Chromogenic detection of aminoglycoside phosphotransferases.
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.
Amoroso
A M
AM
Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Argentina.
Gutkind
G O
GO
D002863:Chromogenic Compounds / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D007525:Isoelectric Focusing; D019868:Kanamycin Kinase / Q000302:isolation & purification* / Q000378:metabolism; D011513:Proteus mirabilis / Q000201:enzymology*
D002863:Chromogenic Compounds; D019868:Kanamycin Kinase