Title

Rna-Cleaving Deoxyribozyme Sensor For Nucleic Acid Analysis: The Limit Of Detection

Keywords

Deoxyribozyme sensor; DNA; Dna enzyme; Fluorescence; Limit of detection

Abstract

Along with biocompatibillty, chemical stability, and simplicity of structural prediction and modification, deoxyribozymebased molecular sensors have the potential of an improved detection limit due to their ability to catalytically amplify signal. This study contributes to the understanding of the factors responsible for the limit of detection (LOD) of RNA-cleaving deoxyribozyme sensors. A new sensor that detects specific DNA/ RNA sequences was designed from deoxyribozyme OA-II [Chiuman, W.; Li, Y. (2006) J. Mol. Biol. 357, 748-754]. The sensor architecture allows for a unique combination of high selectivity, low LOD and the convenience of fluorescent signal monitoring in homogeneous solution. The LOD of the sensor was found to be ~1.6x10 -10M after 3 h of incubation. An equation that allows estimation of the lowest theoretical LOD using characteristics of parent deoxyribozymes and their fluorogenlc substrates was derived and experimentally verified. According to the equation, quot;catalytically perfectquot; enzymes can serve as scaffolds for the design of sensors with the LOD not lower than 2x10-15M after 3 h of incubation. A new value termed the detection efficiency (DE) is suggested as a time-independent characteristic of a sensor's sensitivity. The expressions for the theoretical LOD and DE can be used to evaluate nucleic acid and protein enzymes for their application as biosensing platforms. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Publication Date

4-12-2010

Publication Title

ChemBioChem

Volume

11

Issue

6

Number of Pages

811-817

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1002/cbic.201000006

Socpus ID

77950675373 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/77950675373

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