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Published on: October 20, 2012
Analysis of myeloperoxidase activity in wound fluids as a marker of infection
A Hasmann1, E Wehrschuetz-Sigl, A Marold
1Department of Environmental Biotechnology, Graz University of Technology, Petersgasse 12, Graz, Austria.
Background:
Neutrophilic polymorphonuclear leukocytes play a crucial role in the host defence against bacterial and fungal infections. They participate in the inflammatory response through the liberation of peptides and enzymes like myeloperoxidase (MPO). Therefore, MPO has a potential as a marker enzyme for the diagnosis of wound infection.
Methods:
Substrate specificities and reaction pathways of MPO were investigated for new MPO substrates: crystal violet, leuco crystal violet, fast blue RR (4-benzoylamino-2,5-dimethoxybenzenediazonium chloride hemi(zinc chloride) salt) and various systematically substituted model substrates based on 2,7-dihydroxy-1-(4-hydroxyphenylazo)naphtalene-3,6-disulphonic acid. In addition, fast blue RR was covalently bound to siloxanes allowing immobilization of the substrate, while cellobiosedehydrogenase was integrated for generation of hydrogen peroxide required by MPO.
Results:
Elevated concentrations of MPO were found in infected wounds compared with non-infected wounds (92.2 ± 45.0 versus 1.9 ± 1.8 U/mL). Various soluble and immobilized substrates were oxidized by MPO in wound samples and the influence of substrate structure and reaction pathways were elucidated for selected compounds.
Conclusions:
Incubation of different MPO substrates with infected wound fluid samples resulted in a clear colour change in the case of elevated MPO concentrations, thus allowing early diagnosis of wound infection.
Insights
Myeloperoxidase (MPO) levels in wound fluid can diagnose infections early. Elevated MPO concentrations in infected wounds lead to a clear color change with specific substrates, enabling rapid detection.
Area of Science:
- Biochemistry
- Clinical Diagnostics
- Medical Microbiology
Background:
- Neutrophilic polymorphonuclear leukocytes are vital for host defense against infections.
- Myeloperoxidase (MPO), an enzyme released by leukocytes, plays a role in inflammation.
- MPO shows potential as a diagnostic marker for wound infections.
Purpose of the Study:
- To investigate substrate specificities and reaction pathways of MPO.
- To explore new MPO substrates for potential diagnostic applications.
- To evaluate MPO as a marker for early wound infection diagnosis.
Main Methods:
- Investigated MPO activity using novel substrates including crystal violet and fast blue RR.
- Developed immobilized MPO substrates for enhanced detection capabilities.
- Utilized cellobiosedehydrogenase for hydrogen peroxide generation required by MPO.
Main Results:
- Significantly higher MPO concentrations were observed in infected wounds (92.2 U/mL) versus non-infected wounds (1.9 U/mL).
- MPO oxidized various soluble and immobilized substrates in wound samples.
- Elucidated the influence of substrate structure and reaction pathways on MPO activity.
Conclusions:
- A clear color change occurred upon incubation of MPO substrates with infected wound fluid.
- Elevated MPO concentrations correlate with wound infection presence.
- This method allows for early diagnosis of wound infection.
