Hospital-acquired infections are to date a major challenge in the patient safety. The proliferation of pathogens such as Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae is often reported in connection with textiles, which represent a significant source of transmission. This leads most often to the contamination and cross-contamination of the hospitalized patient and the hospital staff. A promising approach and the immediate objective of this research is the application of probiotics to a textile fabric. They provide preliminary evidence in being able to inhibit pathogenic bacteria growth through their competitive mechanism. During this study, screen-printing was used as a method to apply probiotics on a polyester fabric. The viability of probiotics on the fabric was evaluated in the agar plate test method. Samples that exhibited a growth of grown out colonies were further tested regarding their efficacy towards the abovementioned pathogens. This was determined in a competition test, that included the individually inoculation of the samples with the different bacteria strains. Contact-angle measurements and abrasion resistance as well as the durability were tested in order to investigate the applicability of the fabric and scanning electron microscope images were taken to detect probiotics and to evaluate the quality of the print on the fabric. Major findings included that probiotics were able to survive on the polyester fabric and that these viable probiotics are further successful in the growth inhibition of pathogenic bacteria. Thereby their efficacy against pathogens is related to the viability the probiotics. As neither the probiotics used in this study, nor the other ingredients are considered as hazardous, this process is environment-friendly. This work increases the understanding of probiotics mechanism and their survival and competition behaviour on a textile material. It opens a way in the reduction of hospital-acquired infections through the beneficial effects of probiotics.