A closed-tube methylation-sensitive high resolution melting assay (MS-HRMA) for the semi-quantitative determination of CST6 promoter methylation in clinical samples.
A closed-tube methylation-sensitive high resolution melting assay (MS-HRMA) for the semi-quantitative determination of CST6 promoter methylation in clinical samples.
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DOI：
10.1186/1471-2407-12-486
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发表时间：
2012-10-22
期刊：
影响因子：
3.8
通讯作者：
Lianidou ES
中科院分区：
文献类型：
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作者：
Dimitrakopoulos L;Vorkas PA;Georgoulias V;Lianidou ES
CST6 promoter is highly methylated in cancer, and its detection can provide important prognostic information in breast cancer patients. The aim of our study was to develop a Methylation-Sensitive High Resolution Melting Analysis (MS-HRMA) assay for the investigation of CST6 promoter methylation. We designed primers that amplify both methylated and unmethylated CST6 sequences after sodium bisulfate (SB) treatment and used spiked control samples of fully methylated to unmethylated SB converted genomic DNA to optimize the assay. We first evaluated the assay by analyzing 36 samples (pilot training group) and further analyzed 80 FFPES from operable breast cancer patients (independent group). MS-HRMA assay results for all 116 samples were compared with Methylation-Specific PCR (MSP) and the results were comparable. The developed assay is highly specific and sensitive since it can detect the presence of 1% methylated CST6 sequence and provides additionally a semi-quantitative estimation of CST6 promoter methylation. CST6 promoter was methylated in 39/80 (48.75%) of FFPEs with methylation levels being very different among samples. MS-HRMA and MSP gave comparable results when all samples were analyzed by both assays. The developed MS-HRMA assay for CST6 promoter methylation is closed tube, highly sensitive, cost-effective, rapid and easy-to-perform. It gives comparable results to MSP in less time, while it offers the advantage of additionally providing an estimation of the level of methylation.
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影响因子：
3.8
作者：
Krypuy M;Newnham GM;Thomas DM;Conron M;Dobrovic A
通讯作者：
Dobrovic A
DOI：
10.1186/bcr2783
发表时间：
2010
期刊：
Breast cancer research : BCR
影响因子：
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作者：
Ko E;Park SE;Cho EY;Kim Y;Hwang JA;Lee YS;Nam SJ;Bang S;Park J;Kim DH
通讯作者：
Kim DH
影响因子：
19.7
作者：
Kundel, HL;Polansky, M
通讯作者：
Polansky, M
影响因子：
11.2
作者：
Lin HJ;Zuo T;Lin CH;Kuo CT;Liyanarachchi S;Sun S;Shen R;Deatherage DE;Potter D;Asamoto L;Lin S;Yan PS;Cheng AL;Ostrowski MC;Huang TH
通讯作者：
Huang TH
DOI：
10.1038/labinvest.2008.66
发表时间：
2008-09
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
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作者：
通讯作者：
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