Chromogenic Detection of Aminoglycoside Phosphotransferases

Acquired resistance to aminoglycosides is most frequently due to the presence of the so-called aminoglycoside modifying enzymes (AGME) (1 ) able to catalyze one or more of three general reactions: N-acetylation, O-nucleotidylation and O-phosphorylation (2 ). Although resistance phenotype (to different (substrate or not for enzymatic modification) may serve as an approach for identifying actual enzymes present in a given isolate (3 ), results can be obscured or confusing, particularly when several different enzymes (4 ) (even, isoenzymes with different affinities) are superimposing their action in a single microorganism with potential “permeability” or target alterations. Thus, identification of the AGME content of a given strain also requires screening at the DNA level using probes specific to all the known AGME (5 ). However, the complete set of probes is available only to a few laboratories around the world, making surveillance for the appearance of novel enzymes, or the unlikely evolution of those already known, a relatively nonfeasible goal, as search for new enzymes may begin only after failing to hybridize to all known probes.