An extraction-free, lyophilized one-pot RAA-CRISPR assay for point-of-care testing of <i>Haemophilus influenzae</i>.

ABSTRACT: Timely and accurate diagnosis is essential for the effective treatment of Haemophilus influenzae (HI) infection. Notably, a rapid point-of-care testing (POCT) method for the diagnosis of HI DNA is still lacking. The aim of this study was to establish a one-pot detection device for HI DNA detection using CRISPR/Cas13a that would be applicable for POCT. We established a two-step polymerase chain reaction (PCR)-CRISPR assay, a two-step recombinase-aided amplification (RAA)-CRISPR assay, and a one-pot RAA-CRISPR assay. Additionally, reagent lyophilization, rapid lysis technology, and a one-pot detection device were integrated to construct an extraction-free one-pot detection device, named EFORCA (extraction-free one-pot RAA-CRISPR/Cas13a assay). Validation was performed on 90 simulated

SUBMITTER: Cao Y 

PROVIDER: S-EPMC12607747 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature