Method for the identification of microorganisms with at least two chromogens

• US 5,962,251 A
• Filed: 01/26/1996
• Issued: 10/05/1999
• Est. Priority Date: 07/28/1993
• Status: Expired due to Term

• Current Assignee: Alain Rambach
• Original Assignee: Alain Rambach
• Inventors: Rambach, Alain
• Primary Examiner(s): Dees, Jose G.
• Assistant Examiner(s): JONES, DAMERON LEVEST
• Application Number:
  US08/591,611
• Time in Patent Office: 1,348 Days
• Field of Search: 435/4, 435/7.2, 435/29, 435/30, 435/34
• US Class Current: 435/34
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: C12Q 1/10 Enterobacteria
• CPC Class Codes: C12Q 2334/50 Indoles
• CPC Class Codes: C12Q 2334/52 5-Bromo-4-chloro-3-indolyl, i.e. BCI

Abstract

A method for revealing the presence or absence of a particular microorganism strain in a culture medium, wherein at least two strain enzyme substrate chromogens are added to the culture medium, said chromogens being selected so that the presence of said strain in the culture medium is revealed by a third color.

Claims

1. A method for determining the presence or absence of a particular strain of microorganism in a culture medium, said method comprising:

introducing at least two chromogens which are enzyme substrates for said strain into said culture medium, said chromogens selected so that, in said culture medium, the presence of said strain is revealed by an accessory color, said accessory color having a dominant wavelength different than a dominant wavelength of the chromophore corresponding to each of the chromogens and different than a dominant wavelength of the mixture of the chromophore of each of the chromogens; and

identifying the presence of said strain by identifying said accessory color.

2. The method of claim 1, wherein said chromogens are compounds of the same chemical family.

3. The method of claim 1 wherein said chromogens liberate on hydrolysis at least two different chromophores which undergo a coupling reaction to produce said accessory color.

4. The method of claim 1 wherein said chromogens are of the indoxyl family.

5. The method of claim 3, wherein said chromophore is selected from the indoxyl derivatives bromoindoxyl, chloroindoxyl, dichloroindoxyl, chlorobromoindoxyl, trichloroindoxyl and methylindoxyl.

6. The method of claim 1 wherein said microorganisms are yeasts.

7. The method of claim 6, wherein said yeasts are of the genus Candida.

8. The method of claim 1 wherein said microorganisms are bacteria.

9. The method of claim 8, wherein said bacteria are one of the genus Strertococcus, Klebsiella, Enterobacter, Escherichia, Citrobacter, Staphylococcus, Listeria, Clostridium and Proteus.

10. The method of claim 1, wherein said culture medium possesses a carbohydrate concentration in the range of 10 to 30 grams/liter.

11. The method of claim 10, wherein said carbohydrate concentration is a glucose concentration in a peptone-based medium.

12. The method of claim 1 wherein said chromogen is a substrate for phosphatase.

13. The method of claim 1, wherein said medium comprises a phosphate concentration in the range of 1 to 3 grams/liter.

14. The method of claim 3, wherein said chromophore is selected from the group consisting of 6-chloroindoxyl, 5-bromoindoxyl, 3-bromoindoxyl, 4,6-dichloroindoxyl, 6,7-dichloroindoxyl, 5-bromo-4-chloroindoxyl, 5-bromo-6-chloroindoxyl and 4,6,7-trichloroindoxyl.