用于灵敏检测 HbA1c 的 CRISPR 扩增无标记电化学适配传感器

IF 3.5 Q2 CHEMISTRY, ANALYTICAL Sensors & diagnostics Pub Date : 2024-07-16 DOI:10.1039/D4SD00193A
Jianfeng Ma, Youwei Zheng, Yaoyao Xie, Dan Zhu, Lianhui Wang and Shao Su
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摘要

糖化血红蛋白(HbA1c)是监测和早期诊断糖尿病的重要生物标志物。CRISPR-Cas 系统具有高特异性、高效性、灵活性和定制性等特点,在下一代生物传感器中具有广阔的应用前景。本文设计了一种无标记的电化学适配体传感器,将适配体的特异性识别能力与CRISPR-Cas12a系统的信号放大效应相结合,用于检测HbA1c。在 HbA1c 存在的情况下,Cas12a 蛋白的顺反裂解能力被激活,导致预先形成的探针 DNA 被大量裂解,电化学信号增加。通过 CRISPR 辅助信号放大,所开发的电化学适配传感器可以检测到低至 0.84 纳克/毫升的 HbA1c。此外,由于该传感器具有高选择性、可重复性和长期稳定性,它还能检测 50%人体血清中 10 ng/mL HbA1c 的含量,低于人体血样中的生理水平。所有结果都证明,该传感器在糖尿病的早期诊断和长期监测方面具有广阔的应用前景。
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A CRISPR-amplified label-free electrochemical aptasensor for the sensitive detection of HbA1c†

Glycated hemoglobin (HbA1c) is a pivotal biomarker for the monitoring and early diagnosis of diabetes. The CRISPR-Cas system has fascinating application prospects in the next generation of biosensors due to its high specificity, efficiency, flexibility, and customization. Herein, a label-free electrochemical aptasensor was designed for the detection of HbA1c by combining the specific recognition ability of aptamers with the signal amplification effect of the CRISPR-Cas12a system. In the presence of HbA1c, the cistrans cleavage ability of Cas12a protein was activated, causing the pre-formed probe DNA to be heavily cleaved and the electrochemical signal to increase. With CRISPR-assisted signal amplification, the developed electrochemical aptasensor can detect as low as 0.84 ng mL−1 HbA1c. Moreover, this aptasensor can detect 10 ng mL−1 HbA1c in 50% human serum due to its high selectivity, reproducibility, and long-term stability, which is lower than its physiological level in human blood samples. All results proved that the proposed aptasensor has a promising application in the early diagnosis and long-term monitoring of diabetes.

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