Dai Xiaoqi, Meng Changle, Huang Songfeng, Wang Yuye, He Jianan, Chen Zhi, Ji Yinyue, Tai Jiali, Zhang Jinming, Ni Hailong, Zhuang Zhuolun, Chen Jiajie, Zhang Han, Qu Junle, Shao Yonghong
Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Physics and Optoelectronics Engineering, Shenzhen University, Shenzhen 518060, China.
Shenzhen International Travel Health Care Center (Shenzhen Customs District Port Outpatient Clinics), Shenzhen Customs District, Shenzhen 518045, China.
Anal Chem. 2025 Aug 5;97(30):16296-16303. doi: 10.1021/acs.analchem.5c01772. Epub 2025 Jul 23.
Driven by the growing need for real-time, amplification-free, and label-free nucleic acid detection in clinical diagnostics and pathogen surveillance, traditional methods often fall short due to limited sensitivity, a narrow dynamic range, and difficulties in detecting low-concentration nucleic acids and single-nucleotide mutations. To address these challenges, we developed a clustered regularly interspaced short palindromic repeats (CRISPR) enhanced Phase-interrogation Surface Plasmon Resonance imaging (CRISPR-PSPRi) sensor that employs phase delay modulation for highly sensitive extraction of SPR phase signals and a wavelength scanning strategy to extend its dynamic range. By harnessing CRISPR-Cas12a for target DNA recognition and activating -cleavage to cleave ssDNA-linked gold nanoparticle probes, our platform converts extremely weak signals from low-concentration DNA into readily detectable cleavage signals. Achieving a sensitivity of 1.436 × 10 RIU and a dynamic range of 0.0111 RIU, this system successfully detects specific DNA from the SARS-CoV-2 Omicron BA.2 variant and monkeypox virus, and it can detect single-nucleotide mutations down to 1 aM. This breakthrough offers a real-time, high-throughput, and ultrasensitive nucleic acid detection approach, promising significant advancements in clinical diagnostics and pathogen monitoring.
在临床诊断和病原体监测中，对实时、无扩增和无标记核酸检测的需求日益增长，传统方法往往因灵敏度有限、动态范围窄以及检测低浓度核酸和单核苷酸突变存在困难而难以满足需求。为应对这些挑战，我们开发了一种成簇规律间隔短回文重复序列（CRISPR）增强的相位询问表面等离子体共振成像（CRISPR-PSPRi）传感器，该传感器采用相位延迟调制来高度灵敏地提取SPR相位信号，并采用波长扫描策略来扩展其动态范围。通过利用CRISPR-Cas12a识别靶DNA并激活切割以切割与单链DNA连接的金纳米颗粒探针，我们的平台将来自低浓度DNA的极其微弱的信号转化为易于检测的切割信号。该系统实现了1.436×10 RIU的灵敏度和0.0111 RIU的动态范围，成功检测出严重急性呼吸综合征冠状病毒2（SARS-CoV-2）奥密克戎BA.2变体和猴痘病毒的特异性DNA，并且能够检测低至1 aM的单核苷酸突变。这一突破提供了一种实时、高通量和超灵敏的核酸检测方法，有望在临床诊断和病原体监测方面取得重大进展。
