title: Development and application of a quantitative real-time PCR assay to detect feline leukemia virus RNA.
abstract: We previously defined four categories of feline leukemia virus (FeLV) infection, designated as abortive, regressive, latent, and progressive. To determine if detectable viral DNA is transcriptionally active in the absence of antigenemia, we developed and validated a real-time viral RNA qPCR assay. This assay proved to be highly sensitive, specific, reproducible, and allowed reliable quantitation. We then applied this methodology, together with real-time DNA qPCR and p27 capsid antigen capture ELISA, to examine cats challenged with FeLV. We found that circulating viral RNA and DNA levels were highly correlated and the assays were almost in perfect agreement. This indicates that the vast majority of viral DNA is transcriptionally active, even in the absence of antigenemia. The real-time qPCR assays are more sensitive than the most commonly used FeLV diagnostic assay, the p27 capsid antigen capture ELISA. Application of qPCR assays may add greater depth in understanding of FeLV-host relationships.
authors.affiliation: Department of Microbiology, Immunology, and Pathology, 1619 Campus Delivery, Colorado State University, Ft. Collins, CO 80523-1619, USA.
authors.forename: Andrea N
authors.initials: AN
authors.lastname: Torres
authors.forename: Kevin P
authors.initials: KP
authors.lastname: O'Halloran
authors.forename: Laurie J
authors.initials: LJ
authors.lastname: Larson
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Schultz
authors.forename: Edward A
authors.initials: EA
authors.lastname: Hoover
mesh_terms: D000818:Animals; D002415:Cats; D004279:DNA, Viral / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005256:Leukemia Virus, Feline / Q000235:genetics* / Q000302:isolation & purification; D016582:Leukemia, Feline / Q000097:blood / Q000821:virology*; D012367:RNA, Viral / Q000097:blood* / Q000737:chemistry / Q000235:genetics; D011897:Random Allocation; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral