Detection of feline coronavirus RNA in feces, tissues, and body fluids of naturally infected cats by reverse transcriptase PCR.
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.
Herrewegh
A A
AA
Department of Infectious Diseases and Immunology, Veterinary Faculty, University Utrecht, The Netherlands.
de Groot
R J
RJ
Cepica
A
A
Egberink
H F
HF
Horzinek
M C
MC
Rottier
P J
PJ
D000818:Animals; D001483:Base Sequence; D001826:Body Fluids / Q000821:virology; D002353:Carrier State / Q000821:virology; D002415:Cats; D002478:Cells, Cultured; D016765:Coronavirus, Feline / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology; D016766:Feline Infectious Peritonitis / Q000175:diagnosis / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D012367:RNA, Viral