Amplification- free, OR- gated CRISPR- Cascade reaction for pathogen detection in blood samples

• 成果类型: Article
• 署名作者: Lim, Jongwon; Van, An Bao; Koprowski, Katherine; Wester, Matthew; Valera, Enrique; Bashir, Rashid
• 署名单位: University of Illinois System; University of Illinois Urbana-Champaign; University of Illinois System; University of Illinois Urbana-Champaign; University of Illinois System; University of Illinois Urbana-Champaign; University of Illinois System; University of Illinois Urbana-Champaign; University of Illinois System; University of Illinois Urbana-Champaign; University of Illinois System; University of Illinois Urbana-Champaign; Chan Zuckerberg Initiative (CZI)
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-9225
• DOI: 10.1073/pnas.2420166122
• 发表日期: 2025-03-18
• 关键词: nucleic-acid detection; point; pcr

摘要

Rapid and accurate detection of DNA from disease- causing pathogens is essential for controlling the spread of infections and administering timely treatments. While traditional molecular diagnostics techniques like PCR are highly sensitive, they include nucleic acid amplification and many need to be performed in centralized laboratories, limiting their utility in point- of- care settings. Recent advances in CRISPR- based diagnostics (CRISPR- Dx) have demonstrated the potential for highly specific molecular detection, but the sensitivity is often constrained by the slow trans- cleavage activity of Cas enzymes, necessitating preamplification of target nucleic acids. In this study, we present a CRISPR- Cascade assay that overcomes these limitations by integrating a positive feedback loop that enables nucleic acid amplification- free detection of pathogenic DNA at atto- molar levels and achieves a signal- to- noise ratio greater than 1.3 within just 10 min. The versatility of the assay is demonstrated through the detection introduce a multiplexing OR- function logic gate, further enhancing the potential Our findings highlight the ability of the CRISPR- Cascade assay to provide highly sensitive and specific molecular detection, paving the way for advanced applications in point- of- care diagnostics and beyond.