A polymerase chain reaction (PCR) assay for the detection of bovine herpesvirus 1 (BHV1) in selectively digested whole bovine semen |
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| Authors: | L. H. A. Wagter R. D. Glas N. Bleumink-Pluym F. A. C. Van Engelenburg F. A. M. Rijsewijk D. J. Houwers |
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| Affiliation: | (1) Animal Health Service in the North Netherlands, PO Box 361, 9200 AJ Drachten, The Netherlands;(2) College of Veterinary Medicine, Veterinary Microbiological Diagnostic Centre, University of Utrecht, PO Box 80165, 3508 TD Utrecht, The Netherlands;(3) Department of Bovine Virology, DLO-Institute for Animal Science and Health, PO Box 65, 8200 AB Lelystad, The Netherlands |
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| Abstract: | A PCR assay for the detection of bovine herpesvirus type 1 (BHV1) DNA in selectively digested whole bovine semen was developed and evaluated. A brief treatment with proteinase-K was used to lyse free virus, virus present in non-sperm cells and virus adhering to the spermatozoa. Genomic bovine DNA was not released by this treatment. Primers and probes were based on the nucleotide sequence of the gD gene. BHV1 virus-spiked split samples were used as positive controls and the PCR products were detected by eye in ethidium bromide-stained agarose gels. Sequentially collected non-extended semen samples from experimentally infected bulls were used to compare this assay with virus isolation. Of a total of 162 ejaculates, 51 were found positive by virus isolation, whereas PCR detected BHV1 DNA in 73. PCR detected BHV1 DNA for a longer period after infection and reaction. Apart from its superior sensitivity, this PCR assay also has the advantage of being a relatively simple procedure, providing results within 24 h.Abbreviations AI artificial insemination - BHV1 bovine herpesvirus type 1 - PCR polymerase chain reaction - TCID50 tissue culture infective dose, 50% |
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| Keywords: | BHV1 bovine herpesvirus diagnosis PCR polymerase chain reaction semen virus isolation |
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