Abstract
This report describes a feline calicivirus (FCV) p30 gene‐based real‐time SYBR Green I reverse transcriptase polymerase chain reaction (RT‐PCR) assay that is capable of detecting low virus concentrations and a broad range of FCV isolates. The assay consisted of a 1‐step RT‐PCR reaction with primers delineating a 126‐base‐pair (bp) region of the FCV p30 gene. Sensitivity of the RT‐PCR assay was determined to be equivalent to a FCV titer of 1.2 × 101 to 1.2 × 102 TCID50/mL. The assay was linear over a wide range of template concentrations and had a reaction efficiency of 95%. Specific FCV amplification products were detected from 51 wild‐type FCV isolates, whereas specific products were not detected from a canine calicivirus, a rabbit calicivirus, and a bovine calicivirus. The primers used in this study amplified a large number of North American FCV isolates and further confirm the diagnostic utility of p30 gene‐based real‐time RT‐PCR for detection of FCV.
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