Biometric Metabolic Identification Using Fourier-Transform Infrared Spectroscopy
Keywords:
Fourier-Transform infrared (FTIR),, spectroscopy, high-throughput screeningAbstract
The rate of new antibiotic discoveries has recently slowed to a crawl. Antibiotic discovery has shifted back to phenotypic screening, but identifying mechanisms of action (MOA) remains a big challenge. Consequently, there is an urgent need for metabolic fingerprinting techniques like Fourier-Transform Infrared (FTIR) spectroscopy that can both deduce MOAs and screen complete cells at high throughput. To uncover the metabolic fingerprint caused by fifteen antibiotics on the metabolism of Escherichia coli, a bioassay based on high-throughput whole-cell FTIR spectroscopy was created. Spectra were swiftly obtained in the high-throughput mode after cells were exposed for a short period of time to a concentration four times greater than the minimum inhibitory concentration. Partially least squares discriminant and principal component analyses followed optimization of the preprocessing steps. Using either analysis algorithm, the biochemical fingerprints acquired using FTIR spectroscopy were highly specific enough to distinguish between several antibiotics across three separate cultures. These fingerprints aligned with the known modes of action (MOA) of all the antibiotics that were studied. This includes instances of antibiotics that target cell wall production, DNA, RNA, and proteins.
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