POLYMERASE CHAIN-REACTION AMPLIFICATION OF WILDEBEEST-ASSOCIATED AND CERVINE-DERIVED MALIGNANT CATARRHAL FEVER VIRUS-DNA

A polymerase chain reaction (PCR) assay was developed for the detectio n of alcelaphine herpesvirus 1 (AHV1), a causative agent of malignant catarrhal fever (MCF) of ruminants. A pair of 20-base primers was cons tructed based on the published nucleotide sequence of gene A of the WC 11 isolate of AHV1 and was used to amplify a DNA fragment of 413 base pairs. The optimised PCR assay was highly sensitive, i.e. it detected 10 fg of genomic DNA of AHV1 (WC11 isolate). The amplified fragment wa s shown to be specific for AHV1. DNA by (i) cleavage with XbaI which y ielded 2 subfragments of approximately 140 and 280 base pairs and (ii) chemiluminescence Southern blot hybridisation with a digoxigenin-labe lled 25-base internal probe. The PCR assay also amplified AHV1 gene se quences in tissue samples from deer and rabbits experimentally infecte d with materials derived from deer with clinical sheep-associated MCF.