In vitro detection of equine arteritis virus from seminal plasma for identification of carrier stallions.
Equine arteritis virus (EAV) was readily isolated in RK-13 cell monolayers by plaque assay from seminal plasma of experimental carrier stallions when they contained high titers of virus regardless of the presence of non-viral cytotoxicity in the seminal plasma. The cytotoxicity interfered with virus isolation from seminal plasma which contained virus at titers less than 10 PFU/ml. However, it was possible to detect the virus in seminal plasma pretreated with PEG (#6000). EAV was consistently identified by RT-PCR from crude seminal plasma which contained virus at titers of more than 10(2.7) PFU/ml. In vitro detection of EAV by virus isolation supplemented with RT-PCR using seminal plasma was proved to be an effective alternative to the standard test mating as a diagnostic method for carrier stallions.
Fukunaga
Y
Y
Epizootic Research Station, Equine Research Institute, Japan Racing Association, Kokubunji, Tochigi.
Wada
R
R
Sugita
S
S
Fujita
Y
Y
Nambo
Y
Y
Imagawa
H
H
Kanemaru
T
T
Kamada
M
M
Komatsu
N
N
Akashi
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D001167:Arteritis / Q000175:diagnosis / Q000517:prevention & control / Q000662:veterinary*; D002353:Carrier State / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D002460:Cell Line; D004279:DNA, Viral / Q000737:chemistry; D004197:Disease Reservoirs / Q000662:veterinary; D007240:Equartevirus / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D006734:Horse Diseases / Q000175:diagnosis* / Q000635:transmission / Q000821:virology; D006736:Horses; D008297:Male; D011092:Polyethylene Glycols / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012661:Semen / Q000821:virology*; D013739:Testosterone / Q000008:administration & dosage / Q000097:blood; D010948:Viral Plaque Assay; D017201:Virus Shedding
D000911:Antibodies, Monoclonal; D004279:DNA, Viral; D011092:Polyethylene Glycols; D013739:Testosterone