Rapid assay for testing overall oxidative stress
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IPC분류정보
국가/구분 |
United States(US) Patent
등록
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국제특허분류(IPC7판) |
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출원번호 |
US-0766522
(2001-01-19)
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발명자
/ 주소 |
- Obayan, Adebola
- Juurlink, Bernhard H.
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대리인 / 주소 |
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인용정보 |
피인용 횟수 :
1 인용 특허 :
2 |
초록
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The invention relates to an assay for testing oxidative stress of a subject by measurement of oxidants in biological fluids such as urine, plasma, bioreactor medium and respiratory aspirants. There is provided a method of determining oxidative stress in a mammalian subject. The method comprises: obt
The invention relates to an assay for testing oxidative stress of a subject by measurement of oxidants in biological fluids such as urine, plasma, bioreactor medium and respiratory aspirants. There is provided a method of determining oxidative stress in a mammalian subject. The method comprises: obtaining a sample of a biological fluid from the subject; mixing the biological fluid with a ferrous reaction reagent; incubating the biological fluid and the reaction reagent; and detecting a colored reaction product.There is further provided a ferrous reaction reagent suitable for use in assaying oxidative stress, said reaction reagent comprising 2-deoxyglucose, TBA, EDTA, and ferrous sulfate, and being substantially free of ascorbic acid.
대표청구항
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1. A method of detecting hydrogen peroxides and organic peroxides in a mammalian subject said method comprising:a. obtaining a sample of a biological fluid from the subject; b. mixing the biological fluid with a ferrous reaction reagent comprising 2-deoxyglucose and a ferrous (Fe2+) compound; c. inc
1. A method of detecting hydrogen peroxides and organic peroxides in a mammalian subject said method comprising:a. obtaining a sample of a biological fluid from the subject; b. mixing the biological fluid with a ferrous reaction reagent comprising 2-deoxyglucose and a ferrous (Fe2+) compound; c. incubating the biological fluid and the reaction reagent; and d. detecting a coloured reaction product. 2. The method of claim 1 wherein the reaction reagent comprises a solution of 2-deoxyglucose, TBA, EDTA and ferrous sulfate.3. The method of claim 2 wherein the reaction reagent is substantially free of ascorbic acid.4. The method of claim 2 wherein the reaction reagent comprises 2-deoxyglucose in a concentration of between about 30 and 400 mM.5. The method of claim 2 wherein the reaction reagent comprises 2-deoxyglucose in a concentration of between about 75 and 150 mM.6. The method of claim 2 wherein the reaction reagent comprises TBA in a concentration of between about 10 and 200 mM.7. The method of claim 2 wherein the reaction reagent comprises EDTA in a concentration of between about 0.5 and 3 mM.8. The method of claim 2 wherein the reaction reagent comprises ferrous sulphate in concentration of between about 0.5 and 2.0 mM.9. The method of claim 2 wherein the reaction reagent comprises an excess of Fe2+.10. The method of claim 2 wherein the reaction reagent comprises 100 mM 2-deoxyglucose, 50 mM TBA, 1.4 mM EDTA, and 1 mM ferrous sulphate.11. The method of claim 1 wherein the biological fluid is selected from the group consisting of: urine, plasma, bioreactor material and respiratory aspirate.12. The method of claim 1 wherein one part biological fluid is mixed with between about 5 and 15 parts of the reaction reagent.13. The method of claim 1 wherein the mixture of the biological fluid and the reaction reagent is incubated at between 20 and 45 degrees Centrigrade.14. The method of claim 1 wherein the mixtures is incubated for between about 5 and 30 minutes.15. The method of claim 1 wherein the ferrous reaction reagent is absorbed to a solid support.16. A method of identifying a mammalian subject in need of medical treatment comprising:a. obtaining a sample of biological fluid from the subject; b. determining oxidant level in the biological fluid by mixing the fluid with a reagent comprising 2-deoxyglucose, and a ferrous (Fe2+) compound; c. incubating the fluid and the reagent and determining the presence of oxidative stress within the subject by detecting a colorimetric change in the reaction product by comparing the reaction product with a reference standard and correlating the presence of oxidative stress with a difference between the colorimetric properties of the product and the standard, thereby determining the need from medical treatment within the subject. 17. The method of claim 16 wherein the determination of oxidant level in the biological fluid comprises detecting hydrogen peroxides and organic peroxides.18. The method of claim 16 wherein the biological fluid is selected from the group consisting of: urine, plasma, bioreactor fluid and respiratory aspirant.19. The method of claim 16 wherein the subject is a human.20. A ferrous reaction reagent suitable for use in assaying oxidative stress, said reaction reagent comprising 2-deoxyglucose, TBA, EDTA, and ferrous sulfate, and being substantially free of ascorbic acid.21. The reaction reagent of claim 20 comprising 2-deoxyglucose in a concentration of between about 30 and 400 mM.22. The reaction reagent of claim 20 comprising TBA in a concentration of between about 10 and 200 mM.23. The reaction reagent of claim 20 comprising EDTA in a concentration of between about 0.5 and 3 mM.24. The reaction reagent of claim 20 comprising ferrous sulphate in a concentration of between about 0.5 and 2.0 mM.25. The reaction reagent of claim 20 comprising an excess of Fe2+.26. The reaction reagent of claim 20 comprising 100 mM 2-deoxyglucose, 50 mM TBA, 1.4 mM EDTA, and 1 mM ferrous sulphate.27. The reaction reagent of claim 20 absorbed on a solid support.28. A kit suitable for use in assaying oxidative stress from a biological fluid, said kit comprising:a. a ferrous reaction reagent comprising 2-deoxyglucose and a ferrous (Fe2+) compound; and b. a reference standard indicating oxidant levels. 29. The kit of claim 28 further comprising instructions for detecting hydrogen peroxides and organic peroxides wherein the detecting comprises the steps of:(a) obtaining a sample of a biological fluid from a subject (b) mixing the biological fluid with the ferrous reaction reagent comprising 2-deoxyglucose and a ferrous (Fe2+) compound; (c) incubating the biological fluid with the reaction reagent; and (d) detecting a coloured reaction product. 30. The kit of claim 28 wherein the reaction reagent comprises 2-deoxyglucose, TBA, EDTA, and ferrous sulfate.31. The kit of claim 30 wherein the reaction reagent is substantially free of ascorbic acid.32. The kit of claim 28 wherein the reaction reagent is absorbed to a solid support.33. The kit of claim 28 wherein the ferrous reaction reagent comprises 100 mM 2-deoxyglucose, 50 mM TBA, 1.4 mM EDTA, and 1 mM ferrous sulphate.34. The kit of claim 28 wherein the standard indicating oxidant levels is based on differences in color that correspond to different oxidant levels.35. The method of claim 1 comprising the further step of determining the presence of oxidative stress within the subject, wherein the further step comprises detecting a colorimetric change in the reaction product by comparing the reaction product with a reference standard and correlating the presence of oxidative stress with a difference between the colorimetric properties of the reaction product and the standard.
이 특허에 인용된 특허 (2)
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Pugia Michael (Granger IN), Ascorbate resistant detection of hydrogen peroxide.
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O\Brien Mark R. (Glastonbury CT) Barre Mark S. (Stamford CT) Newton Michael S. (Durham CT), Method and apparatus for residential water test kit.
이 특허를 인용한 특허 (1)
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Hopkins, Christopher E.; Gunaratna, Miluka, Transgenic biosensors.
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