Characterization of RKZ isolate of ovine herpesvirus 1.
Cytopathic effect (CPE) characterized mainly by foci of rounded cells was observed in cultures of primary plexus choroideus cells from healthy lamb following cryopreservation. It was possible to transmit the infectious agent to other primary cells of ovine origin by co-cultivation with infected cells. By indirect immunofluorescence microscopy it was found that high percentage of sheep (65-80% in 3 different herds from Slovakia) are infected with this infectious agent. Electron microscopy of cells with CPE revealed the presence of herpesvirus particles. Viral DNA was isolated from infected cells using pulse-field gel electrophoresis and further used as probe in Southern blot analysis. The probe reacted specifically only with DNA from cells infected with Ovine herpesvirus 1 (OvHV-1) but not with DNA of other ruminant herpesviruses. Some of the HindIII restriction fragments of DNA of the obtained OvHV-1 isolate denominated RKZ were cloned. Part of the H9 clone was sequenced identifying a gene that encoded a polypeptide homologous to conserved herpesvirus VP23 structural protein. From comparison of the sequence of this clone with VP23 sequences of other herpesviruses it was deduced that OvHV-1 might be classified within the Rhadinovirus genus of the Gammaherpesvirinae subfamily. The sequencing of the H9 clone of DNA of RKZ isolate enabled establishment of sensitive and highly specific polymerase chain reaction (PCR) assay for detection of OvHV-1.
Kopácek
J
J
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, 842 45 Bratislava, Slovak Republic.
Rejholcová
O
O
Koptidesová
D
D
Ciampor
F
F
Rijsewijk
F M
FM
Reid
H W
HW
Pastorek
J
J
Zelník
V
V
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002213:Capsid / Q000235:genetics; D018920:Coculture Techniques; D046508:Culture Techniques; D003588:Cytopathogenic Effect, Viral; D017931:DNA Primers; D015247:Deoxyribonuclease HindIII; D006564:Herpesviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D006566:Herpesviridae Infections / Q000662:veterinary*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D018144:Rhadinovirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood / Q000453:epidemiology / Q000821:virology*; D018154:Slovakia / Q000453:epidemiology; D014412:Tumor Virus Infections / Q000662:veterinary*
D000914:Antibodies, Viral; D017931:DNA Primers; D015247:Deoxyribonuclease HindIII