Molecular typing of a BHV-4 (bovine herpesvirus 4) field isolate.
A new BHV-4 (bovine herpesvirus 4) isolated from a case of bovine interdigital dermatitis was characterized by PCR and restriction enzyme analysis. To determine whether the new isolate (PR/1) belonged to BHV-4, DNA from infected cells was specifically amplified by PCR. We used a set of primers spanning a large 2.571 kb conserved region of the BHV-4 genome, including the 3' end of ORF1 (homologous to the EBV BVRF1 gene), ORF2 (homologous to the EBV BXRF1 gene), ORF3 (TK gene) and ORF4 (gH gene) 5' end, respectively. The identity of the amplified product was confirmed by HindIII restriction enzyme digestion and Southern hybridization. No product was observed from the DNA of other bovine herpesviruses tested. The restriction patterns of the PR/ 1 genome compared to DN 599, MOVAR 33/63 and LVR BHV-4 reference strains showed two kinds of differences, either related or not related to the prDNA (polyrepetitive DNA). Taken together, these data show that PR/ 1 is a new BHV-4. We would consider that the present report provides a scheme of work for diagnosis and typing of BHV-4 isolates.
Donofrio
G
G
Istituto di Malattie Infettive Profilassi e Polizia Veterinaria, Facoltà di Medicina Veterinaria, Università di Parma, Italy. donofrio@unipr.it
Cavirani
S
S
Flammini
C F
CF
Scatozza
F
F
D000818:Animals; D015139:Blotting, Southern / Q000662:veterinary; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology; D003588:Cytopathogenic Effect, Viral; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification; D015247:Deoxyribonuclease HindIII / Q000737:chemistry; D003872:Dermatitis / Q000662:veterinary / Q000821:virology; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D006564:Herpesviridae / Q000737:chemistry / Q000145:classification* / Q000235:genetics; D006566:Herpesviridae Infections / Q000662:veterinary* / Q000821:virology; D007558:Italy; D016133:Polymerase Chain Reaction / Q000662:veterinary; D015183:Restriction Mapping / Q000662:veterinary; D017422:Sequence Analysis, DNA / Q000662:veterinary
D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral; D015247:Deoxyribonuclease HindIII