Detection of bovine herpesvirus-1 infection in breeding bull semen by virus isolation and polymerase chain reaction.
Serum samples from 51 apparently healthy breeding bulls were screened for bovine herpesvirus-1 (BHV-1) antibodies using an avidin-biotin enzyme-linked immunosorbent assay, revealing a sero-positive prevalence rate of 45.09%. Semen samples were then collected from 12 of the sero-positive and 12 of the sero-negative bulls and tested for BHV-1 antigen using both a virus isolation assay and a polymerase chain reaction (PCR) assay; PCR was applied to detect BHV-1 deoxyribonucleic acid by using primers selected from the relatively conserved sequence of the gl glycoprotein gene to amplify a 468 base pair fragment. The PCR-amplified products were confirmed as BHV-1 by restriction enzyme, Dde 1, which produced fragments of predictable sizes, namely 340 and 128 base pairs. Positive virus isolation test results, confirmed by virus neutralisation, found BHV-1 antigen in the semen of five sero-positive and six sero-negative bulls. In comparison, positive PCR results found BHV-1 genome in the semen of six sero-positive and eight sero-negative bulls. From the 24 semen samples tested, 14 were shown to be positive by PCR and 11 by virus isolation. The sensitivity and specificity of virus isolation were 57.14% and 70% respectively, and were significantly lower than PCR. In the semen samples taken from sero-negative bulls, BHV-1 was detected more often by PCR methods than by virus-isolation, suggesting that PCR is a more sensitive method for BHV-1 screening in bulls.
Deka
D
D
Department of Veterinary Microbiology, College of Veterinary Science, Punjab Agricultural University, Ludhiana 141 004, India.
Ramneek
Maiti
N K
NK
Oberoi
M S
MS
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D001947:Breeding; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000453:epidemiology; D004269:DNA, Bacterial / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards / Q000662:veterinary; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary*; D007242:Herpesvirus 1, Bovine / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D007194:India / Q000453:epidemiology; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D015203:Reproducibility of Results; D012661:Semen / Q000821:virology*; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies
D000914:Antibodies, Viral; D004269:DNA, Bacterial