The following is a summary of “Rapid, sensitive, and user-friendly detection of Pseudomonas aeruginosa using the RPA/CRISPR/Cas12a system,” published in the April 2024 issue of Infectious Disease by Zhang et al.

Researchers conducted a retrospective study to assess trends in identifying Pseudomonas aeruginosa (P. aeruginosa) to improve diagnostic strategies for this multi-threat bacterium.

They chose the lasB gene of P. aeruginosa as their detection target. RPA primers and crRNA were designed to target specific regions within the lasB gene. The specificity of the RPA/CRISPR/Cas12a detection platform was tested using 15 strains. The detection limit was determined with a pseudo-dilution series of P. aeruginosa DNA. Practical applicability was validated by comparing it with qPCR on 150 samples, including 35 processed meat product samples, 55 cold seasoned vegetable dishes, and 60 bottled water samples.

The results showed that the RPA/CRISPR/Cas12a detection platform had high specificity, with no cross-reactivity observed (with non-P. aeruginosa strains). The assay demonstrated remarkable sensitivity, with a limit of detection (LOD) of 10⁰ copies/µL for fluorescence assay and 10¹ copies/µL for the LFTS method. The RPA/CRISPR/Cas12a detection platform also performed comparably to qPCR, with benefits including shorter reaction times, simplified operation, and reduced equipment needs.

Investigators concluded that the RPA/CRISPR/Cas12a platform emerged as a promising tool for straightforward, accurate, and early detection of P. aeruginosa.

Source: bmcinfectdis.biomedcentral.com/articles/10.1186/s12879-024-09348-3