Colorimetric detection of both total genomic and loci-specific DNA methylation from limited DNA inputs

Clinical Epigenetics

Matt Trau³  Thu Ha Ngo²  Eugene J. H. Wee¹ 

[1] Centre for Personalized NanoMedicine, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, St Lucia, Queensland, Australia;Faculty of Biotechnology, Vietnam National University of Agriculture, Hanoi, Vietnam;School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Queensland, Australia

关键词: GSTP1;    Electrochemical;    Colorimetric;    DNA methylation;    MBD enrichment;   

Others  :  1220622
DOI  :  10.1186/s13148-015-0100-6

 received in 2015-05-12, accepted in 2015-06-25,  发布年份 2015

【 摘 要 】

Background

Aberrant DNA methylation marks are potential disease biomarkers, and detecting both total genomic and gene-specific DNA methylation can aid in clinical decisions. While a plethora of methods exist in research, simpler, more convenient alternatives are needed to enhance both routine diagnostics and research.

Results

Herein, we describe colorimetric assays using methyl-binding domain (MBD) proteins for rapid and convenient evaluation of total genomic and gene-specific methylation from 50 ng or less DNA input in under 2 h. As little as 5 % methylation differences can be detected and are enhanced by a novel MBD protocol for improved specificity. Our assays could differentiate naïve from de-methylating drug-treated cells and detect the presence of a methylated prostate cancer biomarker in the urine. Finally, the assay was evolved onto disposable screen-printed electrodes for convenient detection of gene-specific methylation in urine.

Conclusions

Rapid MBD-based colorimetric and electrochemical approaches to detect DNA methylation from limited samples were successfully demonstrated and applied to clinical samples. We envision that the ease, low sample requirements and speed of these assays could have both clinical and research-wide applications.

【 授权许可】

   
2015 Wee et al.

【 图 表 】

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