Rapid Antibiotic Susceptibility Determination for Yersinia pestis Using Flow Cytometry Spectral Intensity Ratio (SIR) Fluorescence Analysis
| Autor: | David Gur, Eran Zahavy, Moshe Aftalion, Shahar Rotem, Raphael Ber, Ronit Aloni-Grinstein |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Microbiological culture Time Factors Sociology and Political Science medicine.drug_class Yersinia pestis Clinical Biochemistry Population Antibiotics Microbial Sensitivity Tests Bacterial growth Biochemistry Flow cytometry Microbiology Antibiotic susceptabilty test 03 medical and health sciences Minimum inhibitory concentration medicine education Spectroscopy education.field_of_study medicine.diagnostic_test biology Chemistry Bacterial fluorescence biology.organism_classification Anti-Bacterial Agents Clinical Psychology 030104 developmental biology Spectrometry Fluorescence Original Article Law Social Sciences (miscellaneous) Bacteria |
| Zdroj: | Journal of Fluorescence |
| ISSN: | 1573-4994 |
| Popis: | Rapid antimicrobial susceptibility tests (ASTs) are essential tool for proper treatment of patients infected by Yersinia pestis (Y. pestis), the causative agent of plague, or for post-exposure prophylaxis of a population exposed to a naturally acquired or deliberately prepared resistant variant. The standard AST of Y. pestis is based on bacterial growth and requires 24–48 h of incubation in addition to the time required for prior isolation of a bacterial culture from the clinical or environmental sample, which may take an additional 24–48 h. In this study, we present a new and rapid AST method based on a fluorescence determination of the minimum inhibitory concentration (MIC). Our method includes the incubation of bacteria with an antibiotic, followed by staining of the bacteria with oxonol dye (SynaptoGreen C4/FM1–43), which enables the rapid detection of an antibiotic’s effect on bacterial viability. We show that stained, non-viable bacteria exhibit a spectral redshift and an increase in fluorescence intensity compared to intact control bacteria. Based on these criteria, we developed a rapid flow cytometer measurement procedure and a unique spectral intensity ratio (SIR) analysis that enables determination of antibiotic susceptibility for Y. pestis within 6 h instead of the 24 to 48 h required for the standard AST. This new rapid determination of antibiotic susceptibility could be crucial for reducing mortality and preventing the spread of disease. |
| Databáze: | OpenAIRE |
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