Analytical specificity, sensitivity and variability testing:
Analytical specificity of the assay was determined using DNA extracted
from cell culture amplified PRV strains Bartha, Becker, Shope, Bristol
C61, and JS-2012, common swine viral pathogens encountered in modern
swine herds, and bovine herpesvirus-1 (BHV-1), a related herpesvirus.
For evaluating the intra-assay variability, 20 replicates of a single
sample were tested in a single run. The inter-assay variability was
calculated by running the same assay three times over 3 days. The
analytical sensitivity of the triplex assay was determined using known
concentration of PRV plasmid with net copy number of
7.34x1011. The limit of detection is interpreted as
the lowest copy number that can be detected by the assay.