title: Feline coronavirus quantitative reverse transcriptase polymerase chain reaction on effusion samples in cats with and without feline infectious peritonitis.
abstract: Objectives The aim of the study was to determine whether feline coronavirus (FCoV) RNA in effusion samples can be used as a diagnostic marker of feline infectious peritonitis (FIP); and in FCoV RNA-positive samples to examine amino acid codons in the FCoV spike protein at positions 1058 and 1060 where leucine and alanine, respectively, have been associated with systemic or virulent (FIP) FCoV infection. Methods Total RNA was extracted from effusion samples from 20 cats with confirmed FIP and 23 cats with other diseases. Feline coronavirus RNA was detected using a reverse transcriptase quantitative polymerase chain reaction assay (qRT-PCR), and positive samples underwent pyrosequencing of position 1058 with or without Sanger sequencing of position 1060 in the FCoV spike protein. Results Seventeen (85%) of the effusion samples from 20 cats with FIP were positive for FCoV RNA, whereas none of the 23 cats with other diseases were positive. Pyrosequencing of the 17 FCoV-positive samples showed that 11 (65%) of the cats had leucine and two (12%) had methionine at position 1058. Of the latter two samples with methionine, one had alanine at position 1060. Conclusions and relevance A positive FCoV qRT-PCR result on effusions appears specific for FIP and may be a useful diagnostic marker for FIP in cats with effusions. The majority of FCoVs contained amino acid changes previously associated with systemic spread or virulence (FIP) of the virus.
authors.affiliation: 1 The Feline Centre, Langford Veterinary Services and School of Veterinary Sciences, University of Bristol, Langford, Bristol, UK.
authors.forename: Louise
authors.initials: L
authors.lastname: Longstaff
authors.affiliation: 2 School of Veterinary Sciences, University of Bristol, Langford, Bristol, UK.
authors.forename: Emily
authors.initials: E
authors.lastname: Porter
authors.affiliation: 1 The Feline Centre, Langford Veterinary Services and School of Veterinary Sciences, University of Bristol, Langford, Bristol, UK.|Current address: Clinical Sciences and Services, Royal Veterinary College, Royal College Street, London, UK.
authors.forename: Victoria J
authors.initials: VJ
authors.lastname: Crossley
authors.affiliation: 2 School of Veterinary Sciences, University of Bristol, Langford, Bristol, UK.
authors.forename: Sophie E
authors.initials: SE
authors.lastname: Hayhow
authors.affiliation: 3 Molecular Diagnostic Unit, Langford Veterinary Services, Langford, Bristol, UK.
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Helps
authors.affiliation: 1 The Feline Centre, Langford Veterinary Services and School of Veterinary Sciences, University of Bristol, Langford, Bristol, UK.|3 Molecular Diagnostic Unit, Langford Veterinary Services, Langford, Bristol, UK.
authors.forename: Séverine
authors.initials: S
authors.lastname: Tasker
mesh_terms: D000818:Animals; D002415:Cats; D016765:Coronavirus, Feline / Q000302:isolation & purification*; D016766:Feline Infectious Peritonitis / Q000175:diagnosis*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*
chemical_list: D012367:RNA, Viral