MARTIN LEWIS AGAR WITH LINCOMYCIN, PILL POCKET

Cat. no. E31 Martin Lewis with Lincomycin, 15x100mm Pill Pocket Plate, 24ml 10 plates/bag

INTENDED USE

Hardy Diagnostics Martin Lewis with Lincomycin is a selective medium for the recovery of Neisseria gonorrhoeae from both genital and oropharyngeal specimens. It is used in qualitative procedures for the isolation of Neisseria gonorrhoeae with suppression of most other gram-negative diplococci, gram-negative bacilli, gram-positive organisms and yeast. The CO 2 enriched environment allows for the growth of pathogenic Neisseria spp.

SUMMARY

Several successive media have been developed for the isolation of pathogenic Neisseria from specimens containing mixed flora. Each successive formula, Thayer Martin Selective Agar, Modified Thayer Martin and Martin Lewis Agar, provides greater inhibition of contaminating flora, however, each has been shown to be inhibitory to certain vancomycin sensitive strains of Neisseria spp.

Martin Lewis with Lincomycin provides a rich, selective growth media with decreased vancomycin. These plates are supplied with a CO 2 tablet (sodium bicarbonate and citric acid) that fits into a molded well in the petri dish. When sealed inside the ziplock bag, moisture in the air slowly activates the tablet to produce a CO 2 enriched atmosphere that is conducive to the growth of gonococci.

FORMULA

Ten CO 2 generating tablets (sodium bicarbonate and citric acid) and 10 ziplock bags are included with each bag of Martin Lewis with Lincomycin, Pill Pocket media.

Ingredients per liter of deionized water:*

Hemoglobin, Bovine 10.0gm
Pancreatic Digest of Casein 7.5gm
Selected Meat Peptone 7.5gm
Sodium Chloride 5.0gm
Dipotassium Phosphate 4.0gm
Monopotassium Phosphate 1.0gm
Corn Starch 1.0gm
Colistin 7.5mg
Trimethoprim Lactate 5.0mg
Amphotericin B 1.5mg
Vancomycin 1.0mg
Lincomycin 1.0mg
Koenzyme Enrichments 10.0ml
Agar 10.0gm

Final pH 7.2 +/- 0.2 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-8ºC. away from direct light. Media should not be used if there are any signs of deterioration (shrinking, cracking, or discoloration), hemolysis, contamination, or if the expiration date has passed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

Method of Use: Bring media to room temperature before use. Inoculate media by swabbing the surface from side to side while rolling in a large "z" pattern to sufficiently transfer the specimen. Remove the CO 2 generating tablet from the foil wrapper and immediately place the tablet into the circular well of the petri plate. The tablet will be activated by the moisture in the medium. Do not place a drop of water on the tablet. There is adequate moisture generated in the bag to slowly release the CO 2 . Place the plate in the ziplock bag and seal. The completeness of the seal is essential to maintain CO 2 environment for growth. Invert plates and incubate at 35ºC. for 24-48 hours. Some strains may require up to 72 hours to appear.

If the plate is to be sent to a laboratory after inoculation, incubate them under appropriate conditions before shipment. Specimens should be submitted directly to the laboratory without delay and protected from excessive heat and cold. Specimens must be transported at ambient temperatures (15-30ºC.). Do not refrigerate.

INTERPRETATION OF RESULTS

Neisseria gonorrhoeae appears as small, grayish-white to colorless mucoid colonies. N. meningitidis forms similar colonies to N. gonorrhoeae , but larger and blue-gray.

An oxidase test may be performed from the primary medium for presumptive identification.

LIMITATIONS

This medium is intended for transport and primary isolation. Some diagnostic tests may be performed with the primary media. However, additional tests including Gram stain and biochemical testing should be performed on pure cultures for complete identification. For more information, see appropriate references.

The agents in selective media may inhibit some strains of desired species or permit the growth of species they were designed to inhibit. Therefore, specimens cultured on selective media should also be cultured on non-selective media to obtain additional information and to help insure recovery of potential pathogens.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, other culture media, swabs, applicator sticks, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Inoculation Method* Incubation Results
Time Temperature Atmosphere
Neisseria gonorrhoeae
ATCC ® 43069
A 24-48hr 35°C CO 2 ** Growth
Neisseria meningitidis
ATCC ® 13090***
A 24-48hr 35°C CO 2 ** Growth
Proteus mirabilis
ATCC ® 43071
B 24-48hr 35°C CO 2 ** Partial to complete inhibition; no swarming
Staphylococcus epidermidis
ATCC ® 12228
B 24-48hr 35°C CO 2 ** Partial to complete inhibition
Escherichia coli
ATCC ® 25922***
B 24-48hr 35°C CO 2 ** Partial to complete inhibition
Candida albicans
ATCC ® 60193***
B 24-48hr 35°C CO 2 ** Partial to complete inhibition
Neisseria sicca
ATCC ® 9913***
B 24-48hr 35°C CO 2 ** Inhibited

** Atmosphere of incubation is enriched with 5-10% CO 2 .

*** To be used only by commercial media manufacturers.

USER QUALITY CONTROL

Physical Appearance

Martin Lewis with Lincomycin, Pill Pocket should appear opaque, and smooth brown in color.

P. mirabilis inhibited on Martin Lewis Agar with Lincomycin

Neisseria gonorrhoeae (ATCC ® 43069) colonies growing on Martin Lewis Agar with Lincomycin, Pill Pocket (Cat. no. E31). Incubated in CO 2 for 48 hours at 35ºC.

P. mirabilis inhibited on Martin Lewis Agar with Lincomycin

Shown with ziplock bag and CO 2 tablet.
Neisseria gonorrhoeae (ATCC ® 43069) colonies growing on Martin Lewis Agar with Lincomycin, Pill Pocket (Cat. no. E31). Incubated in CO 2 for 48 hours at 35ºC.

P. mirabilis inhibited on Martin Lewis Agar with Lincomycin

Proteus mirabilis (ATCC ® 43071) growth inhibited on Martin Lewis Agar with Lincomycin, Pill Pocket (Cat. no. E31). Incubated in CO 2 for 48 hours at 35ºC.

REFERENCES

1. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

2. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

3. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

4. Anderson, N.L., et al. Cumitech 3B; Quality Systems in the Clinical Microbiology Laboratory, Coordinating ed., A.S. Weissfeld. American Society for Microbiology, Washington, D.C.

5. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.


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032116gr