Overexpression of prostate specific membrane antigen by canine hemangiosarcoma cells provides opportunity for the molecular detection of disease burdens within hemorrhagic body cavity effusions
Fig 5
Optimization of qualitative PCR methodology and dynamic range of detection for PSMA.
(A) Differential PSMA amplicon generation by qualitative PCR with the lowly expressing PSMA cHSA cell line (FITZ; red) compared with CAoEC (green) across different thermocycles ranging from 24 to 30. Determination of qualitative PCR sensitivity for detecting PSMA amplicons at 27 cycles with a logarithmic titration of (B) FITZ cells only or (C) when FITZ cells are spiked into whole blood. Lowest cell density capable that produces faintly visible amplicons is 104 FITZ cells, alone or in addition to whole blood nucleated cells. Amplicon production expressed as a ratio of PSMA/β actin pixels/area. Abbreviations FITZ (FZ; red) and CAoEC (CE; green).