Beta 2-microglobulin detection kit

A detection kit and microglobulin technology, which is applied in the field of medical immunology, can solve the problems of false positive test results, antibody shedding, false elevation, etc., and achieve the effects of high accuracy, reduced dosage, and low production cost

Active Publication Date: 2012-07-18
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used method of coating β2-microglobulin antibody on latex particles is physical adsorption, which has poor stability and the antibody is easy to fall off from latex particles; and it is easily affected by rheum

Method used

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  • Beta 2-microglobulin detection kit

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Experimental program
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Effect test

Embodiment 1

[0030] The preparation of embodiment 1β2-microglobulin detection kit

[0031] The kit of the present invention relates to the main raw materials of reagents as follows:

[0032] 1. Antibody to β2-microglobulin: the titer was determined by indirect ELISA method to be 1:100000.

[0033] 2. Latex: The present invention only exemplarily uses polystyrene latex particles with a diameter of 200-300 nm and carboxyl groups to carry out experiments.

[0034] The preparation of the main reagent of this embodiment is as follows:

[0035] Reagent R1: 0.2M ammonium chloride buffer containing 1.0% PEG6000 (polyethylene glycol 6000), 0.09% sodium azide, 0.2% BSA, 0.8% NaCl, 0.3% Tween 20. The reagent is a colorless transparent solution.

[0036] Reagent R2: 0.1M Tris buffer containing 0.2% β2-microglobulin antibody sensitized polystyrene latex particles, 1% BSA, 0.2% NaN 3 . The reagent is a milky white solution. Specific steps are as follows:

[0037] 1. Take 1ml (100mg / ml) latex, was...

Embodiment 2

[0047] Determination of embodiment 2β2-microglobulin

[0048] Detection tool: Hitachi 7060 automatic analyzer.

[0049] Analysis method: two-point endpoint method; main wavelength: 546nm, secondary wavelength: 700nm: sample volume: 3.0ul; reagent R1: 240ul; reagent R2: 60ul; calibration method: spline function Spline; reaction direction: rising; measurement temperature: 37°C; after mixing the sample and reagent R1, read the absorbance A at 30 seconds 1 ; Add reagent R2 at 5 minutes, read absorbance A at 5 minutes 2 . The absorbance of the reaction was calculated as A 2 with A 1 calibration difference.

[0050] Calculation method: multi-point calibration, using spline function as the calculation mode, and making a dose / response curve according to the absorbance and reference serum value, and the sample content can be calculated on the dose / response curve according to its absorbance value.

[0051] Reference range: 1.0~3.0mg / L

Embodiment 3

[0052] Example 3 Testing of various performance indicators of the β2-microglobulin detection kit

[0053] 1. Sensitivity test

[0054] Measure the sample of 6 kinds of different β2-microglobulin contents, each sample measures 10 times, by calculating mean value and SD value (standard deviation value), as seen from Table 1, the sensitivity of detection kit of the present invention is 0.08mg / L.

[0055] Table 1

[0056] β2-MG(mg / L)

[0057] 2. High value linear determination

[0058] Measure 10 samples of different β2-microglobulin content, each sample is measured 2 times, as can be seen from Table 2, the highest detection range of the detection kit of the present invention can reach 18mg / L, judge basis r 2 ≥0.990.

[0059] Table 2

[0060] Theoretical value of β2-MG (mg / L)

[0061] 10

[0062] 3. Precision test

[0063] Two human serum samples with different β2-microglobulin contents were used to determine the intra-assay precision of the ...

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Abstract

The invention provides a beta 2-microglobulin detection kit comprising a reagent R1 and a reagent R2, wherein the reagent R1 comprises a buffer solution, a preservative, a stabilizing agent, an electrolyte, a surfactant and the balance of purified water; and the reagent R2 comprises polystyrene latex grains sensitized by the beta 2-microglobulin antibody, a buffer solution, a preservative, a stabilizing agent and a residual amount of purified water. The polystyrene latex grains sensitized by the beta 2-microglobulin antibody are directionally coupled. A preparation method comprises the following steps of: activating the polystyrene latex grains; oxidizing the beta 2-microglobulin antibody; and coupling the beta 2-microglobulin antibody and the polystyrene latex grains. The kit provided by the invention has the advantages of high sensitivity, good specificity, good accuracy, strong anti-jamming capability and low production cost.

Description

technical field [0001] The invention belongs to the field of medical immunology, and relates to an immunodetection reagent, and further, the invention relates to a β2-microglobulin detection kit. Background technique [0002] β2-microglobulin is a small molecular globulin produced by lymphocytes, platelets, and polymorphonuclear leukocytes, with a molecular weight of 11.8KD. It is the β chain (light chain) part of the human lymphocyte antigen (HLA) on the cell surface (for a single-chain polypeptide), widely exists in plasma, urine, cerebrospinal fluid, saliva and colostrum, the content is very low. The synthesis rate of normal human β2-microglobulin and the amount released from the cell membrane are quite constant, β2-microglobulin can be freely filtered from the glomerulus, and 99.9% is absorbed in the proximal renal tubule, so the determination of blood β2-microglobulin Protein can be used as an indicator to reflect glomerular filtration rate. [0003] The commonly used...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531
Inventor 邹炳德邹继华石春林
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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