Visual Detection of Labeled Oligonucleotides Using Visible-Light-Polymerization-Based Amplification

• Published in: Biomacromolecules 2008-01, Vol.9 (1), p.355-362
• Main Authors: Hansen, Ryan R, Sikes, Hadley D, Bowman, Christopher N
• Format: Article
• Language: English
• Subjects: Application fields; Applied sciences; Base Sequence; Biological and medical sciences; Biosensors; Biotechnology; DNA Primers; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Light; Methods. Procedures. Technologies; Oligonucleotides - analysis; Polymer industry, paints, wood; Polymers - chemistry; Sensitivity and Specificity; Technology of polymers; Various methods and equipments
• ISSN: 1525-7797; 1526-4602
• DOI: 10.1021/bm700672z

DNA biochip technology holds potential for highly parallel, rapid, and sensitive genetic diagnostic screening of target pathogens and disease biomarkers. A primary limitation involves a simultaneous, sequence-specific identification of low copy number target polynucleotides using a clinically appropriate detection methodology that implements only inexpensive detection instrumentation. Here, a rapid (20 min), nonenzymatic method of signal amplification utilizing surface-initiated photopolymerization is presented in glass microarray format. Visible light photoinitiators covalently coupled to streptavidin were used to bind biotin-labeled capture sequences. Amplification was achieved through subsequent contact with a monomer solution and the appropriate light exposure to generate 20−240-nm-thick hydrogel layers exclusively from spots containing the biotin-labeled DNA. An amplification factor of 106 to 107 was observed as well as a detectable response generated from as low as ∼104 labeled oligonucleotides using minimal instrumentation, such as an optical microscope or CCD camera.