A differential fluorescent receptor for nucleic acid analysis.

A differential fluorescent receptor for nucleic acid analysis.
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用于核酸分析的差异荧光受体。

DOI:
10.1002/cbic.201300657
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发表时间:
2014-01-24
期刊:
影响因子:
3.2
通讯作者:
Kolpashchikov, Dmitry M.
Kolpashchikov, Dmitry M.
中科院分区:
生物学3区
文献类型:
--
作者:
Bengtson, Hillary N.;Kolpashchikov, Dmitry M.

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差异受体使用传感器阵列来识别分析物。阵列中的每个传感器可以识别不是一个,而是具有不同速率的几个分析物,因此单个分析物触发阵列中的几个传感器的响应。因此,受体产生对于每个分析物唯一的信号模式,从而能够通过产生“指纹”模式来识别特定分析物。我们应用这种方法分析了结核分枝杆菌菌株的DNA序列,这些菌株的差异在于81 bp热点区域的单核苷酸取代,这赋予了利福平耐药性。该技术利用了新的多组分自组装传感器,该传感器在特定DNA序列存在的情况下产生荧光信号。差异荧光受体(DFR)包含三个这样的传感器阵列,并区分至少八个DNA序列。该方法只需要一个分子信标样荧光报告基因,可以被所有三个传感器使用。本研究中开发的DFR代表了使用荧光杂交探针的分子诊断技术的一种具有成本效益的替代方案。
Differential receptors use an array of sensors to recognize analytes. Each sensor in the array can recognize not one, but several analytes with different rates, so a single analyte triggers a response of several sensors in the array. The receptor thus produces a pattern of signals that is unique for each analyte, thereby enabling identification of a specific analyte by producing a “fingerprint” pattern. We applied this approach for the analysis of DNA sequences ofMycobacterium tuberculosisstrains that differ by single nucleotide substitutions in the 81‐bp hot‐spot region that imparts rifampin resistance. The technology takes advantage of the new multicomponent, selfassembling sensor, which produces a fluorescent signal in the presence of specific DNA sequences. A differential fluorescent receptor (DFR) contained an array of three such sensors and differentiated at least eight DNA sequences. The approach requires only one molecular‐beacon‐like fluorescent reporter, which can be used by all three sensors. The DFR developed in this study represents a cost‐efficient alternative to molecular diagnostic technologies that use fluorescent hybridization probes.
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