4. The mucolytic reagent can be used within 24 hours of

preparation, if stored at 2–8oC.

Processing of Specimen

1. Take approximately 2.5 mL of the specimen in a clean

sterile 15–25 mL universal container.

2. Add 2.5 mL of the mucolytic Reagent and close the

container tightly with a screw cap fitted with an intact

liner.

3. Mix well by gently vortexing at every 5 minutes interval

for 20 minutes.

4. After 20 minutes, unscrew the cap of the container

carefully and add 5 mL of reagent C.

5. Close again the container tightly as in step 2.

6. Mix well and centrifuge for 25 minutes at 3000–4000 g.

7. After centrifugation unscrew the cap of the container

with the content carefully and discard the supernatant

gently in an activated 2% glutaraldehyde solution,

taking care as not to disturb the pellet at the bottom.

8. To the pellet at the bottom, add 1 mL distilled water

and resuspend the contents.

9. Use this suspended material for microscopy (acid fast

bacilli), acid fast bacilli culture or polymerase chain

reaction.