XXXX marker panel with single cell sensitivity in urine.

@entity1 @entity5435 @entity263 @entity5 @entity6639 @entity25792 @entity21416
BACKGROUND: Over one million @entity1 undergo prostate biopsies annually in the United States, a majority of whom due to elevated serum PSA. More than half of the biopsies turn out to be negative for @entity263 (CaP). The limitations of both the PSA test and the biopsy procedure have led to the development for more precise CaP detection assays in urine (e.g., @entity21416 , @entity5435 -ERG) or blood (e.g., PHI, 4K). Here, we describe the development and evaluation of the Urine CaP Marker Panel (UCMP) assay for sensitive and reproducible detection of CaP cells in post-digital rectal examination (post-DRE) urine. METHODS: The cellular content of the post-DRE urine was captured on a translucent filter membrane, which is placed on Cytoclear slides for direct evaluation by microscopy and immuno-cytochemistry (ICC). Cells captured on the membrane were assayed for PSA and @entity25792 expression to identify prostate epithelial cells, and for ERG and @entity6639 to identify @entity263 cells. Immunostained cells were analyzed for quantitative and qualitative features and correlated with biopsy positive and negative status for @entity5 . RESULTS: The assay was optimized for single cell capture sensitivity and downstream evaluations by spiking a known number of cells from established CaP cell lines, LNCaP and VCaP, into pre-cleared control urine. The cells captured from the post-DRE urine of subjects, obtained prior to biopsy procedure, were co-stained for ERG, @entity6639 (CaP specific), and @entity25792 or PSA (prostate epithelium specific) rendering a whole cell based analysis and characterization. A feasibility cohort of 63 post-DRE urine specimens was assessed. Comparison of the UCMP results with blinded biopsy results showed an assay sensitivity of 64% (16 of 25) and a specificity of 68.8% (22 of 32) for CaP detection by biopsy. CONCLUSIONS: This pilot study assessing a minimally invasive CaP detection assay with single cell sensitivity cell-capture and characterization from the post-DRE urine holds promise for further development of this novel assay platform. Prostate 75: 969-975, 2015.    2015 The Authors. The Prostate, published by Wiley Periodicals, Inc.
