Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method and detection kit for identifying nephritis biomarkers

A technology of biomarkers and kits, 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
View PDF0 Cites 0 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 expensive and not Suitable for routine purposes; large serum sample volume required
Currently, there is no literature reporting the simultaneous detection of free mannose and glucose in the serum of patients with nephritis by pre-column 1-phenyl-5-methylpyrazolone (PMP)-derived HPLC

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 and detection kit for identifying nephritis biomarkers
  • Method and detection kit for identifying nephritis biomarkers
  • Method and detection kit for identifying nephritis biomarkers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] (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;

[0051] (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;

[0052] (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;

[0053] (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;

[0054] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloro...

Embodiment 2

[0078] (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;

[0079] (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;

[0080] (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;

[0081] (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;

[0082] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, keep the supernatant, repeat 3 times;

[0083] (6) Centrifuge the sample at 13000r / min for 10min, take 80μL su...

Embodiment 3

[0105] (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;

[0106] (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;

[0107] (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;

[0108] (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;

[0109] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, keep the supernatant, repeat 3 times;

[0110] (6) Centrifuge the sample at 130...

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
phaseaaaaaaaaaa
Login to View More

Abstract

The invention provides a method and a detection kit for identifying nephritis biomarkers. The biomarkers is free mannose, which is obtained by subjecting serum to pre-column 3-methyl-1-phenyl-2-pyrazolin-5-one (PMP) derivatization high-performance liquid chromatography, and a concentration ratio of glucose to mannose. A detection method is pre-column PMP derivatization high-performance liquid chromatography. According to the technical scheme, the method and the detection kit for identifying the nephritis biomarkers 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 and a nephritis patientcan be distinguished by just blood sampling, moreover, the required serum amount is very less, the amount of sampled blood is less than 1 mL, and the like. An obtained result shows that by means of the analysis method, the free mannose and the free glucose in the serum of the nephritis patient can be fast quantified, so that the method and the detection kit for identifying the nephritis biomarkershave significant meaning in studying a relation between the free mannose and the free glucose in the serum and nephritis and finding a novel nephritis 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 biomarkers of nephritis and a detection kit thereof. Background technique [0002] The kidney is an important organ of the urinary system. Its main function is to produce and excrete urine, and to excrete the metabolic waste of the human body. generation and bone growth etc. Therefore, kidney disease has a great impact on the function of the human body. Nephritis is actually a group of diseases, and its diagnosis generally requires urine tests, blood tests, imaging tests and pathological tests, combined with clinical manifestations for diagnosis. These diagnostic methods will bring certain disadvantages. For example, renal biopsy plays a very important role in the diagnosis and prognosis of glomerulonephritis, but renal biopsy is an invasive diagnostic method that requires postoperative care. Therefore, it is very necessary to find a conv...

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
Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/067
Inventor 张丽娟何燕利邢广群
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com