Direct use of cell lysates in PCR-based diagnosis of bovine leukemia virus infection
Fechner, H.; Kurg, A.; Blankenstein, P.; Mewes, G.; Geue, L.; Albrecht, C.; Ebner, D.
Berliner und Münchener Tierärztliche Wochenschrift 109(11-12): 446-450
1996
ISSN/ISBN: 0005-9366 PMID: 8956539 Document Number: 459800
The use of non-purified DNA and highly-purified DNA were compared in the polymerase chain reaction (PCR) based diagnosis of bovine leukaemia virus (BLV) infection. DNA extracted from whole blood by chloroform extraction (CP-DNA) and DNA prepared only by osmotic shock, washing, heating and freezing procedures (RPoS-DNA) were utilized. 15 cattle were examined for BLV-provirus with PCR using these 2 DNA preparations. With both methods all animals, but one, were correctly identified. It was estimated that in case of CP-DNA PCR, 10 BLV-provirus copies were sufficient to obtain a positive result. The sensitivity of RPoS-DNA PCR was similar. The latter technique was used in a field study with 10 cattle. All serologically positive animals were correctly identified by the PCR. In addition one seronegative animal was found to carry BLV-provirus. It is concluded that RPoS-DNA PCR might be a good tool for the routine diagnosis of BLV-infected cattle.