BORDET GENGOU METHICILLIN AGAR

Technical Data #1910a / 2000.11.01

Bordet Gengou Methicillin Agar is a selective medium used for the isolation of Bordetella pertussis and Bordetella parapertussis from contaminated clinical specimens. Potato infusion, casein peptone and glycerol are the essential nutrients for Bordetella growth. Defibrinated sheep blood permit the evaluation of hemolytic zone around colonies. Methicillin is the inhibitor.

FORMULA
in grams per liter purified filtered water

Potato Infusion Solids	4,5
Sodium Chloride	5,5
Agar	20,0
Methicillin	0,0025
Glycerol	10,0 ml
Defibrinated Sheep Blood	150,0 ml

pH 7,4 +/- 0,2 à 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 30 g in 850 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121° C for 15 minutes. Cool to 45-50° C and aseptically add 150 ml of Defibrinated Sheep Blood (4106) and 2,5 mg of sterile Methicillin solution. Dispense into sterile Petri dishes.


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

1. Inoculate plates, as soon as possible, with specimens and streak for isolation. Bordetella Pertussis Transport Medium (2555) should be used if inoculation is delayed.
2. Incubate at 35° C under a humid atmosphere for up to 7 days.
3. Examine the plates daily.
   
   

QUALITY CONTROL
Expected results after 3 days at 35ºC under humid atmosphere

Organisms	ATCC	Growth
Bordetella pertussis	8467	+
Bordetella pertussis	18323	+
Streptococcus pyogenes	19615	-
Streptococcus pneumoniae	6305	-
Staphylococcus aureus	25923	-

Colonies of Bordetella pertussis are tiny, smooth, with a characteristic compact, glistening appearance.

LIMITATIONS OF METHOD
As this selective medium may inhibit growth of Bordetella and colonies may appear after 4-6 days, a non- selective Bordet Gengou Agar should be inoculated simultaneously. This medium is only a part of the identification. Other tests may be required.


REFERENCES

1. Regan, J and Lowe, F. 1977 Enrichment Medium for the Isolation of Bordetella. J. Clin. Microbiol. 6: 303-309.
2. 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.
3. Gilligan,P.H., and M.C. Fisher. 1984 Importance of culture in laboratory diagnosis of Bordetella pertussis infections. J. Clin. Microbiol. 20:891-893
4. Mac Faddin, J.F. 1985 Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol.1 Williams and Wilkins, Baltimore.
5. Stauffer,L.R., D.R. Brown & R.E. Sandstrom 1983 Cephalexin-Supplemented Jones-Kendrick Charcoal Agar for Selective Isolation of Bordetella pertussis: Comparaison with Previously Described Media. J. Clin. Microbiol. 17:60-62.
6. Kurzynski,T., D.M.Boehm, J.A.Rott-Petri, R.F.Schell, P.E. Allison 1988 Comparaison of Modified Bordet Gengou & Modified Regan-Lowe Media for the Isolation of Bordetella pertussis & Bordetella parapertussis. J. Clin. Microbiol. 26:2661-2663

CATALOGUE NUMBERS

Dehydrated media	500 grams	QB-39-0126
Defibrinated Sheep Blood	200 ml	4106
Prepared media
Plate 100x15 mm	10/pkg	1910
Mono-Plate 100x15 mm	10/pkg	1910M
B. pertussis Transport Medium	10/pkg	2555

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