Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for isolation and cultivation of fastidious organisms.
Composition**
| Ingredients | g/L |
|---|---|
| Peptone, special | 23.000 |
| Corn starch | 1.000 |
| Sodium chloride | 5.000 |
| Agar | 15.000 |
| Defibrinated Sheep Blood | 50ml |
| Final pH (at 25°C) | 7.3±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate.
Principle And Interpretation
Columbia Blood Agar Base was devised by Ellner et al (1). This medium contains special peptone which supports rapid and luxuriant growth of fastidious and non-fastidious organisms. Also, this medium promotes typical colonial morphology; better pigment production and more sharply defined haemolytic reactions. Fildes found that Nutrient Agar supplemented with a digest of sheep blood supplied both of these factors and the medium would support the growth of H. influenzae (2,3). The inclusion of bacitracin makes the enriched Columbia Agar Medium selective for the isolation of Haemophilus species from clinical specimens, especially from upper respiratory tract (4). Columbia Agar Base is used as the base for the media containing blood and for selective media formulations in which different combinations of antimicrobial agents are used as additives. Corn starch serves as an energy source and also neutralizes toxic metabolites. Sheep blood permits the detection of haemolysis and also provides heme (X factor) which is required for the growth of many bacteria. However it is devoid of V factor (Nicotinamide adenine dinucleotide) and hence Haemophilus influenzae which needs both the X and V factors, will not grow on this medium.
Type of specimen
Clinical samples - throat swabs, pus, etc.
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (5,6). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
In Vitro diagnostic use only. For professional use only. Read the label before opening the container. Wear protective gloves/protective clothing/eye protection/face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling clinical specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations
- As this medium have a relatively high carbohydrate content, beta-haemolytic Streptococci may exhibit a greenish haemolytic reaction which may be mistaken for the alpha haemolysis.
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality Control
Appearance: Sterile Columbia Agar W/5% sheep blood in 90mm disposable plates with smooth surface and absence of black particles/ cracks/bubbles
Colour of medium: Red coloured medium
Quantity of medium: 25 ml of medium in 90mm disposable plates.
pH: 7.10-7.50
Sterility Check: Passes release criteria
Growth Promotion Test
In accordance with the harmonized method of USP/EP/BP/JP.
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours.
| Organism | Inoculum (CFU) | Growth | Recovery | Haemolysis |
|---|---|---|---|---|
| Neisseria meningitidis ATCC 13090 | 50-100 | luxuriant | >=70% | none |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | 50-100 | luxuriant | >=70% | beta / gamma |
| Staphylococcus epidermidis ATCC 12228 (00036*) | 50-100 | luxuriant | >=70% | gamma |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | 50-100 | luxuriant | >=70% | beta / gamma |
| Streptococcus pneumoniae ATCC 6303 | 50-100 | luxuriant | >=70% | alpha |
| Streptococcus pyogenes ATCC 19615 | 50-100 | luxuriant | >=70% | beta |
| Clostridium sporogenes ATCC 19404 (00008*) | 50-100 | luxuriant | >=50% | |
| Clostridium sporogenes ATCC 11437 | 50-100 | good-luxuriant | >=50% | |
| Clostridium perfringens ATCC 13124 (00007*) | 50-100 | luxuriant | >=50% | |
| Clostridium perfringens ATCC 12934 | 50-100 | luxuriant | >=50% |
Key: (*) Corresponding WDCM numbers.
Storage and Shelf Life
On receipt store between 2-8°C. Use before expiry date on the label. Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (5,6).
| Product Name | Columbia 5% Sheep Blood Agar Plate |
|---|---|
| SKU | MP144 |
| Customized Product Available | No |
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