A programmable sensitive platform for pathogen detection based on CRISPR/Cas12a -hybridization chain reaction-poly...

Area of Science:

• Biotechnology
• Molecular Biology
• Nanotechnology

Background:

• Rapid and sensitive detection of pathogenic bacteria is vital for public health.
• CRISPR/Cas12a systems show potential for bacterial diagnosis but face sensitivity limitations.

Purpose of the Study:

• To develop a versatile and highly sensitive pathogen sensing platform (HTCas12a).
• To improve the sensitivity and reduce the cost of CRISPR-based bacterial detection.

Main Methods:

• Utilized the CRISPR/Cas12a system combined with hybridization chain reaction (HCR).
• Employed a Poly T-copper fluorescence nanoprobe for signal amplification.
• Validated specificity against various pathogen nucleic acid fragments.

Main Results:

• Achieved enhanced sensitivity through HCR and the Poly-T-Cu reporter probe.
• Demonstrated specific recognition of target nucleic acid fragments.
• Established a linear correlation between fluorescence and target quantity with low detection limits (23.36 fM for DNA, 4.17 CFU/mL for S. aureus).
• Reduced experiment cost to under one dollar per sample.

Conclusions:

• The HTCas12a system provides a sensitive, specific, and cost-effective platform for pathogen detection.
• This technology has broad applications in environmental monitoring, clinical diagnostics, and food safety.