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Antimicrobial Agents and Chemotherapy, February 1998, p. 228-230, Vol. 42, No. 2
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Chromogenic Detection of Aminoglycoside Phosphotransferases

Ana M. Amoroso and Gabriel O. Gutkind*

Laboratorio de Resistencia Microbiana, Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina

Received 14 May 1997/Returned for modification 14 August 1997/Accepted 18 November 1997

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.

* Corresponding author. Mailing address: Laboratorio de Resistencia Microbiana, Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 954, Piso 8, 1113 Buenos Aires, Argentina. Phone: 54 1 964 8285. Fax: 54 1 962 5341. E-mail: ggutkind{at}huemul.ffyb.uba.ar.

Antimicrobial Agents and Chemotherapy, February 1998, p. 228-230, Vol. 42, No. 2
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.




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