Veterinary Research · 2014 · 108 citations · 28 references
Veterinary MedicineInfectious DiseasesMicrobiologyDiagnostic MicrobiologyMolecular VirologyVirologyAnimal Infectious DiseasesAnimal VirusesVeterinary Diagnostic MicrobiologyVeterinary VirologyZoonotic Disease DetectionCoronavirus ZoonosesFeline Infectious DiseasesFeline VirologyFeline VirusesMolecular Diagnosis From Tissue SamplesQuantitative Polymerase Chain ReactionQuantitative Polymerase Chain Reaction AssaysRna Extraction
Recent evidence suggests that a mutation in the spike protein gene of feline coronavirus (FCoV), which results in an amino acid change from methionine to leucine at position 1058, may be associated with feline infectious peritonitis (FIP). Tissue and faecal samples collected post mortem from cats diagnosed with or without FIP were subjected to RNA extraction and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) to detect FCoV RNA. In cats with FIP, 95% of tissue, and 81% of faecal samples were PCR-positive, as opposed to 22% of tissue, and 60% of faecal samples in cats without FIP. Relative FCoV copy numbers were significantly higher in the cats with FIP, both in tissues (P < 0.001) and faeces (P = 0.02). PCR-positive samples underwent pyrosequencing encompassing position 1058 of the FCoV spike protein. This identified a methionine codon at position 1058, consistent with the shedding of an enteric form of FCoV, in 77% of the faecal samples from cats with FIP, and in 100% of the samples from cats without FIP. In contrast, 91% of the tissue samples from cats with FIP and 89% from cats without FIP had a leucine codon at position 1058, consistent with a systemic form of FCoV. These results suggest that the methionine to leucine substitution at position 1058 in the FCoV spike protein is indicative of systemic spread of FCoV from the intestine, rather than a virus with the potential to cause FIP.
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Primer3 on the WWW for General Users and for Biologist Programmers
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