RESUMO
Naturally occurring infections of Vaccinia virus (VACV) have been recognized in Brazil during the past 10 years. Human Brazilian Vaccinia virus (BVV) infections typically occur as a zoonosis transferred from affected dairy cows to their handlers. Outbreaks have caused notable economic losses to the rural community in the region. The origins of BVV are unclear but previous analyses have shown that at least two distinct clades of BVV exist. The aim of this study was to develop a rapid and inexpensive process for identification and differentiation of BVV that should facilitate epidemiological and ecological investigations including the improved diagnosis of Brazilian Orthopoxvirus infections. A SYBR green quantitative real-time polymerase chain reaction (PCR) targeting the hemagglutinin gene was developed to identify different populations of BVV, VACV vaccine strains used in Brazil during the smallpox eradication campaign (Vaccinia Lister (VACV-LIS) and New York City Board of Health (VACV-NYCBH)), and currently available vaccines (VACV-NYCBH DRYVAX and VACV-NYCBH Acambis 2000). Three primer combinations (one to amplify many orthopoxviruses including all vaccinia viruses described so far; one to differentiate BVV from vaccine strains (VACV-LIS, VACV-NYCBH DRYVAX and VACV-NYCBH Acambis 2000); and one to differentiate BVV clades) were designed to work at the same annealing temperature and reaction conditions. In addition, these methods were able to detect orthopoxvirus viral DNA in lesion biopsy material without the need for DNA extraction.
Assuntos
Doenças dos Bovinos/virologia , Reação em Cadeia da Polimerase/métodos , Vaccinia virus/isolamento & purificação , Vacínia/veterinária , Animais , Sequência de Bases , Brasil/epidemiologia , Bovinos , Doenças dos Bovinos/genética , Primers do DNA , Hemaglutininas Virais/genética , Humanos , Dados de Sequência Molecular , Orthopoxvirus/classificação , Orthopoxvirus/genética , Sensibilidade e Especificidade , Alinhamento de Sequência , Vacínia/transmissão , Vacínia/virologia , Vaccinia virus/classificação , Vaccinia virus/genéticaRESUMO
Carrion's disease is caused by infection with the alpha-proteobacterium Bartonella bacilliformis. Distribution of the disease is considered coincident with the distribution of its known vector, the sand fly Lutzomyia verrucarum. Recent epidemics of B. bacilliformis infections associated with atypical symptomatology in nonendemic regions have raised questions regarding the historic and present distribution of this bacterium and the scope of disease that infection causes. Phylogenetic relationships and genomic diversity of 18 B. bacilliformis isolates (10 isolates from a region where Carrion's disease is epidemic, Cuzco, Peru, and 8 isolates from a region where Carrion's disease is endemic, Caraz, Peru) were assessed using genomic data generated by infrequent restriction site PCR and gene sequence analysis of the flagellin gltA and ialB genes. A population genetic analysis of the genomic diversity suggests that what was once considered an epidemic region of Peru did not result from the recent introduction of B. bacilliformis.