STANDARD OPERATING PROCEDURES FOR STOOL CULTURE

1. SPECIMEN:

1. Place a small quantity of stool in a wide mouth clean bottle. If mucus or flakes present - must be included.

2. CULTURE MEDIA:

1. MacConkey Agar (MA)
2. Selenite 'F' enrichment broth
3. Alkaline peptone water and TCBS agar
4. Deoxycholate Citrate Agar (DCA)
5. Blood Agar (BA)

3. PROCEDURE:

Isolation

1. Direct inoculation on MA and DCA . BA+TCBS if V. cholera is suspected

2. BA inoculated when membranous colitis is suspected

3. Inoculate into Selenite `F' enrichment broth if Salmonella/Shigella suspected.

4. Alkaline peptone water if V. cholera is suspected ( re-examine after 3-4 hrs of incubation.)

24 hours

1. Suspected Salmonella/Shigella colonies are identified and reported

2.Pure growth of any other Gram Neg. Bacteria in Children less than 3 years is identified and

reported.

3. Pure growth of staphylococci is reported

4. Vibrio cholerae is reported. If present.

If not positive for Salmonella/Shigella- Do a subculture from Selenite `F' enrichment broth.

48 hours

If none of the above is positive - Report as “No Salmonella/Shigella/ Vibrio grown.

A comment on normal flora can be made , if relevant.