Rapid detection of biofilm by wound blotting following sharp debridement of chronic pressure ulcers predicts wound healing: A preliminary study

Nakagami, Gojiro; Schultz, Gregory S.; Kitamura, Aya; Minematsu, Takeo; Akamata, Kaname; Suga, Hiraku; Kurita, Masanori; Hayashi, Chieko; Sanada, Hiromi

doi:10.1111/iwj.13256

Cited by 23 publications

(25 citation statements)

References 17 publications

(19 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Because current methods of biofilm diagnosis have their limitations, the development of a rapid, easy, and reliable biofilm detection assay is urgently needed. Considering membrane‐based diagnosing tools are getting popular clinically, 23,30 we aimed to develop a membrane‐based alcian blue staining protocol for biofilm detection.…”

Section: Discussionmentioning

confidence: 99%

“…In the first part of this study, we tried to optimize the alcian blue staining process by selecting a positively charged nylon transfer membrane (Biodyne B Nylon Membrane) because it provided better binding and adhesion for the negatively charged polysaccharides in biofilm EPS. In the literature, Nakagami et al used pre‐wetted nitrocellulose membrane (supported nitrocellulose membrane, 0.2 μm pore size, Bio‐Rad Laboratories., California) for alcian blue staining procedure 23 . We found that the nylon transfer membrane used in our study provided several advantages: (a) it remained positively charged across a wide pH range (up to pH 10), which facilitated improved adhesion of negatively charged polysaccharides; (b) it provided a high signal‐to‐noise ratio and less background noise after staining; and (c) it provided a higher binding capacity, up to 100 to 200 μg/cm 2 , compared to that of supported nitrocellulose, 80 to 100 μg/cm 2 31,32…”

Section: Discussionmentioning

confidence: 99%

“…The washing solution employed in our staining protocol, cationic detergent, was applied immediately after sampling with the nylon transfer membrane to facilitate blocking and decrease nonspecific binding during staining. Because the cationic detergent used in the staining methods proposed by Nakagami et al was not described clearly, 23 we examined five types of cationic detergents for use as washing solutions. Among the washing solutions examined, CTAC provided the best enhancement of polysaccharide staining and the lowest background noise.…”

Section: Discussionmentioning

confidence: 99%

“…Tissue samples were cultured with tryptic soy agar plates and stained with alcian blue as per our modified staining protocol. Additionally, we collected information on the outcomes of these wounds at a 1‐month follow‐up visit according to the previous literature, which correlated biofilm presence with wound outcome 23 . Our modified alcian blue staining protocol showed high sensitivity with adequate specificity, indicating that it was suitable as a first‐line screening tool for early biofilm infection detection and could facilitate further biofilm‐based wound therapy.…”

Section: Discussionmentioning

confidence: 99%

“…Nakagami et al introduced a wound blotting method to provide a noninvasive approach to evaluate the chronic wounds 22‐24 . This novel method specifically stained polysaccharides in the biofilm EPS matrix using ruthenium red or alcian blue dyes to visualize blotted biofilm on a nitrocellulose membrane and established a process that could be used to map the location of biofilm as well as guide debridement surgery and other therapies to improve wound healing.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Rapid detection of biofilm with modified alcian blue staining: In‐vitro protocol improvement and validation with clinical cases

Lee

Liao

et al. 2020

Wound Repair Regeneration

View full text Add to dashboard Cite

For chronic wounds, biofilm infection is a critical issue because it can tip the scales toward an unhealing state. Biofilm‐based wound therapy has been extensively advocated. However, point‐of‐care biofilm diagnosis still largely relies on clinical judgment. In this study, we aimed to develop a rapid tool for diagnosing wound biofilm presence by alcian blue staining. First, we sought to optimize alcian blue staining using a colorimetric‐based approach to detect the biofilm, specifically targeting polysaccharides in the extracellular polymeric substances. Among examined transfer membranes and cationic detergents at various concentrations, we selected a positively charged nylon transfer membrane for sample loading, and 1% cetyl trimethyl ammonium chloride (CTAC) as the blocking solution. After sample loading and blocking, the membrane was immersed in alcian blue solution for staining, followed by immersion in 1% CTAC to decrease background noise. Each step required only 30 seconds, and the whole procedure was completed within a few minutes. In the second part of this study, we enrolled 31 patients with chronic wounds to investigate the predictive validity of biofilm detection for unhealed wounds at a 1‐month follow‐up visit. Among the 18 cases with positive wound biofilm staining, 15 wounds (83.3%) were not healed at the 1‐month follow‐up visit. Only three unhealed wounds (30%) produced in negative staining cases. This finding indicates that biofilm infection is associated with poor healing outcome for chronic wounds. Moreover, our staining results correlated well with the clinical microbiological culture assessment (83.9% consistency; 95.2% sensitivity, and 60% specificity). In conclusion, the modified alcian blue staining protocol used here represents a rapid and sensitive procedure for detecting biofilm in chronic wounds. This technique provides a practical point‐of‐care approach for detection of wound biofilm, the implementation of which may improve clinical outcomes for chronic wound patients. Additional studies are required to validate this method.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations