Diphtheria Virulence HiVeg™ Agar Base

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MV882
Used for determining toxigenicity of Corynebacterium diphtheriae.


Intended Use

Diphtheria Virulence HiVeg Agar Base with supplement is used for testing toxigenicity of Corynebacterium diphtheriae.

Composition

Ingredients Grams/Litre
HiVeg peptone No. 3 20.0
Sodium chloride 2.5
Agar 15.0

Final pH (at 25°C) 7.8 ± 0.2

Formula adjusted, standardized to suit performance parameters.

Directions

Suspend 37.5 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 55-60°C. Aseptically add 2ml sterile Diphtheria Virulence Supplement (FD072) and 0.5 ml sterile 1% Potassium Tellurite (FD052) to a 100 mm petri plate and quickly add 10 ml of the sterile Diphtheria Virulence HiVeg Agar Base. Before the medium solidifies, place a filter paper strip saturated with potent Diphtheria antitoxin across the diameter of the plate. Allow the strip to sink to the bottom of the plate. Inoculate the plate with heavy inoculum across the strip.

Principle and Interpretation

Diphtheria Virulence HiVeg Agar Base is prepared by using HiVeg peptone No. 3 which is free of BSE/TSE risks in place Proteose peptone in the conventional medium. Diphtheria Virulence HiVeg Agar Base is the modification Diphtheria Virulence Agar Base formulated as per the Hermann et al (1) to support luxuriant growth of Corynebacterium diphtheriae. Elek (2) demonstrated an agar diffusion technique for testing virulence of Corynebacterium diphtheriae in-vitro. King et al (3) standardized the medium by comparing with animal inoculation tests to obtain consistent results. However, it was found that serum of different animals gives different results. Hermann et al (1) developed medium to overcome these difficulties. This medium is prepared on the same lines except that vegetable peptone is used. Upon incubation of the inoculated plate, a line of precipitin is observed for toxigenic strains. Potassium tellurite inhibits most gram-negative bacteria except Corynebacterium species, Streptococcus mitis, Streptococcus salivarius and Enterococci. Staphylococcus epidermidis may exhibit growth.

Product Profile

Vegetable based (Code MV)

  • MV882
  • HiVeg peptone No. 3

Animal based (Code M)

  • M882
  • Proteose peptone

Recommended for : Testing toxigenicity of Corynebacterium diphtheriae.

Reconstitution : 37.5 g/l

Quantity on preparation (500g) : 13.33 L

pH (25°C) : 7.8 ± 0.2

Supplement : Diphtheria Virulence Supplement (FD072), Potassium Tellurite (FD052)

Sterilization : 121°C / 15 minutes.

Storage : Dry Medium - Below 30°C, Prepared Medium 2 - 8°C.

Quality Control

Appearance of powder
Light yellow coloured, may have slightly greenish tinge, homogeneous, free flowing powder.

Gelling
Firm, comparable with 1.5% Agar gel.

Colour and Clarity
Medium amber coloured, slightly opalescent gel forms in petri plates.

Reaction
Reaction of 3.75% w/v aqueous solution is pH 7.8 ± 0.2 at 25°C

Cultural Response
Cultural characteristics observed after an incubation of 24 - 72 hours at 35 - 37°C with added Diptheria Virulence Supplement (FD072) and 1% Potassium tellurite solution (FD052).

Organisms (ATCC)

Organisms (ATCC) Growth Line of precipitin
Bacillus subtilis (6633) inhibited -
Corynebacterium diphtheriae type gravis luxuriant +
Corynebacterium diphtheriae type intermedius luxuriant +
Corynebactrium diphtheriae type mitis luxuriant +
Staphylococcus epidermidis (12228) none-poor -

References

  1. Hermann, Moore and Parsons, 1958, Am. J. Clin. Path., 29:181.
  2. Elek, 1948, Brit. Med. J., 1:493.
  3. King, Frobisher and Parsons, 1949, Am. J. Pub. Health, 39:1314.
More Information
Product Name Diphtheria Virulence HiVeg™ Agar Base
SKU MV882
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. Collee J. G., Fraser A. G., Marmion B. P., Simmons A., (Eds.), Mackie and McCartney, Practical Medical Microbiology,1996, 14th Edition, Churchill Livingstone2.Elek S. D., 1948, Br. Med. J., 1:493.3.King E. O., Frobisher M. and Parsons E. I., 1949, Am. J. Public Heal th, 39:1314.4.Hermann G. J., Moore M. S., and Parsons E. I., 1958, Am. J. Clin. Pathol., 29:181.5.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria. Vol. I, Williamsand Wilkins, Baltimore.6.Murray P. R., Baron E. J., Jorgensen J. H., Pfaller M. A., Yolken R. H., (Eds.), 8th Ed., 2003, Manual of ClinicalMicrobiology, ASM, Washington, D.C.7.Branson, 1972, Methods in Clinical Bacteriology, Charles C. Thomas,Springfield, III
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