title: Validation of methylation-sensitive high-resolution melting (MS-HRM) for the detection of stool DNA methylation in colorectal neoplasms.
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.
authors: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Guangdong Provincial Key Laboratory of Gastroenterology, Guangzhou 510515, China.
authors: Zhujun
authors: Z
authors: Xiao
authors: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Department of Gastroenterology, Huizhou First Hospital, Huizhou 516003, China.
authors: Bingsheng
authors: B
authors: Li
authors: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Guangdong Provincial Key Laboratory of Gastroenterology, Guangzhou 510515, China.
authors: Guozhen
authors: G
authors: Wang
authors: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Guangdong Provincial Key Laboratory of Gastroenterology, Guangzhou 510515, China.
authors: Weisi
authors: W
authors: Zhu
authors: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Guangdong Provincial Key Laboratory of Gastroenterology, Guangzhou 510515, China.
authors: Zhongqiu
authors: Z
authors: Wang
authors: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Guangdong Provincial Key Laboratory of Gastroenterology, Guangzhou 510515, China.
authors: Jinfeng
authors: J
authors: Lin
authors: Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Huizhou Medicine Institute, Huizhou 516003, China. Electronic address: angao62@21cn.com.
authors: Angao
authors: A
authors: Xu
authors: 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.
authors: Xinying
authors: X
authors: Wang
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics; D015179:Colorectal Neoplasms / Q000175:diagnosis*; D019175:DNA Methylation / Q000235:genetics*; D005243:Feces / Q000737:chemistry; D005260:Female; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D008745:Methylation; D008875:Middle Aged; D009780:Occult Blood; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D013447:Sulfites / Q000737:chemistry; D014746:Vimentin / Q000235:genetics; D000094742:Secreted Frizzled-Related Proteins
chemical_list: D014408:Biomarkers, Tumor; D008565:Membrane Proteins; D013447:Sulfites; D014746:Vimentin; C109618:SFRP2 protein, human; D000094742:Secreted Frizzled-Related Proteins; C009279:sodium bisulfite
keywords: Advanced adenoma; Cancer screening; Colorectal cancer; DNA methylation; Methylation-sensitive high-resolution melting (MS-HRM); SFRP2/VIM