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 detection of alcelaphine herpesvirus 1 (AHV1), a causative agent of malignant catarrhal fever (MCF) of ruminants. A pair of 20-base primers was constructed based on the published nucleotide sequence of gene A of the WC11 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 was shown to be specific for AHV1 DNA by (i) cleavage with XbaI which yielded 2 subfragments of approximately 140 and 280 base pairs and (ii) chemiluminescence Southern blot hybridisation with a digoxigenin-labelled 25-base internal probe. The PCR assay also amplified AHV1 gene sequences in tissue samples from deer and rabbits experimentally infected with materials derived from deer with clinical sheep-associated MCF.
Tham
K M
KM
Virology Section, Central Animal Health Laboratory, MAF Quality Management, New Zealand.
Ng
K
K
Young
L W
LW
D000818:Animals; D000866:Antelopes / Q000821:virology*; D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis* / Q000737:chemistry; D003670:Deer / Q000821:virology*; D004587:Electrophoresis, Agar Gel; D016679:Genome, Viral; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D008198:Lymph Nodes / Q000821:virology*; D008304:Malignant Catarrh / Q000821:virology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D011817:Rabbits / Q000821:virology
D017931:DNA Primers; D004279:DNA, Viral; D015345:Oligonucleotide Probes