25-hydroxyvitamin D kit for detecting peripheral blood, and preparation method and application thereof

A hydroxyvitamin and kit technology, which can be applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of large demand for blood samples, difficult to repeatedly draw, and complicated operations, and achieves short sample processing time, small sample consumption, and preparation. simple effect

Inactive Publication Date: 2019-11-22
BEIJING ELCOTEQ BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the commonly used clinical methods for the detection of 25-OH-D include chemiluminescence, colloidal gold immunochromatography, enzyme-linked immunoassay, and electrochemiluminescence. Most of these detection methods require the processing of collected whole blood samples, which is time-consuming. Long, cumbersome operation, the blood samples used are in large demand and all are venous blood
However, for infants, children, and patients with extensive burns, it is difficult to collect enough blood samples and it is not easy to repeatedly draw blood samples.

Method used

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  • 25-hydroxyvitamin D kit for detecting peripheral blood, and preparation method and application thereof
  • 25-hydroxyvitamin D kit for detecting peripheral blood, and preparation method and application thereof
  • 25-hydroxyvitamin D kit for detecting peripheral blood, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: Preparation of 25-hydroxyvitamin D (25-OH-D) kit

[0041] 1.1 Preparation of 25-hydroxyvitamin D (25-OH-D) immunochromatographic detection card

[0042] 1) Goat anti-25-hydroxyvitamin D (25-OH-D) monoclonal antibody labeled latex fluorescent microspheres

[0043] Take 1 mL of latex fluorescent microspheres (purchased from Merck Millipore) and add 1 mg of carbodiimide (EDC), 1 mg of N-hydroxysuccinimide (NHS) and stir at room temperature for 3 h, then add 0.1 mg of goat anti-25-hydroxyvitamin D monoclonal Antibody, stirred at room temperature for 1 h, then added 10 mg of BSA blocking solution, and continued to stir for 1 h. Centrifuge at 11000r / min for 30min at 2-8°C, and remove the supernatant. Finally, take 1 mL of 0.2M phosphate buffer solution (pH=7.4) and add it to the solid precipitate obtained after centrifugation, then add 1 μL of Proclin300, shake on a 40 Hz ultrasonic oscillator for 1 min to mix well, and then incubate at 4 °C Save for later us...

Embodiment 2 Embodiment 1

[0059] The detection of the 25-hydroxyvitamin D kit of embodiment 2 embodiment 1

[0060] 2.1 Detection process (see image 3 )

[0061] 1) Dilute the collected blood sample 21 such as fingertip blood with the release agent 22 at a volume ratio of 3:10;

[0062] 2) Take 60 μL of the blood sample 23 diluted in step 1) and drop it into the sample hole (corresponding to the sample pad 9 of the test strip) of the 25-hydroxyvitamin D test card 24, and let it stand at room temperature for 15 minutes to perform the immune layer. The analysis reaction is performed, and then the 25-hydroxyvitamin D detection card 24 is inserted into the fluorescent immunoassay analyzer 25 for detection, and the detection result can be obtained immediately.

[0063] 2.2. Establishment of standard curve

[0064] Prepare 25-hydroxyvitamin D standard substances (0, 5, 15, 30, 60, 120ng / mL) of different concentrations, and use the 25-hydroxyvitamin D kit prepared in Example 1 of the present invention to ...

Embodiment 3

[0067] Embodiment 3 comparison experiment

[0068] The 25-hydroxyvitamin D test kit prepared in Examples 1 and 2 was compared with the Siemens 25-hydroxyvitamin D detection kit. Detect by collecting 20 patients' fingertip peripheral blood and venous blood (serum, plasma, whole blood), to determine whether the 25-hydroxyvitamin D kit prepared in the application of the present invention is applicable to 25-hydroxyvitamin D in fingertip peripheral blood For the detection of D, the fingertip blood detection was repeated twice. The test results are shown in Table 2 below.

[0069] Table 2 Detection data

[0070]

[0071]

[0072] In the embodiment of the present invention 1, 25-OH-D kit detects fingertip peripheral blood and Siemens 25-OH-D kit detects the detection result of venous blood (serum) and carries out correlation analysis such as Figure 5 As shown, the regression equation is y=1.0113x+0.3081, R 2 =0.9982, n=20, showing that the test results of the 25-OH-D kit ...

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Abstract

The invention relates to a 25-hydroxyvitamin D kit for detecting peripheral blood, and a preparation method and an application thereof. The 25-hydroxyvitamin D kit comprises a 25-hydroxyvitamin D immunochromatographic detection card and a release agent, wherein the 25-hydroxyvitamin D immunochromatographic detection card comprises a test strip, the test strip comprises a detection line and a quality control line, the detection line is coated with a 25-hydroxyvitamin D complete antigen, and the quality control line is coated with a goat-anti-rabbit polyclonal antibody. The 25-hydroxyvitamin D kit provided by the invention is high in sensitivity, high in stability and wide in linear range, and has excellent accuracy and precision.

Description

technical field [0001] The invention belongs to the field of in vitro diagnosis, and relates to a 25-hydroxyvitamin D kit for detecting peripheral blood, a preparation method and application thereof. Background technique [0002] Vitamin D is a steroid derivative, which is a fat-soluble vitamin. It can regulate calcium and phosphorus metabolism, affect cell proliferation and differentiation, and is an essential steroid hormone for maintaining human health. Vitamin D in the human body is mainly synthesized through sunlight exposure, and only a small part is taken in from food. The main form of vitamin D in the body is 25-hydroxyvitamin D. The main function of vitamin D is to regulate phosphorus and calcium metabolism, promote the absorption of phosphorus and calcium in the intestine, and reabsorb phosphorus and calcium in the renal tubules to increase the level of phosphorus and calcium in the blood. The method of calcification, and calcitonin, Thyroid hormone, along with ot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/533G01N33/543
CPCG01N33/533G01N33/54313G01N33/558
Inventor 林斯
Owner BEIJING ELCOTEQ BIO TECH
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