Evaluation of a p30 gene-based real-time reverse transcriptase polymerase chain reaction assay for detection of feline caliciviruses.
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.
Scansen
Brian A
BA
Department of Small Animal Clinical Sciences, Michigan State University, College of Veterinary Medicine, East Lansing, MI 48824-1314, USA.
Wise
Annabel G
AG
Kruger
John M
JM
Venta
Patrick J
PJ
Maes
Roger K
RK
D000818:Animals; D017250:Caliciviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D017927:Calicivirus, Feline / Q000235:genetics* / Q000302:isolation & purification; D002371:Cat Diseases / Q000175:diagnosis* / Q000821:virology; D002415:Cats; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D011237:Predictive Value of Tests; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral