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Quantitative measurement kit for myeloperoxidase antibody (MPO-Ab) and detection method thereof

A technology for quantitative determination of myeloperoxidase, which is applied in the field of quantitative determination of myeloperoxidase antibody kits and its detection, can solve the problems of inability to distinguish non-specific recognition, expensive fluorescent microscope, and inability to perform quantitative determination, etc. To achieve the effect of fast and reliable reaction process, high detection sensitivity and linear range, service life and detection performance guarantee

Active Publication Date: 2015-03-25
北京贝尔医疗设备有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method cannot distinguish non-specific recognition according to the size of the molecular weight when analyzing the results; the operation is relatively complicated and requires a more expensive fluorescence microscope, which can only be used for qualitative detection and cannot be used for quantitative determination

Method used

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  • Quantitative measurement kit for myeloperoxidase antibody (MPO-Ab) and detection method thereof
  • Quantitative measurement kit for myeloperoxidase antibody (MPO-Ab) and detection method thereof
  • Quantitative measurement kit for myeloperoxidase antibody (MPO-Ab) and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1, the preparation of magnetic separation reagent:

[0057] 1. The preparation process of magnetic particle buffer, taking the preparation of 1L as an example:

[0058] 1. Measure 800mL of purified water into a container, weigh 12.1g of Tris and 8.5g of NaCl into the container, stir well until completely dissolved;

[0059] 2. Weigh 5g of BSA, 50mL of newborn bovine serum, and 0.2 mL of Proclin300 into the container, stir well until completely dissolved;

[0060] 3. Use 4M HCl to adjust the pH, and control the pH between 7.9-8.1;

[0061] 4. Finally, dilute to 1L with purified water and store at 2-8°C until use.

[0062] 2. Preparation process of magnetic separation reagent

[0063] 1. Take magnetic particles containing carboxyl groups (COOH) active groups, the carboxyl content per gram (g) of magnetic particles (dry weight) is not less than 0.3 millimoles (mmoL), with superparamagnetic properties, and the diameter is between 0.5-2 μm;

[0064] 2. MPO ant...

Embodiment 2

[0071] Embodiment 2, the preparation of enzyme labeling reagent:

[0072] 1. The preparation process of enzyme labeling reagent diluent, taking the preparation of 1L as an example:

[0073] 1. Measure 800mL of purified water into a container, weigh 6.06g of Hepes and 8.5g of NaCl into the container, stir well until completely dissolved;

[0074] 2. Weigh BSA 5g, Zncl 2 0.1g, Proclin-300 0.2mL and MgCl 2 0.1g in the container, fully stirred until completely dissolved;

[0075] 3. Use 4M HCl to adjust the pH, and control the pH between 7.5-8.0;

[0076] 4. Finally, dilute to 1L with purified water and store at 2-8°C until use.

[0077] 2. Coupling of alkaline phosphatase (ALP) to MPO antibody

[0078] 1. MPO antibody, purchased from Beijing Jiufeng Runda Biotechnology Co., Ltd., with a purity of more than 95%;

[0079] 2. The purity of alkaline phosphatase should exceed 95%, the specific activity should exceed 1000U / mg, and the concentration should exceed 5mg / mL;

[008...

Embodiment 3

[0084] Embodiment 3, the preparation of calibrator:

[0085] one, Preparation of calibrator dilutions:

[0086] 1. Measure 600mL of purified water in a container, weigh 200mL of newborn bovine serum and add it to the container, stir well and mix evenly;

[0087] 2. Weigh 3.4g of dipotassium hydrogen phosphate, 0.36g of potassium dihydrogen phosphate, and 0.02 mL of Proclin300 into the container, stir well until completely dissolved;

[0088] 3. Use 4M HCl to adjust the pH, and control the pH between 7.0-7.5;

[0089] 4. Finally, dilute to 1L with purified water and store at 2-8°C until use.

[0090] 2. Preparation of calibrator:

[0091] 1. Prepare the MPO antibody at 0, 5, 20, 50, 150, and 300 U / mL with the calibrator diluent, a total of 6 concentration points.

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Abstract

The invention belongs to the technical field of clinical immunological detection, and discloses a quantitative measurement kit for a myeloperoxidase antibody (MPO-Ab). The kit comprises a magnetic separation reagent, an enzyme labeling reagent, a calibrator, a cleaning concentrate, a substrate solution and a stabilizer. The invention also discloses a preparation method of the kit. The invention also discloses a detection method of the kit. The prepared kit is good in precision, sensitivity and stability, low in cost and simple to operate, so the kit has a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of clinical immunology detection, and in particular relates to a myeloperoxidase antibody (MPO-Ab) quantitative assay kit and a detection method thereof. Background technique [0002] Vasculitis is a group of inflammatory diseases with vascular inflammation and necrosis as the main pathological changes, including inflammatory cell infiltration in and around blood vessels, accompanied by vascular damage, including cellulose deposition, collagen fiber degeneration, endothelial cells and muscle Cell necrosis, also known as vasculitis. The clinical manifestations vary according to the type, size, location and pathological characteristics of the blood vessels involved. It often involves multiple systems of the whole body (such as skin, kidney, lung, nervous system, etc.), causing dysfunction of multiple systems and organs, but it can also be limited to a certain organ. The pathogenic factors act directly on t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535
CPCG01N33/531G01N33/535G01N33/543G01N2800/328
Inventor 于大为杨晓勇
Owner 北京贝尔医疗设备有限公司
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