Universal test systems and methods of use thereof for identifying multiple families of microorganisms

• US 5,888,760 A
• Filed: 04/10/1997
• Issued: 03/30/1999
• Est. Priority Date: 04/10/1997
• Status: Expired due to Term

• Current Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Original Assignee: Dade Microscan, Inc. (Siemens AG)
• Inventors: Nothaft, Daniel M., Godsey, James H.
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US08/843,634
• Time in Patent Office: 719 Days
• Field of Search: 435/34, 435/14, 435/21, 435/23, 435/24, 435/19, 435/12, 435/968, 435/882, 435/885, 435/843, 435/853, 435/851, 435/871
• 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: Y10S 435/843 Corynebacterium
• CPC Class Codes: Y10S 435/851 Haemophilus
• CPC Class Codes: Y10S 435/853 Lactobacillus
• CPC Class Codes: Y10S 435/871 Neisseria
• CPC Class Codes: Y10S 435/882 Staphylococcus
• CPC Class Codes: Y10S 435/885 Streptococcus
• CPC Class Codes: Y10S 435/968 High energy substrates, e.g. fluorescent, chemiluminescent, radioactive

Abstract

The present invention relates to a universal test systems and methods of use thereof for identifying a microorganism among at least two groups of widely divergent microorganisms. The universal test system comprises a predetermined combination of non-redundant biochemical tests comprising a substrate for at least one enzyme wherein the substrate, if acted on by the enzyme results in formation of a detectable product. Detectable products from the combination of biochemical tests are then used to identify the microorganism.

Claims

1. A test system for identifying a microorganism in a sample, wherein the test system is capable of identifying that microorganism from among at least two groups of widely divergent microorganisms which may be present in such a sample and wherein the test system comprises:

a predetermined combination of non-redundant biochemical tests disposed in a predetermined number of reaction chambers, wherein each biochemical test comprises a substrate for an enzyme or a group of enzymes, and further wherein the substrate, if acted on by the enzyme or group of enzymes, results in the formation of a detectable product in the reaction chamber; and

wherein the detectable products from the combination of biochemical tests are used to identify the microorganism in the sample.

2. The test system according to claim 1, wherein identifying a microorganism comprises classifying the microorganism to a genus or a species of microorganism or both.

3. The test system according to claim 1, wherein the predetermined combination of non-redundant tests, comprises fluorescent tests, colorimetric tests or a combination thereof.

4. The test system according to claim 3, wherein the fluorescent tests are performed in a fluorogenic or fluorometric format.

5. The test system according to claim 1, wherein the predetermined combination of non-redundant tests are performed at a temperature of between about 25°

to about 37°

C.

6. The test system according to claim 5, wherein the colorimetric tests are read visually or by a calorimeter.

7. The test system according to claim 4, wherein the fluorescent tests are read using a fluorometer.

8. The test system according to claim 1, comprising at least one test for detecting a peptidase, glycosidase, a sugar fermentation enzyme, a urease, a decarboxylase, an esterase, a carbon utilization enzyme, a phosphatase, or a tryptophanase.

9. The test system according to claim 8, comprising a test for at least one peptidase and at least one glycosidase.

10. The test system according to claim 9, further comprising a test for at least one sugar fermentation enzyme.

11. The test system according to claim 10, further comprising a test for at least one urease.

12. The test system according to claim 11, further comprising a test for at least one decarboxylase.

13. The test system according to claim 12, further comprising a test for at least one esterase.

14. The test system according to claim 13, further comprising a test for at least one carbon assimilation enzyme.

15. The test system according to claim 1, wherein the predetermined combination of non-redundant of biochemical tests is capable of identifying the microorganism among at least two of enteric bacteria, nonfermenting bacteria, anaerobic bacteria, yeast, Staphylococcus sp., Streptococcus sp., Enterococcus sp., and fastidious bacteria.

16. The test system according to claim 1, wherein the predetermined combination of non-redundant of biochemical tests is capable of identifying the microorganism among least one of Staphylococcus sp. , Streptococcus sp., Enterococcus sp. , Corynebacteria sp., Lactobacillus sp., Pediococcus sp., Leuconostoccus sp., Alloicoccus sp., Vagococcus sp., Kluyvera sp., Leminorella sp., Haemophilus sp., Neisseria sp., Moraxella sp., Salmonella sp., Clostridia sp., and Listeria sp.

17. The test system according to claim 1, wherein the predetermined combination of non-redundant biochemical tests is capable of identifying the microorganisms among at least one of anaerobes, yeast or fastidious bacteria.

18. The test system according to claim 17, wherein the predetermined combination of non-redundant biochemical tests is capable of identifying the microorganisms among anaerobes and yeast or fastidious bacteria.

19. The test system according to claim 18, wherein the predetermined combination of non-redundant biochemical tests is capable of identifying the microorganism among yeast and fastidious bacteria.

20. The test system according to claim 1, wherein the predetermined combination of non-redundant biochemical tests is capable of identifying the microorganisms among at least one of Staphylococcus, Streptococcus, or Enterococcus and at least one of anaerobes, yeast, enterics, nonfermentors or fastidious bacteria or combinations thereof.

21. The test system according to claim 1, wherein the predetermined combination of non-redundant biochemical tests is capable of identifying the microorganisms among Staphylococcus, Streptococcus, and Enterococcus.

22. The test system according to claim 1, wherein the test system is capable of identifying among yeast, anaerobic bacteria and fastidious bacteria.

23. The test system according to claim 1, wherein the test system is capable of identifying Enterococcus, Staphylococci, Streptococci, anaerobes, yeast and fastidious bacteria.

24. The test system according to claim 1, wherein the substrates comprise lysine (AMC), leucine (AMC), methionine (AMC), glycyl-proline (AMC), Isoleucine (AMC), Trehalose, maltose, L-tryptophan (7-AMC), 4-MeU-phosphate, 4-MeU-phosphate, tyrosine (AMC), 4-MeU-β

-D-galactosidase, mannose, sucrose, and proline (AMC).

25. The test system according to claim 24, wherein the substrates further comprise fructose, glycerol, L-histidine 7-AMC, pyroglutamic acid (AMC) and 4-MeU-β

-D-glucoside.

26. A method for identifying a microorganism in a sample from among at least two groups of widely divergent microorganisms which may be present in such a sample by use of a test system according to claim 1, wherein the method comprises:

a) adding the sample to each reaction chamber comprising a substrate;

b) allowing the enzyme, if present, to react with the substrate;

c) determining the presence of the enzyme in the sample by detecting the detectable product in a test; and

d) comparing results of the combination of predetermined tests with at least one predetermined standard to identify the microorganism in the sample.

27. A test for detecting carbon source utilization by a microorganism, wherein the test comprises at least one carbon source and at least one fluorometric indicator, wherein the microorganism acts on the carbon source to produce a pH change which causes a change in fluoresence of the indicator, the change in fluoresence being indicative of carbon source utilization by the microorganism.

28. A test system for identifying a microorganism in a sample, wherein the test system is capable of identifying that microorganism from among at least two groups of widely divergent microorganisms which may be present in such a sample and wherein the test system comprises:

a predetermined combination of non-redundant biochemical tests comprising fluorescent tests, colorimetric test or a combination thereof, the test being disposed in a predetermined number of reaction chambers, wherein each biochemical test comprises a substrate for an enzyme or a group of enzymes, and further wherein the substrate, if acted on by the enzyme or group of enzymes, result in the formation of a detectable product in the reaction chamber, and

wherein the detectable products from the combination of biochemical tests are used to identify the microorganism, wherein the predetermined combination of non-redundant tests are preformed within about 15 minutes to about 8 hours.