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Method and detection kit for identifying psoriasis biomarker

A biomarker and psoriasis technology, applied in the field of medicine, can solve the problems of being unsuitable for routine purposes, cumbersome processing, and large volume of serum samples, and achieve the effects of simplified operation, short analysis time, and simple pretreatment

Inactive Publication Date: 2019-03-01
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, when using enzymatic method, gas-liquid chromatography and high-resolution liquid chromatography, in order to avoid the influence of high-concentration glucose, it needs to be removed, resulting in cumbersome pretreatment process; gas-liquid chromatography-mass spectrometry instruments are expensive and not Suitable for routine purposes; large serum sample volume required

Method used

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  • Method and detection kit for identifying psoriasis biomarker
  • Method and detection kit for identifying psoriasis biomarker
  • Method and detection kit for identifying psoriasis biomarker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1) Precisely weigh mannose (Man), glucosamine (GlcN), galactosamine (GalN), glucuronic acid (GlcUA), glucose (Glc), galactose (Gal), xylose (Xyl), rock Add appropriate amount of algalose (Fuc), add deionized water to prepare two mixed standard solutions containing the above monosaccharide 0.1mg / mL, and prepare immediately for use;

[0053] (2) Create an alkaline environment: Take 40 μL of the mixed monosaccharide standard in a 1.5mL EP tube, add 40 μL of 0.3mol / L sodium hydroxide, and vortex to mix;

[0054] (3) PMP derivatization: Add 60 μL 0.5mol / L 1-phenyl-5-methylpyrazolone (PMP) to each sample, vortex and mix well, and react in an oven at 70°C for 1 h after centrifugation;

[0055] (4) Neutralization reaction by adding acid: Take out the samples in the oven, let them cool down, add 40 μL of 0.3mol / L hydrochloric acid to each sample, and vortex to mix;

[0056] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloro...

Embodiment 2

[0080] (1) Accurately weigh the appropriate amount of mannose (Man), rhamnose (Rha) and glucose (Glc), add deionized water to prepare 5 parts of the same mixed standard solution containing the above monosaccharide 0.1mg / mL, and use it immediately match;

[0081] (2) Create an alkaline environment: Take 40 μL of the mixed monosaccharide standard in a 1.5mL EP tube, add 40 μL of 0.3mol / L sodium hydroxide, and vortex to mix;

[0082] (3) PMP derivatization: Add 60 μL 0.5mol / L PMP to each sample, vortex and mix well, and react in an oven at 70°C for 1 h after centrifugation;

[0083] (4) Neutralization reaction by adding acid: Take out the samples in the oven, let them cool down, add 40 μL of 0.3mol / L hydrochloric acid to each sample, and vortex to mix;

[0084] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, keep the supernatant, repeat 3 times;

[0085] (6) Centrifuge the sample at 13000r / min for 10min, take 80μL su...

Embodiment 3

[0107] (1) Accurately weigh the appropriate amount of mannose and glucose, add deionized water to prepare the above monosaccharides containing 0.5mg / mL, 0.25mg / mL, 0.1mg / mL, 0.05mg / mL, 0.01mg / mL, 0.005mg / mL, 0.0025mg / mL, 0.001mg / mL, 0.0005mg / mL mixed standard solution, ready-to-use;

[0108] (2) Create an alkaline environment: Take 40 μL of the mixed monosaccharide standard in a 1.5mL EP tube, add 40 μL of 0.3mol / L sodium hydroxide, and vortex to mix;

[0109] (3) PMP derivatization: Add 60 μL 0.5mol / L PMP to each sample, vortex and mix well, and react in an oven at 70°C for 1 h after centrifugation;

[0110] (4) Neutralization reaction by adding acid: Take out the samples in the oven, let them cool down, add 40 μL of 0.3mol / L hydrochloric acid to each sample, and vortex to mix;

[0111] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, keep the supernatant, repeat 3 times;

[0112] (6) Centrifuge the sample at 130...

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Abstract

The invention provides a method and a detection kit for identifying a psoriasis biomarker. The biomarker is a ratio of free glucose to free mannose, wherein the free glucose and the free mannose are obtained by subjecting serum to pre-column 3-methyl-1-phenyl-2-pyrazolin-5-one (PMP) derivatization high-performance liquid chromatography. A detection method is a pre-column PMP derivatization high-performance liquid chromatography. According to the technical scheme, the method and the detection kit for identifying the psoriasis biomarker have the advantages that the pre-treatment is simple, the analysis time is short, the instrument price is reasonable, the conventional use is met, the operation steps are simple and easy to learn, the detection result is high in accuracy, a normal person anda psoriasis patient can be distinguished by just blood sampling, moreover, the required serum amount is very few, the amount of sampled blood is less than 1 mL, and the like. An obtained result showsthat by means of the analysis method, the free mannose and the free glucose in the serum of the psoriasis patient can be fast quantified, so that the method and the detection kit for identifying the psoriasis biomarker have significant meaning in studying a relation between free glucose and the free mannose in the serum and psoriasis and finding a novel psoriasis clinical detection marker.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for identifying psoriasis biomarkers and a detection kit thereof. Background technique [0002] Psoriasis, commonly known as "psoriasis", is a common chronic relapsing inflammatory skin disease, and its prevalence varies significantly among different populations around the world. The course of psoriasis is long, stubborn, and easy to relapse, which seriously affects the physical and mental health and quality of life of patients. The current diagnostic methods for psoriasis mainly include observation of skin lesions, dermoscopy, and imaging examinations. The observation steps of skin lesions are cumbersome, there are many observation items, and misdiagnosis is prone to occur. Imaging examinations include skin CT, X-ray films, etc., but imaging examinations contain radiation, which will have adverse effects on the human body. Therefore, it is very necessary...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/067
Inventor 张丽娟杨丹丹王君
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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