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Stable 25-hydroxyvitamin D (25-OH VD) chemiluminescence immunoassay kit

A technology for chemiluminescence immunity and hydroxyvitamin, which is applied in biological testing, measuring devices, scientific instruments, etc., can solve the problems of inconvenient storage and transportation, destruction and poor stability of the kit, so as to achieve convenient storage and transportation and high detection sensitivity. , the effect of a wide linear range

Pending Publication Date: 2020-01-17
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] When detecting 25-hydroxyvitamin D, it is necessary to use 25-hydroxyvitamin D antigen derivatives. However, 25-hydroxyvitamin D antigen derivatives are substances with poor stability and are very sensitive to light and heat. When hot, it will lose the α-hydrogen atom connected to the double bond to become an allyl radical, and at the same time, the 22 carbon in VD is easily oxidized and broken to destroy its structure
Therefore, due to the instability of the 25-hydroxyvitamin D antigen derivative, the assay kit cannot be stored stably for a long time
In addition, the disadvantage of inconvenient storage and transportation of the kit due to poor stability also affects the use and test results of the test reagent, which is the biggest problem in the current 25-hydroxyvitamin D detection technology

Method used

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  • Stable 25-hydroxyvitamin D (25-OH VD) chemiluminescence immunoassay kit
  • Stable 25-hydroxyvitamin D (25-OH VD) chemiluminescence immunoassay kit
  • Stable 25-hydroxyvitamin D (25-OH VD) chemiluminescence immunoassay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 125

[0062] Example 1 Preparation of 25-hydroxyvitamin D chemiluminescent immunoassay kit

[0063] R1 reagent composition:

[0064] TRIS buffer 25mmol / L

[0065] NaCl 150mmol / L

[0066] Mass ratio of streptavidin magnetic particles 0.072%

[0067] The particle diameter of the magnetic particles is 3 μm.

[0068] R2 reagent composition:

[0069]

[0070] Chemiluminescent marker-labeled polyclonal antibody to 25-hydroxyvitamin D 0.15 μg / mL

[0071] The chemiluminescent label is acridinium ester, and the labeling ratio of antibody to label is 1:3.

[0072] R3 reagent composition:

[0073]

[0074] The concentration of small molecule antigen derivatives of biotin-conjugated vitamins was 0.05 μg / mL.

[0075] R4 reagent composition:

[0076] The buffer is selected from 0.5M ethylene carbonate, and the reducing agent is selected from dithiothreitol (DTT).

[0077] R5 reagent composition:

[0078] 0.3M sodium hydroxide.

[0079] Kit preparation:

[0080] (1) Preparation o...

Embodiment 2

[0096] Example 2 Preparation of 25-hydroxyvitamin D chemiluminescent immunoassay kit

[0097] R1 reagent composition:

[0098] HEPES buffer 10mmol / L

[0099] KCl 100mmol / L

[0100] Mass ratio of streptavidin magnetic particles 0.01%

[0101] The particle diameter of the magnetic particles is 0.05 μm.

[0102] R2 reagent composition:

[0103]

[0104] Chemiluminescent marker-labeled polyclonal antibody to 25-hydroxyvitamin D 0.15 μg / mL

[0105] The chemiluminescence label is luminol, and the labeling ratio of antibody to label is 1:10.

[0106] R3 reagent composition:

[0107]

[0108]

[0109] The concentration of small molecule antigen derivatives of biotin-conjugated vitamins was 0.05 μg / mL.

[0110] R4 reagent composition:

[0111] The buffer is selected from 0.1M ethylene carbonate, and the reducing agent is selected from dithiothreitol (DTT).

[0112] R5 reagent composition:

[0113] 0.1 M sodium hydroxide.

[0114] Kit preparation:

[0115] (1) Prepa...

Embodiment 3

[0126] Example 3 Preparation of 25-hydroxyvitamin D chemiluminescent immunoassay kit

[0127] R1 reagent composition:

[0128] Disodium hydrogen phosphate-citric acid buffer 30mmol / L

[0129] MgCl 2 300mmol / L

[0130] Mass ratio of streptavidin magnetic particles 1%

[0131] The particle diameter of the magnetic particles is 1 μm.

[0132] R2 reagent composition:

[0133]

[0134] Chemiluminescent marker-labeled polyclonal antibody to 25-hydroxyvitamin D 0.15 μg / mL

[0135] The chemiluminescence label is ruthenium terpyridine, and the labeling ratio of antibody to label is 1:20.

[0136] R3 reagent composition:

[0137]

[0138] The concentration of small molecule antigen derivatives of biotin-conjugated vitamins was 0.05 μg / mL.

[0139] R4 reagent composition:

[0140] The buffer is selected from 0.1M ethylene carbonate, and the reducing agent is selected from dithiothreitol (DTT).

[0141] R5 reagent composition:

[0142] 0.5M sodium hydroxide.

[0143] Ki...

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Abstract

The invention discloses a stable 25-hydroxyvitamin D (25-OH VD) chemiluminescence immunoassay kit. The kit includes a reagent R1, a reagent R2, a reagent R3, a reagent R4 and a reagent R5. R1 is a buffer containing streptavidin magnetic particles. R2 is a buffer containing chemiluminescence marker-labeled 25-hydroxyvitamin D polyclonal antibodies and a protective agent. R3 is a buffer containing asmall molecule antigen derivative of biotin-bound vitamin and a protective agent. R4 and R5 are sample pretreatment reagents, and are used to dissociate 25-hydroxyvitamin D existing in a binding protein form in serum or plasma. The kit of the invention uses casein micelles as the protective agents to add the same in the reagent buffers, and is used to detect the content of the 25-hydroxyvitamin Din the serum or the plasma, effectively improves stability of the antigen derivative and accuracy of a testing result, solves the defect problems existing in the prior art, and facilitates storage and transportation of the reagents.

Description

technical field [0001] The invention relates to the technical field of in vitro detection, in particular to a stable 25-hydroxyvitamin D chemiluminescence immunoassay kit. Background technique [0002] Vitamin D is a steroid derivative and is a fat-soluble vitamin. Vitamin D is mainly synthesized by human skin after ultraviolet radiation, and a small part is ingested from food or supplements. Vitamin D not only affects calcium and phosphorus metabolism, but also has a wide range of physiological effects. It is an essential substance for maintaining human health, cell growth and development, and is closely related to many diseases. [0003] There are two forms of vitamin D in the human body, vitamin D3 (cholecalciferol) and vitamin D2 (ergocalciferol), and vitamin D is converted into 25-hydroxyvitamin D (25-OH VD) by hydroxylation in the liver, It is then converted to active 1,25-dihydroxyvitamin D in the kidneys. Other metabolites of vitamin D, circulating at concentratio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/82G01N33/543G01N33/531
CPCG01N33/82G01N33/54326G01N33/531
Inventor 张丹丹孟令敏韩美玉王凯高威孙成艳何浩会
Owner DIRUI MEDICAL TECH CO LTD
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