Detection of BHV-1 in a naturally infected bovine fetus by a nested PCR assay.
Bovine herpesvirus 1 (BHV-1) is frequently associated with abortion in naturally and experimentally infected cattle. Most of the virus isolation and immunofluorescent antibody protocols described in the literature for detecting BHV-1 in bovine foetuses are rather laborious, costly and time-consuming. The detection is described of BHV-1 in the tissues of a naturally aborted bovine foetus by a nested PCR assay with no further hybridization procedures. Optimal results were achieved by filtering the foetal tissues on a chromatography column before DNA extraction, by using two pairs of primers in a nested PCR and by evaluating the amplification products on silver-stained polyacrylamide gels. This nested PCR was faster and easier to perform than the virus isolation test. To our knowledge, this is the first time that BHV-1 has been detected in the tissues of a naturally infected bovine foetus by means of a nested PCR. The test seems to be a practical alternative for rapid detection of BHV-1 in bovine foetus.
Rocha
M A
MA
Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Barbosa
E F
EF
Guedes
R M
RM
Lage
A P
AP
Leite
R C
RC
Gouveia
A M
AM
D000034:Abortion, Veterinary / Q000821:virology*; D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology*; D002460:Cell Line; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D005260:Female; D006566:Herpesviridae Infections / Q000662:veterinary*; D007242:Herpesvirus 1, Bovine / Q000235:genetics / Q000302:isolation & purification*; D007672:Kidney Cortex / Q000196:embryology / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D011247:Pregnancy