Evaluation of an acriflavine disk assay for differentiating Staphylococcus aureus from other staphylococci isolated from bovine milk.
OBJECTIVE: To develop an acriflavine disk assay for identification of Staphylococcus aureus and to test whether the acriflavine disk assay could be used to differentiate S aureus from other staphylococci isolated from bovine milk samples. DESIGN: Prospective study. SAMPLE POPULATION: 882 staphylococcal isolates from bovine milk samples and 3 S intermedius isolates from dogs. PROCEDURE: Paper disks saturated with various amounts of acriflavine were used in a growth inhibition assay to determine the amount of acriflavine that would most reliably differentiate S aureus from other staphylococci. For all isolates, hemolytic pattern, results of tube coagulase tests after 4 and 24 hours of incubation, growth on acriflavine-supplemented media, results of an acriflavine disk assay, and results of an automated identification system were determined. RESULTS: 10 micrograms of acriflavine/disk was determined to be the most appropriate concentration for use in the assay. All 112 isolates identified as S aureus by the automated identification system were resistant to this concentration of acriflavine, and only 1 of 236 isolates identified as non-S aureus staphylococci was resistant. There was substantial agreement between results of using the acriflavine disk assay as a diagnostic criterion for differntiating S aureus isolates from non-S aureus staphylococci and results of the automated identification system. Agreement between results of determining hemolytic pattern and results of other diagnostic tests was only moderate. CLINICAL IMPLICATIONS: The acriflavine disk assay, using 10 micrograms of acriflavine/disk, was a practical, accurate method for differentiating S aureus isolates from non-S aureus staphylococci.