Evaluation of real-time RT-PCR for the quantification of FCoV shedding in the faeces of domestic cats.

ABSTRACT: Faecal samples were taken from cats living in multi-cat households with endemic feline coronavirus (FCoV) infection. Total RNA was extracted from faecal suspensions and FCoV RNA was quantified using a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay. The real-time RT-PCR threshold cycle (C(T)) values were consistently high suggesting that the samples contained very little viral RNA. However, experiments in which RNA extracted from FCoV-infected cell culture supernatants was combined with RNA extracted from faecal suspensions revealed the presence of faecal factors that significantly inhibited the reverse transcription reaction. Consequently, three methods of RNA extraction were investigated and RNA dilution was undertaken to investigate whether the effects of the fa

SUBMITTER: Dye C 

PROVIDER: S-EPMC2582154 | biostudies-literature | 2008 Apr

REPOSITORIES: biostudies-literature