Wayne Sulphatase Agar Base

Wayne Sulphatase Agar Base is recommended for biochemical differentiation of Mycobacteria on the basis of their ability to produce aryl sulphatase. This medium was developed by Wayne (to enable recognition of M.fortuitum ) using a 3-day phenolphthalein sulphatase test. Rapid growing and slow growing species of Mycobacterium can be differentiated, based on the 3 days test or 2 weeks test respectively. Some species of Mycobacteria produce arylsulphatase, an enzyme that attacks the substrate component viz. tripotassium phenolphthalein sulphate, with the resultant release of free phenolphthalein as indicated by a colour change (red) in the medium after addition of sodium bicarbonate reagent. After incubation of 3-14 days, add 0.5 to 1.0 ml of 2N Na2CO3 to each tube and observe the colour change within 30 minutes.