REGAN LOWE AGAR

Technical Data #1485a / 2013.05.12

Regan Lowe Agar a selective solid medium used for the cultivation and isolation of Bordetella pertussis and Bordetella parapertussis from clinical specimens contaminated with respiratory flora. Nicotinic acid is an essential nutrient for Bordetella growth. Charcoal and starch, as absorbents, remove fatty acids. Cephalexine inhibits normal respiratory flora while Amphotericin B prevents yeast or fungal contamination.

 


 

FORMULA
in grams per liter purified filtered water

Beef Extract

10,0

Meat Peptone

10,0

Starch, soluble

10,0

Charcoal, activated

4,0

Sodium Chloride

5,0

Nicotinic Acid

0,001

Agar

15,0

Cephalexin

0,04

Amphotericin B

0,05

Defibrinated Horse Blood

100 ml

pH 7.4 +/- 0,2 25 C

This approximate formula may be adjusted and/or enriched to obtain best results.


PRECAUTIONS
This medium is for IN VITRO diagnostic use only.


STORAGE
Store prepared media at 2-8 C protected from direct light. and dehydrated powder, in a dry place, in tightly-sealed containers at 2-25 C.


SIGN OF DETERIORATION
Media should not be used if the expiry date has passed. Prepared media should not be used if there are signs of contamination or deterioration (shrinking, cracking, evaporation or discoloration). Do not use dehydrated media if it is caked.


DIRECTIONS
Suspend 54 g in 900 ml of purified filtered water. Heat with frequent agitation and boil for one minute. Sterilize at 121 C for 15 minutes. Cool to 45-50 C and aseptically add 100 ml of Defibrinated Horse Blood (4281) and 10 ml of Regan-Lowe Supplement (8715). Dispense into sterile Petri dishes.


PROCEDURE
Prior to inoculate, the prepared medium should be brought to room temperature.

  1. Inoculate plates, as soon as possible, with specimens, streak for isolation. Bordetella Pertussis Transport Medium (2555) should be used if inoculation is delayed.
  2. Incubate at 35 C under a humid atmosphere for up to 7 days.
  3. Examine the plates daily.


QUALITY CONTROL

Expected results after 3 days at 35C under humid atmosphere

Organisms

ATCC

Growth

Bordetella pertussis

8467

+

Escherichia coli

25922

- or partial

Staphylococcus aureus

25923

- or partial

Candida albicans

10231

- or partial

Colonies of Bordetella pertussis are tiny, smooth, with a characteristic compact, glistening appearance.


LIMITATIONS OF METHOD
Regan-Lowe as selective medium may inhibit some luxuriant growth of Bordetella pertussis, a Bordet Gengou Agar should be inoculated simultaneously. This medium is only a part of the identification. Other tests may be required.


REFERENCES

  1. Regan, J and Lowe, F. 1977 Enrichment Medium for the Isolation of Bordetella. J. Clin. Microbiol. 6: 303-309.
  2. Lennette, E.H., Ballows, A., Hausler, W.J.Jr., and Shadomy, H.J. Manual of Clinical Microbiology. 4th ed. 1985 Washington D.C.: American society for Microbiology.
  3. Gilligan,P.H., and M.C. Fisher. 1984 Importance of culture in laboratory diagnosis of Bordetella pertussis infections. J. Clin. Microbiol. 20:891-893
  4. Stauffer,L.R., D.R. Brown & R.E. Sandstrom 1983 Cephalexin- Supplemented Jones-Kendrick Charcoal Agar for Selective Isolation of Bordetella pertussis: Comparaison with Previously Described Media. J. Clin. Microbiol. 17:60-62.
  5. Hoppe J.E., J. Schwaderer 1989 Comparaison of Four Charcoal Media for the Isolation of Bordetella pertussis. J. Clin. Microbiol. 27:1097-1098
  6. Kurzynski,T., D.M.Boehm, J.A.Rott-Petri, R.F.Schell, P.E. Allison 1988 comparaison of Modified Bordet Gengou & Modified Regan-Lowe Media for the Isolation of Bordetella pertussis & Bordetella parapertussis. J. Clin. Microbiol. 26:2661-2663


CATALOGUE NUMBERS

Dehydrated media 500 g QB-39-0911
     
Regan-Lowe Supplement 10x5 ml 8715
     
Defibrinated Horse Blood 100 ml 4281
     
Prepared media    
Plates, 100x15 mm 10/pkg 1485
     
B. Pertussis Transport Medium, Vial 8 ml 10/pkg 2555

 


 





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