title: Comparison of three polymerase chain reaction methods for routine detection of bovine herpesvirus 1 DNA in fresh bull semen.
abstract: Five bulls were inoculated intrapreputially with Bovineherpes virus 1 (BHV 1), in order to compare the relative sensitivity of three polymerase chain reaction (PCR) assays for routine diagnosis of fresh bovine semen for the presence of BHV 1 Semen was collected twice a week up to 107 days post-infection (dpi). To reactivate latent virus, the bulls were treated with dexamethasone from 44 until 48 dpi. All samples were examined before and after cryopreservation treatment using a standard virus isolation (VI) method and three PCR assays: PCR A, PCR B and PCR C. PCR A and PCR C used an internal control plasmid DNA template and PCR B used the split sample method in order to control for false negative results. Of the 149 fresh semen samples that were tested, PCR A detected 45 positive, PCR B detected 39 positive and PCR C detected 66 positive, while virus was isolated from 22 samples. Of the 149 samples treated by cryopreservation, the virus was isolated from 13 samples and PCR C was positive in 21 samples. The results demonstrate that all three PCR assays are more sensitive than virus isolation, particularly during the later phases of infection.
authors.affiliation: Institute for Animal Science and Health (ID-DLO) Department of Mammalian Virology, Lelystad, The Netherlands.
authors.forename: C B
authors.initials: CB
authors.lastname: Smits
authors.forename: C
authors.initials: C
authors.lastname: Van Maanen
authors.forename: R D
authors.initials: RD
authors.lastname: Glas
authors.forename: A L
authors.initials: AL
authors.lastname: De Gee
authors.forename: T
authors.initials: T
authors.lastname: Dijkstrab
authors.forename: J T
authors.initials: JT
authors.lastname: Van Oirschot
authors.forename: F A
authors.initials: FA
authors.lastname: Rijsewijk
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis*; D003907:Dexamethasone / Q000494:pharmacology; D006566:Herpesviridae Infections / Q000662:veterinary* / Q000821:virology; D007242:Herpesvirus 1, Bovine / Q000235:genetics* / Q000302:isolation & purification; D007271:Injections, Intradermal; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods; D012661:Semen / Q000821:virology*; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral; D003907:Dexamethasone