How to Use Thayer Martin Medium Base

Thayer Martin Medium Base

Carpenter and Morton reported an improved medium to isolate Gonococci in 24 hours. Later on the efficiency of GC medium supplemented with haemoglobin and yeast concentrate was demonstrated for isolating gonococci. Subsequently Thayer Martin Medium Base was developed for the primary isolation of Neisseria gonorrhoeae and Neisseria meningitidis from specimens containing mixed flora collected from throat, vagina, rectum an d urethra. Thayer and Martin used Vancomycin, Colistin and Nystatin. Martin and Lester used an additional antibiotic, Trimethoprim to make the medium selective.

PRINCIPLE

Special peptone provides nutrients to the organisms while starch neutralizes the toxic fatty acids if present in the agar. Haemoglobin provides the X factor whereas the V factor (N.A.D.) is provided by the added supplement. Starch neutralizes the toxic fatty acids if present in the agar. Supplement (TS 022) also supplies vitamins, amino acids, coenzymes etc. which enhances the growth of pathogenic Neisseria. Vancomycin and colistin inhibits gram-positive and gram-negative bacteria respectively. Nystatin inhibits fungi. This medium may inhibit Haemophilus species. Some strains of Capnocytophaga species may grow on this medium when inoculated with oropharyngeal specimens.

INSTRUCTION FOR USE

1. Dissolve 42.0 grams in 900 ml distilled water.
2. Heat to boiling to dissolve the medium completely.
3. Sterilize by autoclaving at 15 psi pressure (121°C) for 15 minutes.
4. Cool to 45-50°C.
5. Aseptically add 100 ml of sterile lysed blood and rehydrated contents of one vial of Vitamins     Amino Growth Supplement (Vitamins & Amino Acids Mixture) (TS 022) and V.C.N Supplement (TS 038) or V.C.N.T Supplement (TS 039).
6.  If desired GC Supplement with Antibiotics (TS 036) can be used as a single supplement.
7. Mix well before pouring into sterile Petri plates.
8. If Hemoglobin powder (TS 021) is used suspend 42.0 grams of Thayer Martin Medium Base in 500 ml distilled water.
9. Heat to boiling to dissolve the medium completely.
10. Prepare 500 ml of 2% hemoglobin solution.
11. Sterilize separately by autoclaving at 15 psi pressure (121°C) for 15 minutes.
12. Cool to 45°C.
13. Mix both and add the supplements as above. 

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