| Colorimetric and Ultrasensitive Bioassay Based on a Dual-Amplification System Using Aptamer and DNAzyme. | |
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MedLine Citation:
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PMID: 22533853 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Rapid detection of ultralow amount of biomarkers in a biologically complex mixture remains a major challenge. Herein, we report a novel aptamer-based protein detection assay that integrates two signal amplification processes, namely polymerase-mediated rolling-circle amplification (RCA) and DNA enzyme-catalyzed colorimetric reaction. The target biomarker is captured in a sandwich assay by primary aptamer-functionalized microbeads and secondary aptamer that is connected to a RCA primer/circular template complex. RCA reaction, which amplifies the single biomarker binding events hundreds to thousands of times (the first amplification), produces a long DNA molecule containing multiple DNAzyme units. The peroxidase-like DNAzyme catalyzes the oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (the second amplification), which generates a blue-green colorimetric signal. This new biosensing platform permits the ultrasensitive, label-free, colorimetric detection of biomarker in real time. Using platelet-derived growth factor B-chain (PDGF-BB) as a model system, we demonstrated that our assay can detect protein marker specifically in serum-containing medium, at a concentration as low as 0.2 pg/mL in ~ 2 h which rivals traditional assays such as ELISA. We anticipate this simple methodology for biomarker detection can find utility in point-of-care applications. |
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Authors:
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Longhua Tang; Yang Liu; Md Monsur Ali; Dong Ku Kang; Weian Zhao; Jinghong Li |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2012-4-25 |
Journal Detail:
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Title: Analytical chemistry Volume: - ISSN: 1520-6882 ISO Abbreviation: - Publication Date: 2012 Apr |
Date Detail:
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Created Date: 2012-4-26 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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