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: We used 50 mL of voided urine for each of the ALA-treated sediments and ALA-untreated sediments. After the treatment with ALA in serum-free media, urine sediments were obtained by centrifugation and resuspended in 500 μL of PBS. The resuspended sediments in PBS were transferred in triplicate (100 μL each) to a flat-bottom transparent 96-well plate. In each urine sample, ALA-treated sediments and untreated sediments were subjected to the spectrophotometric assay. Fluorescence emission spectra were recorded using a microplate spectrophotometer (Infinite 200M PRO; Tecan, Männedorf, Switzerland) equipped with i-control version 1.8 software. The excitation wavelength was 400 nm, and the photomultiplier tube voltage was fixed. A series of spectra obtained from each specimen (Fig.3) and the fluorescence intensity at 635 nm was measured. The values of ALA-treated sediments and those of the ALA-untreated control were compared. If all of the three intensity values of ALA-treated samples were higher than any values of ALA-untreated controls, the sample was considered positive for the spectrophotometric assay.
Affiliation: Department of Urology, Nara Medical University, Kashihara-shi, Nara, Japan.