Animal experiments:
To generate clinical samples for validation of the triplex assay, a
total of 19 Landrace x large white crossbred pigs (11 piglets at three
weeks of age and eight weaner pigs at seven weeks of age) were purchased
from a commercial swine farm in Southern Manitoba and transported to the
National Centre for Foreign Animal Disease (NCFAD) animal facility.
After seven days of acclimatization, pigs were randomly assigned to
three groups: Group 1 (eight 7-weeks old pigs for PRV-JS-2012, Pig#
5-12), Group 2 (eight 3 weeks-old piglets for PRV-JS-2012, pig # 13-20)
and Group 3 (three 3 weeks old piglets for PRV-Bristol, Pig # 24-26).
Each group was housed in an individual pen. Each pig in Group 1 and 2
received 2 ml of PRV Strain JS-2012 (105 TCID50/ml, 1
ml in each nostril) and each pig in Group 3 received 2 ml of PRV Bristol
strain (105 TCID50/ml, 1 ml in each nostril). After
the challenge, all animals were monitored twice daily for clinical signs
(fever, neurological, respiratory etc.); and oral fluid, nasal and
oropharyngeal swabs were collected every other day starting at 2 days
post infection (dpi) until 14 dpi. Pigs that reached humane endpoint
were euthanized and tissue samples (tonsils, spleen, lung, brain,
trigeminal ganglia, kidney, and lymph nodes) were collected. The
remaining animals were euthanized at 14 days post infection and similar
samples as above were collected. All the samples were stored at -80ÂșC
until tested. The animal experiment was approved by the Animal Care
Committee at the Canadian Science Centre for Human and Animal Health.
The guidelines from the Canadian Council for Animal Care were observed
during all procedures with animals.