Bisulfite conversion-specific and methylation-specific PCR: a sensitive technique for accurate evaluation of CpG methylation

Biochem Biophys Res Commun. 2003 Sep 19;309(2):305-9. doi: 10.1016/j.bbrc.2003.08.005.

Abstract

Methylation-specific PCR (MSP) is frequently used to distinguish methylated alleles in the genome. Sequences that have been incompletely converted during bisulfite treatment are frequently co-amplified during MSP. For accurate MSP, it is important to detect methylated sequences in a background of unconverted DNA with a high level of sensitivity. We report here sensitive techniques, bisulfite conversion-specific MSP (BS-MSP) to accurately evaluate CpG methylation. BS-MSP provides accurate results across a wide spectrum of bisulfite conversion levels. BS-MSP is also confirmed to be a useful technique for the routine analysis of clinical tumor specimens that were paraffin-embedded and microdissected. BS-MSP thus provides the powerful features of ease of use and compatibility with paraffin sections. We recommend that methylation analysis should include a step to eliminate unconverted DNA to avoid overestimation of the DNA methylation level in the samples.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.
  • Validation Study

MeSH terms

  • Cadherins / genetics*
  • Cadherins / metabolism
  • CpG Islands / genetics*
  • DNA Methylation*
  • DNA, Neoplasm / genetics*
  • Polymerase Chain Reaction / methods*
  • Reference Values
  • Reproducibility of Results
  • Restriction Mapping / methods*
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods
  • Sulfites
  • Urinary Bladder Neoplasms / genetics
  • Urinary Bladder Neoplasms / metabolism*

Substances

  • Cadherins
  • DNA, Neoplasm
  • Sulfites
  • sodium bisulfite