MIDDLEBROOK 7H9 BROTH

Technical Data #2488a / 2012.03.15

Middlebrook 7H9 Broth is used for the cultivation of Mycobacterium sp. Various cations and asparagine enhance the distinctive cord like growth of Mycobacterium sp.


FORMULA
in grams per liter purified filtered water

Potassium Phosphate, monobasic

1,0

Sodium Phosphate, dibasic

2,5

Sodium Glutamate

0,5

Sodium Citrate

0,1

Ammonium Sulfate

0,5

Ferric Ammonium Sulfate

0,04

Magnesium Sulfate

0,05

Copper Sulfate

0,001

Pyridoxine

0,001

Zinc Sulfate

0,001

Biotin

0,0005

Calcium Chloride

0,0005

Enrichments

 

Albumin Fraction V

5,0

Dextrose

2,0

Catalase

0,003

Glycerol

2,0 ml

pH 6,6 +/- 0,2 à 25º C

This approximate formula may be adjusted and/or enriched to obtain best results.


PRECAUTIONS
This medium is for IN VITRO diagnostic use only.


STORAGE
Store prepared media at 2-8° C protected from direct light and dehydrated powder, in a dry place in tightly-sealed containers at 2-25° C.


SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination (evaporation or discoloration). Do not use dehydrated media if it is caked.


DIRECTIONS
Suspend 4,7 g in 900 ml of purified filtered water containing 2 ml of glycerol (#8467). Heat with frequent agitation and boil for one minute. Sterilize at 121° C for 15 minutes. Cool to 45-50° C. Add 100 ml of ADC Enrichment (#8671). Mix thouroughly and dispense into sterile culture tubes.


PROCEDURE
Bring prepared medium at room temperature before inoculation.

  1. Inoculate specimens after decontaminating treatment.
  2. Incubate at 35° C for 3 weeks. Examine weekly for growth.
  3. Subculture growth on selective Middlebrook 7H11 Mitchison Agar Modified (#2851) and not selective Middlebrook 7H11 Agar (#2044) or Middlebrook 7H10 Agar (#2049).


QUALITY CONTROL

Expected results after 14 days at 35ºC under CO2 atmosphere

Organisms

ATCC

GROWTH

Mycobacterium tuberculosis H3Ra

25177

+

Mycobacterium kansasii Group 1

12478

+

Mycobacterium scrofulaceum Group II

19981

+

Mycobacterium fortuitum Group IV

6841

+


LIMITATIONS OF METHOD
This is only a part of the identification. Other tests may be required.


REFERENCES

  1. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American Society for Microbiology.
  2. Sommers and MacClatchy, Cumitech 16, Laboratory diagnosis of the mycobacterioses Coord. ed. Morello, ASM Washington D.C. 1983
  3. Finegold and Baron, Bailey and Scottís 19866 Diagnostic Microbiology 7th, Ed., The CV Mosby Co., St. Louis


CATALOGUE NUMBERS

Dehydrated media 500 grams QB-39-2950
     
Glycerol 100 ml 8467
     
Middlebrook ADC Enrichment 100 ml 8671
     
Prepared media    
Tubes 20x125 mm, 10 ml 10/pkg 2488


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