MIDDLEBROOK 7H11 AGAR

Technical Data #1380a / 2013.08.18

The Middlebrook 7H11 Agar is used for isolation and cultivation of Mycobacterium sp. The addition of Casein Peptone enhances Mycobacterium growth more than on Middlebrook 7H10. This medium may be used also for the Mycobacterium susceptibility.

 


 

FORMULA
in grams per liter purified filtered water

Casein Peptone

1,0

Ammonium Sulfate

0,5

Glutamic Acid

0,5

Sodium Citrate

0,4

Potassium Phosphate, mono-basic

1,5

Sodium Phosphate, dibasic

1,5

Magnesium sulfate

0,05

Ferric Ammonium citrate

0,04

Copper Sulfate

0,001

Biotin

0,0005

Pyridoxine

0,001

Zinc Sulfate

0,001

Malachite Green

0,00025

Calcium chloride

0,0005

Agar

13,5

Glycerol

5,0 ml

OADC Supplement

 

Albumin Fraction V

5,0 g

Dextrose

2,0

Sodium Chloride

0,85

Catalase

0,004

Oleic Acid

0,06 ml

pH 6,9 +/- 0,2 à 25º C

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


PRECAUTIONS
This medium is for laboratory use only.


STORAGE
Store prepared media at 2-8º C protected from direct light. Medium must not be used if there are signs of contamination or dehydratation or if the expiry date has passed. Store dehydrated powder in a dry place, in thightly-sealed containers at 2-25° C. Do not use dehydrated media if it is caked or if expiry date has passed.


DIRECTIONS
Suspend 19 g in 900 ml of purified water containing 5 ml of Glycerol. Heat with frequent agitation and boil for one minute. Sterilize at 121° C for 15 minutes. Cool to 45-50° C and, aseptically, add 100 mL of OADC Supplement. Dispense into sterile Petri dishes or culture tubes and let solidify in slant position.


PROCEDURE
Bring prepared medium at room temperature before inoculation. As usual, decontamination, homogenizing, grinding should be done before inoculation.

  1. Inoculate specimen as soon as possible.
  2. Incubate at 35° C in 5-10% CO2 for up to 3 weeks. Incubate tubes with loosened caps.


QUALITY CONTROL

Expected results after 21 days at 35ºC 5-10% CO2

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 medium 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. Middlebrook G., and M.L. Cohn Am. J. Publ. Health, 48 844
  3. Isenberg, 1992 Clinical microbiology procedures handbook vol 1 Americain Society for Microbiology, Washington, D.C.
  4. Forbes, Sahm and Weissfeld 1998 Bailey and Scott’s Diagnostic microbiology 10th ed Mosby Inc.


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-2974
     
Glycerol 500 ml 8466
OADC Supplement 100 ml 8673
     
Prepared media    
Plates, 100x15 mm 10/pkg 1380
Tubes, 20x125 mm 10/pkg 2044

 


 

 

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