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开发一种新型简单的生物传感器,用于检测胱抑素 C,作为肾小球滤过率的一个有吸引力的标志物,采用 DNase I 辅助循环扩增策略。

Developing a novel and simple biosensor for Cystatin C as a fascinating marker of glomerular filtration rate with DNase I-aided recycling amplification strategy.

机构信息

NMPA Key Laboratory for Bioequivalence Research of Generic Drug Evaluation, Shenzhen Institute for Drug Control, Shenzhen 518057, China.

NMPA Key Laboratory for Bioequivalence Research of Generic Drug Evaluation, Shenzhen Institute for Drug Control, Shenzhen 518057, China.

出版信息

J Pharm Biomed Anal. 2021 Sep 5;203:114230. doi: 10.1016/j.jpba.2021.114230. Epub 2021 Jun 24.

DOI:10.1016/j.jpba.2021.114230
PMID:34182410
Abstract

Cystatin C (Cys C) has been proposed as a fascinating glomerular filtration rate (GFR) marker for early detection of acute kidney injury and chronic kidney disease. However, most of traditional methods for Cys C detection are immunoassays, which was tedious to perform and unfriendly for economics. In this work, a novel and simple biosensor for the sensitive measurement of Cys C via DNase I-aided recycling amplification strategy was successfully constructed based on the graphene oxide (GO) and fluorophore-labelled aptamer, which can be used to the early prediction of kidney injury. The fluorescence of fluorophore-labelled aptamer was quenched by GO based on the Fluorescence Resonance Energy Transfer (FRET) and recovered with the existence of Cys C. In addition, the DNase I enzyme would digest the fluorophore-labelled aptamer and dissociate the Cys C to launch the next reaction, resulting in an increase of signal amplification. Hence, the limit of detection is found to be 0.16 ng mL, which is almost 3 times lower than that without DNase I. Consequently, the developed biosensor offers a novel approach towards simple and rapid detection of Cys C based on the integration of GO and aptamer. Conceivably, this strategy holds a wide scope in the application of numerous other analytes if corresponding aptamers are available.

摘要

半胱氨酸蛋白酶抑制剂 C(Cys C)已被提议作为一种很有前途的肾小球滤过率(GFR)标志物,用于早期检测急性肾损伤和慢性肾病。然而,大多数传统的 Cys C 检测方法都是免疫测定法,操作繁琐,不经济。在这项工作中,基于石墨烯氧化物(GO)和荧光标记的适体,成功构建了一种用于通过 DNase I 辅助循环放大策略灵敏测量 Cys C 的新型简单生物传感器,可用于早期预测肾损伤。荧光标记的适体的荧光基于荧光共振能量转移(FRET)被 GO 猝灭,并在存在 Cys C 的情况下恢复。此外,DNase I 酶会消化荧光标记的适体并将 Cys C 解离以启动下一个反应,从而导致信号放大增加。因此,检测限被发现为 0.16ng/mL,比没有 DNase I 时低约 3 倍。因此,所开发的生物传感器基于 GO 和适体的集成,为基于简单和快速检测 Cys C 提供了一种新方法。可以想象,如果有相应的适体,这种策略在许多其他分析物的应用中具有广泛的应用前景。

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