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Updated: Aug 29, 2026
Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses
Published on: May 2, 2019
Evaluation of a p30 gene-based real-time reverse transcriptase polymerase chain reaction assay for detection of
Brian A Scansen1, Annabel G Wise, John M Kruger
1Department of Small Animal Clinical Sciences, Michigan State University, College of Veterinary Medicine, East Lansing, MI 48824-1314, USA.
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 x 10(1) to 1.2 x 10(2) 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.
