Biomarker for detecting nephritis and appliction thereof

A biomarker, nephritis technology, applied in the field of medicine, to achieve the effect of less dosage, less blood usage, and high sensitivity

Inactive Publication Date: 2019-10-18
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the CEM Discover single-mode microwave protein hydrolysis instrument introduced in this patent combines the advantages of high-efficiency microwave energy and the vacuum / gas exchange port of the Pico-Tag workstation to achieve a more efficient effect in blood sample degradation, and this This method has not been reported for monosaccharide composition analysis of blood samples

Method used

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  • Biomarker for detecting nephritis and appliction thereof
  • Biomarker for detecting nephritis and appliction thereof
  • Biomarker for detecting nephritis and appliction thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1. The effect of different acids on serum microwave acid hydrolysis

[0052] (1) Randomly take 10 patient serums, take 2 of each serum, take 2μL to the microwave hydrolysis tube, add deionized water to 10μL, mix well and then add 10μL 6mol / L HCl or 4mol / L TFA;

[0053] (2) Under the conditions of power 100w and temperature 100℃, use a single-mode microwave proteolysis instrument for microwave acid hydrolysis for 10 minutes;

[0054] (3) Transfer the sample after microwave acid hydrolysis with 20 μL deionized water to a 1.5 mL centrifuge tube, repeat three times, and centrifuge to concentrate and remove the acid;

[0055] (4) Add 100μL methanol to each tube, centrifuge to concentrate to remove residual HCl, repeat three times;

[0056] (5) Each tube of the obtained dry sample is dissolved in 150μL deionized water at a speed of 13000r / min, centrifuged for 5min, and the supernatant is taken and used for anion exchange chromatographic analysis. After the analysis is complete, rinse t...

Embodiment 2

[0076] 1. Drawing the standard curve of monosaccharide

[0077] Accurately weigh the appropriate amount of fucose, galactosamine, glucosamine, galactose, glucose, mannose, add deionized water to prepare the above monosaccharides each containing 0.05mg / mL, 0.01mg / mL, 0.005mg / mL, 0.001 mg / mL, 0.0005mg / mL, 0.0001mg / mL, 0.00005mg / mL solution; take 80μL to the sample bottle for anion exchange chromatography analysis.

[0078] The chromatographic conditions are as follows:

[0079] Analytical column: Thermo Scientific Dionex Carbo PAC TM PA10, 4.0mm×250mm,

[0080] Guard column: Thermo Scientific Dionex Carbo PAC TM PA10, 4.0mm×50mm,

[0081] Eluent: 18mM NaOH for 0-18min; flow rate: 1.0mL / min, injection volume: 10μL, column temperature: 30℃,

[0082] Detector: electrochemical detector, gold electrode (P / N 061875), standard sugar potential, running time: 18min. Obtain the anion exchange chromatograms of 6 monosaccharide standards, see Image 6 A.

[0083] From Image 6 A can see that the 6 ...

Embodiment 3

[0096] Take 2μL of blood from 216 patients with diagnosed nephritis, and the rest of the steps are the same as in Example 2. Statistical test results, see Figure 7 .

[0097] will Figure 7 The results are summarized in Table 1, and the analysis results of the 6 monosaccharide concentrations are shown in Table 2.

[0098] Table 1. The relative trend of the 6 monosaccharides obtained after blood degradation in patients with nephritis and those without nephritis

[0099] Fucose Galactosamine Glucosamine Galactose glucose Mannose Nephritis ↑*** ↑*** ↑*** ↑** ↑** ↑***

[0100] Note: "NS" means no change compared with people without nephritis, "***p <0.001", "**p <0.01" and "*p "<0.05" respectively represent different significant differences compared with those without nephritis, and "↑" and "↓" respectively represent an increase or decrease compared with those without nephritis.

[0101] Table 2. Contrast results of 6 monosaccharide concentrations (μmol / L) in blood of patients with...

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Abstract

The invention provides a blood biomarker for detecting nephritis and appliction thereof. The biomarker includes fucose, galactosamine, glucosamine and mannose that are obtained by processing blood based on microwave acid hydrolysis and anion exchange chromatography with pulsed amperometric detection. The degradation is fast, the batch throughput and compatibility are high, the acid consumption isreduced, and the operation steps are simple and are easy to learn; and the sensitivity and accuracy are high. The obtained results show that the degradation analysis method can rapidly degrade the blood sample, analyze the monosaccharide composition in the blood sample, and calculate the concentration of the monosaccharide; and the person without nephritis and a patient with nephritis can be distinguished to some extents.

Description

Technical field [0001] The invention belongs to the field of medicine, and specifically relates to a biomarker for detecting nephritis and its application. 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 body's metabolic wastes. It plays an important role in maintaining the stability of the body's environment. In addition, the kidney is also an endocrine organ, mainly regulating blood pressure and red blood cells. Generation and bone growth, etc. Therefore, kidney disease has a great impact on human body functions. Nephritis is actually a group of diseases with different causes. Generally speaking, it may be related to factors such as genetics, infection, immunity, metabolism, tumors, etc., but some of the causes are still unclear. Therefore, it is particularly important to find an early diagnosis method. [0003] Diagnosis of kidney disease includes urine tests (proteinur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/66G01N33/531
CPCG01N33/531G01N33/66G01N2800/347
Inventor 张丽娟何燕利
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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