Detection of feline immunodeficiency virus by a nested polymerase chain reaction.
A specific and sensitive polymerase chain reaction (PCR) procedure for the detection of feline immunodeficiency virus (FIV) in peripheral blood mononuclear cells (PBMC) was developed. PBMC from both blood samples and cultures were digested by proteinase K in a lysis buffer, and after heat inactivation of the proteinase, the resultant material was used in a two step amplification protocol using nested sets of primers. Two independent amplifications, from the gag and pol genes respectively, were performed in each tube. The PCR was positive for six of 14 samples from FIV seropositive adult cats, while all 36 samples from seronegative cats were negative. In comparison with an antigen-capturing ELISA procedure, the PCR detected FIV infection in PBMC cultures on average two days earlier.
Rimstad
E
E
Department of Microbiology and Immunology, Norwegian College of Veterinary Medicine, Oslo.
Ueland
K
K
D000818:Animals; D002415:Cats; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016181:Feline Acquired Immunodeficiency Syndrome / Q000175:diagnosis*; D016089:Immunodeficiency Virus, Feline / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*