Development of TaqMan® Real-time Reverse Transcription- Polymerase Chain Reaction for the Quantification of Feline Leukemia Virus Load

Abstract

Feline leukemia virus (FeLV), a gamma retrovirus of the domestic cat, is not only of veterinary interest, but is also an important model for the study of pathogenesis of tumors and AIDS in many animals. After initial infection, disease progression has been associated with cellular and humoral immune response and FeLV proviral load. Moreover, an earlier study showed that not only the proviral load, but also the plasma viral loads are important parameters, which are associated with progression of the disease. In this study, the TaqMan^® real-time reverse transcription-polymerase chain reaction (RT-PCR) was developed for the quantification of FeLV loads. The assay was developed to amplify a 131 base pairs conserved domain within the unique region (U3) of a long terminal repeat (LTR). The detection limit of this assay was 8.3 copies of RNA standard template or 4.15 viruses per reaction that is equivalent to 1,778 viruses per ml of plasma or serum.