Culture medium containing a spore germination inhibiting or delaying compound

Patent metadata

Field	Value
Publication number	US-9347083-B2
Application number	US-99549809-A
Country	US
Kind code	B2
Filing date	Jun 23, 2009
Priority date	Jun 26, 2008
Publication date	May 24, 2016
Grant date	May 24, 2016

Abstract

A medium for the culture and detection of target microorganisms, having at least one natural or synthetic specific substrate configured to detect at least one enzyme activity or metabolic activity of the target microorganisms and at least one compound that inhibits or delays the germination of spores of microorganisms, other than the target microorganisms, that are capable of interfering with the culture and detection of the target microorganisms.

First claim

The invention claimed is:

1. A liquid medium for the culture and detection of coagulase-positive staphylococci, comprising:
peptones;
at least one metabolic substrate capable of being degraded by a metabolic activity of the coagulase-positive staphylococci to cause the pH of the medium to vary;
a pH indicator capable of detecting pH variation of the medium due to degradation of the metabolic substrate, the pH indicator being a chromophore or fluorophore; and
D-alanine in an amount from 0.05 mol/L to 1 mol/L in the liquid culture medium.

2. The culture medium as claimed in claim 1 , wherein the pH indicator is a chromophore.

3. The culture medium as claimed in claim 1 , wherein the pH indicator is a fluorophore.

4. A method for determining whether coagulase-positive staphylococci are present in a sample, comprising:
a) bringing said sample into contact with a liquid culture medium as claimed in claim 1 ,
b) incubating said culture medium under conditions suitable for growth of the coagulase-positive staphylococci, and
c) detecting whether the coagulase-positive staphylococci are present in the culture medium in order to determine whether the coagulase-positive staphylococci were present in the sample.

5. The method as claimed in claim 4 , further comprising:
adding to said culture medium, prior to step a), a solution containing D-alanine so that D-alanine is present in the culture medium in step a).

6. The method as claimed in claim 4 , wherein step c) comprises measuring a variation in the level of fluorescence in the culture medium using a fluorescence reader, the variation corresponding to a variation in the pH in said culture medium.

7. The method as claimed in claim 4 , wherein the sample is a food or environmental sample.

8. The method as claimed in claim 4 , further comprising counting the coagulase-positive staphylococci.

9. A method of preparing a liquid culture medium, the method comprising:
mixing together in a liquid medium:
peptones;
at least one metabolic substrate capable of being degraded by a metabolic activity of coagulase-positive staphylococci to cause the pH of the medium to vary;
a pH indicator capable of detecting pH variation of the medium due to degradation of the metabolic substrate, the pH indicator being a chromophore or fluorophore; and
D-alanine so that D-alanine is in an amount from 0.05 mol/L to 1 mol/L in the liquid culture medium.

10. The culture medium as claimed in claim 1 , wherein the amount of D-alanine in the culture medium is from 0.05 mol/L to 0.1 mol/L.

11. The culture medium as claimed in claim 1 , wherein the culture medium has a sensitivity of at least 10 colony forming units per gram (CFU/g) for the detection of the coagulase-positive staphylococci.

12. The culture medium as claimed in claim 1 , wherein the peptones are animal and plant peptones.

13. The culture medium as claimed in claim 1 , further comprising selective agents that permit the growth of the coagulase-positive staphylococci under culture conditions when present in a sample contacted with the medium.

14. The culture medium as claimed in claim 13 , further comprising growth supplements and a buffer system.

15. The culture medium as claimed in claim 1 , wherein the at least one metabolic substrate is one or more sugars.

16. A liquid culture medium for detecting coagulase-positive staphylococci, comprising:
peptones;
at least one sugar capable of being degraded by a metabolic activity of the coagulase-positive staphylococci;
a fluorescent pH indicator for detecting pH variation of the medium;
selective agents that permit the growth of the coagulase-positive staphylococci under culture conditions when present in a sample contacted with the medium; and
D-alanine in an amount from 0.5 mol/L to 1 mol/L in the liquid culture medium.

17. The culture medium as claimed in claim 16 , wherein the amount of D-alanine is from 0.05 mol/L to 0.1 mol/L.

18. The culture medium as claimed in claim 16 , further comprising growth supplements and a buffer system.

19. A method for determining whether coagulase-positive staphylococci are in a sample, comprising:
inoculating the liquid culture medium as claimed in claim 16 with the sample;
incubating the inoculated culture medium under conditions suitable for growth of the coagulase-positive staphylococci; and
detecting whether the coagulase-positive staphylococci are present in the culture medium in order to determine whether the coagulase-positive staphylococci were present in the sample.

20. The method as claimed in claim 19 , further comprising forming the liquid culture medium, before inoculation, using a solution including D-alanine.

Assignees

• Costa Aurelien
• Biomerieux Sa

Inventors

• Costa Aurelien

Classifications

• C12Q1/045 Primary

  Culture media therefor · CPC title

• C12N2500/34

  Sugars · CPC title

• C12Q1/14 Primary

  Streptococcus; Staphylococcus · CPC title

• C12N2501/998

  Proteins not provided for elsewhere · CPC title

• C12N2500/32

  Amino acids · CPC title