Claims
- 1. A method to detect disease in a patient comprising:
digesting a phospholipid in a sample of bodily fluid from the subject with a first enzyme to produce substrate; reacting the substrate with a second enzyme in an enzymatic cycling reaction to produce a detectable product; determining the concentration of phospholipid by measuring the detectable product; and correlating the concentration of phospholipid to the disease condition by comparison to a normal concentration.
- 2. The method of claim 1, wherein said first enzyme is selected from the group consisting of phospholipase B, lysophospholipase, phospholipase A1, and phospholipase A2.
- 3. The method of claim 1, wherein the second enzyme is selected from the group consisting of glycerol-3-phosphate dehydrogenase, glycerol-3-phosphate oxidase, glycerokinase and glycerol dehydrogenase.
- 4. The method of claim 1, wherein the substrate is glycerol-3-phosphate.
- 5. The method of claim 1, wherein the detectable product is hydrogen peroxide.
- 6. The method of claim 5, wherein the step of determining the concentration of phospholipid by measuring detectable product comprises measuring an increase in hydrogen peroxide by colorimetry.
- 7. The method of claim 1, wherein the detectable product is NADH.
- 8. The method of claim 7, wherein said step of determining the concentration of the phospholipid by measuring the detectable product comprises measuring oxidation of NADH.
- 9. The method of claim 1, wherein the step of reacting the substrate in an enzyme cycling reaction comprises reacting G-3-P with glycerol-3-phosphate dehydrogenase and glycerol-3-phosphate oxidase.
- 10. The method of claim 1, wherein, the sample of bodily fluid is selected from the group consisting of plasma, serum, urine, saliva, ascites, cerebral spinal fluid and pleural fluid.
- 11. The method of claim 1, further comprising the step of extracting lipids from the sample of bodily fluid.
- 12. The method of claim 1, further comprising the step of comparing the concentration of phospholipid with an earlier concentration from the same subject.
- 13. The method of claim 1, wherein an increase or decrease in the concentration of phospholipid relative to normal subjects indicates the presence of the disease condition.
- 14. The method of claim 1, wherein the disease condition is gynecological cancer or ovarian cancer.
- 15. The method of claim 1, wherein the disease condition is a blood disorder associated with alteration in the level of phospholipid.
RELATED APPLICATION INFORMATION
[0001] This application is a continuation-in-part of U.S. patent application Ser. No. 09/176,813 filed Oct. 12, 1998. The priority of the prior application is expressly claimed, and the disclosure of this prior application is hereby incorporated by reference in its entirety.
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
09176813 |
Oct 1998 |
US |
Child |
09852066 |
May 2001 |
US |