RESUMO
Using the RT-PCR with primers that anneal to the 5' and the 3' extremities of the genome segments of bunyaviruses and internal primers that anneal to the S segment of Simbu serogroup viruses in a nested PCR it was possible to amplify the Oropouche virus (ORO) genome from the sera of three patients. These results show that this RT-nested-PCR is a useful tool for rapid diagnosis of Oropouche fever infections.
Assuntos
Infecções por Bunyaviridae/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vírus Simbu/isolamento & purificação , Sequência de Bases , Primers do DNA , DNA Viral/análise , Humanos , Dados de Sequência Molecular , RNA Viral/isolamento & purificação , Análise de Sequência de DNA , Vírus Simbu/genéticaRESUMO
The nucleotide sequence of the S RNA segment of the Oropouche (ORO) virus prototype strain TRVL 9760 was determined and found to be 754 nucleotides in length. In the virion-complementary orientation, the RNA contained two overlapping open reading frames of 693 and 273 nucleotides that were predicted to encode proteins of 231 and 91 amino acids, respectively. Subsequently, the nucleotide sequences of the nucleocapsid genes of 27 additional ORO virus strains, representing a 42 year interval and a wide geographical range in South America, were determined. Phylogenetic analyses revealed that all the ORO virus strains formed a monophyletic group that comprised three distinct lineages. Lineage I contained the prototype strain from Trinidad and most of the Brazilian strains, lineage II contained six Peruvian strains isolated between 1992 and 1998, and two strains from western Brazil isolated in 1991, while lineage III comprised four strains isolated in Panama during 1989.
Assuntos
Genes Virais , Nucleocapsídeo/genética , Vírus Simbu/genética , Animais , Sequência de Bases , Primers do DNA/genética , Humanos , Dados de Sequência Molecular , Fases de Leitura Aberta , Orthobunyavirus/classificação , Orthobunyavirus/genética , Filogenia , RNA Viral/genética , Vírus Simbu/classificação , Vírus Simbu/isolamento & purificação , América do SulRESUMO
An outbreak of a febrile illness characterized by headache, ocular pain, myalgia, and arthralgia occurred during June 1994 among Peruvian army troops in Northern Peru. On June 14-16, 1994, clinical data and blood samples were obtained from eight soldiers with a febrile illness, and from 26 others who had a history of febrile illness during the past three months. A follow-up blood sample was obtained 107 days later from four of the febrile and seven of the afebrile soldiers. Serum samples were tested for dengue (DEN), Oropouche (ORO), and Venezuelan equine encephalitis (VEE) IgM and IgG antibodies by an enzyme-linked immunosorbent assay (ELISA). Virus isolation was performed by inoculation of newborn mice and Vero cell cultures. Viral isolates were identified by immunofluorescence, ELISA, and nucleotide sequencing. A VEE virus infection was confirmed in three of the eight febrile soldiers, two by virus isolation, and one by serology. Antigenic analysis indicated that one of the virus isolates was similar to VEE subtype I, variety ID, viruses previously isolated in Colombia and Venezuela. Nucleotide sequence data showed that both viral isolates were identical to one another and closely related to VEE ID viruses previously isolated in Peru, Colombia, and Venezuela. Serologic results showed that two of 26 afebrile soldiers had IgM antibody to VEE and four had IgG antibody to VEE; two febrile soldiers had IgG antibody in their first serum samples. Oropouche-specific IgM antibody was detected in one of the eight febrile and five of the afebrile soldiers, and 18 of the 34 soldiers had low titers of ORO IgG antibody titers, which did not meet the diagnostic criteria for confirmed cases. All soldiers were negative for DEN IgM antibody, and 10 had flavivirus IgG antibody that reacted with DEN antigens. These data indicated that VEE ID virus was one of the causes of illness among Peruvians soldiers and that this was the first association of this VEE subtype with human disease in Peru.
Assuntos
Infecções por Bunyaviridae/epidemiologia , Surtos de Doenças , Encefalomielite Equina Venezuelana/epidemiologia , Adolescente , Adulto , Animais , Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/virologia , Vírus da Encefalite Equina Venezuelana/imunologia , Vírus da Encefalite Equina Venezuelana/isolamento & purificação , Encefalomielite Equina Venezuelana/virologia , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Camundongos , Orthobunyavirus , Peru/epidemiologia , Estudos Soroepidemiológicos , Vírus Simbu/imunologia , Vírus Simbu/isolamento & purificaçãoRESUMO
Os autores descrevem a ocorrência de epidemias causadas pelo vírus Oropouche (Oro) nos Estados dpo Maranhäo (MA) e Goiás (GO) em 1988. 36 amostras de vírus foram obtidas a partir da inoculaçäo do sangue de 120 pacientes em camundongos recém nascidos. A doença foi caracterizada por febre, cefaléia, dores musculares, articulares, fotofobia, dor retro ocular, náuseas e tontura. 128 das 197 pessoas examinadas em Porto Franco, MA, tinham anticorpos inibidores da hemaglutinaçäo (IH) para o agente e, em 106 foram detectados anticorpos IGM por MACELISA. Todos os grupos etários foram infectados, embora a incidência tenha sido mais elevada entre aqueles com 10 a 19 anos de idade. Quanto ao sexo, a infecçäo ocorreu igualmente em ambos os sexos. Recorrência dos sintomas foi observada em 56% dos casos positivos estudados. A inoculaçäo em camundongos Swiss recém nascidos de 3.624 Culicoides paraensis (Ceratopogonidae) e 1.970 Culex (Culex) quinquefasciatus (Culicidae), coletados em Porto Francos-MA, resultou em um único isolamento do vírus ORO a partir dos Culicoides. Essa é a primeira descriçäo de casos confirmados de infecçäo pelo vírus Oropouche nos Estados do Maranhäo e Goiás, Brasil
Assuntos
Humanos , Animais , Masculino , Feminino , Criança , Adulto , Pessoa de Meia-Idade , Adolescente , Infecções por Bunyaviridae/epidemiologia , Surtos de Doenças , Anticorpos Anti-Idiotípicos/análise , Anticorpos Antivirais/análise , Brasil , Culex/microbiologia , Ensaio de Imunoadsorção Enzimática , Imunoglobulina M/análise , Vírus Simbu/imunologia , Vírus Simbu/isolamento & purificaçãoRESUMO
The authors describe the occurrence of outbreaks caused by Oropouche virus (ORO) in the states of Maranhão and Goiás, Brazil in 1988. 36 strains of the virus were obtained from the intracerebral inoculation of the blood of 120 patients into 2-3 day-old infant mice. The illness was characterized by headache, fever, pain in the muscles, joints and back, photophobia, retrobulbar pain, nausea and dizziness. 128 of 197 people examined in Porto Franco, MA, had hemagglutination-inhibiting antibodies to the agent, while 106 of them had IgM antibodies by MAC ELISA test. All age groups were infected, although the incidence was higher among who had 10 to 19 years old. There was no difference, in relation to sex infections. Recurrence of symptoms was reported in 56% of sick people. Mice inoculated with 3624 Culicoides paraensis (Ceratopogonidae) and 1970 Culex (Cux.) quinquefasciatus (Culicidae) collected in Porto Franco resulted in one single isolation of ORO virus, from the Culicoides. These are the first confirmed cases of ORO infection in Maranhão and Goiás states.
Assuntos
Infecções por Bunyaviridae/epidemiologia , Surtos de Doenças , Adolescente , Adulto , Animais , Anticorpos Anti-Idiotípicos/análise , Anticorpos Antivirais/análise , Brasil , Criança , Culex/microbiologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina M/análise , Masculino , Pessoa de Meia-Idade , Vírus Simbu/imunologia , Vírus Simbu/isolamento & purificaçãoRESUMO
The isolation and laboratory studies of a new virus isolated from a human fever case in Trinidad, West Indies, are described. The virus has been named Oropouche virus after the region from which it was obtained. Oropouche virus has been found to be related to Simbu virus, an agent isolated in South Africa. Neutralising antibodies were found in the blood of a few forest workers, 8 of 26 native cebus monkeys in the Nariva Swamp and 9 of 26 howler monkeys widely distributed over the island (Summary)