RESUMO
BACKGROUND: Granada virus belongs to the genus Phlebovirus within the Naples serocomplex and was detected for the first time in sand flies from Spain in 2003. Seroprevalence studies have revealed that Granada virus may infect humans with most cases being asymptomatic. Moreover, recent studies in vector samples revealed that the related Massilia and Arrabida phleboviruses could be also circulating in Spain. The objective of this study was to develop and assess a new sensitive real-time RT-PCR assay for Granada virus diagnosis able to detect the related phleboviruses Massilia and Arrabida. METHODS: Two specific primers and one unique probe to detect Granada, Massilia and Arrabida viruses, without differentiating between them, were designed targeting the conserved L-segment of their genome. Sensitivity was assessed using 10-fold serial dilutions of quantified in vitro DNA samples. Specificity was evaluated by testing different genomic RNA extracted from other representative phleboviruses. The new assay was used for virus detection in sand flies collected in 2012 from the Balearic Archipelago, a touristic hotspot in the Mediterranean. RESULTS: The real-time RT-PCR assay exhibited a sensitivity per reaction of 19 copies for Granada and Arrabida, and 16 copies for Massilia. No other related phleboviruses were detected. From the 37 pools of sand fly samples studied from four different Balearic Islands, we detected one positive in the island of Cabrera. CONCLUSIONS: To our knowledge, the method described here is the first real-time RT-PCR designed to detect Granada virus and the related Massilia and Arrabida phleboviruses. The study demonstrated that this is a rapid, robust and reliable assay for the accurate diagnosis of human infections as well as for virus surveillance in vectors.
Assuntos
Phlebovirus/classificação , Phlebovirus/isolamento & purificação , Psychodidae/virologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Primers do DNA/genética , Feminino , Genoma Viral , Masculino , Filogenia , RNA Viral/genética , Sensibilidade e Especificidade , EspanhaRESUMO
Chikungunya virus emerged on Saint-Martin Island in the Caribbean in late 2013. Since then in July of 2104 Venezuela reported autochthonous cases. This study reports the first phylogenetic characterization of CHIKV autochthonous cases in Venezuela, 2014. The phylogenetic analysis showed that the CHIKV circulating in Venezuela (Aragua state) belong to the Asian genotype (Caribbean clade) and it is related to viruses that circulated in the same year in the Caribbean.
Assuntos
Febre de Chikungunya/virologia , Vírus Chikungunya/classificação , Vírus Chikungunya/genética , Variação Genética , Genótipo , Humanos , Filogenia , VenezuelaRESUMO
We evaluated the risk for the Spanish Olympic Team acquiring Zika virus in Rio de Janeiro, Brazil, during 2016. We recruited 117 team members, and all tested negative for Zika virus. Lack of cases in this cohort supports the minimum risk estimates made before the Games.
Assuntos
Esportes , Viagem , Infecção por Zika virus/epidemiologia , Infecção por Zika virus/virologia , Zika virus , Aniversários e Eventos Especiais , Atletas , Brasil , Surtos de Doenças/prevenção & controle , Saúde Global , Humanos , Medição de Risco , EspanhaRESUMO
El virus chikungunya (CHIKV) es un Alfavirus causante de la fiebre chikungunya (CHIKF). En Venezuela, una región desprovista de inmunidad contra CHIKV y con presencia de Aedes aegypti y Aedes albopictus, el primer caso importado fue reportado por las autoridades sanitarias en junio de 2014. Por la relevancia del hecho, se analizaron 94 muestras de pacientes febriles que acudieron a los centros de salud públicos y privados del estado Aragua entre enero y diciembre de 2014, mediante la detección de los fragmentos de los genes nsP4 (Alfavirus) y E1 (CHIKV) utilizando técnicas moleculares, como Transcripción Reversa acoplada a Reacción en Cadena de la Polimerasa (RT-PCR) y/o secuenciación nucleotídica. Los resultados indicaron positividad en 19,2 % de las muestras analizadas. Se vieron afectados pacientes con edades entre 6 y 66 años, con predominio del sexo femenino (12/18). Clínicamente, todos los pacientes positivos a CHIKV manifestaron signos y síntomas asociados a CHIKF, tales como fiebre (18/18), artralgia (18/18) y erupción (16/18), entre otros. A pesar de que la positividad puede considerarse baja con relación a lo reportado en otras comunidades, este estudio representa el primer reporte local de detección molecular de CHIKV en Venezuela (estado Aragua) durante el año 2014.
Chikungunya virus is an Alphavirus that causes chikungunya Fever (CHIKF). In Venezuela, a region devoid of immunity against CHIKV and presence of Aedes aegypti and Aedes albopictus. The first imported case was reported by health authorities in June 2014. The relevance of the fact, 94 samples of febrile patients who came to the centers of public and private health Aragua state between january and december for detection of the nsP4 (Alphavirus) and E1 (CHIKV) fragments were analyzed by molecular techniques (Reverse Transcriptase Polymerase Chain Reaction and/or nucleotide sequencing). The results showed 19.2 % of positivity by CHIKV. Clinically all CHIKV positive patients showed signs and symptoms related with CHIKF, such as fever (18/18), arthralgia (18/18) and rash (16/18), among others. Were affected patients between the ages of 6 and 66 years with a predominance of the female sex (12/18). Although the positivity may be considered low compared to those reported in other communities, this represents the first local report of molecular detection of CHIKV in Venezuela (Aragua state) during 2014.
RESUMO
Zika virus (ZIKV) and chikungunya virus (CHIKV) are currently circulating in overlapping areas in the American continents and may both be transmitted by Aedes spp. mosquitoes. The first documented case, to the authors' knowledge, of sequential CHIKV and ZIKV infections diagnosed in a nonendemic area in a returning traveler is reported. The implications for heightened clinical surveillance for these infections and specific patient recommendations are emphasized.
Assuntos
Febre de Chikungunya/diagnóstico , Infecção por Zika virus/diagnóstico , Adulto , Aspartato Aminotransferases/sangue , Proteína C-Reativa/metabolismo , Febre de Chikungunya/complicações , Febre de Chikungunya/virologia , Vírus Chikungunya , Feminino , Honduras , Humanos , Zika virus , Infecção por Zika virus/complicaçõesRESUMO
Since the first documented autochthonous transmission of chikungunya virus in the Caribbean island of Saint Martin in 2013, the infection has been reported within the Caribbean region as well as North, Central and South America. The risk of autochthonous transmission of chikungunya virus becoming established in Spain may be elevated due to the large numbers of travellers returning to Spain from countries affected by the 2013 epidemic in the Caribbean and South America, as well as the existence of the Aedes albopictus vector in certain parts of Spain. We retrospectively analysed the laboratory diagnostic database of the National Centre for Microbiology, Institute of Health Carlos III (CNM-ISCIII) from 2008 to 2014. During the study period, 264 confirmed cases, of 1,371 suspected cases, were diagnosed at the CNM-ISCIII. In 2014 alone, there were 234 confirmed cases. The highest number of confirmed cases were reported from the Dominican Republic (n = 136), Venezuela (n = 30) and Haiti (n = 11). Six cases were viraemic in areas of Spain where the vector is present. This report highlights the need for integrated active case and vector surveillance in Spain and other parts of Europe where chikungunya virus may be introduced by returning travellers.
Assuntos
Febre de Chikungunya/diagnóstico , Vírus Chikungunya/isolamento & purificação , Febre/etiologia , Viagem , Aedes/virologia , Animais , Febre de Chikungunya/epidemiologia , Febre de Chikungunya/virologia , Vírus Chikungunya/genética , Surtos de Doenças , República Dominicana , Feminino , Haiti , Humanos , Insetos Vetores/virologia , Masculino , RNA Viral , Estudos Retrospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vigilância de Evento Sentinela , Espanha/epidemiologia , VenezuelaRESUMO
Chikungunya virus (CHIKV) is currently spreading in the Caribbean and America. Lymphadenopathy, described in infections with other alphaviruses, is not commonly reported in CHIKV infections. Painful lymphadenopathy was found in three of the first six CHIKV infections from the current outbreak diagnosed at a reference center in Madrid, Spain.
Assuntos
Febre de Chikungunya , Vírus Chikungunya/isolamento & purificação , Surtos de Doenças , Doenças Linfáticas/etiologia , Neutropenia/etiologia , Viagem , Adulto , Febre de Chikungunya/sangue , Febre de Chikungunya/diagnóstico , Febre de Chikungunya/etiologia , Febre de Chikungunya/fisiopatologia , Febre de Chikungunya/terapia , República Dominicana/epidemiologia , Feminino , Haiti/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Espanha/epidemiologiaRESUMO
Venezuelan equine encephalitis viruses (VEEV) are emerging pathogens of medical and veterinary importance circulating in America. Argentina is a country free from epizootic VEEV activity, with circulation of enzootic strains belonging to Rio Negro virus (RNV; VEEV subtype VI) and Pixuna virus (PIXV, VEEV subtype IV). In this work, we aim to report the sequencing and phylogenetic analyses of all Argentinean VEE viruses, including 7 strains previously isolated from mosquitoes in 1980, 5 sequences obtained from rodents in 1991 and 11 sequences amplified from mosquitoes between 2003 and 2005. Two genomic regions, corresponding to the non-structural protein 4 (nsP4) and the protein E3/E2 (PE2) genes were analyzed, but only 8 samples could be amplified in the last one (longer and more variable fragment of 702 bp). For both genomic fragments, phylogenetic trees showed the absence of lineages within RNV group, and a close genetic relationship between Argentinean strains and the prototype strain BeAr35645 for PIXV clade. The analysis of nsP4 gene opens the possibility to propose a possible geographic clustering of strains within PIXV group (Argentina and Brazil). Coalescent analysis performed on RNV sequences suggested a common ancestor of 58.3 years (with a 95% highest posterior density [HPD] interval of 16.4-345.7) prior to 1991 and inferred a substitution rate of 9.8×10(-5)substitutions/site/year, slightly lower than other enzootic VEE viruses. These results provide, for the first time, information about genetic features and variability of all VEEVs detected in Argentina, creating a database that will be useful for future detections in our country. This is particularly important for RNV, which has indigenous circulation.
Assuntos
Vírus da Encefalite Equina Venezuelana/genética , Encefalomielite Equina Venezuelana/epidemiologia , Evolução Molecular , Doenças dos Cavalos/epidemiologia , Filogenia , Animais , Argentina/epidemiologia , Análise por Conglomerados , Culicidae/virologia , Vírus da Encefalite Equina Venezuelana/classificação , Encefalomielite Equina Venezuelana/transmissão , Encefalomielite Equina Venezuelana/virologia , Genes Virais , Doenças dos Cavalos/transmissão , Doenças dos Cavalos/virologia , Cavalos , Humanos , RNA Viral , Análise de Sequência de DNARESUMO
A new member of the phlebovirus genus, tentatively named Granada virus, was detected in sandflies collected in Spain. By showing the presence of specific neutralizing antibodies in human serum collected in Granada, we show that Granada virus infects humans. The analysis of the complete genome of Granada virus revealed that this agent is likely to be a natural reassortant of the recently described Massilia virus (donor of the long and short segments) with a yet unidentified phlebovirus (donor of the medium segment).
Assuntos
Febre por Flebótomos/virologia , Psychodidae/virologia , Vírus da Febre do Flebótomo Napolitano/classificação , Vírus da Febre do Flebótomo Napolitano/genética , Animais , Anticorpos Antivirais/sangue , Sequência de Bases , Chlorocebus aethiops , Feminino , Genoma Viral , Humanos , Insetos Vetores/virologia , Masculino , Dados de Sequência Molecular , Febre por Flebótomos/epidemiologia , Filogenia , Vírus Reordenados , Vírus da Febre do Flebótomo Napolitano/imunologia , Estudos Soroepidemiológicos , Células VeroRESUMO
Venezuelan Equine Encephalitis complex viruses cause epidemics and epizootics periodically in some regions of the Americas. In Argentina, only enzootic Rio Negro virus (AG80-663) (RNV) has been isolated. To survey and identify activity of viruses that belong to Venezuelan Equine Encephalitis complex in a template region of the country, a generic Alphavirus RT-Nested PCR was performed in 99 mosquito pools collected in Chaco province. Five pools were positive, and amplicons were sequenced: four of them clustered with RNV(AG80-663) and one with Pixuna virus. This is the first report of the circulation of Pixuna virus in Argentina, and it confirms enzootic and endemic activity of RNV(AG80-663) in neotropical regions of this country.
Assuntos
Culicidae/virologia , Vírus da Encefalite Equina Venezuelana/classificação , Vírus da Encefalite Equina Venezuelana/genética , Animais , Argentina , Filogenia , Clima TropicalRESUMO
Twenty-six years after it was last detected, Saint Louis encephalitis virus (SLEV) genotype III reemerged in 2005 in C6rdoba, Argentina, where it caused an outbreak. Two genotype III SLEV strains were isolated from Culex quinquefasciatus. A 71.43% prevalence for neutralizing antibodies was found in domestic fowl in the homestead of a patient with encephalitis.