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Diagnostic approach for cancer cells in urine sediments by 5-aminolevulinic acid-based photodynamic detection in bladder cancer.

Miyake M, Nakai Y, Anai S, Tatsumi Y, Kuwada M, Onishi S, Chihara Y, Tanaka N, Hirao Y, Fujimoto K - Cancer Sci. (2014)

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.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Nara Medical University, Kashihara-shi, Nara, Japan.

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Sensitivity of Cellular Fluorescence Analysis Unit (CFAU) in the detection of urothelial carcinoma. (a) T24 cells without ALA treatment were used as simulated non-cancerous cells. The mixed cell suspension (105 cells/mL) composed of untreated T24 cells and ALA-treated T24 cells at indicated ratios was prepared in PBS. Red arrows indicate the detectable fluorescent peaks emitted by ALA-treated cells. (b) T24 cells and urine samples from cases with bladder cancer were subjected to CFAU. Two representative cases are shown. One harbored a pTa/low-grade papillary-growing tumor, and the other harbored a pTis/high grade tumor. Black arrows indicate the background of urine sediments. Red arrows indicate the PPIX-specific fluorescence in comparison with non-ALA control.
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fig05: Sensitivity of Cellular Fluorescence Analysis Unit (CFAU) in the detection of urothelial carcinoma. (a) T24 cells without ALA treatment were used as simulated non-cancerous cells. The mixed cell suspension (105 cells/mL) composed of untreated T24 cells and ALA-treated T24 cells at indicated ratios was prepared in PBS. Red arrows indicate the detectable fluorescent peaks emitted by ALA-treated cells. (b) T24 cells and urine samples from cases with bladder cancer were subjected to CFAU. Two representative cases are shown. One harbored a pTa/low-grade papillary-growing tumor, and the other harbored a pTis/high grade tumor. Black arrows indicate the background of urine sediments. Red arrows indicate the PPIX-specific fluorescence in comparison with non-ALA control.

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.


Diagnostic approach for cancer cells in urine sediments by 5-aminolevulinic acid-based photodynamic detection in bladder cancer.

Miyake M, Nakai Y, Anai S, Tatsumi Y, Kuwada M, Onishi S, Chihara Y, Tanaka N, Hirao Y, Fujimoto K - Cancer Sci. (2014)

Sensitivity of Cellular Fluorescence Analysis Unit (CFAU) in the detection of urothelial carcinoma. (a) T24 cells without ALA treatment were used as simulated non-cancerous cells. The mixed cell suspension (105 cells/mL) composed of untreated T24 cells and ALA-treated T24 cells at indicated ratios was prepared in PBS. Red arrows indicate the detectable fluorescent peaks emitted by ALA-treated cells. (b) T24 cells and urine samples from cases with bladder cancer were subjected to CFAU. Two representative cases are shown. One harbored a pTa/low-grade papillary-growing tumor, and the other harbored a pTis/high grade tumor. Black arrows indicate the background of urine sediments. Red arrows indicate the PPIX-specific fluorescence in comparison with non-ALA control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317833&req=5

fig05: Sensitivity of Cellular Fluorescence Analysis Unit (CFAU) in the detection of urothelial carcinoma. (a) T24 cells without ALA treatment were used as simulated non-cancerous cells. The mixed cell suspension (105 cells/mL) composed of untreated T24 cells and ALA-treated T24 cells at indicated ratios was prepared in PBS. Red arrows indicate the detectable fluorescent peaks emitted by ALA-treated cells. (b) T24 cells and urine samples from cases with bladder cancer were subjected to CFAU. Two representative cases are shown. One harbored a pTa/low-grade papillary-growing tumor, and the other harbored a pTis/high grade tumor. Black arrows indicate the background of urine sediments. Red arrows indicate the PPIX-specific fluorescence in comparison with non-ALA control.
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.

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.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Nara Medical University, Kashihara-shi, Nara, Japan.

Show MeSH
Related in: MedlinePlus