A Molecular Urine Assay to Detect Recurrences During Surveillance of High-Risk Non-Muscle Invasive Bladder Cancer.
Author(s): de Jong JJ, de Jong FC, van der Made ACJ, van Casteren NJ, Roshani H, Oomens EHGM, Pelger RCM, Steyerberg EW, Boormans JL, Bangma CH, Zuiverloon TCM, Zwarthoff EC
Publication: Bladder Cancer, 2024, Vol. 10, Page 233-242
PubMed ID: 39493819 PubMed Review Paper? No
Purpose of Paper
This paper compared the sensitivity and specificity of a urine genotyping assay with cytological results and between case-matched urine specimens collected in the morning with those collected in the evening from patients with histologically or radiologically confirmed bladder cancer.
Conclusion of Paper
Of the 736 matched pairs, one or more mutations were detected in 204 evening urine specimens and 189 morning urine specimens, with negative results in 501 evening and 514 morning specimens. There were a total of 63 detected tumor reoccurrences, 53 (84%) were confirmed histologically and 4 radiologically. The urine assay detected 75% of the reoccurrences, with a specificity of 70%. The sensitivity was marginally higher using evening than morning urine (70% versus 66%), but the specificity was slightly lower (75% for evening urine versus 77% for morning urine). Cytology results were available for 439 of the urine pairs, and the technique had a sensitivity of 45% and specificity of 89% for detecting reoccurrence; notably, the urine genotyping assay had a sensitivity of 87% and specificity of 64% in the same specimens . Assay scores for OTX1 and TERT were associated with tumor reoccurrence in a multivariable generalized mixed effects model (P = 0.005 and P=0.004, respectively), but no significant association was identified for FGFR3. Finally, there was a significant association between a positive result of the urine genotype assay and recurrence during follow-up (Hazard ratio=3.47, P=8.21 x10-8).
Studies
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Study Purpose
This study compared the sensitivity and specificity of a urine genotyping assay with cytological results and between case-matched urine specimens collected in the morning with those collected in the evening from patients with histologically or radiologically confirmed bladder cancer. Multiple matched morning and evening urine specimens were self-collected by 204 patients (742 pairs) with high-risk non-muscle invasive bladder cancer (HR-NMIBC) into containers with Sedi-tect preservative and returned by mail. Upon arrival, urine was centrifuged at 2,000 g for 10 min and the cell pellet was resuspended in phosphate buffered saline (PBS). The specimen was recentrifuged at 6,000 rpm for 5 min and the pellet was stored without fluid at -20°C until DNA extraction with the Puregene DNA Isolation Kit. DNA was quantified by Qubit fluorometry. Mutations in the FGFR3 (R248C, S249C, G372C, Y375C and A393E), and hTERT (C>T at 1295228, 1295242 and 1295250) promoters were identified using the SNaPshot single nucleotide extension reaction. DNA was bisulfite converted before methylation analysis of the OTX1 promoter using SNaPshot.
Summary of Findings:
Of the 736 matched pairs, one or more mutations were detected in 204 evening urine specimens and 189 morning urine specimens, with negative results in 501 evening and 514 morning specimens. There were a total of 63 detected tumor reoccurrences, 53 (84%) were confirmed histologically and 4 radiologically. The urine assay detected 75% of the reoccurrences, with a specificity of 70%. The sensitivity was marginally higher using evening than morning urine (70% versus 66%), but the specificity was slightly lower (75% for evening urine versus 77% for morning urine). Cytology results were available for 439 of the urine pairs, and the technique had a sensitivity of 45% and specificity of 89% for detecting reoccurrence; notably, the urine genotyping assay had a sensitivity of 87% and specificity of 64% in the same specimens . Assay scores for OTX1 and TERT were associated with tumor reoccurrence in a multivariable generalized mixed effects model (P = 0.005 and P=0.004, respectively), but no significant association was identified for FGFR3. Finally, there was a significant association between a positive result of the urine genotype assay and recurrence during follow-up (Hazard ratio=3.47, P=8.21 x10-8).
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA Fluorometry DNA Bisulfite conversion assay DNA SNP assay Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Time of biospecimen collection Morning
Evening
SNP assay Specific Targeted nucleic acid OTX1
TERT
FGFR3
Preaquisition Diagnosis/ patient condition Histologically confirmed reoccurrence
Radiologically confirmed reoccurrence
Positive urine genotyping assay
Cytology confirmed reoccurrence