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Test strip for rapidly detecting vitamin D

A vitamin and test strip technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of rare standard reference objects, unstable samples, complex vitamin D purification process, etc., to achieve convenient use and fill the gap in the market , low-cost testing effect

Active Publication Date: 2014-09-24
西安良升生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection of vitamin D has not been widely used clinically due to the complicated sample purification process, unstable samples, and rare standard reference materials.

Method used

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  • Test strip for rapidly detecting vitamin D
  • Test strip for rapidly detecting vitamin D

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of conjugates of marker-labeled vitamin D (25-hydroxyvitamin D3) and carrier protein

[0037] Step 1. Dissolve 1 mg vitamin D (25OHD33-HS, purchased from Toronto Research Chemicals Company) in 100 μL organic solvent (n-hexane, acetonitrile, dimethyl sulfoxide or 1,4-dioxane), and then Add the dissolved vitamin D dropwise to 10 mL of carrier protein (bovine serum albumin BSA, or chicken ovalbumin OVA) solution with a concentration of 0.1 mg / mL, and then add pentadiene with a concentration of 50% by volume dropwise. Aldehyde aqueous solution until the volume percentage concentration of glutaraldehyde in the solution is 1%. After stirring for 5 minutes, move to 4°C and stir for 16 hours. The reaction solution after the reaction is dialyzed with PBS buffer solution for 8 hours at 4°C. ~3 times, after centrifuging the dialyzate, take the supernatant, and finally filter the supernatant with a 0.45 μm filter membrane to obtain a conjugate of vitamin D and...

Embodiment 2

[0039] Example 2 Preparation of conjugates of marker-labeled vitamin D (25-hydroxyvitamin D2) and carrier protein

[0040] Step 1. Dissolve 5 mg of vitamin D (25OHD2, purchased from Toronto Research Chemicals) in 100 μL of organic solvent (n-hexane, acetonitrile, dimethyl sulfoxide or 1,4-dioxane), and then Add the dissolved vitamin D dropwise to 10 mL of carrier protein (bovine serum albumin BSA, or chicken ovalbumin OVA) solution with a concentration of 2 mg / mL, and then add a 50% aqueous glutaraldehyde solution dropwise The volume percent concentration of glutaraldehyde in the solution is 2%. After stirring for 10 minutes, move to 4°C and stir for 17 hours. After the reaction, the reaction solution is dialyzed with PBS buffer solution at 4°C for 8 hours. During the dialysis, the liquid is changed for 2 to 3 The second time, the supernatant was taken after the dialyzate was centrifuged, and finally the supernatant was filtered with a 0.45 μm filter membrane to obtain a conju...

Embodiment 3

[0042] Example 3 Preparation of conjugates of marker-labeled vitamin D (25-hydroxyvitamin D3) and carrier protein

[0043] Step 1. Dissolve 10 mg of vitamin D (25OHD3, purchased from Toronto Research Chemicals) in 100 μL of organic solvent (n-hexane, acetonitrile, dimethyl sulfoxide or 1,4-dioxane), and then Add the dissolved vitamin D dropwise to 10 mL of carrier protein (bovine serum albumin BSA, or chicken ovalbumin OVA) solution with a concentration of 5 mg / mL, and then add a 50% aqueous glutaraldehyde solution drop by volume The volume percent concentration of glutaraldehyde in the solution is 3%. After stirring for 20 minutes, move to 4°C and stir for 18 hours. After the reaction, the reaction solution is dialyzed with PBS buffer solution at 4°C for 8 hours. The second time, the supernatant was taken after the dialyzate was centrifuged, and finally the supernatant was filtered with a 0.45 μm filter membrane to obtain a conjugate of vitamin D and carrier protein, which wa...

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Abstract

The invention discloses a test strip for rapidly detecting vitamin D. The test strip comprises a supporting plate which does not absorb water. A sample pad, a combination pad, a chromatography film and an absorbent pad are arranged on the supporting plate in sequence. The combination pad is covered with a marker-marked conjugate of the vitamin D and carrier protein. The chromatography film is a nitrocellulose film and provided with a quality control line C and a detection line T, the detection line T is located between the combination pad and the quality control line C, the detection line T is covered with a vitamin D specific antibody, and the quality control line C is covered with a specific antibody capable of being combined with the carrier protein. The test strip is convenient to use, easy to operate, high in detection speed, and capable of being easily popularized in primary medical units, meeting market requirements, and filling gaps in domestic and foreign markets. A result of the test strip is easy to judge and is represented symbolically, and special requirements of vitamin D detection for equipment, instruments and personnel are eliminated.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a test strip for rapidly detecting vitamin D. Background technique [0002] Vitamin D plays an indispensable role in regulating calcium metabolism in the human body, maintaining the health and stability of the musculoskeletal and immune systems, and preventing the occurrence of some malignant tumors. Statistics from the United Nations Health Organization show that about 90% of the world's population suffers from vitamin D deficiency. Due to the molecular characteristics of vitamin D (highly lipophilic), it is more difficult to separate and purify, and the detection conditions are high, which limits the clinical application of vitamin D detection and limits our access to more information about vitamin D. So far, the human serum vitamin D detection methods that have been reported and accepted clinically include kit detection and instrument detection. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/82G01N33/531
CPCG01N33/532G01N33/558G01N33/82
Inventor 韩雅君
Owner 西安良升生物科技有限公司
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