V AGAR

Cat. no. A80 V Agar, 15x100mm Plate, 17ml 10 plates/bag
Cat. no. A81 V Agar with CNA , 15x100mm Plate, 17ml 10 plates/bag
Cat. no. J181 V Agar / Starch Agar, 15x100mm Biplate, 10ml/10ml 10 plates/bag

INTENDED USE

Hardy Diagnostics V Agar is an enriched media used for the isolation and differentiation of Gardnerella vaginalis .

SUMMARY

V Agar is a Columbia Agar Base media with human blood added for differentiation of G. vaginalis from other genitourinary flora by its hemolytic reaction. G. vaginalis exhibits no hemolysis on Sheep Blood Agar, however, it is beta-hemolytic on Human Blood Agar.

V Agar with CNA contains additionally colistin and nalidixic acid for selective isolation of gram-positive cocci and certain gram-negatives such as G. vaginalis .

FORMULA

Ingredients per liter of deionized water:*

Columbia Agar Base 42.5gm
Proteose Peptone No. 3 10.0gm
Human Blood 50.0ml


V Agar with CNA, additionally, contains:
Colistin 10.0mg
Nalidixic Acid 15.0mg

Final pH 7.1 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-8ºC. away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), hemolysis, contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

Specimen Collection: Infectious material should be submitted directly to the laboratory without delay and protected from excessive heat and cold. If there is to be a delay in processing, specimen should be inoculated into an appropriate transport media and refrigerated until inoculation.

Method of Use: Prior to inoculation, the medium should be brought to room temperature. Inoculate media with specimen and streak for isolation using four quadrant technique. For testing an isolated organism, touch the top of a colony with a sterile wire loop and streak for isolation. Incubate in 5-10% CO2 at 35ºC. for 24-48 hours. Examine plate for growth and typical colony morphology and hemolysis.

INTERPRETATION OF RESULTS

Typical colonies of G. vaginalis appear convex, opaque and gray surrounded by a hazy zone of beta-hemolysis after 24-48 hours of incubation.

LIMITATIONS

V Agar is not selective for G. vaginalis and will support growth of other genitourinary organisms. G. vaginalis can be differentiated from other genitourinary organisms by its hemolytic reaction on V Agar.

V Agar with CNA is a semi-selective media for G. vaginalis . V Agar with CNA will support the growth of gram-positive cocci and certain gram-negative organisms such as G. vaginalis and Bacteriodes species. G. vaginalis can be differentiated from other organisms by its hemolytic reaction and biochemical reactions such as catalase, oxidase, hippurate hydrolysis, and antibiotic sensitivities.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms
Inoculation Method*
Incubation
Results
Time
Temperature
Atmosphere
V Agar:
Gardnerella vaginalis
ATCC ® 14018
A 24-48hr 35°C CO 2 ** Growth; pinpoint colonies with hazy beta-hemolysis
Streptococcus agalactiae
ATCC ® 12386
A 24hr 35°C Aerobic Growth; with beta-hemolysis
Additionally for V Agar with CNA:
Proteus mirabilis
ATCC ® 12453


B 24hr 35°C Aerobic Partial to complete inhibition
Escherichia coli
ATCC ® 25922
B 24hr 35°C Aerobic Inhibited

User Quality Control

PHYSICAL APPEARANCE

V Agar and V Agar with CNA should appear opaque, and cherry red in color.

G. vaginalis growing on V Agar

Gardnerella vaginalis (ATCC ® 14018) colonies growing on V Agar (Cat. no. A80). Photographed with backlight to emphasize hemolysis zones. Incubated in CO 2 for 48 hours at 35ºC.

S. agalactiae growing on V Agar

Streptococcus agalactiae (ATCC ® 12386) colonies growing on V Agar (Cat. no. A80). Photographed with backlight to emphasize hemolysis zones. Incubated aerobically for 24 hours at 35ºC.

V Agar

Uninoculated plate of V Agar (Cat. no. A80).

REFERENCES

1. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

2. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

3. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

4. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

5. MacFaddin, J.F. 1985. Media for Isolation, Cultivation, Identification, Maintenance of Bacteria, Vol. I. Williams & Wilkins, Baltimore, MD.

6. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.


ATCC is a registered trademark of the American Type Culture Collection.

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