Differentiation of species of the genus Orthopoxvirus in a dot blot assay using digoxigenin-labeled DNA-probes.
A dot blot assay using five different digoxigenin-labeled probes was established for specific detection and differentiation of four species belonging to the genus Orthopoxvirus. As little as 20 pg orthopoxvirus DNA can be detected, corresponding to approximately 8 x 10(4) DNA molecules. A total of 37 orthopoxvirus strains and isolates of different origin were investigated and could be assigned to the species vaccinia, cowpox, camelpox, and mousepox virus. These findings agree with their identification by biological means. Additionally, a simple procedure to isolate orthopoxvirus DNA directly from scab material was developed. This allows a fast classification without isolating the virus.
Institute of Microbiology, Federal Armed Forces Medical Academy, Munich, Germany.
H
H
Meyer
N
N
Osterrieder
M
M
Pfeffer
D000818:Animals; D002162:Camelus / Q000382:microbiology*; D015608:Cowpox virus / Q000145:classification / Q000302:isolation & purification; D015342:DNA Probes; D004076:Digoxigenin; D015151:Immunoblotting / Q000662:veterinary*; D011212:Poxviridae / Q000145:classification* / Q000302:isolation & purification; D011213:Poxviridae Infections / Q000382:microbiology / Q000662:veterinary*; D017422:Sequence Analysis, DNA / Q000662:veterinary; D014616:Vaccinia virus / Q000145:classification / Q000302:isolation & purification
D015342:DNA Probes; D004076:Digoxigenin