Fig. 5: Validation of the affinity molecular assay for C. albicans detection.

a Performance of AMS technology in detecting simulated C. albicans positive blood samples: 20 plasma samples (blue) and 20 whole blood samples (red). b The anti-interference ability of AMS-qPCR assay. Simulated clinical samples (blood and BAL) were used as blank groups, interfering with C.glabrata, K.pneumoniae, and C. albicans genomic DNA. c LOD of the affinity molecular assay for C. albicans detection from blood and BAL samples, respectively. Direct visualization of fluorescence by naked eyes under blue light. NC: represents reactions without substrate. d–f Linear regression analysis for the assay. d, e Linear relationship observed between fluorescence intensity and negative logarithm of C. albicans concentration in simulated blood samples and BAL samples (3 × 10¹ CFU/mL to 3 × 10⁶ CFU/mL) in the affinity molecular assay. f The linear relationship observed in the AMS-qPCR assay. Mean ± s.d. of n = 20 biological replicates for a, and mean ± s.d. of n =  3 technical replicates for (b, d–f). The statistical significance of a was analyzed using a two-tailed Student’s t-test. g Fungal fluorescence staining of blood and BAL samples with various C. albicans concentrations, showing Calcofluor White staining of chitin-rich cell walls. Scale bars: 20 µm. h–i Affinity molecular detection for simulated clinical samples and clinical samples. h The simulated samples: left, 80 positive samples, 40 blood samples (red) and 40 BAL samples (blue); right, 40 negative samples (20 blood samples and 20 BAL samples). i Clinical samples .25 BAL (red), 6 blood (green), 1 urine (blue), and 1 peritoneal fluid (green). The fluorescence readout was measured at 20 min at 42 °C. j Analysis of ROC curve in clinical sample testing. k SDA plate culture and colony counting technique examined the capture efficiency of MB-PfCBP-A, the experiment was repeated three times, with one representative result shown. Source data are provided as a Source Data file.