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Kit capable of detecting alpha-1-microglobulin in urine and serum samples simultaneously

A technology in microglobulin and samples, applied in the field of biomedicine, can solve the problems of narrow linear range, false positive detection samples, poor specificity, etc., and achieve the effect of strong anti-interference ability, wide linear range and good specificity

Active Publication Date: 2016-04-06
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] An α 1 - Microglobulin detection kit and its preparation, based on polyclonal antibody colloidal gold enhanced immune turbidimetry, has the advantages of high sensitivity, but its detection linear range is only 0-40.0mg / L (see 3 / in the patent specification for details 10 page [0023] paragraph), and there is the problem of narrow linear range, can't satisfy to α 1 -Requirements for the detection of high-value pathological clinical samples with a microglobulin concentration above 40mg / L
The website of the State Intellectual Property Office of China also disclosed a nano-microsphere immunoturbidimetric method for the detection of α 1 - Kit for microglobulin, enhanced immunoturbidimetric assay using polyclonal antibody nanoparticles for alpha in serum 1 - detection of microglobulin (see patent specification [0001] paragraph for details), but can not be used for α in human urine 1 - Detection of microglobulin
[0005] In addition, polyclonal antibodies are less specific than monoclonal antibodies, and patients with abnormal rheumatoid factors are likely to cause false positives and false negatives in test samples, affecting the accuracy of test results

Method used

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  • Kit capable of detecting alpha-1-microglobulin in urine and serum samples simultaneously
  • Kit capable of detecting alpha-1-microglobulin in urine and serum samples simultaneously
  • Kit capable of detecting alpha-1-microglobulin in urine and serum samples simultaneously

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Experimental program
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Effect test

Embodiment 1

[0025] Reagent R1: 100mmol / L Tris-HCl buffer solution, 15g / L polyethylene glycol (PEG), 20g / L sodium chloride, 1g / L sodium azide, 0.8g / L anti-human rheumatism factor antibody.

[0026] Reagent R2: labeled with anti-human alpha 1-Preparation of latex particles of microglobulin monoclonal antibody: latex particles with a particle size of 198nm (produced by JSR, code No.: P0113; polystyrene nanoparticles) were diluted with 50mmol / LMES, pH6.0 buffer solution When the particles reach 3g / L, add EDAC0.5mg to each ml solution, react at room temperature for 1 hour, centrifuge at 20000rpm for 30min, remove the supernatant (supernatant), and suspend the precipitate in 50mmol / L, pH6.0 MES buffer , ultrasonically disperse; centrifuge again at 20,000rpm for 30min, remove the supernatant, suspend the precipitate in 50mmol / L, pH6.0 MES buffer solution, so that the final concentration of latex particles is 6g / L, ultrasonically disperse, and add while stirring Equal volume containing 8mg / ml an...

Embodiment 2

[0028] Reagent R1: 200mmol / L Tris-HCl buffer solution, 20g / L polyethylene glycol (PEG), 9g / L sodium chloride, 1g / L sodium azide, 0.5g / L anti-human rheumatism factor antibody.

[0029] Reagent R2: Dilute latex particles with a particle size of 240nm (produced by JSR, code No.: P0220) to 3g / L with 50mmol / LMES, pH6.0 buffer solution, add 0.5mg of EDAC to each ml of solution, and store at room temperature React for 1 hour, centrifuge at 20,000rpm for 30min, remove the supernatant, suspend the precipitate in 50mmol / LMES, pH6.0 buffer, and disperse by ultrasonic; centrifuge again at 20,000rpm for 30min, remove the supernatant, and suspend the precipitate in 50mmol / LMES , pH 6.0 buffer solution, so that the final concentration of latex particles is 6g / L, ultrasonically disperse, and add an equal volume of 8mg / ml anti-human α 1 - The MES dilution of the microglobulin antibody, mixed and stirred, reacted at room temperature for 2 hours, and the final concentration of latex particles wa...

Embodiment 3

[0031] alpha 1 - Determination of microglobulins:

[0032] Measuring instrument: Hitachi 7060 automatic biochemical analyzer;

[0033] Analysis method: two-point endpoint method;

[0034] Analysis parameters: reagent 1: 240ul, reagent 2: 60ul, sample: 2ul; wavelength: 546nm;

[0035] Determination steps: first add reagent 1 and sample, incubate at 37°C for 5 minutes, then add reagent 2, immediately read the absorbance value of the first point, after timing the reaction for 5 minutes, read the absorbance value of the second point, and calculate the absorbance difference between the two points value.

[0036] Calibration curve creation: take the concentration of the calibration solution as the abscissa, and the absorbance difference corresponding to each concentration of the calibration solution as the ordinate, and make a logit-log(4p) function curve.

[0037] Calculation method of sample concentration: According to the difference of absorbance value of the sample, it is su...

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Abstract

A kit capable of detecting alpha-1-microglobulin in urine and serum samples simultaneously comprises a regent 1 and a reagent 2, and is characterized in that the reagent 1 is a buffer solution containing antibodies against human rheumatoid factors and a coagulation accelerator; the reagent 2 is a buffer solution containing large-particle-size polystyrene nano-particles coated with monoclonal antibodies against human alpha-1-microglobulin. The kit has the advantages of high sensitivity, wide linearity, better specificity and high anti-interference capacity and can be used for detecting human urine and the serum.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a kit capable of simultaneously detecting α1-microglobulin in urine and serum samples. Background technique [0002] alpha 1 - Microglobulin (alpha-1-microglobulin, alpha 1 -MG) is a relatively small molecular weight glycoprotein in the human body. It is mainly produced by human liver cells and lymphocytes, widely distributed in various body fluids including blood, urine, saliva, etc., and exists in free and high molecular weight bound forms. alpha in the blood 1 - Microglobulin is mainly in free form α 1 - presence of microglobulins, bound to immunoglobulin IgA, bound to albumin. free form α 1 - Microglobulin can freely pass through the glomerular filtration membrane, and is almost completely reabsorbed and catabolized in the proximal convoluted tubule, while the high molecular weight bound α 1 - Microglobulin cannot be filtered by the glomerulus, so only ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/53
CPCG01N33/53G01N33/577G01N33/68
Inventor 邹炳德邹继华刘献文方亮
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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