RESUMO
Rabies is caused by rabies virus (RV), a RNA virus member of the Lyssavirus genus, family Rhabdoviridae. The aim of this study was to determine antigenic characteristics of a rabies virus isolate (RV183-07) recovered from a stray bitch that died of rabies and to infer the most likely source of contamination, since no urban rabies has been reported in the area in more than 20 years. The virus was identified by direct immunofluorescence and multiplied by one passage in mice. The antigenic profile of the isolate was determined with a panel of monoclonal antibodies to lyssavirus antigens on infected brain tissues. A fragment of the viral genome corresponding to the nucleoprotein (N) gene was submitted to reverse transcription/polymerase chain reaction and the amplicon obtained was subjected to restriction enzyme analysis. A 303 base pair fragment of the N gene was cloned, sequenced and compared to other RV sequences available at Genbank. The isolate RV183-07 displayed antigenic and genomic characteristics of rabies virus variants whose natural reservoir is the non-hematophagous bat Tadarida brasiliensis. Therefore, the most likely source of contamination of the bitch was an incidental contact with an infected bat of that species, common inhabitants of the area. In view of that, the status of urban rabies-free of the area was not compromised.
RESUMO
Rabies is caused by rabies virus (RV), a RNA virus member of the Lyssavirus genus, family Rhabdoviridae. The aim of this study was to determine antigenic characteristics of a rabies virus isolate (RV183-07) recovered from a stray bitch that died of rabies and to infer the most likely source of contamination, since no urban rabies has been reported in the area in more than 20 years. The virus was identified by direct immunofluorescence and multiplied by one passage in mice. The antigenic profile of the isolate was determined with a panel of monoclonal antibodies to lyssavirus antigens on infected brain tissues. A fragment of the viral genome corresponding to the nucleoprotein (N) gene was submitted to reverse transcription/polymerase chain reaction and the amplicon obtained was subjected to restriction enzyme analysis. A 303 base pair fragment of the N gene was cloned, sequenced and compared to other RV sequences available at Genbank. The isolate RV183-07 displayed antigenic and genomic characteristics of rabies virus variants whose natural reservoir is the non-hematophagous bat Tadarida brasiliensis. Therefore, the most likely source of contamination of the bitch was an incidental contact with an infected bat of that species, common inhabitants of the area. In view of that, the status of urban rabies-free of the area was not compromised.
RESUMO
Rabies is caused by rabies virus (RV), a RNA virus member of the Lyssavirus genus, family Rhabdoviridae. The aim of this study was to determine antigenic characteristics of a rabies virus isolate (RV183-07) recovered from a stray bitch that died of rabies and to infer the most likely source of contamination, since no urban rabies has been reported in the area in more than 20 years. The virus was identified by direct immunofluorescence and multiplied by one passage in mice. The antigenic profile of the isolate was determined with a panel of monoclonal antibodies to lyssavirus antigens on infected brain tissues. A fragment of the viral genome corresponding to the nucleoprotein (N) gene was submitted to reverse transcription/polymerase chain reaction and the amplicon obtained was subjected to restriction enzyme analysis. A 303 base pair fragment of the N gene was cloned, sequenced and compared to other RV sequences available at Genbank. The isolate RV183-07 displayed antigenic and genomic characteristics of rabies virus variants whose natural reservoir is the non-hematophagous bat Tadarida brasiliensis. Therefore, the most likely source of contamination of the bitch was an incidental contact with an infected bat of that species, common inhabitants of the area. In view of that, the status of urban rabies-free of the area was not compromised.
RESUMO
Rabies is caused by rabies virus (RV), a RNA virus member of the Lyssavirus genus, family Rhabdoviridae. The aim of this study was to determine antigenic characteristics of a rabies virus isolate (RV183-07) recovered from a stray bitch that died of rabies and to infer the most likely source of contamination, since no urban rabies has been reported in the area in more than 20 years. The virus was identified by direct immunofluorescence and multiplied by one passage in mice. The antigenic profile of the isolate was determined with a panel of monoclonal antibodies to lyssavirus antigens on infected brain tissues. A fragment of the viral genome corresponding to the nucleoprotein (N) gene was submitted to reverse transcription/polymerase chain reaction and the amplicon obtained was subjected to restriction enzyme analysis. A 303 base pair fragment of the N gene was cloned, sequenced and compared to other RV sequences available at Genbank. The isolate RV183-07 displayed antigenic and genomic characteristics of rabies virus variants whose natural reservoir is the non-hematophagous bat Tadarida brasiliensis. Therefore, the most likely source of contamination of the bitch was an incidental contact with an infected bat of that species, common inhabitants of the area. In view of that, the status of urban rabies-free of the area was not compromised.
RESUMO
Rabies is caused by rabies virus (RV), a RNA virus member of the Lyssavirus genus, family Rhabdoviridae. The aim of this study was to determine antigenic characteristics of a rabies virus isolate (RV183-07) recovered from a stray bitch that died of rabies and to infer the most likely source of contamination, since no urban rabies has been reported in the area in more than 20 years. The virus was identified by direct immunofluorescence and multiplied by one passage in mice. The antigenic profile of the isolate was determined with a panel of monoclonal antibodies to lyssavirus antigens on infected brain tissues. A fragment of the viral genome corresponding to the nucleoprotein (N) gene was submitted to reverse transcription/polymerase chain reaction and the amplicon obtained was subjected to restriction enzyme analysis. A 303 base pair fragment of the N gene was cloned, sequenced and compared to other RV sequences available at Genbank. The isolate RV183-07 displayed antigenic and genomic characteristics of rabies virus variants whose natural reservoir is the non-hematophagous bat Tadarida brasiliensis. Therefore, the most likely source of contamination of the bitch was an incidental contact with an infected bat of that species, common inhabitants of the area. In view of that, the status of urban rabies-free of the area was not compromised.
RESUMO
Rabies is caused by rabies virus (RV), a RNA virus member of the Lyssavirus genus, family Rhabdoviridae. The aim of this study was to determine antigenic characteristics of a rabies virus isolate (RV183-07) recovered from a stray bitch that died of rabies and to infer the most likely source of contamination, since no urban rabies has been reported in the area in more than 20 years. The virus was identified by direct immunofluorescence and multiplied by one passage in mice. The antigenic profile of the isolate was determined with a panel of monoclonal antibodies to lyssavirus antigens on infected brain tissues. A fragment of the viral genome corresponding to the nucleoprotein (N) gene was submitted to reverse transcription/polymerase chain reaction and the amplicon obtained was subjected to restriction enzyme analysis. A 303 base pair fragment of the N gene was cloned, sequenced and compared to other RV sequences available at Genbank. The isolate RV183-07 displayed antigenic and genomic characteristics of rabies virus variants whose natural reservoir is the non-hematophagous bat Tadarida brasiliensis. Therefore, the most likely source of contamination of the bitch was an incidental contact with an infected bat of that species, common inhabitants of the area. In view of that, the status of urban rabies-free of the area was not compromised.
RESUMO
Bovine Herpesviruses (BHV) type 1 (BHV-1) and type 5 (BHV-5) were analysed by immunoperoxidase staining with a panel of monoclonal antibodies (Mabs) prepared against BHV antigens. One of the Mabs recognized all BHV isolates tested. The remainder four mabs recognized only BHV-1 samples, including standard laboratory strains. All isolates associated with clinical cases of encephalitis (BHV-5) displayed a pattern of reactivity distinct from that of viruses isolated from syndromes associated with BHV-1 infections. The results obtained indicate that such Mabs allowed the differentiation between BHV-1 and BHV-5, with a perfect correlation between the clinical pictures and the patterns of reactivity in vitro.
Amostras de herpesvírus bovinos (BHV) tipo 1 (Virus da Rinotraqueíte Infecciosa Bovina/Vulvovaginite Pustular Infecciosa; BHV-1) e tipo 5 (Herpesvírus da Encefalite Bovina; BHV-5) tiveram seu perfil de reatividade analisado em testes de imunoperoxidase frente a um painel composto por cinco anticorpos monoclonais (AcM) produzidos contra antígenos de BHV-1. Um dos AcM reconheceu todas as amostras de BHV examinadas. Os quatro AcM restantes reconheceram somente amostras de BHV-1. Todas as amostras isoladas de casos de encefalites (BHV-5) apresentaram um padrão de reação distinto daquelas isoladas de outros síndromes associados à infecção pelo BHV-1. Os resultados obtidos indicam que os AcM avaliados permitem a diferenciação entre amostras de BHV-1 e BHV-5, havendo perfeita correlação entre os quadros clínicos observados com os perfis de reatividade obtidos in vitro.
RESUMO
SUMMARY This review is an update on laboratory methods currently available and internationally recognized for virological and serological diagnosis of classical swine fever/hog cholera (HC). Rapid antigen detection is achieved by examination of cryostat sections of tissues of animals by direct immunofluorescence (IF). The diagnosis must be confirmed by virus isolation in cell culture. Fetection of antibodies to HC virus is based on serum neutralization tests followed by IF, immunoperoxidase (IPX), or enzyme immunoassays. Monoclonal antibcdies are extremely useful in distinguishing CSF from other pestiviruses which may eventually infect swine. Molecular techniques such as hybridization, polimerase chain reaction (PCR) and sequencing are not yet routinely used in HC diagnosis.
RESUMO Neste artigo são revisados os diferentes métodos internacionalmente utilizados no diagnóstico de Peste Suína Clássica (PSC). A detecção rápida de antígenos virais é feita através do exame direto de cortes em criostato dos tecidos de animais, por imunofluorescência direta (IF). O diagnóstico é confirmado pelo isolamento do vírus em cultivos celulares. A detecção de anticorpos contra o vírus é baseada em testes de soroneutralizaçâo seguidos de IF, imunoperoxidase (IPX), ou ensaios imunoenzimáticos. Anticorpos monoclonais são úteis na distinção entre o vírus da PSC e outros pestivírus que podem eventualmente infectar os suínos. Técnicas moleculares, como hibridização, reação de polimerase em cadeia (PCR) e seqüenciamento, ainda não são empregadas rotineiramente no diagnóstico de PSC.
RESUMO
SUMMARY This review is an update on laboratory methods currently available and internationally recognized for virological and serological diagnosis of classical swine fever/hog cholera (HC). Rapid antigen detection is achieved by examination of cryostat sections of tissues of animals by direct immunofluorescence (IF). The diagnosis must be confirmed by virus isolation in cell culture. Fetection of antibodies to HC virus is based on serum neutralization tests followed by IF, immunoperoxidase (IPX), or enzyme immunoassays. Monoclonal antibcdies are extremely useful in distinguishing CSF from other pestiviruses which may eventually infect swine. Molecular techniques such as hybridization, polimerase chain reaction (PCR) and sequencing are not yet routinely used in HC diagnosis.
RESUMO Neste artigo são revisados os diferentes métodos internacionalmente utilizados no diagnóstico de Peste Suína Clássica (PSC). A detecção rápida de antígenos virais é feita através do exame direto de cortes em criostato dos tecidos de animais, por imunofluorescência direta (IF). O diagnóstico é confirmado pelo isolamento do vírus em cultivos celulares. A detecção de anticorpos contra o vírus é baseada em testes de soroneutralizaçâo seguidos de IF, imunoperoxidase (IPX), ou ensaios imunoenzimáticos. Anticorpos monoclonais são úteis na distinção entre o vírus da PSC e outros pestivírus que podem eventualmente infectar os suínos. Técnicas moleculares, como hibridização, reação de polimerase em cadeia (PCR) e seqüenciamento, ainda não são empregadas rotineiramente no diagnóstico de PSC.