Infection is a global problem. The resulting pain, impairment and social isolation lead to reduced quality of life and, in the worst case, hospitalization, and eventually sepsis and death. Infected wound fluids contain several enzymes, produced by the cells involved in the immune response, which can be used as markers of infection. Myeloperoxidase (MPO) is one of the enzymes present in the white blood cells and implicated in the first response of the immune system upon inflammation or injury. MPO substrates are phenolic molecules developing color upon oxidation by this enzyme. Point of care (PoC) devices are well-known for the fast detection of hormones and metabolites associated to diseases or other body conditions. Here we show the initial steps to build up a PoC gadget to detect MPO levels after injury following two different strategies. In a first approach, several substrates for MPO, chosen by the distinguishable products obtained upon reaction, have been immobilized on a nitrocellulose membrane. In a second approach, a specific antibody has been coated on the membrane and upon enzyme binding; MPO is detected by the addition of the substrates. Infected and non-infected wounds activity levels of MPO have been tested on both approaches. Our results show that MPO wicks on the membrane and reacts with the immobilized substrates and binds to the antibody keeping its activity. High activities of MPO showed a characteristic change of color on the test area while lower activities were undetectable. However, MPO wicking is not uniformly distributed through the membrane and new strategies for improvement are needed on this matter. The development of this device will help to distinguish between infected and non-infected wounds reducing unnecessary antibiotic administration and consequently decrease antibiotic resistance.

Point of care device for detection of myeloperoxidase as a wound infection marker

251st National Meeting (2016)

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