Liquid 25-hydroxyvitamin D calibrator diluent

A hydroxyvitamin and calibrator technology, applied in biological testing, material testing products, preparation of samples for testing, etc., can solve the problems of increasing production costs of enterprises, difficult preservation of calibrators, deviation of test results, etc. The effect of production cycle and stability guarantee

Inactive Publication Date: 2019-04-16
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Freeze-dried calibrator has the following disadvantages: the production process is complicated, which increases the production cost of the enterprise; it needs to be reconstituted before use, which brings inconvenien

Method used

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  • Liquid 25-hydroxyvitamin D calibrator diluent
  • Liquid 25-hydroxyvitamin D calibrator diluent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Preparation of liquid 25-hydroxyvitamin D calibrator dilution

[0017] Weigh a certain amount of 7-dehydrocholesterol, then dissolve it in a small amount of absolute ethanol, then weigh a certain proportion of human serum albumin and Proclin 300, add it to 100mmol / L PBS buffer, stir for 30min, that is Prepare a 25-hydroxyvitamin D calibrator dilution, which contains human serum albumin 3% w / v, Proclin 300 0.2% w / v, and 7-dehydrocholesterol 2.0mmol / L.

Embodiment 2

[0018] Example 2 Preparation of relevant reagents for the detection of 25-hydroxyvitamin D content in human serum samples

[0019] 1. Preparation of 25-hydroxyvitamin D series calibrator

[0020] Take a certain amount of 25-hydroxyvitamin D antigen, add respectively to the calibration product dilution prepared in Example 1, and prepare the labeled concentration as 0ng / ml, 10ng / ml, 20ng / ml, 40ng / ml, 80ng / ml, 140ng / ml of a series of calibration products, the color of the bottle cap is white, yellow, green, blue, red, purple, black;

[0021] 2. Preparation of magnetic particle suspension coated with anti-25 hydroxyvitamin D antibody

[0022] Take an appropriate amount of carboxyl magnetic particles according to the amount used, and activate them under acidic conditions with excess EDC and NHS. The activation time is 30 minutes. After the activation is completed, add a magnetic field and let it stand for 5 minutes to separate the magnetic particles from the liquid. Discard the s...

Embodiment 3

[0029] Example 3 Detection of 25-hydroxyvitamin D content in human serum samples

[0030] The relevant reagents prepared in Example 2 are used to detect the content of 25-hydroxyvitamin D in human serum samples, and the specific steps are:

[0031] 1. Take out a certain amount of reaction containers and number them. Add 50 μl of calibration / quality control / clinical serum according to the experimental requirements;

[0032] 2. Shake the magnetic particle suspension, add 20 μl to each well;

[0033] 3. Add 50 μ of enzyme marker to each well;

[0034] 4. Mix the solution in the reaction vessel evenly and incubate at 37°C for 15 minutes;

[0035] 5. Using magnetic separation and washing equipment, wash the magnetic particles in the reaction vessel with washing liquid for 5 times;

[0036] 6. The reaction vessel after washing is fully oscillated to disperse the magnetic particles;

[0037] 7. Add 50 μl each of luminescent substrate A and luminescent substrate B to each well, s...

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Abstract

The invention discloses a liquid 25-hydroxyvitamin D calibrator diluent, which comprises the following components: a buffer solution, a protein stabilizer, a preservative and a cyclopentane polyhydrophenanthrene stabilizer. The main innovation of the invention lies in that a) 7-dehydrocholesterol is applied to the 25-hydroxyvitamin D calibrator for the first time for improving the stability of 25-hydroxyvitamin D under a liquid aerobic condition; b) all the components in the calibrator diluent are optimal formulas, so that the stability of the 25-hydroxyvitamin D calibrator is powerfully guaranteed, and the 25-hydroxyvitamin D calibrator can be stably stored at 8 DEG C for 12 months; and c) the liquid 25-hydroxyvitamin D calibrator is applied to detection of 25-hydroxy vitamin D in the serum sample, the calibrator does not need to be redissolved during detection, and the repeatability of a calibration curve which is calibrated for multiple times is superior to that of the calibration curve of the freeze-dried calibrator.

Description

technical field [0001] The invention relates to in vitro diagnostic technology, in particular to a stable liquid 25-hydroxyvitamin D calibrator diluent. Background technique [0002] Vitamin D is a steroid hormone of two main types: vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol), which are similar in structure but differ in their side chains. Vitamin D in the human body mainly comes from the conversion of 7-dehydrocholesterol in the skin to vitamin D3 by ultraviolet radiation, and also includes vitamin D3 ingested from fish, meat and other foods and vitamin D2 in dietary supplements. Regardless of the form of vitamin D, it is combined with binding proteins in the blood and transported to the liver, where it is converted into 25-hydroxyvitamin D, and then converted into 1,25-dihydroxyvitamin D in the kidney. Is the bioactive ingredient that vitamin D plays a role in. However, the content of 1,25-dihydroxyvitamin D in the circulation is very small, and the half...

Claims

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Application Information

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IPC IPC(8): G01N1/38G01N33/82
CPCG01N1/38G01N33/82
Inventor 任永茂靳增明张玉娇付光宇吴学炜
Owner AUTOBIO DIAGNOSTICS CO LTD
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