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REF 43 411 08492 F - en -2010/10

Gardnerella Agar (GAR) Selective isolation of Gardnerella vaginalis

SUMMARY AND EXPLANATION Gardnerella agar is a selective isolation medium for the detection of Gardnerella vaginalis in genital specimens. Gardnerella vaginalis, alone or associated with certain anaerobic bacteria (Mobiluncus, Bacteroides and Prevotella), is responsible for various urogenital infections (1).

PRINCIPLE The presence of human blood favors the growth of the species being tested for and produces ß hemolysis around the colonies (2, 3). The antibiotics included in the medium inhibit most Gram (-) contaminants and yeasts (4).

CONTENT OF THE KIT

Ready-to-use media REF 43 411 Pack of 2 x 10 plates (90 mm) GAR *

* printed on each plate

COMPOSITION Theoretical formula This medium can be adjusted and/or supplemented according to the performance criteria required: Casein and meat peptone (bovine and porcine) ................................10 g Hydrolyzed animal protein (bovine or porcine)...................................10 g Heart peptone (bovine or porcine) .......................................................3 g Corn starch..........................................................................................1 g Sodium chloride...................................................................................5 g Agar................................................................................................13.5 g Tween 80......................................................................................0.075 g Human erythrocytes* ....................................................................... 50 ml Nalidixic acid.................................................................................0.015 g Colistin..........................................................................................0.010 g Amphotericin B .............................................................................0.002 g Purified water........................................................................................1 l

pH 7.1

* This product has been tested and shown to be negative for HBs antigen, antibodies to HIV1, HIV2 and HCV. However, since no existing test method can totally guarantee their absence, this product must be treated as potentially infectious. Therefore, usual safety procedures should be observed when handling.

MATERIAL REQUIRED BUT NOT PROVIDED • Controlled atmosphere generators. • Jars. • Bacteriology incubator. • Or thermoregulated chambers with a controlled

atmosphere.

POSSIBLE ADDITIONAL REAGENTS • Columbia agar + 5% sheep blood (Ref. 43 041).

WARNINGS AND PRECAUTIONS • For in vitro diagnostic use only. • For professional use only. • This kit contains products of human origin. No

known analysis method can totally guarantee the absence of transmissible pathogenic agents. It is therefore recommended that these products be treated as potentially infectious and handled observing the usual safety precautions (see Laboratory biosafety manual - WHO - Geneva - latest edition).

• This kit contains products of animal origin. Certified knowledge of the origin and/or sanitary state of the animals does not totally guarantee the absence of transmissible pathogenic agents. It is therefore recommended that these products be treated as potentially infectious, and handled observing the usual safety precautions (do not ingest or inhale).

• All specimens, microbial cultures and inoculated products should be considered as infectious and handled appropriately. Aseptic technique and usual precautions for handling the bacterial group studied should be observed throughout this procedure. Refer to "CLSI® M29-A, Protection of Laboratory Workers From occupationally Acquired Infections; Approved Guideline – Current Revision". For additional information on handling precautions, refer to "Biosafety in Microbiological and Biomedical Laboratories – CDC/NIH – Latest Edition " or the current regulations in the country of use.

• Culture media should not be used as manufacturing material or components.

• Do not use reagents past the expiration date. • Do not use reagents if the packaging is damaged. • Do not use contaminated or hemolyzed plates, or plates

that exude moisture. • The performance data presented were obtained using

the procedure indicated in this package insert. Any change or modification in the procedure may affect the results.

• Interpretation of the test results should be made taking into consideration the patient's history, the source of the specimen, colonial and microscopic morphology and, if necessary, the results of any other tests performed.

STORAGE CONDITIONS • The plates can be stored in their box at 2-8°C until

the expiration date. • If not in the box, plates can be stored for 2 weeks at

2-8°C in the cellophane sachet.

SPECIMENS The medium is inoculated directly from genital specimens. Good laboratory practices for collection and transport should be respected.

IVD

Gardnerella Agar (GAR) 08492 F - en -2010/10

bioMérieux SA English - 2

INSTRUCTIONS FOR USE

1. Allow the plates to come to room temperature. 2. Inoculate the specimen and then place the plate in a

suitable atmosphere (anaerobiosis), if necessary using a controlled atmosphere generator.

3. Incubate with the cover bottom side at 37°C. The user is responsible for choosing the appropriate incubation temperature for the intended use; in accordance with current standards. The cultures are examined after 48 hours of incubation. In certain cases, it may be necessary to prolong incubation.

Note: In order to compare the hemolysis on reading, it is recommended that a Columbia agar with sheep blood be inoculated in parallel.

READING AND INTERPRETATION • After incubation, observe the bacterial growth. • Record the presence of characteristic ß hemolysis: clear

zone around or under the colony. ß hemolysis obtained solely on human blood agar is highly presumptive of G. vaginalis.

• This indication must be confirmed by performing direct examination and taking into consideration the patient's history.

• Identification of suspect colonies must be followed by biochemical tests.

QUALITY CONTROL

Protocol : The medium can be tested using the following strains: • Gardnerella vaginalis ATCC® 14018

(incubation in an anaerobic atmosphere). • Candida albicans ATCC ® 60193

Range of expected results:

Strain Results at 33-37°C Gardnerella vaginalis

ATCC ® 14018 Growth after

48 hours ß hemolysis

Candida albicans ATCC ® 60193

Partial inhibition within 48 hours

Note : It is the responsibility of the user to perform Quality Control taking into consideration the intended use of the medium, and in accordance with any local applicable regulations (frequency, number of strains, incubation temperature... ).

LIMITATIONS OF THE METHOD • Growth depends on the requirements of each individual

microorganism. It is therefore possible that certain strains of G. vaginalis which have specific requirements may not develop.

PERFORMANCE Performance was evaluated at 37°C using 39 bacterial strains (G. vaginalis, Lactobacillus, corynebacteria, other Gram (+) and Gram (-) bacteria and anaerobic bacteria) and 3 yeasts (Candida and Saccharomyces).

Nutrient capacity : The 11 strains of G. vaginalis grew within 24 hours as did 14 of the other 15 Gram (+) bacteria.

Selectivity : The 8 Gram (-) strains tested and the 3 yeasts were inhibited within 48 hours. Four out of the 5 anaerobic strains tested did not grow within 48 hours.

Hemolysis : All the strains of G. vaginalis showed β hemolysis within 48 hours. Note: β hemolysis is not characteristic of this species: certain other Gram (+) bacteria also produced β hemolysis.

WASTE DISPOSAL Dispose of used as well as any other contaminated disposable material following procedures for infectious or potentially infectious products. It is the responsibility of each laboratory to handle waste and effluents produced according to their nature and degree of hazardousness and to treat and dispose of them (or have them treated and disposed of) in accordance with any applicable regulations.

LITERATURE REFERENCES 1. CASIN I. – Diagnostic bactériologique de Gardnerella

vaginalis - Feuillets de Biologie, 1986, vol. 27, n°149, p. 25-30.

2. LEFEVRE J.C., PRERE M.F., LARENG M.B. – Isolement et identification de Gardnerella vaginalis en pratique courante - Rev. Méd. Toulouse, 1983, vol. 19, p. 117-119.

3. PIOT P., VAN DYCK E., TOTTEN P.A. and al. – Identification of Gardnerella (Haemophilus) vaginalis - J. Clin. Microbiol., 1982, vol. 15, n°1, p. 19-24.

4. TOTTEN P.A., AMSEL R., HALE J. and al. – "Selective differential human blood bilayer media for isolation of Gardnerella (Haemophilus) vaginalis - J. Clin. Microbiol., 1982, vol. 15, n°1, p. 141-147.

Gardnerella Agar (GAR) 08492 F - en -2010/10

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INDEX OF SYMBOLS

Symbol Meaning

Catalogue number

In Vitro Diagnostic Medical Device

Manufacturer

Temperature limitation

Use by

Batch code

Consult Instructions for Use

Contains sufficient for <n> tests

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