nucleic acid sandwich hybridization assay with quantum dot-induced fluorescence resonance energy transfer for pathogen detection三明治核酸杂交分析和量子dot-induced荧光共振能量转移的病原体检测.pdfVIP

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nucleic acid sandwich hybridization assay with quantum dot-induced fluorescence resonance energy transfer for pathogen detection三明治核酸杂交分析和量子dot-induced荧光共振能量转移的病原体检测.pdf

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nucleic acid sandwich hybridization assay with quantum dot-induced fluorescence resonance energy transfer for pathogen detection三明治核酸杂交分析和量子dot-induced荧光共振能量转移的病原体检测

Sensors 2012, 12, 16660-16672; doi:10.3390/s121216660 OPEN ACCESS sensors ISSN 1424-8220 /journal/sensors Article Nucleic Acid Sandwich Hybridization Assay with Quantum Dot-Induced Fluorescence Resonance Energy Transfer for Pathogen Detection Cheng-Chung Chou * and Yi-Han Huang Department of Life Science and Center for Nano Bio-Detection, National Chung-Cheng University, Chia-Yi 62102, Taiwan; E-Mail: je091410show@ * Author to whom correspondence should be addressed; E-Mail: bioccc@.tw; Tel.: +886-5-272-0411 (ext. 66506); Fax: +886-5-272-2871. Received: 22 October 2012; in revised form: 23 November 2012 / Accepted: 30 November 2012 / Published: 4 December 2012 Abstract: This paper reports a nucleic acid sandwich hybridization assay with a quantum dot (QD)-induced fluorescence resonance energy transfer (FRET) reporter system. Two label-free hemagglutinin H5 sequences (60-mer DNA and 630-nt cDNA fragment) of avian influenza viruses were used as the targets in this work. Two oligonucleotides (16 mers and 18 mers) that specifically recognize two separate but neighboring regions of the H5 sequences were served as the capturing and reporter probes, respectively. The capturing probe was conjugated to QD655 (donor) in a molar ratio of 10:1 (probe-to-QD), and the reporter probe was labeled with Alexa Fluor 660 dye (acceptor) during synthesis. The sandwich hybridization assay was done in a 20 μL transparent, adhesive frame-confined microchamber on a disposable, temperature-adjust