title: Real-time, label-free isothermal solid-phase amplification/detection (ISAD) device for rapid detection of genetic alteration in cancers.
abstract: Here, we first present an isothermal solid-phase amplification/detection (ISAD) technique for the detection of single-point mutations that can be performed without labelling in real-time by utilizing both silicon microring-based solid-phase amplification and isothermal recombinase polymerase amplification (RPA). The ISAD technique was performed on a silicon microring device with a plastic chamber containing 10 μL of the reaction mixture, and characterized with an assay for the detection of the HRAS (Harvey RAS) gene single-point mutation. For the solid-phase amplification, the primer of the gene was directly attached to the surface of the device via an amine modification reaction. The amplified DNA was detected, without a label, by measuring the optical wavelength shift of the silicon microring resonator during the reaction. We demonstrated that the sensitivity of the ISAD technique was 100-times higher than that of RPA and conventional PCR methods. Moreover, this technique can be used to distinguish a single-point mutation of the HRAS gene via target amplification. This novel DNA amplification/detection technique will be useful for the detection of sequence alterations such as mutations and single-nucleotide polymorphisms as DNA biomarkers in human diseases.
authors.affiliation: Institute of Microelectronics, A*STAR (Agency for Science, Technology and Research), 11 Science Park Road, Singapore Science Park II, Singapore 117685.
authors.forename: Yong
authors.initials: Y
authors.lastname: Shin
authors.forename: Agampodi Promoda
authors.initials: AP
authors.lastname: Perera
authors.forename: Kyung Woo
authors.initials: KW
authors.lastname: Kim
authors.forename: Mi Kyoung
authors.initials: MK
authors.lastname: Park
mesh_terms: D004273:DNA, Neoplasm / Q000235:genetics; D006801:Humans; D009369:Neoplasms / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*; D045522:Recombinases / Q000378:metabolism; D012825:Silicon / Q000737:chemistry; D013696:Temperature; D013997:Time Factors
chemical_list: D004273:DNA, Neoplasm; D045522:Recombinases; D016283:Proto-Oncogene Proteins p21(ras); D012825:Silicon