STANDARD OPERATING PROCEDURES FOR SPUTUM CULTURE

1. SPECIMEN:

1. “Coughed up" specimen after rinsing the mouth may be sent in a screw capped container.

2. CULTURE MEDIA:

1. MacConkey Agar (MA)
2. Blood Agar (BA)
3. Cholcolate Agar (CA)
4. Thio Glycolate Broth (TGB)

3. PROCEDURE:

1. Inoculate sample into BA, MA and CA where indicated and incubate at 37C (in CO² jar if CA is

put) and also inoculate in TGB.

2. After 24 hours, observe if culture is negative:

i) Reincubate the plates

ii) Subculture from TGB if turbid and observe after another 24 hour

3. If culture is positive:
   1. If pure growth or 2 types - Identify and do antibiotic sensitivity
   2. If scanty growth ( <20 colonies) correlate with smear and report the identity with colony

numbers.

If direct smear shows significant number of Pus cells, then report as mixed flora with predominant colony type.

3. Pure diphtheroids/Coag Neg. Staph/ Viridans strep - Report as normal flora.
4. In case of mixed normal flora growing, correlate with smear and ask to submit a properly

collected sample