Detection of phocine distemper virus using the polymerase chain reaction.
During the fatal seal epizootics in the North and Baltic Seas in summer 1988 a virus was isolated which was shown to be the causal agent. It was subsequently classified as morbillivirus by neutralization assays, reaction with monoclonal antibodies and nucleic acid hybridization studies. The virus (tentatively called Phocine Distemper Virus, PDV) is difficult to grow in culture making rapid diagnosis difficult. We have used the Polymerase Chain Reaction (PCR) as an alternative and fast method to detect the presence of virus-specific nucleic acid and we describe here the amplification of cell culture derived PDV RNA in a "one-tube" reaction using heterologous (Rinderpest Virus cDNA derived) F gene primers. The resulting 370 bp DNA fragment was shown to be morbillivirus derived by Southern blot hybridization using cloned RPV F gene as probe.
Haas
L
L
Institute of Virology, Veterinary School Hannover, FRG.
Barrett
T
T
Harder
T
T
Bostock
C J
CJ
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D005784:Gene Amplification; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010252:Paramyxoviridae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis*
D004279:DNA, Viral; D012367:RNA, Viral