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25-hydroxyvitamin D detection kit, preparation method and detection method

A technology of hydroxyvitamins and detection kits, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of narrow detection range, poor reproducibility, and low sensitivity, so as to improve the accuracy of detection and enhance the activity of antigens and solubility, and the effect of improving detection sensitivity

Inactive Publication Date: 2020-09-29
深圳市爱康试剂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Of course, the test results of the 25-hydroxyvitamin D assay reagent are for clinical reference only, and cannot be used alone as a basis for diagnosing or excluding cases
[0004] However, the currently used chemiluminescent immunoassay for the detection of 25-hydroxyvitamin D still has problems such as low sensitivity, poor specificity, poor reproducibility, and narrow detection range.

Method used

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  • 25-hydroxyvitamin D detection kit, preparation method and detection method
  • 25-hydroxyvitamin D detection kit, preparation method and detection method
  • 25-hydroxyvitamin D detection kit, preparation method and detection method

Examples

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Effect test

Embodiment 1

[0063] A 25-hydroxyvitamin D detection kit, including R1 reagent, R2 reagent, R3 reagent, sample processing reagent, calibrator and quality control product, wherein,

[0064] The R1 reagent includes streptavidin-coated magnetic particle solution, the concentration of the streptavidin-coated magnetic particle is 0.05-1 mg / ml, and the particle diameter of the magnetic particle is 1-4 μm.

[0065] The R2 reagent includes alkaline phosphatase-labeled 25-hydroxyvitamin D monoclonal antibody solution, the concentration of the alkaline phosphatase-labeled anti-25-hydroxyvitamin D monoclonal antibody is 0.2-1.0ng / mL, The labeling ratio of vitamin D monoclonal antibody to alkaline phosphatase is 1:1-10:1.

[0066] The R3 reagent includes biotin-labeled 25-hydroxyvitamin D antigen, and the concentration of the biotin-labeled 25-hydroxyvitamin D antigen is 0.05-0.2%.

[0067] Sample processing reagents include weakly acidic buffer, reducing agent and lower alcohol solvent, which are use...

Embodiment 2

[0101] Detection method of 25-hydroxyvitamin D detection kit

[0102] Step A: Mix the sample to be tested, the sample processing reagent and the R2 reagent, and incubate at 36-38°C for 20-40 minutes to fully dissociate the 25-hydroxyvitamin D in the sample to be tested, and combine with alkaline phosphatase-labeled 25-Hydroxyvitamin D monoclonal antibody is fully combined and washed;

[0103] Step B: Add R1 reagent and R3 reagent, incubate at 36-38°C for 5-15 minutes, and wash;

[0104] Step C: Add a chemiluminescent substrate to make it catalyzed by the enzyme to generate a luminescent signal, which is received by the optical quantum reading system, and the photomultiplier tube measures the number of photons generated by the reaction, and the software automatically calculates the number of photons to be processed based on the optical signal and the Cutoff value. Measure the 25-hydroxyvitamin D content in the sample;

[0105] The chemiluminescent substrate is (3-(2-helicadam...

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Abstract

The invention relates to the technical field of in-vitro diagnosis and detection, and particularly relates to a 25-hydroxyvitamin D detection kit, a preparation method and a detection method. The 25-hydroxyvitamin D detection kit comprises a reagent R1, a reagent R2, a reagent R3 and a sample treatment reagent, wherein the reagent R1 comprises a streptavidin magnetic particle solution; the reagentR2 is prepared from an alkaline phosphatase labeled 25-hydroxyvitamin D monoclonal antibody solution; the R3 reagent is prepared from a biotin labeled 25-hydroxyvitamin D antigen solution; the sampletreatment reagent comprises a weakly acidic buffer solution, a reducing agent and a lower alcohol solvent. The kit provided by the invention has high sensitivity, specificity and a wide detection range, can be used for detection only by using 40 [mu]L of serum or plasma, has good patient experience, can be combined with a full-automatic chemiluminescence analyzer, and realizes automatic monitoring.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnostic detection, in particular to a 25-hydroxyvitamin D detection kit, preparation method and detection method. Background technique [0002] Vitamin D is a fat-soluble sterol derivative, mainly in two forms: vitamin D2 and D3. Vitamin D2 is produced from ergosterol in plants irradiated by ultraviolet rays; vitamin D3 is converted from 7-dehydrocholesterol contained in human epidermis and dermis by ultraviolet irradiation in sunlight. In the human body, vitamin D2 and D3 are combined with vitamin D binding protein in plasma and transported to the liver, where they form 25-hydroxyvitamin D (including 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3). 25-hydroxyvitamin D is transformed into 1,25-dihydroxyvitamin D in the mitochondria of renal proximal tubular epithelial cells by the α-hydroxylase system, which is the largest biological form of vitamin D and can promote the secretion of intestin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/82G01N33/577G01N21/76
CPCG01N33/82G01N33/577G01N21/76
Inventor 高兴科胡贵宾刘芮吴凡李兵古小丹彭燕婷
Owner 深圳市爱康试剂有限公司
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