title: A simple, low-cost, and rapid device for a DNA methylation-specific amplification/detection system using a flexible plastic and silicon complex.
abstract: Abnormal DNA methylation has been associated with the development and progression of several human cancers and is a potential target for treatment. Thus, myriad technologies for the analysis of DNA methylation have been developed over the past few decades. However, most of these technologies are still far from ideal because they are time-consuming, labor-intensive, and complex, and there is the risk of contamination of samples. Here, we present an innovative DNA methylation-specific amplification/detection device for analysis of DNA methylation in cancer-related DNA biomarkers. The assay is based on a microfluidic system that is coupled to a flexible plastic-based on-chip endonuclease digestion device with optimized magnetic field effect and a methylation-specific isothermal solid-phase amplification/detection technique to allow a low-cost, simple, and rapid analysis of DNA methylation status in a label-free and real-time manner. This flexible plastic/silicon-based microfluidic device is relatively simple to fabricate with a flexible thin film and a magnet array by using a laser machine that can overcome the limitations of a PDMS-based microfluidic device. We demonstrated the ability of the methylation analysis based on the proposed flexible device to detect the methylated RARβ gene, which is a common DNA methylation biomarker in several human cancers. The simple platform detected the methylated gene in genomic DNA from human cancer cell lines within 65 min, whereas other methods required at least several hours. Therefore, this simple, low-cost, and rapid methylation analysis platform will be useful for the detection of DNA methylation in point-of-care applications.
authors.affiliation: Institute of Microelectronics, A*STAR (Agency for Science, Technology and Research), 11 Science Park Road, Singapore Science Park II, Singapore 117685. shiny@ime.a-star.edu.sg.
authors.forename: Tae Yoon
authors.initials: TY
authors.lastname: Lee
authors.forename: Yong
authors.initials: Y
authors.lastname: Shin
authors.forename: Mi Kyoung
authors.initials: MK
authors.lastname: Park
mesh_terms: D002460:Cell Line; D004247:DNA / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D019175:DNA Methylation / Q000235:genetics*; D004867:Equipment Design; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation* / Q000379:methods; D010969:Plastics; D018583:Pliability; D012825:Silicon
chemical_list: D010969:Plastics; D004247:DNA; D012825:Silicon