N.T.M. (NEISSERIA TEST MEDIUM) AGAR GC MEDIUM WITHOUT HEMOGLOBIN

Technical Data #1752a / 2013.06.11

NTM Agar is a solid medium used for susceptibility testing of Neisseria gonorrheae. This test differentiate Neisseria gonorrheae by production or no-production lactamase. Routine methods for the rapidly growing pathogens, as described by Bauer et al. are not acceptable. Addition of X and V factors without L-Cysteine to an ordinary GC Medium permit visible growth.

 


 

FORMULA
in grams per liter purified filtered water

Casein Peptone

7,5

Meat Peptone

7,5

Corn Starch

1,0

Potassium Phosphate, dibasic

4,0

Potassium Phosphate, monobasic

1,0

Sodium Chloride

5,0

Agar

12,0

Enrichment

 

Bio-X without L-Cysteine HCl

10,0 ml

pH 7,3 +/- 0,1 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 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 or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.


DIRECTIONS
Suspend 36 g of GC dehydrated media in 1000 ml of purified filtered water and add 2 grams of bacteriological agar. Heat with frequent agitation and boil for one minute. Sterilize at 121 C for 15 minutes. Cool to 45-50 C. Add 10 ml of supplement Bio-X without L-Cysteine HCl. Mix gently and dispense into sterile Petri dishes.


PROCEDURE
Prior to inoculate, the prepared media should be brought to room temperature.

  1. From an overnight chocolate agar plate, suspend the suspected organism in Mueller Hinton Broth and adjust turbidity to be equivalent to that of the 0,5 McFarland standard.
  2. Following established procedures, inoculate plates by swabbing the inoculum on the surface three times.
  3. Place appropriate susceptibility disk on the surface and press gently.
  4. Incubate at 35 C with 5% CO2 for 24 hours.


QUALITY CONTROL

Results after 24 hrs at 35C W CO2

Organisms

ATCC

NCCLS STANDARDS

Neisseria gonorrheae

49226

meets NCCLS Standards

Staphylococcus aureus

25923

meets NCCLS Standards


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


REFERENCES

  1. Biddle, J.W., J.M. Swenson, C. Thornsberry 1978 Disc agar diffusion antimicrobial susceptibility tests with -lactamase producing Neisseria gonorrheae J. Antibiot. 31:352-358
  2. D'Amato, R.F., L. Hochstein 1982 Evaluation of a rapid inoculum preparation method for agar disk diffusion susceptibility tests J. Clin. Microbiol. 15:282-285
  3. Jones, R.N., T.C. Gavan, C. Thornsberry, P.C. Fuchs, E.H. Gerlach, J.S. Knapp, P. Murray, J.A. Washington 1989 Standardization of disk diffusion and agar dilution susceptibility tests for Neisseria gonorrheae Interpretative criteria and quality control guidelines for ceftriaxone, penicillin, spectinomycin and tetracycline. J. Clin. Microbiol. 27:2758-2766.
  4. 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.
  5. NCCLS 1990 Performance Standards for Antimicrobial Disk Susceptibility Tests. Approved Standard 4th ed. Document M2-A4 Vol.10, No.7
  6. NCCLS 1994 Performance Standards for Antimicrobial Susceptibility Testing 5th Informational Supplement Document M100-S5 Vol.14, No.16


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-1906
     
Prepared media    
Plates, 100x15 mm 10/pkg 1752
Plates, 150x15 mm 5/pkg 1852
     
M-H Broth    
Tubes 16x125 mm 10/pkg 2663

 


 





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