LOWENSTEIN JENSEN GRUFT MEDIUM

Technical Data #2161a / 2013.06.23

The selective Lowenstein Jensen Gruft Medium is used for isolation of Mycobacterium sp. from contaminated specimens. Egg supplement enhances Mycobacterium growth. Malachite green, penicillin and nalidixic acid inhibit bacteria Gram negative and Gram positive. Ribonucleic Acid is used as growth factor.

 


 

FORMULA
in grams per liter purified filtered water

L-Asparagine

3,6

Monopotassium Phosphate

2,4

Magnesium Sulfate

0,24

Magnesium Citrate

0,6

Potato Flour

30,0

Malachite Green

0,4

Glycerol

12,0 ml

Eggs

1 L

Penicillin

80,000 UI

Nalidixic Acid

0,056 g

Ribonucleic Acid

0,080 g

pH 7.2 +/- 0,2 25 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. Medium must not be used if there are signs of contamination or dehydration or if the expiry date has passed. Store dehydrated powder in a dry place, in tightly-sealed containers at 2-25 C. Do not use dehydrated media if it is caked or if expiry date has passed. The enrichment must be stored at 2-8 C, in a dark place. Do not use if expiry date has passed or if it is contaminated.


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 or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.


DIRECTIONS
Suspend 37,2 g in 600 ml of purified water containing 12 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 1 L of sterile eggs. Dispense into sterile culture tubes and coagulate at 85 C for 45 minutes in slant position.


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

  1. Inoculate tubes using light inoculum.
  2. Incubate inoculated tubes with loosened caps, at 35 C in 5-10% CO2.
  3. Read weekly for at least 2 months, if there is no growth.


QUALITY CONTROL

Expected results after 21 days at 35C 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

+

Escherichia coli

25922

- (partial)

Staphylococcus aureus

25923

- (partial)


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


REFERENCES

  1. Gruft, H. 1965 Nalidixic acid as a decontamination in Lowenstein Jensen medium. J.Bacteriol. 90:829.
  2. N.C.C.L.S. Quality Assurance for Commercially prepared Microbiological Culture Media. Approved Standard. 1990 Document M22-A.
  3. Sommers, H.M., Mc Clatchy, J.K., Morello, J.A. 1983 Laboratory Diagnosis of the Mycobacteriosis. Cumitech 16. American Society for Microbiology. Washington, D.C.,
  4. Vestal, A.L. 1978 Procedures for the Isolation and Identification of Mycobacterium. DHEW Publication No. CDC 79-8230, CDC Atlanta.


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-2620
Glycerol 500 g QB-39-9012
     
Prepared media    
Tubes 20x125 mm, slant 10/pkg 2161
Tubes 20x125 mm, slant 100/box 2161-H

 


 

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