Glomerular basement membrane antibody (gbm-ab) quantitative assay kit

A technology for quantitative determination of glomerular basement membrane, applied in the field of clinical immunology detection, can solve the problems of inability to distinguish non-specific identification, expensive fluorescence microscope, inability to perform quantitative determination, etc., and achieves fast and reliable reaction process and high detection sensitivity. and the effect of linear range, validity period and detection performance guarantee

Active Publication Date: 2016-03-23
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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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 in 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 3000.2mL of Proclin 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 (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. GBM antigen: it...

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 in a container, weigh 6.06g of Hepes and 8.5g of NaCl into the container, and stir until completely dissolved;

[0074] 2. Weigh BSA5g, Zncl20.1g, Proclin-3000.2mL and MgCl20.1g in a container, stir well until completely dissolved;

[0075] 3. Use 4M HCl to adjust the pH, and control the pH within the range of 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 GBM antibody:

[0078] 1. GBM 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;

[0080] 3. ...

Embodiment 3

[0084] Embodiment 3, the preparation of calibrator:

[0085] 1. Preparation of calibrator diluent:

[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 3000.02mL of Proclin 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 GBM 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

Belonging to the technical field of clinical immunological detection, the invention discloses a quantitative determination kit for a glomerular basement membrane antibody (GBM-Ab). The kit includes a magnetic separation reagent, an enzyme labeled reagent, a calibrator, a cleaning concentrated solution, a substrate solution and a stabilizer. The invention also discloses a preparation method and a detection method of the kit. The kit prepared by the invention has the advantages of good accuracy, sensitivity and stability, low cost, simple operation, and wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of clinical immunology detection, in particular to a kit for quantitative determination of glomerular basement membrane antibody (GBM-Ab) and a detection method thereof. Background technique [0002] Pulmonary hemorrhage-nephritic syndrome is also known as anti-basement membrane glomerulonephritis, Goodpasture syndrome or Goodpasture disease. It is severe damage to the basement membrane of the glomerulus and alveolar wall caused by anti-basement membrane antibodies, clinically manifested as a triad of pulmonary hemorrhage, rapidly progressive glomerulonephritis, and positive serum anti-glomerular basement membrane (GBM) antibodies. Most patients progress rapidly and the prognosis is dire. Goodpasture first reported glomerular injury with pulmonary hemorrhage in a fulminant influenza in 1919. In 1958, Stanton and Tange reported a group of cases of pulmonary hemorrhage combined with crescentic glomerulonephr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/535G01N33/5375G01N2800/347
Inventor 于大为杨晓勇
Owner 北京贝尔医疗设备有限公司
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