Kit and method for quantitative detection of 25-hydroxyvitamin D through TRFIA

A time-resolved fluorescence, hydroxyvitamin technology, applied in biological testing, fluorescence/phosphorescence, analytical materials, etc., can solve the problems of affecting the measurement results, insufficient dissociation, etc., achieving strong fluorescence signal, avoiding fluorescence interference, and wide detection range Effect

Active Publication Date: 2019-07-05
XIAMEN INST OF RARE EARTH MATERIALS
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In addition, most of the 25-OH VD in the blood is combined with the vitamin-binding protein VDBP. Therefore, a sample dissociation solution is required to dissociate 25-OH VD from VDBP for measurement. Most of the existing dissociation methods are insufficiently dissociated and affect the measurement results.

Method used

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  • Kit and method for quantitative detection of 25-hydroxyvitamin D through TRFIA
  • Kit and method for quantitative detection of 25-hydroxyvitamin D through TRFIA
  • Kit and method for quantitative detection of 25-hydroxyvitamin D through TRFIA

Examples

Experimental program
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Effect test

Embodiment 1

[0051] Example 1: Preparation of time-resolved fluorescence quantitative detection kit for 25-hydroxyvitamin D

[0052] The time-resolved fluorescence quantitative detection kit for 25-hydroxyvitamin D uses the principle of competitive immunochromatography to detect trace amounts of 25-hydroxyvitamin D in human blood. It consists of a time-resolved fluorescent test paper card, a sample dissociation solution and an ID card containing a calibration curve.

[0053] Such as figure 1 As shown, in this embodiment, the sample pad 1, the binding pad 2, the coating film 3 and the absorbent paper 4 are sequentially overlapped on the substrate 7, and the sample pad 1 is a sample application area for absorbing the blood test sample to be tested. The binding pad 2 is provided with microsphere lines, and the coating film 3 is provided with detection lines and quality control lines.

[0054] Rare earth vanadate nanoparticles (GdVO) with specific excitation light (285nm) / emission light (618...

Embodiment 2

[0072] Example 2: Quantitative detection of time-resolved fluorescence quantitative detection kit for 25-hydroxyvitamin D

[0073] 1. Drawing of standard curve

[0074] The time-resolved fluorescent test paper card prepared according to Example 1 was added with different concentrations of 25-hydroxyvitamin D antigen quality controls (100ng / mL, 70ng / mL, 50ng / mL, 30ng / mL, 20ng / mL, 10ng / mL, 3ng / mL There are seven concentrations in mL, and each concentration has three repetitions, all of which are obtained by diluting 3.7mg / ml 25-hydroxyvitamin D recombinant antigen with 1% BSA+20mM pH7.4PBS), adding the sample dissociation solution, and after 10min of chromatography, The C and T line fluorescence signals and C / T value were read by the AFS-1000 dry fluorescent immunoassay analyzer produced by Guangzhou Lanbo Biotechnology Co., Ltd. The experimental results and analysis are shown in Table 1:

[0075]

[0076]

[0077] Take the LOG value of the 25-hydroxyvitamin D quality contr...

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Abstract

The invention relates to a 25-hydroxyvitamin D TRFIA (Time-Resolved Fluoroimmunoassay) kit and a detection method, which utilize the sensitivity of TRFIA technology, utilize rare earth vanadate nano fluorescent labeling materials and combine a dry-type immunofluorescence analyzer to realize high sensitivity, accurate quantification, simpleness and convenience. The kit of the invention can accurately and quantitatively detect the content of the 25-hydroxyvitamin D in serum, plasma and whole blood samples of persons, the whole blood samples can be filtered through the treatment of blood filtering membrane sample pads, the fluorescence interference of the samples can be avoided by utilization of a time resolution detection technology, the fluorescent microspheres are rare earth vanadate nanomaterials, thus the advantages of low background, strong fluorescence signals, high signal-to-noise ratio and the like are achieved, rare earth vanadate nano particles are connected with antibody through covalent bonds by labelling, a labeled product is stable, thus the kit has the characteristics of wide detection range (3-100ng/mL), high sensitivity (detection limit is 3ng/mL), high accuracy, quick and convenient detection and the like.

Description

[0001] (1) Technical field [0002] The invention relates to a kit and a detection method for time-resolved fluorescence quantitative detection of 25-hydroxyvitamin D. [0003] (2) Background technology [0004] Vitamin D is a fat-soluble steroid precursor, mainly produced by the skin after exposure to light. Two important forms of vitamin D include vitamin D 3 (cholecalciferol) and vitamin D 2 (ergocalciferol), in humans, vitamin-binding protein and vitamin D 3 and vitamin D 2 Combined and transported to the liver, the two are hydroxylated to produce 25-hydroxyvitamin D. 25-hydroxyvitamin D is a metabolite that can be detected in blood, and most of the 25-hydroxyvitamin D detected in serum is 25-hydroxyvitamin D3, so the status of vitamin D can be determined by detecting 25-OH VD . Vitamin D can promote intestinal absorption of calcium and regulate calcium balance, and is the main element for maintaining bone health. Severe vitamin D deficiency in childhood can lead to s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/82G01N33/58G01N33/577G01N33/533G01N33/558G01N21/64
CPCG01N33/82G01N33/533G01N33/577G01N33/582G01N33/558G01N21/6408Y02A50/30
Inventor 张云张肖宋良
Owner XIAMEN INST OF RARE EARTH MATERIALS
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