Compositions and methods for monitoring oxalate
The present invention features compositions and methods for the detection or measurement of oxalate in a sample. Such compositions include test devices that provide for the rapid and accurate detection of oxalate in a sample from a biological fluid. Advantageously, the compositions can be used to monitor the oxalate levels of a patient at a point of care (e.g., at the patient's home, clinic, physician's office, or other clinical setting).
1. A test device for detecting oxalate in a liquid sample of a subject, the device comprising oxalate decarboxylase, and an indicator system that provides a detectable reaction product in response to formate production, wherein the device detects oxalate in the liquid sample within 10 minutes of obtaining the liquid sample from the subject.
2. The test device of claim 1 , wherein the indicator system comprises phenazine methosulfate (PMS) and iononitro tetrazolium (INT).
3. The test device of claim 1 , wherein the device comprises a liquid permeable substrate.
4. The test device of claim 3 , wherein the liquid permeable substrate is in the form of a strip.
5. The test device of claim 1 , wherein the device comprises the oxalate oxidase or oxalate decarboxylase and the indicator system in powder form, and the device is suitable for containing a liquid.
6. The test device of claim 5 , wherein the device is in the form of a cuvette or other clear liquid container.
7. A test device for detecting oxalate in a liquid sample of a subject, the device comprising a liquid permeable material defining the following portions in capillary communication:
a) a site for application of a liquid sample, comprising a liquid permeable medium;
b) a second portion comprising a liquid permeable medium; and
c) a third portion that is the site for detecting the presence of oxalate at the test site, the third portion comprising a liquid permeable medium having an enzyme system having oxalate specific enzyme activity fixed to the medium at the test site, the enzyme system comprising oxalate specific enzyme, a substance which reacts with a reaction product of the oxalate specific enzyme, and an indicator material which reacts in the presence of the reaction product and changes color, wherein the device detects oxalate in the liquid sample within 10 minutes of obtaining the liquid sample from the subject.
8. The test device of claim 7 , wherein the enzyme system has oxalate decarboxylase activity, the enzyme system comprising an oxalate decarboxylase, a substance having formate dehydrogenase activity, and an indicator material which is reduced in the presence of formate and changes color.
9. The test device of claim 7 , further comprising a fourth portion that acts as a wick, the fourth portion comprising sorbent material.
10. The test device of claim 7 , wherein the second portion comprises an agent that alters the composition of the liquid as it contacts the second portion.
11. The test device of claim 10 , wherein the second portion comprises a liquid permeable material that acts as a filter to remove particulates.
12. The test device of claim 10 , wherein the second portion comprises an agent that alters a physical characteristic of a liquid sample flowing through the second portion.
13. The test device of claim 12 , wherein the agent is a buffer, a surfactant, or a salt.
14. The test device of claim 13 , wherein the altered physical characteristic is pH, salt concentration, or buffering capacity of the liquid sample.
15. The test device of claim 7 , wherein the device further comprises a fourth portion comprising absorbent material that facilitates the flow of liquid through the device.
16. The test device of claim 7 , wherein the liquid sample is blood, urine, or saliva.
17. A method for detecting an analyte in a liquid sample, the method comprising:
a) applying a liquid sample to the device of claim 1 ; and
b) detecting presence or absence of oxalate at a test site, wherein the presence of a detectable reaction product or color change at the test site identifies the presence of oxalate in the sample and the absence of color change at the test site identifies the absence of the oxalate in the sample.
18. A kit comprising the test device of claim 1 , a reaction mix, the reaction mix comprising an enzyme system having oxalate specific enzyme activity, the enzyme system comprising oxalate specific enzyme, a substance which reacts with a reaction product of the oxalate specific enzyme, and an indicator which reacts in the presence of the reaction product and changes color.
19. The kit of claim 18 , wherein the enzyme system has oxalate decarboxylase activity, the enzyme system comprising an oxalate decarboxylase, a substance having formate dehydrogenase activity, and an indicator which is reduced in the presence of formate and changes color.
20. The kit of claim 19 , comprising one or more of a powdered reaction mix, a reconstitution buffer, and dispensing pipette.
21. The test device of claim 1 , wherein the test device detects oxalate in the liquid sample within 5 minutes of obtaining the liquid sample from the subject.
22. The test device of claim 1 , wherein the test device detects oxalate in the liquid sample within 3 minutes of obtaining the liquid sample from the subject.
23. The test device of claim 1 , wherein the test device detects oxalate in the liquid sample within 1 minute of obtaining the liquid sample from the subject.
24. The test device of claim 1 , wherein the test device detects oxalate in the liquid sample at a point of care for the subject.