All fermentation tubes showing gas production in presumptive tests within 48 hours at 350C shall be utilized in the confirmed test Eosin methylene blue( E.M.B ) agar, Endo agar or brilliant green lactose bile broth fermentation tubes may be used in the test A loop-full of culture from each positive fermentation tubes is streaked over the surface of E.M.B agar or Endo agar. Development of typical colonies (nucleated, with or without metallic sheen) or atypical colonies (opaque, nonucleated mucoid, pink) the confirmed test may be considered positive If no colonies develop with in the incubation period the confirmed test may considered negative.
Brilliant green lactose bile broth (BGLBB)
A loop-full of culture from each positive presumptive tube is transferred to brilliant green lactose bile broth fermentation tubes and incubated at appropriate temperature and time
Presence of gas in any amount in the inverted vial within the incubation period may be positive confirmed test
Transfer typical or atypical colonies from E.M.B or Endo agar to lactose fermentations tubes and nutrient agar slants and incubate at appropriate temperature for a period not to exceed 48hours
If Brilliant green lactose bile broth is used in the confirmed test, an E.M.B or Endo agar plate is streaked from each fermentation