Product Specification Sheet
Haemophilus Selective Agar (Chocolate Agar with Vitox, Bacitracin, Vancomycin, Amphotericin B)
Intended Usage: A selective medium for isolation of Haemophilus species from specimen containing mixed flora.
For professional use only.
PO5097A Revision Date: 03 January 2017
Haemophilus Selective Agar (Chocolate Agar with Vitox, Bacitracin, Vancomycin, Amphotericin B)
Thermo Scientific™ Haemophilus Selective Agar (Chocolate Agar with Vitox, Bacitracin, Vancomycin, Amphotericin B)
Form of Product Poured plate Storage 6 – 12°C Filling weight 17 g ± 5 % Packaging 10 plates wrapped in film pH 7.2 ± 0.2 Colour Nut brown, opaque Shelf life 8 weeks Intended Usage A selective medium for isolation of Haemophilus species from specimen containing mixed flora. For professional use only. Technique Depends on the different methods. For information see product information.
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Typical formulation* g/l
Special peptone 15.0
Corn starch 1.0
Sodium chloride 5.0
Dipotassium hydrogen phosphate 4.0
Potassium dihydrogen phosphate 1.0
Glucose 2.0
Defibrinated Horse Blood 100.0 ml
Vitamin B12 0.0002
L-Glutamine 0.2
Adenine 0.02
Guanine 0.0006 p-Aminobenzoic acid 0.00026
L-Cystine 0.022
NAD (coenzyme1) 0.005
Cocarboxylase 0.002
Ferric nitrate 0.0004
Thiamine 0.00006
Cysteine 0.518
Bacitracin 50000 IU
Vancomycin 6000 IU
Amphotericin B 0.008
Agar 10.0 *Adjusted as required to meet performance standards.
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Quality Control 1. Control for general characteristics, labeling and printing.
2. Control for sterility > 72 h @ 25 ± 1°C, aerobic > 72 h @ 36 ± 1°C, aerobic
3. Biological control Inoculum size for productivity: 10 – 100 cfu Inoculum size for selectivity: 104 – 105 cfu
Incubation conditions: 40 – 48 h @ 36 ± 1°C, aerobic, enhanced CO2 atmosphere
Control Strain Growth
Haemophilus influenzae ATCC® 10211™ 3 – 5 mm, cream shiny colonies.
Staphylococcus aureus ATCC® 25923™ Complete inhibition (≤ 10 cfu).
Candida albicans ATCC® 10231™ Complete inhibition (≤ 10 cfu).
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Description Haemophilus influenzae requires for growth the ingredients of erythrocytes designated as X and V factor1, a circumstance that also led to naming of the genus Haemophilus. The X factor corresponds here to the haemin components of the haemoglobin and V factor refers to the NAD (Nicotinamide-adenine-dinucleotide). In addition to a rich nutrient offering, the agar contains both growth factors as well as selective additions for inhibition of the accompanying flora from clinical materials. Bacitracin and Vancomycin suppress primarily gram-positive organisms while Amphotericin B is effective against yeasts and fungi.
Technique 1. Spread the investigation material, such as mucous membrane smears, wound secretions, puncture fluid, bronchial secretions, etc. directly in fractions. 2. Incubate the dishes for 24-48 hours, if necessary up to 72 hours at 36 ± 1°C. For primary cultures, the use of a moist chamber (70% humidity) and a CO2-enriched atmosphere can be advantageous. 3. Confirm presumptive Haemophilus species biochemically and/or serologically.
A CO2-enriched atmosphere can be easily and quickly established with CO2 Gen bags (CD0025A) in the anaerobe pot (AG0025A) or CO2 -Gen Compact (CD0020C) in anaerobe bags.
Literature 1. Holt, L.B. (1962), J. Gen. Microbiol., 27, 317-322.
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