Mapping of porcine parvovirus DNA and development of a diagnostic DNA probe.
Dimeric and monomeric replicative forms of DNA of porcine parvovirus (PPV) strain NADL-2 were isolated and examined by restriction enzyme analysis and reciprocal Southern blot hybridization during development of a DNA probe for PPV. Genomic single stranded PPV DNA was 5.0 kb long, and results substantiated the rolling-hairpin model of parvovirus DNA replication with the primer sequence located in the 3' terminal hairpin loop. An additional finding was the generation of a 4.7 kb species of viral DNA which was considered to be a 0.3 kb deletion variant of genomic PPV DNA. A 3.0 kb DNA fragment obtained by Pst I/Hind III digestion of monomer replicative form DNA was cloned into a plasmid vector, pUC 19. The cloned fragment, recovered from transformed Escherichia coli strain TB1 and labelled with [32P] dCTP, was evaluated by dot hybridization as a probe for PPV in infected cell cultures. The probe was specific for PPV infected cells, and was 100 times more sensitive than the standard hemagglutination test.
Krell
P J
PJ
Department of Microbiology, University of Guelph, Ont., Canada.
Salas
T
T
Johnson
R P
RP
D000818:Animals; D015139:Blotting, Southern; D003001:Cloning, Molecular; D015342:DNA Probes; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D006386:Hemagglutination Tests; D009693:Nucleic Acid Hybridization; D010321:Parvoviridae / Q000235:genetics*; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis*; D014771:Virion / Q000235:genetics
D015342:DNA Probes; D004279:DNA, Viral; D004262:DNA Restriction Enzymes