Journal of Clinical Microbiology · 1995 · 215 citations · 29 references
Veterinary MedicineInfectious DiseasesMicrobiologyDiagnostic MicrobiologyMolecular VirologyVirologyAnimal Infectious DiseasesPathogen DetectionVeterinary Diagnostic MicrobiologyVeterinary VirologyViral DiagnosticsViral PathogenesisAscitic Fluid AnalysisCoronavirus DiagnosticsCoronavirus ZoonosesFeline Infectious DiseasesFeline VirologyFeline Viruses
A nested reverse transcriptase PCR (RT-nPCR) was developed for the detection of feline coronavirus (FCoV) RNA in the feces, tissues, and body fluids of infected cats. The RT-nPCR was targeted to the highly conserved 3'-untranslated region of the viral genome and will detect most, if not all, feline coronaviruses in the field. With the RT-nPCR, FCoV RNA was detected in plasma samples from experimentally infected cats as early as 2 days postinoculation. FCoV RNA was also detected in serum, plasma, or ascitic fluid samples from 14 of 18 cats (78%) with naturally occurring feline infectious peritonitis (FIP). The use of RT-PCR for FIP diagnosis is limited because of the occurrence of apparently healthy FCoV carriers. These asymptomatic cats shed the virus in the feces and, in a number of cases, also had detectable virus in the plasma. Because of the nature of FCoV infections, our RT-PCR assay with plasma or serum cannot be used to establish a definite diagnosis of FIP. However, this assay does provide a new means to identify asymptomatic FCoV carriers. As such, RT-nPCR will be of use to screen cats before their introduction into FCoV-free catteries. Moreover, this assay provides an important tool to study the epidemiology of FCoV.
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Rapid and simple method for purification of nucleic acids
René Boom, C. J. A. Sol, M M Salimans et al. · Journal of Clinical Microbiology · 1990 · 5.3K citations · Full text