Fig. 5: ActCRISPR-TB LFA analysis of self-collected tongue swab specimens.

a Sample collection/processing, ActCRISPR-TB, and LFA readout workflow. Created in BioRender. Huang, Z. (2025) https://BioRender.com/0silymt. ActCRISPR-TB LFA images and results for b no-template control (NTC) and positive control (PC) samples, c IS6110 DNA concentration standards, and d lysates of four Mtb complex (MTBC), seven non-tuberculous mycobacteria (NTM), and three non-mycobacterial (NM) species. e Inter- and intra-assay coefficients of variation for ActCRISPR-TB LFA results of spiked tongue swab samples (10 and 100 copies/μL) tested with two test strip batches. f ActCRISPR-TB LFA images and results for postprandial tongue swabs collected before and after oral rinse from healthy donors (HD, n = 3) and then spiked with IS6110 DNA (10 copies/μL). Created in BioRender. Huang, Z. (2025) https://BioRender.com/execr4a. g ActCRISPR-TB LFA images and ActCRISPR-TB and Xpert Ultra results for self-collected tongue swabs of 35 adults diagnosed with (N = 24) and without (N = 11) TB (Table S13). All TB patients were stratified into four categories based on their semi-quantitative sputum Xpert MTB/RIF grades as a proxy for sputum Mtb burden: high (Ct < 16), medium (Ct 16–22), low (Ct 22–28), and very low (Ct > 28). Pink triangles denote weakly positive LFA results, and red and gray dots indicate positive and negative ActCRISPR-TB and Xpert Ultra results. Bar graphs represent mean ± SD results calculated from three independent LFA reads, and a p value calculated with a two-sided Wilcoxon signed-rank test. IntDen (Integrated Density) indicates the colorimetric intensity of the T-line, as determined by the total pixel intensity detected within this region of the test strip when measured by ImageJ. Source data are provided as a Source Data file Abbreviations: RT room temperature.