• Author: Hornyák, Ákos

    Lund University

  • Author: Bálint, Ádám

    Department of Virology, Central Agricultural Office Veterinary Diagnostic Directorate, Hungary

  • Author: Farsang, Attila

    Central Agricultural Office Directorate of Veterinary Medicinal Products, Hungary

  • Author: Balka, Gyula

    Department of Pathology and Forensic Veterinary Medicine, Faculty of Veterinary Science, Szent Istvan University, Hungary

  • Author: Hakhverdyan, Mikhayil

    Lund University

  • Author: Rasmussen, Thomas Bruun

    Sektion for Eksotiske Virussygdomme, Division of Virology, National Veterinary Institute, Technical University of Denmark, Lindholm, 4771, Kalvehave, Denmark

  • Author: Blomberg, Jonas

    Uppsala University, Sweden

  • Author: Belák, Sándor

    Lund University

View graph of relations

Feline infectious peritonitis is one of the most severe devastating diseases of the Felidae. Upon the appearance of clinical signs, a cure for the infected animal is impossible. Therefore rapid and proper diagnosis for both the presence of the causative agent, feline coronavirus (FCoV) and the manifestation of feline infectious peritonitis is of paramount importance. In the present work, a novel real-time RT-PCR method is described which is able to detect FCoV and to determine simultaneously the quantity of the viral RNA. The new assay combines the M gene subgenomic messenger RNA (sg-mRNA) detection and the quantitation of the genome copies of FCoV. In order to detect the broadest spectrum of potential FCoV variants and to achieve the most accurate results in the detection ability the new assay is applying the primer-probe energy transfer (PriProET) principle. This technology was chosen since PriProET is very robust to tolerate the nucleotide substitutions in the target area. Therefore, this technology provides a very broad-range system, which is able to detect simultaneously many variants of the virus(es) even if the target genomic regions show large scale of variations. The detection specificity of the new assay was proven by positive amplification from a set of nine different FCoV strains and negative from the tested non-coronaviral targets. Examination of faecal samples of healthy young cats, organ samples of perished animals, which suffered from feline infectious peritonitis, and cat leukocytes from uncertain clinical cases were also subjected to the assay. The sensitivity of the P-sg-QPCR method was high, since as few as 10 genome copies of FCoV were detected. The quantitative sg-mRNA detection method revealed more than 10–50,000 times increase of the M gene sg-mRNA in organ materials of feline infectious peritonitis cases, compared to those of the enteric FCoV variants present in the faeces of normal, healthy cats. These results indicate the applicability of the new P-sg-QPCR test as a powerful novel tool for the better detection and quantitation of FCoV and for the improved diagnosis of feline infectious peritonitis, this important disease of the Felidae, causing serious losses in the cat populations at a global scale.
Original languageEnglish
JournalJournal of Virological Methods
Publication date2012
Volume181
Issue2
Pages155-163
ISSN0166-0934
DOIs
StatePublished
CitationsWeb of Science® Times Cited: 1

Keywords

  • Feline infectious peritonitis, Sub-genomic messenger RNA, PriProET, Quantitative real-time PCR
Download as:
Download as PDF
Select render style:
APAAuthorCBEHarvardMLAStandardVancouverShortLong
PDF
Download as HTML
Select render style:
APAAuthorCBEHarvardMLAStandardVancouverShortLong
HTML
Download as Word
Select render style:
APAAuthorCBEHarvardMLAStandardVancouverShortLong
Word

ID: 7727080