A study into the mechanism of the Crystal Violet reaction in Staphylococcus aureus

The mechanism of the Crystal Violet (CV) reaction, a trait which has been related to biotype, source and pathogenicity in Staphylococcus aureus, was investigated in agar and broth studies. White reactions could be converted to purple and vice versa by altering the incubation conditions on agar. Broth reactions examined macroscopically and by spectrophotometry revealed that both white and purple human biotype strains take up CV but the former then progressively modify the dye more quickly than the latter. A cell-associated product of CV was detected in white and purple strains by reverse-phase thin-layer chromatography of methanol extracts. White strains appear to produce a second additional product from CV. The white reaction was not inhibited by chloramphenicol or azide but did depend on viable cells with a nutrient source. CV MICs and MBCs for 10 white and 10 purple reactors showed no gross differences in susceptibility, while a standardized assay for the rate of CV modifying activity (52 strains) demonstrated that the two categories comprise discrete populations which alter CV at different rates. Although most white strains belong to either or both of phage-typing groups V and II, purple strains with this pattern of susceptibility and white strains without it both occur. The capacity to modify CV slowly or rapidly appears to subdivide human biotype strains independently of their phage group and is associated in the former case with their capacity to produce hospital-acquired and invasive infections.