H.B.T. (HUMAN BLOOD TWEEN) AGAR FOR GARDNERELLA

Technical Data #1280a / 2013.05.04

HBT for Gardnerella Agar is a selective layered medium for the isolation of Garderella vaginalis from clinical specimens. Antimicrobial agents, colistin, nalidixic acid and amphotericin B suppress vaginal normal flora growth. Addition of human blood permits differentiation by producing -hemolysis around colonies. Proteose peptone # 3 and polysorbate 80 are the enrichments.

 


 

FORMULA
in grams per liter purified filtered water

Casein Peptone

10,0

Meat Peptone

5,0

Yeast Extract

5,0

Beef Extract

3,0

Corn Starch

1,0

Sodium Chloride

5,0

Agar

13,5

Enrichments

 

Proteose Peptone #3

10,0

Polysorbate 80

0,08

Antibiotics  
Colistin

0,010

Nalidixic Acid

0,015

Amphotericin B

0,002

Human Blood

50 ml

pH 7,3 +/- 0,2 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 2-8C absolutely 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 42,5 g of dehydrated media in 950 ml of purified filtered water, add 10 g of Proteose peptone #3 and Polysorbate 80. Prepare twice. Heat with frequent agitation and boil for one minute. Sterilize at 121 C for 15 minutes. Cool to 45-50 C. Aseptically add 2x5 ml Antibiotics Supplements (8707) to one of sterile basal medium, mix gently and dispense into sterile Petri dishes. Let solidify. Then, aseptically add 2x5 ml Antibiotics Supplements (8707) and 50 ml of Human Blood to the other sterile basal medium. Mix gently and dispense overlaying the same sterile Petri dishes.


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

  1. Directly, inoculate plate as soon as possible with swab specimen and streak for isolation.
  2. Incubate at 35 C under 5-10% CO2 for 48-72 hours.
  3. Examine for characterestic haemolytic colonies.


QUALITY CONTROL

Results after 48 hrs at 35C w/ CO2

Organisms

ATCC

Growth

Hemolysis

Gardnerella vaginalis

14018

+

beta

Staphylococcus aureus

25923

+

-

Escherichia coli

25922

-

 
Proteus mirabilis

12453

-

 
Candida albicans

10231

- or partial

 


LIMITATIONS OF METHOD
This medium becomes unsuitable promptly if not properly stored causing premature lysis of red cells. This medium is only a part of the identification. Other tests may be required.


REFERENCES

  1. Greenwood, J.R., M.J. Pickett 1979 Salient features of Haemophilus vaginalis J. Clin. Microbiol. 9:200-204
  2. Greenwood, J.R. 1981 Gardnerella vaginalis Clin. Microbiol. Newsletter 3:23-25
  3. 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.
  4. Totten, P.A., R. Amsel, J. Hale, P. Piot, K.K. Holmes 1982 Selective differential human blood bilayer media for isolation of Gardnerella (Haemophilus) vaginalis J. Clin. Microbiol. 15(1):141-147.


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-1006
     
Polysorbate 80 100 ml 8465
     
Prepared media    
Plates, 100x15 mm 10/pkg 1280
Mono-Plates, 100x15 mm 10/pkg 1280M
     
Antibiotics Supplements: 5 ml 10/pkg 8707

 


 

 

2325, Dandurand, Suite 300 Phone Fax Internet
Montreal (Quebec) (514) 277-2558 (514) 277-4714 http://www.quelab.com
Canada H2G 1Z9 1 (800) 579-0998 1 (800) 579-8177 info@quelab.com