QUAD PLATE: ENTEROCOCCI H.L.A.R. (HIGH LEVEL AMINOGLYCOSIDES RESISTANCE)

Technical Data #1851a / 98.06.15

The Enterococci H.L.A.R. Quad Plate, is used to detect enterococci resistance to high level of aminoglycosides and therapeutic synergy with ß-lactam positive antibiotics and glycopeptidic antibiotics. Aminoglycosides and glycopeptides may stop an ordinary enterococci infection but can not stop it if there is an highly resistance to aminoglycosides. Streptomycin, gentamycin and vancomycin are added to a Brain Heart Infusion Agar base.

FORMULA
in grams per liter purified filtered water

Brain Heard Infusion	10,0
Casein/Meat Peptone	17,5
Dextrose	2,0
Sodium Phosphate, dibasic	2,5
Sodium Chloride	5,0
Agar	15,0
QUAD I: Control X
QUAD II: Gentamycin	0,5
QUAD III: Streptomycin	2,0
QUAD IV: Vancomycin	0,006

pH 7,4 +/ñ 0,2 at 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.

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).

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

1. Suspend 2-3 similar colonies from a 24 hours growth into a B.H.I. Broth. Adjust turbidity to 0,5 McFarland Standard.
2. Inoculate each quadrant with 10 µl of this dilution. Let it dry.
3. Incubate at 35º C for 24-48 hours.

QUALITY CONTROL
Results after 24 hrs at 35ºC

Organisms	ATCC	QUAD I	QUAD II	QUAD III	QUAD IV
Enterococcus faecalis	29212	+	-	-	-
Enterococcus faecalis	49533	+	-	+	+
Enterococcus faecalis	51299	+	+	+	+

LIMITATIONS OF METHOD
Media should be reincubated for 24 hours more to detect a possible resistance. This medium is only a part of the identification. Other tests may be required.

REFERENCES

1. Balows, A., Hausler, W.J., Herrmann, K.L., Isenberg, H.D., Shadow, H.J. 1991 Manual of Clinical Microbiology, 5th ed. Washington D.C.: American Society for Microbiology
2. Sahm, D.F., S. Boonlayangoor, P.C. Iwen, J.L. Baade, G.L. Woods. 1991 Factors influencing determination of High-Level Aminoglycosides resistance in Enterococcus faecalis J. Clin. Microbiol. 29:1934-1039
3. Sahm, D.F., C. Torres, 1988 Effects of medium and inoculum variations on screening for high-level aminoglycoside resistance of Enterococcus faecalis. J. Clin. Microbiol. 26:250-256
4. Sahm, D.F., C. Torres 1988 High content aminoglycoside disk for determinating aminoglycoside-penicillin synergy against Enterococcus faecalis. J. Clin. Microbiol. 26:257-260

CATALOGUE NUMBERS

Prepared media
Plates, 100x15 mm	10/pkg	1851
Mono-Plates, 100x15 mm	10/pkg	1851M
BHI Broth
Tubes, 2ml, 13X100mm	10/pkg	2784

2331, Dandurand	Phone	Fax	Internet
Montreal (Quebec)	(514) 277-2558	(514) 277-4714	http://www.quelab.com
Canada H2G 3C5	1 (800) 579-0998	1 (800) 579-8177	info@quelab.com