A real-time RT-PCR assay for molecular identification and quantitation of feline morbillivirus RNA from biological specimens

J Virol Methods. 2018 Aug:258:24-28. doi: 10.1016/j.jviromet.2018.05.002. Epub 2018 May 3.

Abstract

The aim of this study was to develop a real-time RT-PCR to detect and quantitate feline morbillivirus (FeMV) RNA in biological samples. Primers and probe were targeted on a conserved region of FeMV P/V/C gene. To validate the assay with field samples, a total number of specimens of cats have been recruited including 264 urine and blood samples and compared with a generic RT-PCR targeting the L protein encoding gene of morbilliviruses. In addition, 385 tissue samples from 35 carcasses of cats have been also employed. RNA titres were low in all tested samples. Results also indicated the absence of cross-reaction with related morbilliviruses and existing pathogens of cats. In tissues with low levels of FeMV RNA, the presence of viral antigen was also evidenced by immunohistochemistry targeting the N viral protein. This newly described assay allows for a rapid, accurate and reliable quantitative detection of FeMV RNA that can be applied for diagnostics and research studies.

Keywords: Cats; Feline morbillivirus; Immunohistochemistry; Real-time RT-PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cat Diseases / diagnosis*
  • Cat Diseases / virology*
  • Cats
  • DNA Primers / genetics
  • Molecular Diagnostic Techniques / methods*
  • Morbillivirus / genetics
  • Morbillivirus / isolation & purification*
  • Morbillivirus Infections / diagnosis
  • Morbillivirus Infections / veterinary*
  • Morbillivirus Infections / virology
  • Oligonucleotide Probes / genetics
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Time Factors

Substances

  • DNA Primers
  • Oligonucleotide Probes
  • RNA, Viral

Supplementary concepts

  • Feline morbillivirus