Wound blotting is a novel biotechnology that predicts wound outcomes and localizes biofilms on wound surfaces by determining the distribution pattern of tumor necrosis factor-alpha (TNF-α) and biofilm mucopolysaccharides. The rapid and objective analysis offered by this technique may assist clinicians in treating chronic wound biofilms.

Current clinical and preclinical diagnostic techniques fail to accurately identify pathogens and the precise location of biofilms over wound surfaces, rendering timely medical or surgical intervention to eradicate biofilms elusive.

The impact of biofilms on delayed wound healing has drawn increasing attention. Their importance led to the establishment of biofilm-based wound care where chronic wounds are treated using multipronged strategies to remove biofilms over wound beds to facilitate the recovery of epithelial integrity.

Chronic non-healing wounds have become a major worldwide healthcare burden. The impact of biofilms on chronic wound infection is well established. Despite increasing understanding of the underlying mechanism of biofilm formation in chronic wounds, current strategies for biofilm diagnosis in chronic wounds are still far from ideal. In this review, we briefly summarize the mechanism of biofilm formation and focus on current diagnostic approaches of chronic wound biofilms based on morphology, microbiology, and molecular assays. Innovative biotechnological approaches, such as wound blotting and transcriptomic analysis, may further shed light on this unmet clinical need. The continuous development of these sophisticated diagnostic approaches can markedly contribute to the future implementation of point-of-care biofilm detection in chronic wound care.

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A combined pharmacodynamic quantitative and qualitative model reveals the potent activity of daptomycin and delafloxacin against Staphylococcus aureus biofilms.

Visualization of tumor necrosis factor-α distributions within pressure ulcer tissue using the wound blotting method: a case report and discussion.

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Detection and identification of specific bacteria in wound biofilms using peptide nucleic acid fluorescent in situ hybridization (PNA FISH).

DNA mimics for the rapid identification of microorganisms by fluorescence in situ hybridization (FISH).

Use of 16S ribosomal DNA PCR and denaturing gradient gel electrophoresis for analysis of the microfloras of healing and nonhealing chronic venous leg ulcers.

Use of dithiothreitol to improve the diagnosis of prosthetic joint infections.

The importance of the viable but non-culturable state in human bacterial pathogens.

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Inflammatory cells during wound repair: the good, the bad and the ugly.

The multicellular morphotypes of Salmonella typhimurium and Escherichia coli produce cellulose as the second component of the extracellular matrix.

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Community analysis of chronic wound bacteria using 16S rRNA gene-based pyrosequencing: impact of diabetes and antibiotics on chronic wound microbiota.

Glossary

a congregation of surface-attached microorganisms enclosed in extracellular polymeric substance.

a treatment strategy for chronic nonhealing wounds that aims to remove biofilms over wound beds with multiple treatment modalities.

a technique that separates DNA sequences with the same length but different sequences according to the exposed DNA denaturants (usually urea and formaldehyde) by exploiting the difference in the stability of G–C pairing as opposed to A–T pairing.

natural polymers secreted by microorganisms into the environment. EPS was previously used as an abbreviation for ‘extracellular polysaccharides’, but further studies revealed that extracellular substances in fact contain proteins, nucleic acids, lipids, and other biopolymers, such as humic substances, which led to the changed usage of EPS to refer to extracellular polymeric substances.

an approach to studying the diversity of the genetic materials of an environmental sample. Through targeting a partial 16S rRNA sequence, amplicons are produced via PCR then subcloned into vectors and transformed into a competent bacterial strain. The bacteria are then cultured, and the vectors are extracted for further sequencing.

a combination of techniques that leverages the benefit of partial 16S rRNA amplification and pyrosequencing in identifying the bacterial composition of a flora. Pyrosequencing is a next-generation sequencing technique that determines the sequences of samples via detecting specific nucleotides incorporated by DNA gyrase.

an approach to studying the diversity of the genetic material of an environmental sample. Through targeting a partial 16S rRNA sequence, amplicons are produced via PCR and then separated through denaturing gradient gel electrophoresis. Each band on the electrophoresis gel is then separated and further analyzed with Sanger sequencing.

combines peptide nucleic probes (which are sturdier and bind more quickly to target sites) along with FISH to improve the sensitivity and specificity of the FISH technique.

analyzes the presence and quantity of RNA in a biological sample through next-generation sequencing.

a state where a microorganism fails to be cultured in a condition that usually supports its growth while maintaining functions that are indicative of viability.