RAPID and accurate detection of bloodstream pathogens is essential for effective disease management, but current molecular diagnostics often require nucleic acid amplification, adding both time and cost. A new study presents a CRISPR-Cascade assay that eliminates this need, enabling highly sensitive, multiplexed detection of pathogens within minutes.
The newly developed CRISPR-Cascade assay integrates a positive feedback loop that enhances signal amplification without the need for nucleic acid preamplification. Unlike conventional PCR-based methods, which require complex thermal cycling and centralised laboratory facilities, this approach streamlines the diagnostic process. Additionally, the incorporation of an OR-gate logic function allows for the simultaneous detection of multiple pathogens, improving efficiency and versatility in clinical settings. By leveraging these mechanisms, the assay provides a rapid, user-friendly alternative to traditional molecular diagnostics.
The study demonstrated the assay’s effectiveness in detecting bloodstream infection pathogens, including methicillin-sensitive Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, and hepatitis B virus (HBV) spiked into whole blood samples. The CRISPR-Cascade assay achieved attomolar sensitivity, detecting target DNA within just 10 minutes while maintaining a signal-to-noise ratio exceeding 1.3. These findings highlight the assay’s capability to provide highly sensitive, specific, and rapid pathogen identification, significantly reducing diagnostic turnaround times and improving early disease detection.
By eliminating the need for amplification, this new technology simplifies molecular diagnostics, making it a viable solution for point-of-care and decentralised healthcare applications.
Ada Enesco, EMJ
Reference
Lim J et al. Amplification-free, OR-gated CRISPR-Cascade reaction for pathogen detection in blood samples. Proc Natl Acad Sci U S A. 2025;18;122(11):e2420166122.
