countries worldwide. They affect mainly children less than 24 months of age. Presently, we adapted human rotavirus strains from field materials in cell culture, as well as, studied their properties by the analyses of the cytopathic effect, immunofluorescence features, plaque properties and virus genome stability. The study was carried out with 53 rotavirus-positive fecal specimens, collected in the city of Ponta Grossa, Parana, before the introduction of vaccination program in Brazil. The samples were submitted to at least 3 blind passages in MA-104 cell culture before discarded as non-cultivable. The adaptation process was monitored by the development of the cytopathic effect, immunofluorescence assay , plaque assay and genome stability by reverse transcriptase-polymerase chain reactions amplification. Fourteen human rotavirus strains were adapted to grow in cell culture. The isolation was demonstrated by the development of typical cytopathic effect. Immunofluorescence assay revealed granular inclusions with specific fluorescence mainly at the perinuclear area of the infected cells. Plaque assay allowed the demonstration of plaques with varying diameter depending upon the strains. Reverse transcriptase-polymerase chain reactions resulted in amplification and genotyping, demonstrating the stability of the genome through cell culture adaptation. DOI: http://dx.doi.org/10.17525/vrr.v19i1.97

10.17525/vrr.v19i1.97

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The effect of concanavalin A on the replication of rotavirus (SA-11) in cell culture

Brazilian Archives of Biology and Technology (2002)

Beatriz S. Hasenack Maria Valéria Jacobucci Botelho Flávio Lauretti Fernando L. Melo J.M. Orlandi

Rotavirus (RV) strain SA-11 was studied with respect to its infectivity in MA-104 cell cultures and the effect of concanavalin A (ConA). Receptors for ConA at the surface of MA-104 cell were determined by fluorescence assay and specifically inhibited by D-mannose. The kinetics of virus growth was carried out by plaque assay. Electron microscopy and polyacrylamide gel electrophoresis were used for monitoring the experiments. It was concluded that RV replication was not affected consistently by ConA, however it interfered with the development of cytopathic effect (CPE) without altering virus yields.

Infectivity

Cytopathic effect

Virus quantification

10.1590/s1516-89132002000200003

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Citations (3)

Dengue vaccines: what we know, what has been done, but what does the future hold?

Revista de Saúde Pública (2015)

Emiliana Pereira Abrão Danillo Lucas Alves Espósito Flávio Lauretti Benedito Antônio Lopes da Fonseca

Dengue, a disease caused by any of the four serotypes of dengue viruses, is the most important arthropod-borne viral disease in the world in terms of both morbidity and mortality. The infection by these viruses induces a plethora of clinical manifestations ranging from asymptomatic infections to severe diseases with involvement of several organs. Severe forms of the disease are more frequent in secondary infections by distinct serotypes and, consequently, a dengue vaccine must be tetravalent. Although several approaches have been used on the vaccine development, no vaccine is available against these viruses, especially because of problems on the development of a tetravalent vaccine. Here, we describe briefly the vaccine candidates available and their ability to elicit a protective immune response. We also discuss the problems and possibilities of any of the vaccines in final development stage reaching the market for human use.

Dengue vaccine

10.1590/s0034-8910.2015049006146

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Citations (11)

CCR2 Plays a Protective Role in Rocio Virus–Induced Encephalitis by Promoting Macrophage Infiltration Into the Brain

The Journal of Infectious Diseases (2019)

Alberto A. Amarilla Nilton Nascimento Santos-Júnior Mário Luis Garcia de Figueiredo João Paulo Mesquita Luiz Marcílio Jorge Fumagalli

Rocio virus (ROCV) is a highly neuropathogenic mosquito-transmitted flavivirus responsible for an unprecedented outbreak of human encephalitis during 1975–1976 in Sao Paulo State, Brazil. Previous studies have shown an increased number of inflammatory macrophages in the central nervous system (CNS) of ROCV-infected mice, implying a role for macrophages in the pathogenesis of ROCV. Here, we show that ROCV infection results in increased expression of CCL2 in the blood and in infiltration of macrophages into the brain. Moreover, we show, using CCR2 knockout mice, that CCR2 expression is essential for macrophage infiltration in the brain during ROCV infection and that the lack of CCR2 results in increased disease severity and mortality. Thus, our findings show the protective role of CCR2-mediated infiltration of macrophages in the brain during ROCV infection.

Infiltration (HVAC)

CCR2

Flavivirus

Pathogenesis

Viral encephalitis

10.1093/infdis/jiz029

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Citations (23)

Development of a novel plaque reduction neutralisation test for hantavirus infection

Memórias do Instituto Oswaldo Cruz (2015)

Michelly de Pádua William Marciel de Souza Flávio Lauretti Luiz Tadeu Moraes Figueiredo

In the Americas, hantaviruses cause severe cardiopulmonary syndrome (HCPS) with a high fatality rate. Hantavirus infection is commonly diagnosed using serologic techniques and reverse transcription-polymerase chain reaction. This paper presents a novel plaque reduction neutralisation test (PRNT) for detecting antibodies to Brazilian hantavirus. Using PRNT, plaque detection was enhanced by adding 0.6% of dimethyl sulfoxide into the overlay culture medium of the infected cells. This procedure facilitated clear visualisation of small plaques under the microscope and provided for easy and accurate plaque counting. The sera from 37 HCPS patients from the city of Ribeirão Preto, Brazil was evaluated for the Rio Mamoré virus (RIOMV) using PRNT. Six samples exhibited neutralising antibodies; these antibodies exhibited a low titre. The low level of seropositive samples may be due to fewer cross-reactions between two different hantavirus species; the patients were likely infected by Araraquara virus (a virus that has not been isolated) and RIOMV was used for the test. This assay offers a new approach to evaluating and measuring neutralising antibodies produced during hantavirus infections and it can be adapted to other hantaviruses, including viruses that will be isolated in the future.

Hantavirus Infection

10.1590/0074-02760150102

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Citations (8)

COMPARISON OF DIFFERENT METHODS TO PRODUCE VIRUS FREE STONE FRUITS

Acta Horticulturae (1992)

M. Barba Luciana Santos da Silva Martino Flávio Lauretti

10.17660/actahortic.1992.309.57

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Citations (6)

Incidence of group A rotavirus in urban and rural areas of the city of Londrina-Brazil, from 1995 to 1997

Brazilian Archives of Biology and Technology (2001)

Ângelo Cesar Meneghetti Andrea Bolognini Flávio Lauretti Rosa Elisa Carvalho Linhares Norma Santos

Rotaviruses are common pathogens and the causal agents of acute diarrhea among children and young animals. The involvement of rotavirus in human diarrheal disease among population of urban and rural areas of the city of Londrina, Parana was evaluated. Nine hundred and five fecal specimens from persons with diarrhea were studied, being 686 and 219 from urban and rural areas, respectively. Thirty-eight samples (4,2%) were positive for rotavirus by polyacrylamide gel electrophoresis of viral RNA and latex agglutination test of which 36 were from urban and two from rural areas. Out of the positive specimens, 17 strains were further characterized by RT-PCR typing assay, resulting in 16 strains of G1 genotype while one sample was found to be a mixture of G1 and G3 genotypes.

Acute diarrhea

10.1590/s1516-89132001000300006

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Citations (5)

Development of an enzyme immunoassay for poliovirus antigens

Brazilian Archives of Biology and Technology (2007)

Newton Hashimoto Fabrı́cio José Benati Flávio Lauretti Rosa Elisa Carvalho Linhares Carlos Nozawa

An indirect solid-phase enzyme immunoassay (EIA) was developed for the detection of poliovirus antigen. Virus antigen was obtained in LLC-MK2 cell cultures and used to prepare antibodies in rabbit and guinea pig. Antibodies were evaluated by double immunodiffusion and neutralization test. Optimal concentrations of guinea pig and rabbit immunoglobulins were determined by checkerboard titration. Microtitre plates were coated with 15.0 µg/ml guinea pig anti-polio immunoglobulin and rabbit anti-polio immunoglobulin at the concentration of 7.94 µg/ml was used as detecting antibody. The standard curve with eight different antigen concentrations in eight replicates resulted in a coefficient of variation (CV) between 2.1% to 7.8%. The dose-response relationship was determined by simple linear regression with a coefficient of correlation (R²) equal to 96.4%. The assay detected a minimum of 2.3 µg/ml poliovirus antigen.

Immunodiffusion

Radial immunodiffusion

10.1590/s1516-89132007000100005

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Citations (1)