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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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Related in: MedlinePlus

ALA-based fluorescence detection assay protocol. Urine samples were subjected to urinary tests such as urianalysis and conventional cytology. The remaining urine samples and detached T24 cells were analyzed in comparison with the untreated samples. Because photobleaching of protoporphyrin IX can be observed during light exposure, the experimental process for the 5-ALA treatment was performed in the dark. ALA, aminolevulinic acid; BTA, bladder tumor antigen; NMP22, nuclear matrix protein 22.
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fig01: ALA-based fluorescence detection assay protocol. Urine samples were subjected to urinary tests such as urianalysis and conventional cytology. The remaining urine samples and detached T24 cells were analyzed in comparison with the untreated samples. Because photobleaching of protoporphyrin IX can be observed during light exposure, the experimental process for the 5-ALA treatment was performed in the dark. ALA, aminolevulinic acid; BTA, bladder tumor antigen; NMP22, nuclear matrix protein 22.

Mentions: Before TURBT, 200–300 mL of voided urine was obtained for conventional cytology, a urine BTA test, a urine NMP22 test and fluorescence cytology. Fluorescence cytology was performed within 2 h after sample collection using the following procedures (Fig.1). Voided urine samples obtained from 20 benign cases were used as controls. Voided urine was centrifuged for 5 min at 400g. The cell pellet was washed using PBS. After centrifugation, the pellet was resuspended in serum-free culture medium, RPMI-1640 containing 1 mM of ALA. The cell suspension was then incubated for 2 h at 37°C to allow cells to produce and accumulate intracellular PPIX. After treatment with ALA, subsequent procedures were performed under light-protected conditions.


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)

ALA-based fluorescence detection assay protocol. Urine samples were subjected to urinary tests such as urianalysis and conventional cytology. The remaining urine samples and detached T24 cells were analyzed in comparison with the untreated samples. Because photobleaching of protoporphyrin IX can be observed during light exposure, the experimental process for the 5-ALA treatment was performed in the dark. ALA, aminolevulinic acid; BTA, bladder tumor antigen; NMP22, nuclear matrix protein 22.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig01: ALA-based fluorescence detection assay protocol. Urine samples were subjected to urinary tests such as urianalysis and conventional cytology. The remaining urine samples and detached T24 cells were analyzed in comparison with the untreated samples. Because photobleaching of protoporphyrin IX can be observed during light exposure, the experimental process for the 5-ALA treatment was performed in the dark. ALA, aminolevulinic acid; BTA, bladder tumor antigen; NMP22, nuclear matrix protein 22.
Mentions: Before TURBT, 200–300 mL of voided urine was obtained for conventional cytology, a urine BTA test, a urine NMP22 test and fluorescence cytology. Fluorescence cytology was performed within 2 h after sample collection using the following procedures (Fig.1). Voided urine samples obtained from 20 benign cases were used as controls. Voided urine was centrifuged for 5 min at 400g. The cell pellet was washed using PBS. After centrifugation, the pellet was resuspended in serum-free culture medium, RPMI-1640 containing 1 mM of ALA. The cell suspension was then incubated for 2 h at 37°C to allow cells to produce and accumulate intracellular PPIX. After treatment with ALA, subsequent procedures were performed under light-protected conditions.

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