Evaluation of cell-free DNA in urine as a marker for bladder cancer diagnosis

Matelda Zancan, Francesca Galdi, Fulvio Di Tonno, Chiara Mazzariol, Claudio Orlando, Francesca Malentacchi, Marco Agostini, Michela Maran, Paola Del Bianco, Aline S C Fabricio, Bruno Murer, Carlo Pianon, Massimo Gion

Research output: Contribution to journalArticlepeer-review

Abstract

The diagnosis and follow-up of bladder cancer are mainly based on cystoscopy, an invasive method which could be negative in case of flat malignancies such as carcinoma in situ. Other noninvasive diagnostic methods have not yet given satisfactory results. There is a need for a reliable yet noninvasive method for the detection of bladder cancer. Our aim was to investigate whether cell-free DNA quantified in urine (ucf-DNA) could be a useful marker for the diagnosis of bladder cancer. A standard urine test was performed in 150 naturally voided morning urine samples that were processed to obtain a quantitative evaluation of ucf-DNA. Leukocyturia and/or bacteriuria were found in 18 subjects, who were excluded from the study. Statistical analysis was performed on 45 bladder cancer patients and 87 healthy subjects. Ucf-DNA was extracted from urine samples by a spin column-based method and quantified using four different methods: GeneQuant Pro (Amersham Biosciences, Pittsburg, PA, USA), Quant-iT™ DNA high-sensitivity assay kit (Invitrogen, Carlsbad, CA, USA), Real-Time PCR (Applied Biosystems, Foster City, CA, USA), and NanoDrop 1000 (NanoDrop Technologies, Houston, TX, USA). Median free DNA quantification did not differ statistically between bladder cancer patients and healthy subjects. A receiver-operating characteristic (ROC) curve was developed to evaluate the diagnostic performance of ucf-DNA quantification for each method. The area under the ROC curve was 0.578 for GeneQuant Pro, 0.573 for the Quant-iT™ DNA highsensitivity assay kit, 0.507 for Real-Time PCR, and 0.551 for NanoDrop 1000, which indicated that ucf-DNA quantification by these methods is not able to discriminate between the presence and absence of bladder cancer. No association was found between ucf-DNA quantification and tumor size or tumor focality. In conclusion, ucf-DNA isolated by a spin column-based method and quantified by GeneQuant Pro, Quant-iT™ DNA high-sensitivity assay kit, Real-Time PCR or NanoDrop 1000 does not seem to be a reliable marker for the diagnosis of bladder cancer.

Original languageEnglish
Pages (from-to)147-155
Number of pages9
JournalInternational Journal of Biological Markers
Volume24
Issue number3
Publication statusPublished - 2009

Keywords

  • Bladder cancer
  • Diagnosis
  • ucf-DNA
  • Urine

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cancer Research
  • Oncology
  • Pathology and Forensic Medicine

Fingerprint

Dive into the research topics of 'Evaluation of cell-free DNA in urine as a marker for bladder cancer diagnosis'. Together they form a unique fingerprint.

Cite this