Diagnostic approach for cancer cells in urine sediments by 5-aminolevulinic acid-based photodynamic detection in bladder cancer.
Bottom Line: The overall sensitivities of conventional cytology, BTA, NMP22, fluorescence cytology, fluorescent spectrophotometric assay and CFAU assay were 48%, 33%, 40%, 86%, 86% and 87%, respectively.Three different ALA-based assays showed high sensitivity and specificity.Development of a rapid and automated device for ALA-based photodynamic assay is necessary to avoid the variability induced by troublesome steps and low stability of specimens.
Affiliation: Department of Urology, Nara Medical University, Kashihara-shi, Nara, Japan.Show MeSH
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Mentions: To determine the detection sensitivity of CFAU, we carried out a preliminary experiment using T24 cells. Non-cancerous cells are supposed to be negative for red fluorescence after ALA exposure. In this experiment, untreated T24 cells were used as simulated non-cancerous cells without red fluorescence. ALA-treated T24 cells were used as fluorescence-positive cells (i.e. cancerous cells). The cells were suspended in PBS at a concentration of 105 cells/mL. Series of cell suspensions consisting of fluorescence-negative cells (untreated T24 cells) and fluorescence-positive cells (ALA-treated T24 cells) at various ratios were prepared as follows: 100:0, 50:50, 25:75, 10:90, 5:95 and 0:100. The cell suspensions were subjected to CFAU assay, and the spectra of emitted fluorescence were recorded by the equipped computer. No peak at the 635-nm wavelength was observed in a sample containing only untreated T24 cells (Fig.5a). However, an apparent peak at the wavelength of 635 nm could be detected by CFAU in samples containing more than 1.0 × 104 cells/mL of fluorescence-positive cells in a final concentration of 105 cells/mL. Subtle peaks were observed in samples containing 0.5 × 104 cells/mL of fluorescence-positive cells. According to the preliminary experiments using the cell line, the lower detection limit ranged from 0.5 to 1.0 × 104 cells/mL.
Affiliation: Department of Urology, Nara Medical University, Kashihara-shi, Nara, Japan.