FIGURE 5.
Performance of Cas12a-UPTLFA on detection of the Y. pestis genomic DNA or Y. pestis bacteria in the spiked blood samples. (A) Components of the Cas12a-UPTLFA platform. The sample pad, conjugation pad, analytical membrane, and absorbent pad were mounted on a lamination card with proper overlaps. All antibodies for detection were fixed in advance, including the UCP-labeled with anti-FAM antibody in the conjugation pad, streptavidin for the T line, and goat anti-mouse IgG for the C line. (B) Cas12a-UPTLFA detection relies on the cleavage of a FAM-biotin-labeled ssDNA reporter by the collateral activity of the Cas12 enzymes upon target recognition, allowing the detection on UPT strips. (C) Detection results of Y. pestis genomic DNA samples at concentration ranging from 3 × 10− 2 to 3 × 103 aM. Data are represented as mean with SD (n = 3). (D) The table shows the summarized results for 112 blood samples detected by Cas12a-UPTLFA platform and RPA-Cas12a fluorescent read-out assay. P, the number of positive samples; N, the number of negative samples; RPA, recombinase polymerase amplification; UPT, up-converting phosphor technology.