Real-time reverse transcription polymerase chain reaction assay to detect a broad range of feline calicivirus isolates

This report describes a real-time reverse transcription polymerase chain reaction (RT-PCR) assay with SYBR® Green targeting the VP2 (ORF 3) of feline caliciviruses. All of the 44 feline calicivirus isolates tested were detected, whereas neither feline herpesvirus, feline panleukopenia virus, feline immunodeficiency virus and feline leukemia virus nor other calicivirus like rabbit hemorrhagic disease virus and a canine calicivirus isolate showed specific amplification products. The sensitivity of the SYBR® Green reaction was shown to be equivalent to 5 × 101 to 5 × 102 copies/reaction and the overall sensitivity equivalent to a feline calicivirus titer of 100.6 TCID50/100 μl in Crandell Reese Feline Kidney (CRFK) cells.