Comparison of a prototype reverse hybridization assay and MethyLight for detection of SFRP2 promotor methylation in fecal DNA



This study aimed to evaluate the diagnostic performance of a novel nonquantitative methylation-specific reverse hybridization (MSRH) assay to detect secreted frizzled-related protein 2 (SFRP2) promotor methylation in fecal DNA.


SFRP2 promoter methylation was investigated in stool DNA isolated from 18 colorectal cancer (CRC) patients and 22 healthy controls using the MSRH assay based on methylation-specific DNA amplification followed by reverse hybridization of biotinylated amplicons to sequence-specific methylation detection probes, with MethyLight serving as a reference method.


SFRP2 promotor methylation as determined by MSRH vs. MethyLight showed a sensitivity and specificity of 61.1% and 86.3% vs. 77.7% and 77.3%, respectively. Moderate agreement (ĸ = 0.54, 95% confidence interval [95% CI], 0.29-0.80, p<0.001) was observed between the 2 methods. However, the differences in SFRP2 promotor methylation observed between CRC patients and healthy individuals by both assays were statistically significant (p<0.001).


Our findings, although limited by the small sample size, do not support the use of the MSRH assay for CRC screening in stool.

Int J Biol Markers 2017; 32(4): e467 - e470




Gernot Kriegshäuser, Dietmar Enko, Matthias Zitt, Michael Oberwalder, Christian Oberkanins, Dietmar Öfner, Robert Zeillinger, Hannes Maximilian Müller

Article History


Financial support: No grants or funding have been received for this study.
Conflict of interest: None of the authors has any financial interest related to this study to disclose.

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  • Institute of Clinical Chemistry and Laboratory Medicine, General Hospital Steyr, Steyr - Austria
  • Clinical Institute of Medical and Laboratory Diagnostics, Medical University of Graz, Graz - Austria
  • Department of Surgery, General Hospital Dornbirn, Dornbirn - Austria
  • Department of Visceral, Transplant and Thoracic Surgery, Center of Operative Medicine, Medical University Innsbruck, Innsbruck - Austria
  • ViennaLab Diagnostics GmbH, Vienna - Austria
  • Department of Surgery, University Hospital Salzburg, Salzburg - Austria
  • OncoLab Diagnostics GmbH, Vienna - Austria
  • Department of Surgery, General Hospital Schwaz, Schwaz - Austria

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