Related Experiment Videos
[High-pressure liquid chromatography versus a radiometric method in the identification of aminoglycoside-acetylating
Background:
High Pressure Liquid Chromatography and a radiometric method for detecting and iso-enzyme identification of aminoglycoside-acetylation enzymes were compared. HPLC determines the acetylating activity of an enzyme by using its reaction product.
Material And Methods:
Three reference strains that produce AAC (3)-I, AAC (6')-IV and AAC (2') and 15 strains isolated from patients. Identification was performed using radiometric and HPLC method in all cases.
Results:
References strains: The enzymes produced by these strains were correctly identified with HPLC. Patient's strains: All give positive acetylase screening results with the chromatographic method, with holding time peaks that allows the identification of at least three different compounds. Identification at isozyme level was possible in all cases. In 14 out of 15 strains the results were the same using either technique. Radiometric method could not identify the enzyme of one strain.
Conclusion:
HPLC is a useful technique for isoenzyme identification of aminoglycoside-acetylation enzymes. The results were easily interpreted when compared to those of radiometric method. The group identification was possible with sisomycin.