B.H.I.A. VANCOMYCIN AGAR

Technical Data #1950a / 98.06.15

B.H.I.A. Vancomycin Agar, a selective medium, is used to screen vancomycin resistance on all isolated Enterococcus spp. from hospitalized patients. Vancomycin, the selective agent, permits the growth of vancomycin resistant bacteria only, such as Pediococcus, Leuconostoc and Enterococcus Vancomycin resistant. Enterococcus sp. shows positive results of both PYR and LAP Tests.

FORMULA
in grams per liter purified filtered water

Brain Heart Infusion	10,0
Casein Peptone	8,5
Meat Peptone	8,5
Sodium Chloride	5,0
Dextrose	2,5
Disodium Phosphate	2,5
Agar	15,0
Supplement
Vancomycin	0,006

pH 7,4 +/- 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 2-8º C protected from direct light and dehydrated powder, in a dry place in tightly-sealed containers at 2-25º 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 (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.

DIRECTIONS
Suspend 52 g in 1000 ml of purified water. Heat with frequent agitation and boil for one minute. Sterilize at 121º C for 15 minutes. Cool to 45-50º C and aseptically, add 2x5 ml of # 8736 VRE Vancomycin Supplement. Mix thouroughly and dispense into Petri dishes.

PROCEDURE
Bring prepared medium at room temperature before inoculation.

1. Inoculate a sterile saline tube with 4-5 similar colonies from an enterococci growth and adjust to 0,5 Mc Farland Standard.
2. Inoculate plates by a straight line on the surface.
3. Incubate at 35º C for 24 hours.

QUALITY CONTROL
Results after 24 hrs at 35ºC

Organisms	ATCC	Growth
Enterococcus faecalis	29212	-
Enterococcus faecalis	51299	+
Enterococcus faecalis	49533	+
Enterococcus gallinarum	clinical	+

LIMITATIONS OF METHOD
Gram positive cocci, catalase negative, bile esculin positive should be inoculated on this medium. This is only a part of the identification. Others tests may be required.

REFERENCES

1. Leclercq, R., E. Derlot, J. Duval, P. Courvalin 1988 Plasmid-mediated resistance to vancomycin and teicoplanin in Enterococcus faecium N. Engl. J. Med. 319:157-161.
2. Ballows, A., W.J. Hausler Jr., K.L. Herrmann, H.D. Isenberg, H.J. Shadomy, 1991 Manual of Clinical Microbiology. 5th ed. Washington D.C.: American society for Microbiology.
3. Facklam, R., D. Hollis, M.D. Collins 1989 Identification of gram-positive coccal and coccobacillary vancomycin-resistant bacteria. J. Clin. Microbiol. 27:724-730.
4. Swenson, J.M., N.C. Clark, M.J. Ferraro, D.F. Sahm, G. Doern, M.A. Pfaller, L.B. Reller, M.P. Weinstein, R.J. Zabransky, F.C. Tenover 1994 Development of a Standardized Screening Method for Detection of Vancomycin-Resistant Enterococci J. Clin. Microbiol. 32:1700-1704
5. Tenover, F.C., J. Tokars, J. Swenson, S. Paul, K. Spitalny, W. Jarvis 1993 Ability of clinical laboratories to detect antimicrobial agent-resistant enterococci J. Clin. Microbiol. 31:1695-1699.

CATALOGUE NUMBERS

Dehydrated media	500 g	QB-39-0206
VRE Vancomycin Supplement	10 vials/pkg	8736
Prepared media
Plates, 100x15 mm	10/pkg	1950
Mono-Plates, 100x15 mm	10/pkg	1950M

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