Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended Use
Recommended for direct Escherichia coli identification under UV light (366 nm).
Composition
C.L.E.D. Agar w/Bromo Thymol Blue
| Ingredients | g/L |
|---|---|
| Peptone | 4.000 |
| Tryptone | 4.000 |
| HM Peptone B# | 3.000 |
| Lactose | 10.000 |
| L-Cystine | 0.128 |
| Bromothymol blue | 0.020 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.3±0.2 |
**Formula adjusted, standardized to suit performance parameters
#- Equivalnet to Beef extract
MUG MacConkey Agar
| Ingredients | g/L |
|---|---|
| Peptone | 20.000 |
| Lactose | 10.000 |
| Bile salts mixture | 1.500 |
| Sodium chloride | 5.000 |
| Neutral red | 0.030 |
| Crystal violet | 0.001 |
| 4-Methylumbelliferyl B-D-glucuronide (MUG) | 0.100 |
| Agar | 15.000 |
| Final pH (at 25°C) | 7.1±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
- Surfaces: Loosen cap and remove HiDip™ slide from container taking care not to touch agar surfaces. Check for dehydration or contamination. Gently lower the slides and press agar to touch the test surface by bending the scull around the hinge line. Apply even and firm pressure for 15-20 seconds. Take care not to smudge agar over the test surface. Repeat procedure using the second agar surface on an area adjacent to the initial test side. Return the slide to the container and close tightly. Incubate in an up right position at indicated temprature.
- Liquids: Loosen cap and remove the HiDip™ slide from container. Check for dehydration or contamination. Dip slide into test fluid for upto 15-20 seconds so that agar surface becomes totally covered. (In case of inadequate liquid sample availability, pour sample over the surface of the slide). Allow to drain. Tab it gently to remove excess fluid from surface. Return the slide to the container and close tightly. Incubate in an upright position at indicated temprature. Label the container for sample number, source, date and time etc. for reference.
Principle And Interpretation
C.L.E.D.Agar is recommended for isolation, enumeration and identification of urinary pathogens on the basis of lactose fermentation.
MUG MacConkey Agar is used for selective isolation and detection of lactose fermenting coliform organisms by flurogenic procedure. The medium helps to detect the presence of an enzyme ẞ-glucuronidase and thereby rapidly identifying E.coli in mixed clinical specimens. MUG is cleaved by the enzyme to release an end product 4- methylumbelliferone which produces a visible greenish-blue fluorescence under long wave ultra-violet light. Growth indicators aids in easier observation and enumeration of colonies.
Type of specimen
Clinical sample : Urine; Food and dairy samples, Water samples.
Specimen Collection and Handling
Refer Directions.
Warning and Precautions
In Vitro diagnostic 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
C.L.E.D.Agar
- This medium is recommended for urine infection. Low urine count may be a result of antibiotic therapy, low pH of urine.
- Recovery depends on the urine count.
- Inoculate the medium immediately after urine collection.
MacConkey Agar:
- The plates are exposed to ammonia fumes to increase fluorescence as suggested by Freir and Hartman (1).
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
The HiDip™ slide containing combination of sterile CLED Agar and MUG- MacConkey Agar Medium on separate individual surfaces.
Colour
Colour of CLED Agar
Green coloured gel.
Colour of MUG-MacConkey Agar
Red coloured gel with purplish tinge
Quantity of medium
2.5ml of medium per surface
Sterility Check :
Passes release criteria
pH of CLED Agar
pH Range: 7.10-7.50
pH of MUG-MacConkey Agar
pH Range: 6.90-7.30
Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 18-48 hours.
CLED Agar
| Organism | Growth | Colour of colony | Fluorescence under UV |
|---|---|---|---|
| Escherichia coli ATCC 25922 (00013*) | good-luxuriant | yellow, opaque, centre slightly deeper yellow | - |
| Enterococcus faecalis ATCC 29212 (00087*) | good-luxuriant | slight yellowish or greenish | - |
| ## Proteus hauseri ATCC 13315 | good-luxuriant | blue | - |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | good-luxuriant | deep yellow | - |
MUG-MacConkey Agar
| Organism | Growth | Colour of colony | Fluorescence under UV |
|---|---|---|---|
| Salmonella Typhi ATCC 6539 | good-luxuriant | bluish | - |
| Klebsiella pneumoniae ATCC 13883 (00097*) | good-luxuriant | yellow to whitish blue | - |
| Escherichia coli ATCC 25922 (00013*) | luxuriant | - | positive |
| # Klebsiella aerogenes ATCC 13048 (00175*) | luxuriant | - | negative |
Key :
- * Corresponding WDCM numbers
- ## Formerly known as Proteus vulgaris
- # Formerly known as Enterobacter aerogenes
Storage and Shelf Life
On receipt store between 15-30°C. Use before expiry date on the label. Product performance is best if used within stated expiry period.
Disposal
Used HiDip™ slides should be handled carefully, as it contains live microorganisms. These slides can be best disposed off either by or by immersing in a suitable disinfectant solution (i.e. dettol, phenyl etc.) over night or by autoclaving them after loosening the cap.
| Product Name | HiDip™ CLED-MUG Mac Medium |
|---|---|
| SKU | HD005 |
| Customized Product Available | No |
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