RNA-cleaving deoxyribozyme sensor for nucleic acid analysis: the limit of detection.
RNA-cleaving deoxyribozyme sensor for nucleic acid analysis: the limit of detection.
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DOI：
10.1002/cbic.201000006
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发表时间：
2010-04-12
期刊：
影响因子：
3.2
通讯作者：
Kolpashchikov, Dmitry M.
中科院分区：
文献类型：
--
作者：
Gerasimova, Yulia V.;Cornett, Evan;Kolpashchikov, Dmitry M.
沿着生物相容性、化学稳定性以及结构预测和修饰的简单性，基于脱氧核酶的分子传感器由于其催化放大信号的能力而具有改善检测极限的潜力。本研究有助于了解负责的RNA切割脱氧核酶传感器的检测限（LOD）的因素。从脱氧核酶OA-II设计了一种检测特异性DNA/RNA序列的新传感器[Chiuman，W.; Li，Y.等人（2006）J. Mol. 357，748 - 754]。该传感器结构允许高选择性、低LOD和在均匀溶液中监测荧光信号的便利性的独特组合。孵育3 h后，传感器的LOD为~1.6 × 10 m。一个方程，允许估计的最低理论LOD使用的父母脱氧核酶和它们的荧光底物的特性推导和实验验证。根据该方程，"催化完美"的酶可以作为设计传感器的支架，孵育3小时后LOD不低于~2 × 10 − 15 m。一个新的值被称为检测效率（DE）建议作为一个时间无关的传感器的灵敏度特性。理论LOD和DE的表达式可用于评估核酸和蛋白质酶作为生物传感平台的应用。
Along with biocompatibility, chemical stability, and simplicity of structural prediction and modification, deoxyribozyme-based 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 ~1.6 10×10 m after 3 h of incubation. An equation that allows estimation of the lowest theoretical LOD using characteristics of parent deoxyribozymes and their fluorogenic substrates was derived and experimentally verified. According to the equation, “catalytically perfect” enzymes can serve as scaffolds for the design of sensors with the LOD not lower than ~2×10−15 m 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.
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影响因子：
15
作者：
Kolpashchikov, Dmitry M.
通讯作者：
Kolpashchikov, Dmitry M.
影响因子：
5.6
作者：
Chiuman, W;Li, YF
通讯作者：
Li, YF
DOI：
10.1007/s00018-008-8029-y
发表时间：
2008-07
期刊：
Cellular and molecular life sciences : CMLS
影响因子：
--
作者：
Baum DA;Silverman SK
通讯作者：
Silverman SK
影响因子：
--
作者：
BREAKER, RR;JOYCE, GF
通讯作者：
JOYCE, GF
影响因子：
15
作者：
Mokany, Elisa;Bone, Simon M.;Young, Paul E.;Doan, Tram B.;Todd, Alison V.
通讯作者：
Todd, Alison V.