RESUMO
AIM: Detection-and identification of Venezuelan equine encephalomyelitis (VEE) virus RNA in biological samples by reverse-transcription polymerase chain reaction (RT-PCR) and RT-PCR in real time (rRT-PCR). MATERIALS AND METHODS: VEE, Sindbis, West Nile, Japanese and tick-borne encephalitis viruses were studied. Cell culture of chicken fibroblasts, outbred mice and rats, Javanese macaques were used in the experiments. Biological activity determination of the running culture of causative agents used in the experiments was carried out by negative colony method in monolayer cell culture under agar coating. and using intra-cerebral infection of mice. Reagent kits developed in the 48th Central Research Institute and Institute of Analytical Instrument Engineering were used during execution of experiments of VEE virus RNA detection by RT-PCR and rRT-PCR. RESULTS: VEE virus was detected in biological samples by various methods. Data from RT-PCR and rRT-PCR are in accordance with the results of virus detection in samples using sensitive animals. CONCLUSION: Use of molecular-diagnostics methods for detection in biological samples of a causative agent of a dangerous infectious disease is important for procuring biological safety of Russian Federation.
Assuntos
Vírus da Encefalite Japonesa (Espécie)/genética , Vírus da Encefalite Equina Venezuelana/genética , Vírus da Encefalite Transmitidos por Carrapatos/genética , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sindbis virus/genética , Vírus do Nilo Ocidental/genética , Infecções por Alphavirus/diagnóstico , Infecções por Alphavirus/virologia , Animais , Animais não Endogâmicos , Galinhas , Vírus da Encefalite Japonesa (Espécie)/isolamento & purificação , Vírus da Encefalite Equina Venezuelana/isolamento & purificação , Vírus da Encefalite Transmitidos por Carrapatos/isolamento & purificação , Encefalite Japonesa/diagnóstico , Encefalite Japonesa/virologia , Encefalite Transmitida por Carrapatos/diagnóstico , Encefalite Transmitida por Carrapatos/virologia , Encefalomielite Equina Venezuelana/diagnóstico , Encefalomielite Equina Venezuelana/virologia , Fibroblastos/patologia , Fibroblastos/virologia , Humanos , Macaca mulatta , Camundongos , RNA Viral/isolamento & purificação , Ratos , Kit de Reagentes para Diagnóstico/normas , Sindbis virus/isolamento & purificação , Febre do Nilo Ocidental/diagnóstico , Febre do Nilo Ocidental/virologia , Vírus do Nilo Ocidental/isolamento & purificaçãoRESUMO
Epidemiologic analysis of epidemic outbreaks caused by American equine encephalitis causative agents is carried out in the review. Eastern equine encephalomyelitis (EEE), Western equine encephalomyelitis (WEE) and Venezuela equine encephalomyelitis (VEE) viruses are etiologic agents of dangerous transmissive diseases that are usually accompanied by fever and neurologic symptoms. Among the New World alphaviruses, VEE virus has the most potential danger for humans and domestic animals. Currently, enzootic strains of VEE play an increasing role as etiologic agents of human diseases. Most of the VEE cases in humans in endemic regions during inter-epidemic period are caused by infection with VEE subtype ID virus. A possibility of emergence of novel epidemic outbreaks of VEE is determined by mutations of ID subtype strains into IC subtype, and those currently pose a potential threat as an etiologic agent of the disease. Despite low morbidity, EEE and WEE are a problem for healthcare due to a relatively high frequency of lethal outcomes of the disease.
Assuntos
Vírus da Encefalite Equina do Oeste/patogenicidade , Encefalomielite Equina/epidemiologia , Encefalomielite Equina/virologia , Animais , Vírus da Encefalite Equina do Leste/genética , Vírus da Encefalite Equina do Leste/patogenicidade , Vírus da Encefalite Equina Venezuelana/genética , Vírus da Encefalite Equina Venezuelana/patogenicidade , Vírus da Encefalite Equina do Oeste/genética , Encefalomielite Equina/transmissão , Encefalomielite Equina/veterinária , Cavalos/virologia , Humanos , Estados UnidosRESUMO
The article presents data concerning analysis of bioassays under suspicion of influenza A(H1N1)pdm09. The technique of back transcription polymerase chain reaction and sequencing was applied. The separation of native influenza virus A was implemented using passaging through developing chick embryos.