RESUMEN
The method allowing the detection of BLV proviral DNA in the peripheral blood leukocytes of cattle is reported. Cell DNA from leukocytes used without preliminary cultivation was dot-hydridized with 32P-labeled plasmid that included a fragment of BLV proviral DNA. In parallel, sera from the cattle under study were tested by immunodiffusion assay (ID). The results indicate that dot-hybridization assay is more sensitive as a diagnostic test than ID because it detects BLV infection in apparently normal cattle which was seronegative by ID.
Asunto(s)
ADN Viral/genética , ADN/genética , Virus de la Leucemia Bovina/aislamiento & purificación , Leucocitos/microbiología , Hibridación de Ácido Nucleico , Provirus/aislamiento & purificación , Retroviridae/aislamiento & purificación , Animales , Anticuerpos Antivirales/análisis , Bovinos , Enfermedades de los Bovinos/diagnóstico , Estudios de Evaluación como Asunto , Inmunodifusión/veterinaria , Leucemia/diagnóstico , Leucemia/veterinaria , Virus de la Leucemia Bovina/genética , Virus de la Leucemia Bovina/inmunología , Leucocitos/inmunología , Plásmidos , Provirus/genética , Provirus/inmunologíaRESUMEN
The vaccine L-IVP strain of vaccinia virus (VV) was used to construct the recombinant viral clones containing the influenza A hemagglutinin gene. The recombinant T plasmid was obtained with HA gene inserted in the vector pGS-20 (B. Moss) under the 7.5 K promoter of VV. A homologous recombination technique was used to insert the gene with the flanking TK sequences into vaccinia virus genome. The recombinant clones were selected by dot-hybridization with [32P]-labeled HA-probe. These recombinants were analysed for HA gene expression by the indirect immunoperoxidase method in situ using the peroxidase conjugate of the staphylococcal A-protein. This technique allows to obtain stable stained preparation and analyse the protein behavior at the ultrastructural level.