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Research ArticleExperimental Studies

SPMSP: A Solid-phase PCR with Colorimetric Read-out for Quantitative Methylation Analysis

SONJA SIEVERS, CARLA FRITZSCH, CORNELIUS KUHNEN and OLIVER MÜLLER
Anticancer Research July 2008, 28 (4B) 2055-2060;
SONJA SIEVERS
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  • For correspondence: sonja.sievers@mpi-dortmund.mpg.de
CARLA FRITZSCH
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CORNELIUS KUHNEN
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OLIVER MÜLLER
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Abstract

Background: As DNA methylation has been determined as an important diagnostic biomarker in cancer management, methods for the detection of quantitative DNA methylation which are high-throughput and low-cost are needed. Materials and Methods: In this work, we introduce the solid-phase methylation-specific PCR (SPMSP) as a method for quantitative methylation analysis. SPMSP combines methylation-specific DNA amplification on a solid phase with subsequent colorimetric detection in an ELISA-based format. Results: In contrast to existing methods for quantitative methylation analysis, SPMSP can be carried out with standard laboratory equipment, i.e. a thermocycler and an ELISA reader. With this method, DNA methylation in the promoter of the APC tumour suppressor gene was quantified in a set of colorectal carcinoma samples. Conclusion: As SPMSP can be modified to the analysis of other promoter sequences and is also adaptable to a high-throughput system, SPMSP offers a platform for methylation profiling with widespread applications in cancer research and management.

  • Solid-phase PCR
  • PCR-ELISA
  • quantitative DNA methylation analysis
  • promoter hypermethylation
  • quantitative PCR

Footnotes

  • Abbreviations: APC, adenomatous polyposis coli; ELISA, enzyme-linked immunosorbent assay; MSP, methylation-specific PCR; SPMSP, solid-phase methylation-specific PCR; qPCR, quantitative real-time PCR.

  • Received January 24, 2008.
  • Revision received April 8, 2008.
  • Accepted April 21, 2008.
  • Copyright© 2008 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved
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Anticancer Research: 28 (4B)
Anticancer Research
Vol. 28, Issue 4B
July-August 2008
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SPMSP: A Solid-phase PCR with Colorimetric Read-out for Quantitative Methylation Analysis
SONJA SIEVERS, CARLA FRITZSCH, CORNELIUS KUHNEN, OLIVER MÜLLER
Anticancer Research Jul 2008, 28 (4B) 2055-2060;

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SPMSP: A Solid-phase PCR with Colorimetric Read-out for Quantitative Methylation Analysis
SONJA SIEVERS, CARLA FRITZSCH, CORNELIUS KUHNEN, OLIVER MÜLLER
Anticancer Research Jul 2008, 28 (4B) 2055-2060;
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