Aptamer-based rolling circle amplification coupled with graphene oxide-based fluorescence resonance energy transfer for sensitive detection of cardiac troponin I,Analytical Methods

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Aptamer-based rolling circle amplification coupled with graphene oxide-based fluorescence resonance energy transfer for sensitive detection of cardiac troponin I
Analytical Methods ( IF 2.7 ) Pub Date : 2018-03-06 00:00:00 , DOI: 10.1039/c8ay00309b
Yu Li _{1,

2,

3,

4,

5} , Weiqiang Dai _{1,

2,

3,

4,

5} , Xuefei Lv _{1,

2,

3,

4,

5} , Yulin Deng _{1,

2,

3,

4,

5}

Affiliation

Combining the high affinity and specificity of an aptamer for a protein, a highly sensitive strategy for protein detection was developed based on rolling circle amplification (RCA) and graphene oxide based fluorescence resonance energy transfer (FRET). In this strategy, a protein was captured using an antibody-coated microplate, and then a circularized product containing an aptamer–primer and padlock probe was added to form a sandwich structure. Under the influence of DNA polymerase, the RCA reaction extended the primer sequence, and generated hundreds of copies of the circular DNA template. Detection of the RCA products via FRET using graphene oxide allowed the sensitivity enhancement in target protein detection. This method was used to detect the target protein of cardiac troponin I (cTnI) with a detection limit as low as 14.40 pg mL⁻¹. The highly sensitive and specific assay can be used as a promising tool in clinical sample analysis.

中文翻译：

基于适体的滚环扩增结合基于氧化石墨烯的荧光共振能量转移，可灵敏检测心肌肌钙蛋白I

结合适体对蛋白质的高亲和力和特异性，基于滚环扩增（RCA）和基于氧化石墨烯的荧光共振能量转移（FRET），开发了一种高灵敏度的蛋白质检测策略。在这种策略中，使用抗体包被的微孔板捕获蛋白质，然后添加包含适体-引物和挂锁探针的环化产物以形成夹心结构。在DNA聚合酶的影响下，RCA反应扩展了引物序列，并生成了数百个环状DNA模板。在RCA产物的检测通过使用氧化石墨烯的FRET可以提高目标蛋白检测的灵敏度。该方法用于检测心肌肌钙蛋白I（cTnI）的目标蛋白，检出限低至14.40 pg mL ^-1。高灵敏度和特异性的测定可以用作临床样品分析中的有前途的工具。

更新日期：2018-03-06

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