Rapid and sensitive leukemia-derived exosome quantification via nicking endonuclease-assisted target recycling,Analytical Methods

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Rapid and sensitive leukemia-derived exosome quantification via nicking endonuclease-assisted target recycling
Analytical Methods ( IF 2.7 ) Pub Date : 2021-08-11 , DOI: 10.1039/d1ay00854d
Mengyang Zhou ₁ , Chao Li ₁ , Baolong Wang ₂ , Lin Huang ₁

Affiliation

Exosomes as fluid biomarkers hold great promise for noninvasive cancer diagnosis. However, a method for the rapid and convenient detection of exosomes is still a challenge because current analysis processes involve multiple steps and yield low sensitivity. Here, we developed a wash-free fluorescent biosensor for the rapid and sensitive quantification of exosomes by combining aptamer and nicking endonuclease (Nb·BbvCI). In this system, an aptamer–trigger complex was used as the recognition element; the trigger probe could be released, and it hybridized with gold nanoparticles (GNPs)–DNA–FAM conjugates, thereby resulting in Nb·BbvCI-assisted target recycling. As a result, our method allowed the quantification of exosomes with lower analysis time by using a cocktail containing an aptamer–trigger complex, Nb·BbvCI, and GNPs–DNA–FAM. A high sensitivity with a limit of detection (LOD) of 1.0 × 10⁴ particles per μL could be achieved. Besides, this biosensor exhibited potential application for the quantification of exosomes in human plasma, facilitating the development of exosome-based noninvasive cancer diagnosis.

中文翻译：

通过切口内切核酸酶辅助靶标回收快速灵敏地定量白血病衍生的外泌体

外泌体作为流体生物标志物对非侵入性癌症诊断具有很大的前景。然而，一种快速方便地检测外泌体的方法仍然是一个挑战，因为目前的分析过程涉及多个步骤并且灵敏度低。在这里，我们开发了一种免洗荧光生物传感器，用于通过结合适体和切口内切核酸酶 (Nb·BbvCI) 快速灵敏地定量外泌体。在这个系统中，一个适体-触发复合物被用作识别元件；触发探针可以被释放，并与金纳米粒子 (GNPs)-DNA-FAM 结合物杂交，从而导致 Nb·BbvCI 辅助靶标回收。因此，我们的方法允许通过使用含有适体-触发复合物、Nb·BbvCI 和 GNPs-DNA-FAM 的混合物以更短的分析时间对外泌体进行定量。可以达到每 μL^{4 个颗粒。}此外，这种生物传感器在人血浆中外泌体定量方面表现出潜在的应用，促进了基于外泌体的非侵入性癌症诊断的发展。

更新日期：2021-08-24

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