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Paraoxonase diagnostic reagent kit and method for measuring active concentration of paraoxonase

A technology of nitrophenylphosphatase and diethyl, used in the field of medical testing and determination, can solve problems such as blood vessel blockage, heart disease or stroke, and reduced antioxidant effect

Inactive Publication Date: 2008-05-07
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The active concentration of diethyl-p-nitrophenyl phosphatase in smokers is lower than that of non-smokers or ex-smokers, so the antioxidant effect is also reduced, resulting in more cholesterol deposition in blood vessels, and excess Cholesterol can clog blood vessels, eventually leading to heart attack or stroke

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The diethyl-p-nitrophenyl phosphatase diagnostic reagent of this embodiment is a single reagent, comprising:

[0036] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0037] Stabilizer 50mmol / L

[0038] Coenzyme 3mmol / L

[0039] Aryl alcohol dehydrogenase 14000U / L

[0040] Aryl dialkyl phosphoric acid 14mmol / L

[0041] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.

[0042] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test secondary wavelength 405nm, tested diethyl-p-nitrophenyl phosphatase samples and reagents The volume ratio is 1 / 25, the reaction direction is positive reaction (rising reaction), the delay time is about 1 minute, the detection time is about 2 minutes, and the theoretical K value is 4180.

[0043] A...

Embodiment 2

[0045] The diethyl-p-nitrophenyl phosphatase diagnostic reagent of this embodiment is a dual reagent, comprising:

[0046] Reagent 1

[0047] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0048] Stabilizer 50mmol / L

[0049] Coenzyme 3mmol / L

[0050] Aryl dialkyl phosphoric acid 10mmol / L

[0051] Reagent 2

[0052] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0053] Stabilizer 50mmol / L

[0054] Aryl alcohol dehydrogenase 16000U / L

[0055] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0056] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength 340nm, test secondary wavelength 405nm, tested diethyl-p-nitrophenyl phosphatase samples and reagents 1. The volume ratio of reagent 2 is 2 / 20 / 5, the reaction direction is positive reaction (rising reaction), t...

Embodiment 3

[0059] The diethyl-p-nitrophenyl phosphatase diagnostic reagent of the present embodiment is three reagents, comprising:

[0060] Reagent 1

[0061] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0062] Stabilizer 50mmol / L

[0063]Coenzyme 3mmol / L

[0064] Reagent 2

[0065] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0066] Stabilizer 50mmol / L

[0067] Aryl dialkyl phosphoric acid 18mmol / L

[0068] Reagent 3

[0069] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0070] Stabilizer 50mmol / L

[0071] Aryl alcohol dehydrogenase 10000U / L

[0072] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0073] When measuring the activity concentration of diethyl-p-nitrophenyl phosphatase, set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance ≤ 0.1, test main wavelength...

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Abstract

The invention relates to a diethyl nitrophenyl phosphoesteraase diagnosis / determination reagent kit which utilizes an enzyme colorimetry method and an enzyme-linked method technology, simultaneously the invention also relates to a method principle for determining the diethyl nitrophenyl phosphoesteraase activity concentration, the composition and the components of a reagent, belonging to the technical field of the medicine examination and determination. The main components of the reagent kit of the invention comprise buffer solution, coenzyme, aryl dialkyl phosphoric acid, aryl alcohol dehydrogenase, and stabilizer. A sample is mixed with the reagent according to a certain volume to lead to perform a series of enzymatic reaction, a reacting substance is arranged under an ultraviolet / visible light analyzer to examine the rising degree / speed of absorbance at the position of a 340 nm dominant wavelength, thereby detecting and calculating the diethyl nitrophenyl phosphoesteraase concentration value. Through the adoption of the invention, the required determined results can be obtained completely through the ultraviolet / visible light analyzer.

Description

technical field [0001] The invention relates to a diagnostic kit for diethyl-p-nitrophenyl phosphatase, and at the same time, the invention also relates to a method for measuring the activity of diethyl-p-nitrophenyl phosphatase, belonging to the technical field of medical testing. Background technique [0002] Diethyl-p-nitrophenyl phosphatase in serum is mainly expressed in the liver, and its function is to hydrolyze phosphate in tissues. Diethyl-p-nitrophenyl phosphatase in serum is related to the metabolism of high and low density lipoproteins in the human body. Diethyl-p-nitrophenyl phosphatase can protect low-density lipoproteins from oxidative modification, so it can be considered that diethyl-p-nitrophenyl phosphatase is an anti-atherosclerotic and anti-inflammatory agent enzyme. The increase in plasma diethyl-p-nitrophenyl phosphatase activity reduced the level of autoantibodies against oxidatively modified low-density lipoprotein in plasma by 18.8%, while diethyl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/25C12Q1/32C12Q1/44C12Q1/00
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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