Method for identifying biomarkers of gouty arthritis and detection kit thereof

A technology for gouty arthritis and biomarkers, applied in the field of medicine, can solve the problems of unsuitability for routine purposes, cumbersome processing process, large volume of serum samples, etc., and achieve the effects of simplified operation, short analysis time, and simple pretreatment.

Inactive Publication Date: 2019-04-12
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
View PDF0 Cites 1 Cited by
  • 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 expen

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for identifying biomarkers of gouty arthritis and detection kit thereof
  • Method for identifying biomarkers of gouty arthritis and detection kit thereof
  • Method for identifying biomarkers of gouty arthritis and detection kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0052] Example 1

[0053] (1) Precisely weigh mannose (Man), glucosamine (GlcN), galactosamine (GalN), glucuronic acid (GlcUA), glucose (Glc), galactose (Gal), xylose (Xyl), rock Proper amount of fucose (Fuc), add deionized water to prepare two mixed standard solutions containing the above monosaccharides 0.1mg / mL, ready to use;

[0054] (2) Create an alkaline environment: Take 40μL of mixed monosaccharide standard in a 1.5mL EP tube, add 40μL of 0.3mol / L sodium hydroxide, and vortex to mix;

[0055] (3) PMP derivation: add 60μL 0.5mol / L 1-phenyl-5-methylpyrazolone (PMP) to each sample, vortex to mix, centrifuge and react in an oven at 70°C for 1h;

[0056] (4) Acid neutralization reaction: Take out the samples in the oven, leave them to cool, add 40μL 0.3mol / L hydrochloric acid to each sample, vortex and mix;

[0057] (5) Extraction: add 500μL of chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, save the supernatant, repeat 3 times;

[0058] (6) Centrif...

Example Embodiment

[0081] Example 2

[0082] (1) Precisely weigh appropriate amounts of mannose (Man), rhamnose (Rha) and glucose (Glc), and add deionized water to prepare the same 5 portions of the same 0.1 mg / mL mixed standard solution containing the above monosaccharides. Match

[0083] (2) Create an alkaline environment: Take 40μL of mixed monosaccharide standard in a 1.5mL EP tube, add 40μL of 0.3mol / L sodium hydroxide, vortex to mix;

[0084] (3) PMP derivation: add 60μL 0.5mol / L PMP to each sample, vortex to mix, centrifuge and react in an oven at 70°C for 1h;

[0085] (4) Acid neutralization reaction: take out the samples in the oven, leave them to cool, add 40μL 0.3mol / L hydrochloric acid to each sample, vortex and mix;

[0086] (5) Extraction: add 500μL of chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, save the supernatant, repeat 3 times;

[0087] (6) Centrifuge the sample at 13000r / min for 10min, take 80μL of supernatant into a brown sample bottle equipped w...

Example Embodiment

[0108] Example 3

[0109] (1) Precisely weigh an appropriate amount of mannose and glucose, add deionized water to prepare the above monosaccharides 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;

[0110] (2) Create an alkaline environment: Take 40μL of mixed monosaccharide standard in a 1.5mL EP tube, add 40μL of 0.3mol / L sodium hydroxide, vortex to mix;

[0111] (3) PMP derivation: add 60μL 0.5mol / L PMP to each sample, vortex to mix, centrifuge and react in an oven at 70°C for 1h;

[0112] (4) Acid neutralization reaction: take out the samples in the oven, leave them to cool, add 40μL 0.3mol / L hydrochloric acid to each sample, vortex and mix;

[0113] (5) Extraction: add 500μL of chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, save the supernatant, repeat 3 times;

[0114] (6) Centrifuge the sample at 13000 r / min for 10 min, take 80 μL of supernatant into a...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Wavelengthaaaaaaaaaa
Bandwidthaaaaaaaaaa
Login to view more

Abstract

The invention provides a method for identifying biomarkers of gouty arthritis and a detection kit thereof. The biomarkers are free mannose and glucose obtained by high performance liquid chromatography derived from pre-column PMP in serum, and the ratio of glucose to mannose. The detection method is the pre-column 1-phenyl-5-methyl-pyrazolone (PMP) derived high performance liquid chromatography. The method has the advantages that pretreatment is simple, analysis time is short, the instrument price is reasonable, conventional use is conformed to, operation steps are simple and easy to learn, the accuracy of detection results is high, only blood collection is needed, the amount of serum required is extremely small, the blood collection is less than 1 mL, and the like. The results show that the analysis method can quickly quantify the free mannose and glucose in the serum of patients with gouty arthritis, and has great significance to study the relationship between free mannose and glucose in serum and gouty arthritis and to find new clinical detection markers of gouty arthritis.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for identifying biomarkers of gouty arthritis and a detection kit thereof. Background technique [0002] Gouty arthritis is caused by gout. When uric acid is in a supersaturated solution, monosodium urate crystals are formed, and most of them are located in the joint fluid and around the joints. Patients with gouty arthritis will deposit monosodium urate (MUS) in the joints, which is caused by long-term hyperuricemia, leading to acute gouty arthritis, joint deformities, and repeated attacks. The disease often has associated complications (diabetes, cardiovascular disease, chronic kidney disease or obesity), so finding a way to diagnose it early is especially important. [0003] The diagnostic criteria for gout are needle-like monosodium urate (MSU) crystals in tissue or synovial fluid that can exhibit polar birefringence under the microscope, but other join...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/067
Inventor 张丽娟李秀莲李长贵张朦
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap