Hygiene monitoring

• US 20040028608A1
• Filed: 08/01/2003
• Published: 02/12/2004
• Est. Priority Date: 08/01/2000
• Status: Active Grant

• Current Assignee: Charm Sciences, Inc.
• Original Assignee: Charm Sciences, Inc.
• Inventors: Markovsky, Robert J., Saul, Steven J, Francisco, Cheryl B., Salter, Robert S., Charm, Stanley E.
• Granted Patent: US 7,229,783 B2
• Time in Patent Office: Days
• US Class Current: 424/9.1
• CPC Class Codes: C12Q 1/22 Testing for sterility conditions
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: C12Q 1/42 involving phosphatase
• CPC Class Codes: C12Q 1/48 involving transferase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 33/523 the element being adapted for a specific analyte

Abstract

A test method and test device for hygiene monitoring of a test sample from a biological material or a surface with a biological material to detect the presence of phosphate and glucose as a measure of hygiene by employing a swab-type, puncturable-membrane test apparatus or a lateral-flow or capillary-flow test apparatus.

Claims

1. A method for detecting phosphate and glucose as an indication of biological contamination in a sample, which method comprises:

a) adding to said sample a phosphorylase enzyme and a carbohydrate substrate to produce a reaction product of free glucose and phosphate bound to a saccharide; and

b) detecting said free glucose as an indication of said contamination.

2. The method of claim 1, wherein the phosphorylase enzyme comprises maltose phosphorylase, and the carbohydrate substrate comprises maltose.

3. The method of claim 1, further comprising cleaving free phosphate from bound phosphate.

4. The method of claim 3, wherein glucose-1-phosphate is the phosphate-bound saccharide reaction product, and wherein phosphatase enzyme cleaves phosphate from said glucose-1-phosphate.

5. The method of claim 3, wherein a phosphatase enzyme cleaves phosphate from an adenosine phosphate.

6. The method of claim 5, wherein the phosphatase enzyme comprises apyrase.

7. The method of claim 4, wherein the phosphatase enzyme comprises acid phosphatase.

8. The method of claim 4, wherein the phosphatase enzyme comprises glucose phosphatase.

9. The method of claim 4, wherein the phosphatase enzyme comprises alkaline phosphatase.

10. The method of claim 1, further comprising cleaving free glucose from a complex carbohydrates.

11. The method of claim 10, wherein one or more hydrolytic enzymes cleave free glucose from complex carbohydrates.

12. The method of claim 11, wherein the hydrolytic enzyme is β

-galactosidase.

13. The method of claim 1, further comprising releasing intracellular phosphate from a biological cell of said sample.

14. The method of claim 13, wherein intracellular phosphate is released by a lysing reagent.

15. The method of claim 1, further comprising catalyzing the interconversion of α

-D-glucose to β

-D-glucose.

16. The method of claim 1, further comprising adding glucose-phosphate to a sample suspected of containing phosphatase, to produce a reaction product of free glucose and free phosphate.

17. The method of claim 16, wherein the glucose-phosphate is glucose-6-phosphate.

20. The method of any one of claims 1, 18, and 19, which method comprises introducing the sample to the reagents through one or more membrane-puncturable niblets.

21. The method of any one of claims 1, 18, and 19, which method comprises introducing the sample onto a capillary-flow membrane containing reagents for production of glucose and the components for the colorimetric detection of glucose.

22. The method of claim 21, wherein the reagents are not immobilized on the test membrane and which method further comprises preventing flow of reagents after test completion so as to provide a defined area for detecting a color change reaction.

23. The method of claim 22, wherein a wetting agent is included in a sealed area within packaging of a test strip, and said wetting agent is applied to a surface sample prior to introducing said surface sample onto the test strip.

26. The method of claim 1, wherein said sample is a liquid obtained from a surface.

27. The method of claim 11, wherein the hydrolytic enzyme is invertase.

28. The method of claim 11, wherein the hydrolytic enzyme is amylase.

29. The method of claim 11, wherein the hydrolytic enzyme is cellulase.

18. A method for detecting phosphate and glucose as an indication of biological contamination in a sample, which method comprises:

a) adding a phosphorylase enzyme and a carbohydrate substrate to a sample containing phosphate, to produce a reaction product of free glucose and phosphate bound to a saccharide;

b) cleaving free phosphate from bound phosphate;

c) cleaving free glucose from a complex carbohydrate;

d) releasing intracellular phosphate; and

e) changing the configuration of free glucose from α

-D-glucose to β

-D-glucose,

wherein free glucose is detected in a colorimetric reaction as a measure of surface contamination.

19. A method for the rapid, colorimetric determination of hygiene monitoring of surface contamination, including contamination with phosphate and phosphatase, which method comprises:

a) adding to a sample containing phosphate a phosphorylase enzyme and a carbohydrate substrate, to produce a reaction product of free glucose and phosphate bound to a saccharide;

b) adding glucose-phosphate to produce, in the presence of phosphatase, a reaction product of free glucose and free phosphate;

c) cleaving free phosphate from bound phosphate;

d) cleaving free glucose from complex carbohydrates;

e) releasing intracellular phosphate; and

f) changing the configuration of free glucose from α

-D-glucose to β

-D-glucose,

wherein glucose is detected in a colorimetric reaction as a measure of surface contamination.

24. A method for rapid colorimetric determination of hygiene monitoring of surface contamination, which method comprises:

a) adding to a sample containing phosphate;

b) adding to said sample phosphorylase enzyme and carbohydrate substrate to produce a reaction product of bound phosphate and a monosaccharide;

b) phosphatase enzyme to cleave free phosphate from bound phosphate; and

c) detecting said monosaccharide in a colorimetric reaction as a measure of surface contamination.

25. The method of claim 24, wherein the phosphorylase enzyme is maltose phosphorylase, the carbohydrate substrate is maltose, and the phosphatase enzyme is acid phosphatase.

30. A device for detecting biological material in a liquid sample, which device comprises:

a) a test strip comprising a first end and a second end, said first end comprising a liquid absorbent material;

b) reagents for converting said biological material to free glucose; and

c) reagents for the colorimetric detection of said free glucose.

31. The device of claim 30, which further includes a sealed area containing a wetting agent.

32. The device of claim 30, wherein said reagents are at one or more reagent zones on said test strip.

33. The device of claim 32, wherein there are at least two reagent zones on the test strip, and wherein a first reagent zone comprises maltose and acid phosphatase, and a second reagent zone comprises TOOS, glucose oxidase, horseradish peroxidase, 4-aminoantipyrene, maltose phosphorylase, apyrase, and β

-galactosidase.

34. The device of claim 32, wherein there are at least two reagent zones on the test strip, and wherein a first reagent zone comprises maltose and glucose-6phosphate, and the second zone comprises TOOS, glucose oxidase, horseradish peroxidase, 4-aminoantipyrene, maltose phosphorylase, apyrase, and β

-galactosidase.

35. A device for detecting biological material in a test sample, which device comprises:

a) an enzyme reagent composition comprising glucose oxidase, horseradish peroxidase, maltose phosphorylase, apyrase, and β

-galactosidase;

b) a substrate reagent composition comprising 4-aminoantipyrene, TOOS, maltose and a glucose-phosphate; and

c) a buffering reagent composition comprising a buffered solution,

wherein the sample is contacted sequentially with the buffering reagent composition, enzyme reagent composition, and substrate reagent composition, and a color change or lack thereof is observed as a measure of hygiene.

36. A device for detecting biological material in a test sample, which device comprises:

a) enzyme reagents comprising glucose oxidase, horseradish peroxidase, maltose phosphorylase, apyrase, acid phosphatase and β

-galactosidase;

b) substrate reagents comprising 4-aminoantipyrene, TOOS and maltose; and

c) a buffering reagent composition comprising a buffered solution,

wherein the sample is contacted sequentially with the buffering reagents, enzyme reagents, and substrate reagents, and a color change or lack thereof is observed as a measure of hygiene.

37. A device for rapid, colorimetric hygiene monitoring, which device comprises:

a) a lateral, capillary-flow, elongated test strip having two ends, the test strip includes at a first end, a liquid sample absorbing material;

b) a series of spaced-apart reaction zones comprising reagents providing multiple biochemical pathways for conversion of biological material to glucose; and

c) a zone, at the second end, comprising reagents for the colorimetric detection of glucose, wherein a liquid sample from a material is absorbed onto the first end and flows by capillarity through the series of biochemical reaction zones to the zone, which provides a visually detectable readout in the presence of glucose.

38. A device for detecting biological material in a test sample, which test device comprises:

a) a swab to collect the test sample;

b) one or more separate, swab-puncturable, membrane compartments enclosing for one or more reagents;

c) enzyme reagents comprising glucose oxidase, horseradish peroxidase, maltose phosphorylase, apyrase, and β

-galactosidase;

d) substrate reagents comprising 4-aminoantipyrene, TOOS, maltose, and a glucose-phosphate; and

e) liquid reagents comprising a buffering solution and lysing solution,

wherein the sample is contacted sequentially with the liquid reagents, enzyme reagents, and substrate reagents, and a color change or lack thereof is observed as a measure of hygiene.

39. A device for hygiene monitoring, by detecting biological material in a test sample from or on a material, which test device comprises:

a) a swab to collect the test sample;

b) one or more separate, swab-puncturable, membrane compartments enclosing one or more reagents;

c) enzyme reagents comprising glucose oxidase, horseradish peroxidase, maltose phosphorylase, apyrase, acid phosphatase and β

-galactosidase;

d) substrate reagents comprising 4-aminoantipyrene, TOOS, and maltose; and

e) liquid reagents comprising a buffering solution, cofactors, antimicrobial substances and lysing solution,

wherein the sample is contacted sequentially with the liquid reagents, enzyme reagents, and substrate reagents, and a color change or lack thereof is observed as a measure of hygiene.