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LABORATORY PROCEDURES FOR ANIMAL & HUMAN CELL LINES

REFERENCE NO: AHC/1998/3/3.3/2.1


TITLE: BACTERIA AND FUNGI DETECTION BY CULTURE


INTRODUCTION

This protocol describes the detection of bacteria and fungi in cell cultures and reagents by isolation by culture.

PROCEDURE

Notes:

All cultures should be submitted for testing in antibiotic and cryoprotectant free medium. In addition cultures should not be passaged for three days prior to testing.

  1. Inoculate 2 Fluid Thiogylcollate Medium and 2 Soyabean Casein Digest (Tryptone Soya Broth) with 1.5ml of test article.
  2. Inoculate 2 Fluid Thiogylcollate Medium and 2 Soyabean Casein Digest (Tryptone Soya Broth) with 0.1ml of Candida albicans
  3. Inoculate 2 Fluid Thiogylcollate Medium and 2 Soyabean Casein Digest (Tryptone Soya Broth) with Bacillus subtilis
  4. Inoculate 1 Fluid Thiogylcollate Medium (Tryptone Soya Broth) with 0.1ml of Clostridium sporogenes
  5. Leave 2 of each media type un-inoculated as negative controls.
  6. For Tryptone Soya Broths, incubate one broth of each pair at 32+20C for 14 days and one broth of each pair at 22+20C for 14 days
  7. For Fluid Thioglycollate Medium broths, incubate the broth inoculated with Clostridium sporogenes and one broth of each pair at 32+20C for 14 days and one broth of each pair at 22+20C for 14 days
  8. Examine broths for turbidity after 14 days incubation

Record the results on the appropriate worksheet


Guidelines prepared for CABRI by CERDIC, DSMZ, ECACC, INRC, November 1998
Page layout by CERDIC
Copyright CABRI, 1998

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