detection of bovine viral diarrhea virus in bovine semen using nested-pcr

a rapid and sensitive reverse transcription polymerase chain reaction (rt-pcr) and nested-pcr were used to detect bovine viral diarrhea virus 1 (bvdv-1) in bull semen. selected primers could amplify a part of the 5´utr of the bvdv genome. a 294 bp dna fragment was amplified and specificity of the results was confirmed by direct sequencing of the pcr product. prior to rna extraction, the seminal inhibitors were eliminated using a simple dilution method. therefore, a sensitivity of 3×102 50% tissue culture infective dose (tcid50) was achieved when experimentally infected semen was used for rna extraction. in nested-pcr a 160 bp fragment was amplified and sensitivity of the test was increased to 3 tcid50. this technique can be used as a rapid and sensitive method of bvdv-1 detection in bovine semen.