Imperial College London
Alternate

DrPanagiotisVorkas

Faculty of Medicine, Department of Surgery & Cancer

Research Associate
 
 
 
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Contact

 

p.vorkas

 
 
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Location

 

Sir Alexander Fleming BuildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Dimitrakopoulos:2012:10.1186/1471-2407-12-486,
author = {Dimitrakopoulos, L and Vorkas, PA and Georgoulias, V and Lianidou, ES},
doi = {10.1186/1471-2407-12-486},
journal = {BMC Cancer},
pages = {486--486},
title = {A closed-tube methylation-sensitive high resolution melting assay (MS-HRMA) for the semi-quantitative determination of CST6 promoter methylation in clinical samples},
url = {http://dx.doi.org/10.1186/1471-2407-12-486},
volume = {12},
year = {2012}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - BackgroundCST6 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.MethodsWe 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.ResultsThe 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.ConclusionsThe 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.
AU  - Dimitrakopoulos,L
AU  - Vorkas,PA
AU  - Georgoulias,V
AU  - Lianidou,ES
DO  - 10.1186/1471-2407-12-486
EP  - 486
PY  - 2012///
SN  - 1471-2407
SP  - 486
TI  - A closed-tube methylation-sensitive high resolution melting assay (MS-HRMA) for the semi-quantitative determination of CST6 promoter methylation in clinical samples
T2  - BMC Cancer
UR  - http://dx.doi.org/10.1186/1471-2407-12-486
UR  - http://hdl.handle.net/10044/1/21534
VL  - 12
ER  -
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