Development of a rapid and sensitive RPA-CRISPR/Cas12a-based assay for the detection of <i>Brucella melitensis</i>.

ABSTRACT: Brucellosis, a zoonotic disease caused by Brucella species, presents significant public health challenges due to its complex diagnosis and the limited availability of rapid detection methods. To address these challenges, we developed a novel detection method that integrates recombinase polymerase amplification (RPA) with the CRISPR/Cas12a system, enabling dual readout through fluorescence (FL) and lateral flow strip (LFS) detection. The RPA-CRISPR/Cas12a-FL assay demonstrated an impressive detection limit of 1 copy/μL, which is 10 times more sensitive than quantitative polymerase chain reaction, while the RPA-CRISPR/Cas12a-LFS method achieved a detection limit of 10 copies/μL, comparable to nested PCR. Specificity testing confirmed the robustness of the assay, as it produced strong

SUBMITTER: Shen Y 

PROVIDER: S-EPMC12502682 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature