RESUMO
On May 30, 1996, a sick Daubenton's bat (Myotis daubentonii) was recovered from the cellar of a public house in Newhaven, East Sussex. Its condition deteriorated rapidly, and it was euthanased and examined. Positive results, establishing the presence of a rabies or rabies-related virus in its brain, were obtained from the fluorescent antibody test, the rabies tissue culture isolation test, and a hemi-nested reverse-transcription PCR. The complete sequence of the nucleoprotein gene was determined and a phylogenetic analysis, based on the 470 nucleotide bases of the amino terminus of the nucleoprotein, established the genotype of the virus as European bat lyssavirus 2. Bat rabies had not previously been recorded in the UK but does occur in mainland Europe. A study of the back-trajectories of the wind on May 29 and 30, established that the infected bat possibly came from near the Franco-Swiss border.
Assuntos
Quirópteros/virologia , Vírus da Raiva/genética , Vírus da Raiva/isolamento & purificação , Sequência de Aminoácidos , Animais , Sequência de Bases , Encéfalo/virologia , Imunofluorescência , Genótipo , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Reino UnidoRESUMO
A rapid and sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) assay for the detection of classical rabies virus (genotype 1) and the rabies related European bat lyssaviruses (EBLs) (genotypes 5 and 6) was developed. When combined with specific oligonucleotide probes and a PCR-enzyme linked immunosorbent assay (PCR-ELISA), genotype 5 and 6 viruses can be distinguished from each other and from genotype 1 viruses. Ninety-two isolates from the six established genotypes of rabies and rabies-related viruses were screened by RT-PCR and PCR-ELISA to determine the specificity of the assays. All genotype 1, 5 and 6 viruses were detected by RT-PCR while none of the genotype 2, 3 and 4 viruses were detected. All the genotype 5 and 6 viruses were detected by the two PCR-ELISA probes when used in combination while none of the genotype 1-4 viruses were detected. When used individually, the PCR-ELISA probes also distinguished between the genotype 5 and 6 viruses. This new discriminatory test should allow the rapid genotyping of all lyssaviruses likely to be encountered in Europe and as such could provide useful epidemiological information in the event of an outbreak.
Assuntos
Lyssavirus/classificação , Vírus da Raiva/classificação , Raiva/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Quirópteros , Primers do DNA , Ensaio de Imunoadsorção Enzimática , Europa (Continente) , Genótipo , Lyssavirus/genética , Lyssavirus/isolamento & purificação , RNA Viral/análise , Vírus da Raiva/genética , Vírus da Raiva/isolamento & purificação , Sensibilidade e EspecificidadeRESUMO
A method is described to assess RNA template quality by the incorporation of a ribosomal RNA (rRNA) internal (in tube) control into a standard rabies and rabies-related virus specific RT-PCR. Specific virus and rRNA templates were co-amplified in a duplex reaction from RNA extracts derived from 60 isolates representing all six of the established lyssavirus genotypes. To ensure a wide species applicability of this technique we demonstrated that the rRNA assay was capable of functioning using the cells or tissues of 14 different mammals. Parallel studies between the duplex and the unlinked lyssavirus assay demonstrated only a minor reduction in the sensitivity of the former test. The ribosomal and viral targets (unlike beta-actin RNA) were shown to have similar degradation kinetics making rRNA amplification a good control for viral target integrity. As a consequence, the use of this system would reduce the likelihood of obtaining false negative RT-PCR results from lyssavirus infected material.
Assuntos
Lyssavirus/isolamento & purificação , RNA Ribossômico/análise , Vírus da Raiva/isolamento & purificação , Raiva/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Southern Blotting , Encéfalo/virologia , Gatos , Bovinos , Linhagem Celular , Cricetinae , Cães , Humanos , Lyssavirus/genética , Camundongos , RNA Viral/análise , Vírus da Raiva/genética , Padrões de Referência , Sensibilidade e Especificidade , Moldes GenéticosRESUMO
The authors studied 44 outpatients with unipolar depression to determine the association among social support systems, life events, social adjustment, and depressive symptoms. Social support had a reasonably high correlation with outcome measures. Patients with high social support had significantly better scores than those with low social support on the Hamilton Rating Scale for Depression and the Social Adjustment Rating Scale Self-Report. These data corroborate the hypothesis that social support systems play a critical role in the ongoing functioning of a defined group of depressed patients.