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Kit and method for clinical detection of myeloperoxidase

The technology of myeloperoxidase and kit is applied in the field of kits for clinical detection of myeloperoxidase, and can solve the problems such as the inability to use an automatic biochemical analyzer, the high price of ELISA method, and the limited accuracy of manual operation, Achieve the effect of low inhibition constant Ki, high accuracy and simple manufacturing process

Active Publication Date: 2012-10-03
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The above two kits are manual methods, the accuracy of manual operation is limited, and the detection cycle is long, the sample size is large, and it cannot be used in automatic biochemical analyzers. Moreover, the dominant ELISA method is more expensive and expensive. The disadvantages such as complex preparation process and difficulty in industrialized production of the box bring great inconvenience to clinical diagnosis

Method used

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  • Kit and method for clinical detection of myeloperoxidase
  • Kit and method for clinical detection of myeloperoxidase
  • Kit and method for clinical detection of myeloperoxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Reagent 1:

[0070] Tris buffer 200mmol / L

[0071] Phenol 2mmol / L

[0072] Glycerin 10g / L

[0073] Sodium azide 0.1g / L

[0074] Tween 80 1ml / L

[0075] Hydrochloric acid 5ml / L

[0076] Reagent 2:

[0077] Tris buffer 200mmol / L

[0078] h 2 o 2 10mmol / L

[0079] 4-Aminoantipyrine 2mmol / L

[0080] Glycerin 10g / L

[0081] Sodium azide 0.1g / L

[0082] Tween 80 1ml / L

[0083] Hydrochloric acid 5ml / L

[0084] Reagent 3:

[0085] 4-Hydroxybenzohydrazide 25μmol / L

[0086] Glycerin 10g / L

[0087] Sodium azide 0.1g / L

[0088] Tween 80 1ml / L

[0089] When measuring the sample, the rate method is adopted, the temperature is 37°C, the dominant wavelength is 505nm, and the reaction direction is upward (positive reaction). The sample addition steps are as follows:

[0090] Specimen (sample) 25μL

[0091] Reagent 1 140μL

[0092] Mix well and incubate at 37°C for 90 seconds

[0093] Reagent 2 60μL

[0094] Mix well, incubate for 120 seconds,...

Embodiment 2

[0103] Reagent 1:

[0104] Potassium phosphate buffer 100mmol / L

[0105] Guaiacol 5mmol / L

[0106] Mannitol 10g / L

[0107] PC300 1g / L

[0108] Triton X-100 10ml / L

[0109] Hydrochloric acid 10ml / L

[0110] Reagent 2:

[0111] Potassium phosphate buffer 150mmol / L

[0112] h 2 o 2 15mmol / L

[0113] 4-Aminoantipyrine 5mmol / L

[0114] Mannitol 10g / L

[0115] PC300 1g / L

[0116] Triton X-100 10ml / L

[0117] Hydrochloric acid 10ml / L

[0118] Reagent 3:

[0119] 3-methoxybenzohydrazide 40μmol / L

[0120] Mannitol 10g / L

[0121] PC300 1g / L

[0122] Triton X-100 10ml / L

[0123] The assay method of embodiment 2 is identical with embodiment 1.

Embodiment 3

[0125] Reagent 1:

[0126] GOOD'S buffer 100mmol / L

[0127]DHBS 5mmol / L

[0128] BSA 1g / L

[0129] MIT 0.1g / L

[0130] Emulsifier OP 1ml / L

[0131] Hydrochloric acid 20ml / L

[0132] Reagent 2:

[0133] GOOD'S buffer 100mmol / L

[0134] H2O2 7mmol / L

[0135] 4-Aminoantipyrine 5mmol / L

[0136] BSA 1g / L

[0137] MIT 0.1g / L

[0138] OP 1ml / L

[0139] Hydrochloric acid 20ml / L

[0140] Reagent 3:

[0141] 4-Aminobenzohydrazide 30μmol / L

[0142] BSA 1g / L

[0143] MIT 0.1g / L

[0144] Emulsifier OP 1ml / L

[0145] The assay method of embodiment 3 is identical with embodiment 1.

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Abstract

The invention discloses a kit and a method for clinical detection of myeloperoxidase, which belong to the field of biochemical test reagents. The kit provided by the invention consists of three reagents. In the invention, the activity of the myeloperoxidase in a sample is restrained by a myeloperoxidase inhibitor with high specificity, high inhibition and low inhibition constant Ki, the activities of total peroxidase and non-myeloperoxidase are determined respectively, and the difference between the activities of total peroxidase and non-myeloperoxidase is the activity of the myeloperoxidase.The kit provided by the invention has the advantages of low detection cost, small amount of required sample, high determining precision, simple operation and easy popularization.

Description

technical field [0001] The invention belongs to the technical field of biochemistry and testing reagents, in particular to a kit for clinically detecting myeloperoxidase (MPO) and a method thereof. Background technique [0002] Myeloperoxidase (English name: Myeloperoxidase, English abbreviation: MPO, EC: 1.11.1.7) is mainly glycosylated hemoglobin secreted by activated polymorphonuclear neutrophils. It is a peroxidase . MPO is green in color and has a molecular weight of 140kDa. It is a dimer composed of two identical monomers. After the two monomers are separated, they still have enzyme activity. [0003] Studies have confirmed that MPO is a leukocyte enzyme that has pro-inflammatory and atherosclerotic effects, affects the stability of atherosclerotic plaques, and causes acute coronary atherosclerosis syndrome (ACS) by increasing oxidative stress. ). As a marker of activated granulocytes, MPO is a good indicator for ACS risk stratification; MPO also has important detec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/28G01N21/31
Inventor 邹炳德贾江花胡治宝
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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