Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Recommended for the selective isolation of Streptococcus pneumoniae.
Composition
| Ingredients | g/L |
|---|---|
| Proteose peptone | 15.000 |
| HL extract # | 2.500 |
| Yeast extract | 5.000 |
| Sodium chloride | 5.000 |
| Agar | 15.000 |
| Gentamicin | 5mcg/plate |
| Sheep blood | 50.000 ml |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
#-Equivalent to Liver extract
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 CFU) aseptically on the plate
Principle And Interpretation
Blood Agar Base No. 2 is a highly nutritive medium. Microorganisms producing haemolysin give visible haemolytic zones on this medium. Blood Agar with Gentamicin is recommended for the selective isolation of Streptococcus pneumoniae (1). The medium gave increased isolation rates of beta-hemolytic streptococci, other streptococci, Bacteroides, clostridia, and yeasts. It also proved valuable in detecting gentamicin-resistant gram-negative bacilli when these were present in heavy mixed culture (2).
HL extract and yeast extract helps enhance the growth and haemolytic reactions of fastidious organisms like Streptococci and Pneumococci. Proteose peptone serves as the nitrogen source while HL extract and yeast extract provide essential carbon, vitamin, nitrogen and amino acid sources. Sodium chloride maintains the osmotic equilibrium. Blood provides additional growth factors and also serves as basis for determining haemolytic reactions. Gentamicin inhibits other gram positive bacteria. Isolation of Streptococcus pneumoniae from respiratory secretions can be increased by using a 5% sheep blood agar plate supplemented with gentamicin.
Type of specimen
Clinical material : Isolated microorganism
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (3,4).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions
In Vitro diagnostic use. 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
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
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 Blood Agar w/ Gentamicin (5mcg/plate) in 90 mm disposable plate with smooth surface and absence of black particles/cracks/bubbles
Colour of Medium: Red coloured medium.
Quantity of Medium: 25ml of medium in 90 mm disposable plate.
pH: 7.10 -7.50
Sterility Check: Passes release criteria
Cultural Response: Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours (# Incubated anaerobically).
Cultural Response Table
| Organism | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Streptococcus pneumoniae ATCC 6303 | 50-100 | luxuriant | >=50% |
| Clostridium sporogenes ATCC 11437 | 50-100 | luxuriant | >=50% |
| Clostridium perfringens ATCC 13124 | 50-100 | luxuriant | >= 50% |
| Streptococcus pyogenes ATCC 19615 | 50-100 | luxuriant | >=70% |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=103 | inhibited | 0% |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | >=103 | inhibited | 0% |
Key: (*) Corresponding WDCM numbers.
Storage and Shelf Life
Store between 2-8°C. Use before expiry date on the performance is label. Product 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 (3,4).
| Product Name | Blood Agar Plate w/ Gentamicin (5mcg/plate) |
|---|---|
| SKU | MP5304 |
| Customized Product Available | No |
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