Detection of bovine viral diarrhoea virus in bulk milk samples using molecular-genetic methods
1999
Vilcek, S. (University of Veterinary Medicine, Kosice (Slovak Republic)) | Durkovic, B. | Strojny, L. (Research Institute of Veterinary Medicine, Kosice (Slovak Republic)) | Kvokacka, V. (District Office, Presov (Slovak Republic). Department of Veterinary Welfare)
A single RT-PCR method for the detection of BVDV in bulk milk of dairy herds was developed. The total RNA was isolated from milk somatic cells using QIAamp Viral RNA purification kit. The amplification method employed primers 324/326 selected from 5'non-coding region of pestivirus genome. The concentration of Mg sup(2+) and number of amplification cycles of RT-PCR were optimised to obtain a single specific 288 bp DNA fragment. RT-PCR assaz achieved the sensitivity 10E2 TCID sub(50)/ml. The specificity of PCR product was confirmed by a sequencing of DNA fragment in both directions. Six of 24 bulk milk samples, e.g. 25%, collected from dairy herds in the Presov district from January to June 1998 were positive for BVDV, indicating persistently infected animals in the herd. RT-PCR method is a method of choice for the rapid screening of BVDV persistently infected cattle in dairy herds
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