The threat of multidrug resistant bacteria has been increasing steadily in the past century, posing a major health risk to the public. Even though every year, 226 million antibiotics are prescribed in the United States alone, 50% of the prescriptions are inappropriate for the patient’s condition. The increasing abuse of antibiotics in healthcare, as well as agriculture, has resulted in the rise of antibiotic resistance at an alarming rate. Traditional susceptibility testing requires extended cell incubation, delaying timely treatment response to the infection. Herein, we report that a short-pulse fluorescent D-amino acid (FDAA)-based approach provides insight not only into bacterial antibiotic susceptibility, but also into the mechanism of action of the antibiotic. In a clinical setting, fast and accurate recognition of the pathogen allows for the most effective choice of treatment, highlighting the need for a novel, fast, and accurate antibiotic susceptibility testing. Using FDAA-labeling signal as a reflection of peptidoglycan (PG) integrity after treatment with corresponding antibiotics, antibiotics targeting PG biosynthesis resulted in a significant decrease in fluorescence, while antibiotics affecting other cellular targets resulted in no fluorescence changes. This method was tested through fluorescent microscopy and spectrofluorometry, providing fast and accurate results and expanding FDAAs application in the antibiotic susceptibility studies.

Fluorescent D-amino acid-based approach enabling fast and reliable measure of antibiotic susceptibility in bacterial cells

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