Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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HiCrome™ MeReSa HiCynth™ Agar Base
For the isolation and selective identification of Methicillin Resistant Staphylococcus aureus (MRSA) from clinical isolates.
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Intended use
Recommended for the isolation and selective identification of Methicillin Resistant Staphylococcus aureus (MRSA).
Composition
| Ingredients | g/L |
|---|---|
| HiCynth™ Peptone No.2* | 13.000 |
| HiCynth™ Peptone No.5* | 5.000 |
| Sodium chloride | 40.000 |
| Sodium pyruvate | 5.000 |
| Chromogenic mixture | 5.300 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.0±0.2 |
**Formula adjusted, standardized to suit performance parameters
*Chemically Defined peptones
Directions
Suspend 41.65 gram in 500 ml purified / distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Aseptically add sterile rehydrated contents of 1 vial of MeRS Selective Supplement (FD229) & CF Selective Supplement II (FD259) for selectivity. Mix well and pour into sterile Petri plates.
Principle And Interpretation
Staphylococcus aureus is an invasive pathogen that can cause disease in almost any tissue or organ in the human body, primarily in compromised individuals (1). Staphylococcal infections were earlier treated using Penicillin. But over the years resistance to this drug developed. Methicillin was the next drug of choice. While methicillin is very effective in treating most Staphylococcus infections some strains have developed resistance to methicillin and can no longer be killed by this antibiotic. These resistant bacteria are called Methicillin Resistant Staphylococcus aureus (MRSA) (2). Patients with breaks in their skin due to wounds, indwelling catheters or burns are those with certain risk of developing MRSA infection (3). Spread of MRSA infections can be controlled to a great extent by maintaining personal hygiene after interaction with an MRSA infected person (2).
HiCynth™ Peptone No.2 and HiCynth™ Peptone No.5 provides carbonaceous, nitrogenous substances, amino acids, Vitamin B complex and other essential nutrients. This medium is prepared by completely replacing animal or vegetable based peptones with chemically defined peptones. The chromogenic mixture incorporated in the medium is specifically cleaved by Staphylococcus aureus to give bluish green coloured colonies. Sodium pyruvate enhances the growth of Staphylococcus species. Sodium chloride in the medium helps to maintain the osmotic equilibrium of the medium. High concentration of sodium chloride also helps in inhibiting the accompanying microflora. Cefoxitin is recommended to use for selective isolation of MRSA. The medium is made selective for MRSA by the addition of MeRS Selective Supplement (FD229) & CF Selective Supplement II (FD259) in combination.
Type of specimen
Clinical samples - Mouth, skin lesions, intestine, upper respiratory tract of humans, urine, wound samples, etc
Specimen Collection and Handling:
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (4,5). After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions :
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 specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations :
- Certain strains of MRSA which are intermediate may show poor growth. Further incubation upto 48 hours should be carried out.
- Some strains may show poor growth due to varying nutritional requirements.
- Further sensitivity can be carried out to ascertain the extent of resistance.
- Further biochemical tests must be carried out to differentiate between MRSA and MRSE.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within expiry period when stored at the recommended temmperature.
Quality Control
Appearance
Cream to yellow homogeneous free flowing powder
Gelling
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Light yellow coloured, clear to slightly opalescent gel forms in Petri plates
Reaction
Reaction of 8.33% w/v aqueous solution 25°C. pH: 7.0±0.2
pH
6.80-7.20
Cultural Response
Cultural characteristics observed with added MeRS Selective Supplement (FD229) & CF Selective Supplement II (FD259) after an incubation at 35-37°C for 18-48 hours.
| Organism | Inoculum (CFU) | Growth w/ FD229 & FD259 | Recovery w/ FD229 & FD259 | Colour of Colony |
|---|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | >=104 | inhibited | 0% | |
| Enterococcus faecalis ATCC 29212 (00087*) | >=104 | inhibited | 0% | |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=104 | inhibited | 0% | |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | >=104 | inhibited | 0% | |
| Staphylococcus aureus, MRSA ATCC 43300 | 50-100 | luxuriant | >=50% | Light blue -green |
| Staphylococcus epidermidis ATCC 12228 (00036*) | >=104 | inhibited | 0% | |
| Staphylococcus xylosus ATCC 29971 | >=104 | inhibited | 0% |
Key: (*) Corresponding WDCM numbers.
Storage and Shelf Life
Store between 15-25°C in a tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition Seal the container tightly after use. 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 (4,5).
| Product Name | HiCrome™ MeReSa HiCynth™ Agar Base |
|---|---|
| SKU | MCD1674 |
| Product Type | HiCynth™, HiCrome™ |
| Physical Form | Powder |
| Origin | Chemically defined (HiCynth™) |
| Packaging type | HDPE |
| References | 1.DWorkin M et. al 200 |
| Customized Product Available | No |
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