Multi-test format with gel-forming matrix for characterization of microorganisms

• US 5,627,045 A
• Filed: 04/12/1995
• Issued: 05/06/1997
• Est. Priority Date: 04/12/1995
• Status: Expired due to Term

• Current Assignee: Biolog, Inc.
• Original Assignee: Biolog, Inc.
• Inventors: Ralha, Maria C., Bochner, Barry
• Primary Examiner(s): Kight, John
• Assistant Examiner(s): Leary, Louise
• Application Number:
  US08/421,377
• Time in Patent Office: 755 Days
• Field of Search: 435/34, 435/4, 435/29, 435/32, 435/7.32, 435/7.31, 435/240.1, 435/240.2, 435/240.25, 435/810, 435/822, 435/254.1, 435/975
• 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 2304/22 Resazurin; Resorufin
• CPC Class Codes: G01N 2035/00148 Test cards, e.g. Biomerieux or McDonnel multiwell test cards
• CPC Class Codes: Y10S 435/822 using bacteria or actinomycetales
• CPC Class Codes: Y10S 435/975 Kit

Abstract

The present invention is directed to methods and compositions for the characterization of various microorganisms. In particular, the present invention is suited for the characterization of commonly encountered microorganisms (e.g., E. coli, S. aureus, etc.), as well as commercially and industrially important organisms from various and diverse environments. For example, the present invention is particularly suited for the growth and characterization of the actinomycetes and fungi.

Claims

1. A method for testing microorganisms selected from the group consisting of bacteria and fungi, comprising the steps of:

a) providing a testing device comprising a plurality of testing wells, wherein said wells contain a testing substrate and one or more gel-initiating agents;

b) preparing a suspension comprising a pure culture of microorganisms and an aqueous solution comprising a gelling agent, under conditions such that said suspension remains ungelled;

c) introducing said suspension into said wells of said testing device under conditions such that said suspension forms a gel matrix within said well, such that said microorganisms are within said gel matrix; and

d) detecting the response of said microorganisms to said testing substrate.

2. The method of claim 1, wherein said bacteria are members of the Order Actinomycetales.

3. The method of claim 1, wherein said gelling agent is selected from the group consisting of Gelrite™

, carrageenan, and alginic acid.

4. The method of claim 1, wherein said testing substrates are selected from the group consisting of carbon sources and antimicrobials.

5. The method of claim 1, wherein said gel-initiating agent comprises cationic salts.

6. The method of claim 1, further comprising a colorimetric indicator.

7. The method of claim 6, wherein said colorimetric indicator is selected from the group consisting of chromogenic substrates, oxidation-reduction indicators, and pH indicators.

8. The method of claim 1, wherein said testing device is a microplate.

9. A kit for characterizing and identifying microorganisms selected from the group consisting of bacteria and fungi, comprising:

i) a microplate testing device containing a plurality of wells, wherein said wells contain one or more gel-initiating agents and one or more testing substrates; and

ii) an aqueous suspension comprising a gelling agent.

10. The kit of claim 9, wherein said testing substrates are selected from the group consisting of antimicrobials and carbon sources.

11. The kit of claim 9, wherein said gelling agent is selected from the group consisting of Gelrite™

, carrageenan, and alginic acid.

12. The kit of claim 9, wherein said gel initiating agent comprises cationic salts.

13. The kit of claim 9, wherein said testing device further comprises a colorimetric indicator selected from the group consisting of chromogenic substrates, oxidation-reduction indicators, and pH indicators.

14. A method for testing microorganisms, wherein said microorganisms are selected from the group consisting of bacteria and fungi, comprising the steps of:

a) providing a testing device comprising a plurality of testing wells, wherein said wells contain a testing substrate and one or more gel-initiating agents;

b) preparing a suspension comprising a pure culture of microorganisms and an aqueous solution comprising a gelling agent, wherein said gelling agent is selected from the group consisting of gellan gum, carrageenan, and alginic acid, under conditions such that said suspension remains ungelled;

c) introducing said suspension into said wells of said testing device under conditions such that said suspension forms a gel matrix within said well; and

d) detecting the response of said microorganisms to said testing substrate.

15. The method of claim 14, wherein said testing substrates are selected from the group consisting of carbon sources and antimicrobials.

16. The method of claim 14, wherein said gel-initiating agent comprises cationic salts.

17. The method of claim 14, further comprising a colorimetric indicator.

18. The method of claim 17, wherein said colorimetric indicator is selected from the group consisting of chromogenic substrates, oxidation-reduction indicators, and pH indicators.

19. The method of claim 17, wherein said testing device is a microplate.