L.J. Medium Slant

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SKU:
SL001


Intended Use:

Recommended for cultivation of Mycobacterium tuberculosis.

Composition**

Ingredients g/600 ml
L-Asparagine 3.60
Potassium dihydrogen phosphate 2.40
Magnesium sulphate 0.24
Magnesium citrate 0.60
Potato starch, soluble 30.00
Malachite green 0.40
Glycerol 12.00 ml
Whole Egg Emulsion 1000.00 ml

**Formula adjusted, standardized to suit performance parameters

Directions

Inoculate either the sputum sample previously subjected to decontamination and concentration process or the pure culture of Mycobacteria isolated from a clinical sample on the surface of the slants. Incubate the slants at 30-35°C with 5-10% CO2 and examine the slants every week up to 8 weeks.

Principle And Interpretation

Solid media used for isolation and cultivation of Mycobacteria are either egg-based or agar-based. Egg-based media contain whole eggs or egg yolk, potato flour, salts and glycerol and are solidified by inspissation. Of the egg-based media, L.J. Medium is prepared as per the Jensen's (1) modification of the original formulation of Lowenstein (2). The egg base medium supports a wide variety of Mycobacteria and can also be used for niacin testing (3). Glycerol provides fatty acids. Malachite green serves as an inhibitor as well as pH indicator. Formation of blue zones indicates a decrease in pH by Gram-positive contaminants (e.g. Streptoccocci) and yellow zones of dye destruction by Gram-negative bacilli. Proteolytic contaminants cause localized or complete digestion of the medium (4).

Type of specimen

Clinical samples : Sputum

Specimen Collection and Handling

For clinical samples:Refer appropriate references for standard test procedures of decontamination and isolation (1,5-8). 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

  • Certain gram-positive contaminants (e.g. Streptococci) and gram-negative bacilli may grow on the medium.
  • Certain Saprophytes may also grow on the medium.
  • Proteolytic contaminants cause localized or complete digestion of 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

Lowenstein Jensen Medium-a mixture of sterile basal medium and 1000 ml whole egg emulsion, when inspissated coagulates to yield pale bluish green coloured, opaque, smooth slants

Sterility test

Passes release criteria

Cultural response

Cultural characteristics within 2-4 weeks at 35-37°C, with 5-10% CO2(further growth may be observed for 8 weeks)

Organism Inoculum Growth Colony characteristics
Mycobacterium gordonae ATCC 14470 Standardized inoculum giving approximately 1000000 cfu/ml Luxuriant Smooth, yellowish orange colonies
Mycobacterium kansasii ATCC 12478 Standardized inoculum giving approximately 1000000 cfu/ml Luxuriant Smooth-rough, photochromogenic colonies
Mycobacterium smegmatis ATCC 14468 Standardized inoculum giving approximately 1000000 cfu/ml Luxuriant Wrinkled, creamy white colonies
M. tuberculosis H37Rv ATCC 25618 Standardized inoculum giving approximately 1000000 cfu/ml Luxuriant Granular, rough, worty, dry, friable colonies.
M. avium subsp. Avium MTCC 1723 Standardized inoculum giving approximately 1000000 cfu/ml Luxuriant Smooth, non pigmented colonies

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 (7,8).

More Information
Product Name L.J. Medium Slant
SKU SL001
Customized Product Available No
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