Method for measuring 25-hydroxyvitamin D

A hydroxyvitamin and biotinylation technology, applied in the field of detection of 25-hydroxyvitamin D in blood samples, can solve problems such as prolonging detection time, linear influence, and increasing detection steps.

Active Publication Date: 2019-01-11
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, this method requires sample pretreatment, which not only increases the detection steps, but also prolongs the detection time
In addition, due to the addition of two pretreatment solutions, the dilution of the sample and the small amount of detection sample resulted in defects such as low sensitivity and affected linearity of the kit.
In addition, despite the pretreatment, there is still some difference in the final test results between serum and plasma

Method used

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  • Method for measuring 25-hydroxyvitamin D
  • Method for measuring 25-hydroxyvitamin D
  • Method for measuring 25-hydroxyvitamin D

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1 measures the comparison of the results of the same source serum and plasma samples

[0053] Materials and Reagents

[0054] Take serum / plasma (EDTA-K2) pairs from 20 subjects,

[0055] Mike Biological Co., Ltd. 25-hydroxyvitamin D master calibration standard 1 set,

[0056] Mike Biological Co., Ltd. 25-hydroxyvitamin D assay kit (chemiluminescence method) 2 boxes (batch number: 0417011, specification: 100 tests / box),

[0057] Mike Biological Co., Ltd. Washing Buffer (Product No.: IM4202458, Lot No.: 0417141)

[0058] Substrate Solution for Fully Automatic Immunoassay System (Enzymatic Chemiluminescent Substrate Solution (HRP)) of Mike Biological Co., Ltd. (Product No.: IM4202459, Batch No.: 0417031)

[0059] Mike Biological Co., Ltd. IS1200 automatic chemiluminescence immunoassay analyzer (equipment number: 1015-01024).

[0060] 25-Hydroxyvitamin D Assay Kit contains the following reagents:

[0061] The first reagent (R1): 0.2 mol / L acetic acid-sodium ...

Embodiment 2

[0073] The different addition amounts of the second magnetic beads of embodiment 2;

[0074] Materials and Reagents

[0075] The same serum / plasma (EDTA-K2) pair from 20 subjects as in Example 1,

[0076] Mike Biological Co., Ltd. 25-hydroxyvitamin D master calibration standard 1 set,

[0077] Mike Biological Co., Ltd. 25-Hydroxyvitamin D Assay Kit (chemiluminescence method) 3 boxes (batch number: 0417011, specification: 100 tests / box),

[0078] Mike Biological Co., Ltd. Washing Buffer (Product No.: IM4202458, Lot No.: 0417141)

[0079] Substrate Solution for Fully Automatic Immunoassay System (Enzymatic Chemiluminescent Substrate Solution (HRP)) of Mike Biological Co., Ltd. (Product No.: IM4202459, Batch No.: 0417031)

[0080] Mike Biological Co., Ltd. IS1200 automatic chemiluminescence immunoassay analyzer (equipment number: 1015-01024)

[0081] 25-Hydroxyvitamin D Assay Kit contains the following reagents:

[0082] The first reagent (R1): 0.2 mol / L acetic acid-sodium a...

Embodiment 3

[0093] Materials and Reagents

[0094] The same serum / plasma (EDTA-K2) pair from 20 subjects as in Example 1,

[0095] Mike Biological Co., Ltd. 25-hydroxyvitamin D master calibration standard 1 set,

[0096]Mike Biological Co., Ltd. 25-Hydroxyvitamin D Assay Kit (chemiluminescence method) 3 boxes (batch number: 0417011, specification: 100 tests / box),

[0097] Mike Biological Co., Ltd. Washing Buffer (Product No.: IM4202458, Lot No.: 0417141)

[0098] Substrate Solution for Fully Automatic Immunoassay System (Enzymatic Chemiluminescent Substrate Solution (HRP)) of Mike Biological Co., Ltd. (Product No.: IM4202459, Batch No.: 0417031)

[0099] Mike Biological Co., Ltd. IS1200 automatic chemiluminescence immunoassay analyzer (equipment number: 1015-01024)

[0100] 25-Hydroxyvitamin D Assay Kit contains the following reagents:

[0101] The first reagent (R1): 0.2mol / L acetic acid-sodium acetate buffer solution (PH 4.5), 0.2 mg / mL of streptavidin magnetic particles as the firs...

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Abstract

The present application discloses a method for measuring 25-hydroxyvitamin D. The method comprises the following steps: using a first reagent and a second reagent to co-incubate with a biological sample, wherein the first reagent comprises a first magnetic micro particle and a second magnetic micro particle in an acidic buffer solution, wherein the first magnetic micro particle is coated with a streptavidin magnetic micro particle; the second magnetic micro particle is the magnetic micro particle different from the first magnetic micro particle; the second reagent comprises a 25-hydroxyvitaminD antibody marker; further adding a third reagent comprising biotinylated 25-hydroxyvitamin D for co-incubation; separating the magnetic micro particles; adding a luminescent substrate and detectingthe luminescence intensity. The method of the present invention is simultaneously applicable to detection of 25-hydroxyvitamin D in serum and plasma samples without the need of additional sample processing reagents and additional sample processing steps as well.

Description

technical field [0001] The invention relates to a method for detecting 25-hydroxyvitamin D in blood samples. Background technique [0002] Vitamin D is a fat-soluble vitamin that belongs to sterol derivatives. The main function of vitamin D is to regulate the metabolism of calcium and phosphorus in the body, maintain the temperature of plasma calcium and phosphorus levels, and participate in bone development. The vitamin D family mainly includes vitamins D1-D5, the most important of which are vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol). [0003] 25-hydroxyvitamin D is formed by vitamin D in the liver through the action of the monooxygenase system of stem cell microsomes, and is the main storage form of vitamin D in the body. The concentration of 25-hydroxyvitamin D in the blood can reflect the level of vitamin D in the human body, and is the best biochemical indicator for detecting the level of vitamin D in an individual. [0004] Traditional detection me...

Claims

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Application Information

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IPC IPC(8): G01N33/82G01N33/543
CPCG01N33/54326G01N33/82
Inventor 李洪伟吴国平田君喜
Owner SICHUAN MACCURA BIOTECH CO LTD
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