用于临床样品超灵敏蛋白检测的DNA纳米花动力CRISPR/Cas12a生物传感平台

IF 10.7 2区材料科学 Q1 CHEMISTRY, PHYSICAL

Small Methods Pub Date : 2025-03-27 DOI:10.1002/smtd.202402130

Chen Ji, Yongming Han, Junru Li, Jie Wei, Wenzhi Yang, Xiaohan Cai, Siqi Tian, Congshuo Chen, Yuzheng Wang, Peipei Zhao, Shan Cao, Wenhao Zhang, Jinxinyi Xu, Bin Gu, Fengqin Li, Peifeng Liu, Jun Pu

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引用次数: 0

摘要

多种人类细胞分泌的蛋白质标志物为临床疾病的早期诊断和预后评估提供了重要的见解。然而，由于实际临床样品中蛋白质标记物信号扩增效率高，蛋白质标记物的准确、灵敏、快速检测仍然具有很大的挑战性。本文提出了一种DNA纳米花（DNF）驱动的CRISPR/Cas12a生物传感平台（DNF-CRISPR），该平台利用上游的DNF放大蛋白质标记物的输入信号，同时利用下游的CRISPR系统通过反式切割放大输出信号。该上游和下游级联放大传感平台具有高灵敏度（500 fg mL-1），快速（≤2 h）和宽动态范围（2.5 pg mL-1至25 ng mL-1）。作为概念验证，DNF-CRISPR生物传感平台能够定量检测肾损伤患者血液和尿液样本中的中性粒细胞明胶酶相关脂钙蛋白（NGAL）生物标志物，准确率为91%。本研究为临床环境中蛋白质标记物的准确诊断提供了一种强大而通用的方法，促进了基于CRISPR/ cas12的非核酸标记物传感平台的应用。

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DNA Nanoflower-Powered CRISPR/Cas12a Biosensing Platform for Ultrasensitive Protein Detection in Clinical Samples.

Protein markers secreted by various human cells provide crucial insights for the early diagnosis and prognostic assessment of clinical diseases. However, restricted by efficient protein marker signal amplification in real clinical samples with complex compositions, accurate, sensitive, and rapid detection of protein markers remains largely challenging. Herein, a DNA nanoflower (DNF)-powered CRISPR/Cas12a biosensing platform (DNF-CRISPR) is presented that employs the DNF in upstream to amplify input signals for protein markers, while utilizing the CRISPR system in downstream to amplify output signals by trans cleavage. This upstream and downstream cascade amplification sensing platform exhibits high sensitivity (500 fg mL^-1), rapid (≤2 h), and a broad dynamic range (2.5 pg mL^-1 to 25 ng mL^-1). As a proof of concept, DNF-CRISPR biosensing platform enables the quantitative detection of neutrophil gelatinase-associated lipocalin (NGAL) biomarkers in blood and urine samples from kidney injury patients with 91% accuracy. This study provides a powerful and versatile approach for the accurate diagnosis of protein markers in clinical settings, facilitating the application of CRISPR/Cas12a-based sensing platforms for non-nucleic acid markers.

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来源期刊

Small Methods Materials Science-General Materials Science

CiteScore

17.40

自引率

1.60%

发文量

347

期刊介绍： Small Methods is a multidisciplinary journal that publishes groundbreaking research on methods relevant to nano- and microscale research. It welcomes contributions from the fields of materials science, biomedical science, chemistry, and physics, showcasing the latest advancements in experimental techniques. With a notable 2022 Impact Factor of 12.4 (Journal Citation Reports, Clarivate Analytics, 2023), Small Methods is recognized for its significant impact on the scientific community. The online ISSN for Small Methods is 2366-9608.