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Kit for simultaneously detecting beta2-microglobulin in serum and urine specimen

A microglobulin and kit technology, which is applied in the field of biomedicine, can solve the problems of seldom simultaneous measurement of blood and urine, narrow linear range and low detection sensitivity, and achieve biotin-streptavidin immunolabeling and detection. Effects of sensitive, wide linear range and high sensitivity of tracer analysis

Inactive Publication Date: 2018-11-23
宁波海尔施智造有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] CN101813700A discloses a kit for detecting β2-microglobulin by nanometer microsphere immunoturbidimetric method, the kit includes reagent R1, reagent R2 and reference calibrator, and the nanometer of anti-human β2-microglobulin antibody in reagent R2 The particle size of the microspheres is 50-150nm. The combination of the nano-microspheres and the anti-human β2-microglobulin antibody adopts the physical adsorption method. Factor (RF) and heterophilic antibody interference, resulting in false positive or falsely elevated test results
[0008] Various β2-microglobulin detection kits currently on the market usually have some deficiencies and cannot perfectly meet the needs of use, for example, either the detection sensitivity is low, the linear range is narrow, and only β2-microglobulin in serum can be detected. Simultaneous measurement of blood and urine can be achieved, or although it can be measured simultaneously, the production cost of the technical solution based on double particle size is relatively high, etc.

Method used

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  • Kit for simultaneously detecting beta2-microglobulin in serum and urine specimen
  • Kit for simultaneously detecting beta2-microglobulin in serum and urine specimen
  • Kit for simultaneously detecting beta2-microglobulin in serum and urine specimen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] The configuration of embodiment 1 reagent

[0054] 1.1 Configuration of reagent R1

[0055] During the reagent preparation process, prepare according to the concentration in the table below, pass through a 0.22 μm filter membrane after preparation, and store at 4°C for later use.

[0056] components

Matching ratio 1

Matching 2

Matching ratio 3

4-Hydroxyethylpiperazineethanesulfonic acid buffer

50mM

100mM

200mM

Tween-20

0.5mM

1.5mM

2.0mM

Sodium chloride

50mM

100mM

200mM

PEG-6000

4mM

5mM

6mM

Proclin 300

3mM

3mM

3mM

Disodium edetate

5mM

3mM

1.5mM

The rest is purified water

/

/

/

pH

6.5

7.0

8.5

[0057] 1.2 Configuration of reagent R2 buffer

[0058] During the reagent preparation process, prepare according to the concentration in the table below, pass through a 0.22 μm filter membrane after preparation,...

Embodiment 2

[0067] The preparation of embodiment 2 reagent R2

[0068] 2.1 Streptavidin-labeled latex

[0069] 1) Dilute latex particles with particle diameters of 61nm, 145nm, and 220nm (purchased from JSR Life Science, product numbers P0001 / P0112 / P0219 respectively) with 50mM boric acid buffer pH 9.0 to a final concentration of 1%, and react at room temperature for 5 minutes;

[0070] 2) Add EDC to each system obtained in 1), so that EDC / latex = 50 μg / 1 mg, and react at room temperature for 30 minutes;

[0071] 3) Streptavidin was added to 2) to make streptavidin / latex = 50 μg / 1 mg, mixed and stirred, and reacted at room temperature for 1 hour;

[0072] 4) Add BSA to 3) so that the final concentration of BSA is 1%, and block at room temperature for 2 hours;

[0073] 5) Centrifuge at room temperature at 15,000 rpm to remove the supernatant;

[0074] 6) Dissolve the precipitate in 5) with reagent R2 buffer solution to make the final concentration 6-8mg / ml, ultrasonically disperse and p...

Embodiment 3

[0083] Embodiment 3 kit detection process

[0084] 3.1 Test conditions:

[0085] dominant wavelength

600nm

Reaction direction

Forward

temperature reflex

37℃

Response method

endpoint method

[0086] 3.2 Operation process:

[0087]

[0088]

[0089]The reading point is 11-27 on the AU480 automatic biochemical analyzer.

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Abstract

The invention discloses a kit based on single particle diameter for simultaneously detecting beta2-microglobulin in serum and urine specimen. The kit comprises a reagent R1, a reagent R2, a calibration product and a quality control product, wherein the reagent R2 consists of a streptavidin marked latex-biotin marked rabbit anti-human beta2-microglobulin polyclonal antibody and reagent R2 buffer liquid. The kit has the advantages that the reagent sensitivity is improved; the measuring range is greatly increased; the beta2-microglobulin in the blood and urine specimen can be simultaneously detected; meanwhile, most biochemical testing equipment in the market can be compatible; the kit can directly enter the market.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a kit capable of simultaneously detecting β2-microglobulin in serum and urine. Background technique [0002] β2-microglobulin is a small molecular globulin produced by lymphocytes, platelets, and polymorphonuclear leukocytes, with a molecular weight of 11,800 and a single-chain polypeptide composed of 99 amino acids. It is the β chain (light chain) part of the human leukocyte antigen (HLA) on the cell surface. It contains a pair of disulfide bonds and does not contain sugar. It is similar to the structure of the stable region of immunoglobulins and exists widely in a low concentration of free form. In plasma, urine, cerebrospinal fluid, saliva and colostrum. [0003] Free β2-microglobulin is excreted and degraded through glomerular filtration, renal tubular reabsorption. The synthesis rate of normal human β2-microglobulin and the amount released from the cell membrane are q...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6854G01N2333/47
Inventor 柳建敏林威彦张丹丹
Owner 宁波海尔施智造有限公司
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