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Rapid and quantitative detection of CpG-methylation status using TaqMan PCR combined with methyl-binding-domain polypeptide.

Abstract：

OBJECTIVES:To assess the medical applicability of CpG methylation as molecular markers for cancer diagnosis, we established a new system to determine DNA methylation based on TaqMan PCR combined with a methyl-binding-domain polypeptide 2. DESIGN AND METHODS:We evaluated the diagnostic applicability of this approach by examining the methylation status of two tumor suppressor genes, RASSF1A and APC, in 10 paired hepatocellular carcinoma (HCC) and the corresponding non-tumor liver tissues. RESULTS:Methylation levels of total 20 clinical samples measured by the TaqMan PCR assay showed a significantly positive correlation (R=0.814, P<0.0005 for RASSF1A, R=0.736, P<0.00001 for APC) with those calculated by bisulfite sequencing. The methylated DNA amount measured by our TaqMan PCR system precisely replicated the methylation status estimated by direct sequencing. CONCLUSIONS:This suggests our method may serve as a reliable and easy-to-use tool for cancer diagnosis using methylated genes as biomarkers.

journal_name

Clin Biochem

journal_title

Clinical biochemistry

authors

Kimura N,Moribe T,Iizuka N,Miura T,Tamatsukuri S,Ishitsuka H,Hamamoto Y,Oka M

doi

10.1016/j.clinbiochem.2009.03.017

subject

Has Abstract

pub_date

2009-07-01 00:00:00

pages

1113-22

issue

10-11

eissn

0009-9120

issn

1873-2933

pii

S0009-9120(09)00148-9

journal_volume

42

pub_type

杂志文章

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