Evaluation of simple, noninvasive methods for sampling and quantification of skin bacteria
2024
Dziedzic, Kamila
The increased incidence of diabetes mellitus has underscored the importance of effective management strategies, particularly in preventing complications such as diabetic foot ulcers (DFU). Chronic infections associated with DFU pose significant health risks, including lower limb amputations, highlighting the urgent need for non-invasive methods to assess skin microbiota changes. This study aimed to evaluate simple methods of sampling and quantifying skin bacteria, comparing techniques such as Gram staining, DAPI staining, fluorescence in situ hybridization (FISH), and polymerase chain reaction (PCR). Furthermore, the study investigated bacterial abundance variations across different sampling sites on the foot. Skin bacteria were sampled from healthy human volunteers using tape stripping (TPS) and swabbing. Gram-staining of the samples showed that most bacteria were found on the heel of the foot, and only Gram-positive bacteria were found on the skin of healthy study participants. However, Gram-staining showed artifacts in the form of bubbles under the microscope, which interfered with bacteria counting. PCR provided results indicating the presence of Staphylococcal species on the skin of healthy feet. DAPI staining showed images of bacteria like the ones stained with Gram staining. After using FISH-probe it was found that only a few bacteria hybridized with the probe and further optimization of the protocol is required. The study evaluated various techniques for sampling and quantifying skin bacteria and compared the number of bacteria present on the foot of healthy individuals, which may be used to identify infections before they develop into more serious conditions.
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