Work

Detection of waterborne Pseudomonas aeruginosa monovalent serogroup E using the ultrasensitive HDA-CRISPR/Cas12a method

Year: 2024

Type: article

Abstract: The nucleic acid isothermal amplification method allows for the highly sensitive detection of the important pathogen Pseudomonas aeruginosa ( P. aeruginosa ), providing crucial information for controlling pathogen transmission. In this study, we exploited the high GC content of a specific target within the monovalent serogroup E of P. aeruginosa , and through the design of primers and CRISPR RNA (crRNA) sequences, achieved robust exponential amplification of the target DNA. We integrated the HDA detection system with an optimized CRISPR/Cas12a, resulting in the development of the novel “HDA-Cas12aFQ” platform, which enables detection within 2 h. Validation experiments confirmed that the HDA-Cas12aFQ platform exhibits high specificity and sensitivity for detecting P. aeruginosa monovalent serogroup E in serum samples. Specifically, the platform demonstrated a detection limit of 10 2 CFU/mL for pure cultures, a DNA sensitivity of 0.695 pmol/L—nearly 200 times greater than that of the real-time HDA method. Additionally, the positive detection rate of real-world samples was in strong agreement with that of traditional, well-established detection methods. The HDA-Cas12aFQ platform is expected to become a leading method for ensuring drinking water purity due to its exceptional sensitivity and accuracy, thus playing a crucial role in safeguarding public health.

Source: LWT

DOI: 10.1016/j.lwt.2024.117162