Evaluation of a p30 gene-based real-time reverse transcriptase polymerase chain reaction assay for detection of feline caliciviruses

• Published in: Journal of veterinary internal medicine Vol. 18; no. 1; pp. 135 - 138
• Main Authors: Scansen, B.A, Wise, A.G, Kruger, J.M, Venta, P.J, Maes, R.K
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 2004
• Subjects: Animals; Caliciviridae Infections - diagnosis; Caliciviridae Infections - veterinary; Calicivirus; Calicivirus, Feline - genetics; Calicivirus, Feline - isolation & purification; Canine calicivirus; Cat Diseases - diagnosis; Cat Diseases - virology; Cats; cDNA; DNA Primers; DNA, Viral - analysis; Feline calicivirus; Predictive Value of Tests; Quantitative; reverse transcriptase polymerase chain reaction; Reverse Transcriptase Polymerase Chain Reaction - veterinary; RNA; Sensitivity and Specificity; SYBR Green I
• ISSN: 0891-6640; 1939-1676
• DOI: 10.1111/j.1939-1676.2004.tb00150.x

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.