Identification of equine herpesvirus 3 (equine coital exanthema virus), equine gammaherpesviruses 2 and 5, equine adenoviruses 1 and 2, equine arteritis virus and equine rhinitis A virus by polymerase chain reaction.
OBJECTIVE
To develop rapid (< 8 hour) tests using polymerase chain reaction (PCR) for the diagnosis of equine herpesvirus 3 (EHV3; equine coital exanthema virus), equine gammaherpesviruses 2 (EHV2) and EHV5, equine adenovirus 1 (EAdV1), EAdV2, equine arteritis virus (EAV), equine rhinitis A virus (ERAV; formerly equine rhinovirus 1)


METHODS
Either single round or second round (seminested) PCRs were developed and validated.


METHODS
Oligonucleotide primers were designed that were specific for each virus, PCR conditions were defined and the specificity and sensitivity of the assays were determined. The application of the tests was validated using a number of independent virus isolates for most of the viruses studied. The PCRs were applied directly to clinical samples where samples were available.


RESULTS
We developed a single round PCR for the diagnosis of EHV3, a seminested PCR for EHV2 and single round PCRs for EHV5, EAdV1, EAdV2 and RT-PCRs for EAV and ERAV. The PCR primer sets for each virus were designed and shown to be highly specific (did not amplify any recognised non-target template) and sensitive (detection of minimal amounts of virus) and, where multiple virus isolates were available all isolates were detected.


CONCLUSIONS
The development and validation of a comprehensive panel of PCR diagnostic tests, predominantly for viruses causing equine respiratory disease, that can be completed within 8 hours from receipt of clinical samples, provides a major advance in the rapid diagnosis or exclusion diagnosis of these endemic equine virus diseases in Australia.
Dynon
K
K
Centre for Equine Virology, School of Veterinary Science, The University of Melbourne, Parkville, Victoria.
Varrasso
A
A
Ficorilli
N
N
Holloway
S
S
Reubel
G
G
Li
F
F
Hartley
C
C
Studdert
M
M
Drummer
H
H
D000256:Adenoviridae / Q000235:genetics / Q000302:isolation & purification; D000818:Animals; D005537:Aphthovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D017931:DNA Primers; D007240:Equartevirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D004858:Herpesvirus 3, Equid / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006734:Horse Diseases / Q000175:diagnosis* / Q000821:virology; D006736:Horses; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D018144:Rhadinovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D014777:Virus Diseases / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers