Chromogenic Detection of Aminoglycoside Phosphotransferases |
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| Authors: | Ana M. Amoroso and Gabriel O. Gutkind |
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| Affiliation: | Laboratorio de Resistencia Microbiana, Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina |
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| Abstract: | A coupled chromogenic reaction (based on an agar overlay combining NADH, pyruvate kinase, lactate dehydrogenase, phosphoenolpyruvate, ATP, and kanamycin sulfate with thiazolyl blue-phenazine methosulfate for detection of NADH consumption) was optimized for the detection of aminoglycoside phosphotransferases (APHs). When used after analytical isoelectrofocusing of bacterial extracts from APH-producing strains, this method revealed one band in each of two strains with a genetically confirmed APH (3′) I and two bands in another strain with both APH (3′) I and APH (3′) VI, whereas no bands were detected in susceptible control strains or in aminoglycoside-resistant microorganisms without APH genes. |
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