Quantitation of feline leukaemia virus viral and proviral loads by TaqMan real-time polymerase chain reaction.
Feline leukaemia virus (FeLV) infection in cats is not only of veterinary importance but also a well-acknowledged animal model for studying the pathogenesis of retroviral disease. After virus exposure, different courses and outcomes of FeLV infection may prevail; they have been associated with cellular and humoral immune responses and the FeLV proviral load in peripheral blood. We hypothesized that the plasma viral RNA load might be an additional relevant indicator for the infection outcome. To quantify these loads, a real-time reverse transcriptase (RT) polymerase chain reaction (PCR) assay was developed. The assay amplifies FeLV-A, -B, and -C as some naturally infected cats could not be identified with a FeLV-A-based assay previously. The assay was applied to determine plasma FeLV RNA loads in cats infected both naturally and experimentally with FeLV. In addition, an improved real-time PCR assay for quantitation of FeLV proviral loads is described. The assays developed were more sensitive than ELISA and virus isolation in the early phase of infection. In addition, PCR allows the identification of provirus carriers that have overcome antigenaemia. Thus, for most effective detection of FeLV exposure and characterization of the infection in a cat, PCR assays are recommended as diagnostic tools.
Tandon
Ravi
R
Clinical Laboratory, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, CH-8057 Zurich, Switzerland.
Cattori
Valentino
V
Gomes-Keller
Maria Alice
MA
Meli
Marina L
ML
Golder
Matthew C
MC
Lutz
Hans
H
Hofmann-Lehmann
Regina
R
D000818:Animals; D002371:Cat Diseases / Q000175:diagnosis* / Q000821:virology; D002415:Cats; D017931:DNA Primers; D005256:Leukemia Virus, Feline / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification*; D012192:Retroviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D020079:Terminal Repeat Sequences / Q000235:genetics; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D019562:Viral Load
D017931:DNA Primers