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Clin Chim Acta. 2014 Apr 20;431:154-63. doi: 10.1016/j.cca.2014.01.044. Epub 2014 Feb 9.

Validation of methylation-sensitive high-resolution melting (MS-HRM) for the detection of stool DNA methylation in colorectal neoplasms.

Author information

1
Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Guangdong Provincial Key Laboratory of Gastroenterology, Guangzhou 510515, China.
2
Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Department of Gastroenterology, Huizhou First Hospital, Huizhou 516003, China.
3
Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Huizhou Medicine Institute, Huizhou 516003, China. Electronic address: angao62@21cn.com.
4
Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Guangdong Provincial Key Laboratory of Gastroenterology, Guangzhou 510515, China. Electronic address: helenwxy@smu.edu.cn.

Abstract

BACKGROUND:

Methylation-sensitive high-resolution melting (MS-HRM) is a new technique for assaying DNA methylation, but its feasibility for assaying stool in patients with colorectal cancer (CRC) is unknown.

METHODS:

First, the MS-HRM and methylation-specific PCR (MSP) detection limits were tested. Second, the methylation statuses of SFRP2 and VIM were analyzed in stool samples by MS-HRM, and in matching tumor and normal colon tissues via bisulfite sequencing PCR (BSP). Third, a case-control study evaluated the diagnostic sensitivity and specificity of MS-HRM relative to results obtained with MSP and the fecal immunochemical test (FIT). Finally, the linearity and reproducibility of MS-HRM were assessed.

RESULTS:

The detection limits of MS-HRM and MSP were 1% and 5%, respectively. The diagnostic sensitivities of MS-HRM (87.3%, 55/63) in stool and BSP in matching tumor tissue (92.1%, 58/63) were highly consistent (κ=0.744). The MS-HRM assay detected 92.5% (37/40) methylation in CRCs, 94.4% (34/36) in advanced adenomas, and 8.8% (5/57) in normal controls. The results of MS-HRM analysis were stable and reliable and showed fairly good linearity for both SFRP2 (P<0.001, R(2)=0.957) and VIM (P<0.001, R(2)=0.954).

CONCLUSIONS:

MS-HRM shows potential for CRC screening.

KEYWORDS:

Advanced adenoma; Cancer screening; Colorectal cancer; DNA methylation; Methylation-sensitive high-resolution melting (MS-HRM); SFRP2/VIM

PMID:
24518356
DOI:
10.1016/j.cca.2014.01.044
[Indexed for MEDLINE]

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