Biofilm
infection has been identified as a crucial factor of the pathogenesis of chronic
wound, but
wound biofilm diagnosis remains as an unmet clinical need. We previously proposed a modified
wound blotting technique using
Alcian blue staining for biofilm detection that was characterized as being non-invasive, time-saving, non-expansive, and informative for biofilm distribution. In this study, we adapted a novel
Alcian blue grading method as the severity of biofilm
infection for the
wound blotting technique and compared its biofilm detection efficacy with MolecuLight i:X- a point-of-care florescence imaging device to detect bacteria and biofilm in
wounds. Moreover, their predictive value of complete wound healing at 90 days was analyzed. When validated with
wound culture results in the 53 enrolled subjects with chronic
wounds, the modified
wound blotting method showed a strong association with
wound culture, while MolecuLight i:X only exhibited a weak association. In predicting 90-day
wound outcomes, the modified
wound blotting method showed a strong association (Kendall’s tau value = 0.563, p < 0.001), and the
wound culture showed a moderate association (Spearman’s rho = 0.535, p < 0.001), but MolecuLight i:X exhibited no significant association (p = 0.184). In this study, modified
wound blotting with the
Alcian blue grading method showed superior value to MolecuLight i:X both in biofilm detection and predictive validity in 90-day wound-healing outcomes.