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Method for identifying biomarker of esophageal cancer and detection kit thereof

A biomarker and esophageal cancer technology, applied in the field of medicine, can solve the problems of being unsuitable for routine purposes, large volume of serum samples, and expensive instruments, and achieve the effects of simplified operation, short analysis time, and reasonable instrument prices

Inactive Publication Date: 2019-04-09
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 for identifying biomarker of esophageal cancer and detection kit thereof
  • Method for identifying biomarker of esophageal cancer and detection kit thereof
  • Method for identifying biomarker of esophageal cancer and detection kit thereof

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 Proper amount of fucose (Fuc), add deionized water to prepare two mixed standard solutions containing the above monosaccharides 0.1mg / mL, ready to use;

[0053] (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;

[0054] (3) PMP derivatization: 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;

[0055] (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;

[0056] (5) Extraction: add 500μL of chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, save the supernatant, repeat 3 times;

[0057] (6) Centrifuge the sample at ...

Embodiment 2

[0080] (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

[0081] (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;

[0082] (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;

[0083] (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;

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

[0085] (6) Centrifuge the sample at 13000r / min for 10min, take 80μL of supernatant into a brown sample bottle equipped with a 150μL inner ...

Embodiment 3

[0106] (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;

[0107] (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;

[0108] (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;

[0109] (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;

[0110] (5) Extraction: add 500μL of chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, save the supernatant, repeat 3 times;

[0111] (6) Centrifuge the sample at 13000 r / min for 10 min, take 80 μL of supernatant into a brown sample bott...

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Abstract

The invention provides a method for identifying a biomarker of esophageal cancer and a detection kit thereof. The biomarker is the ratio of free glucose and mannose in serum obtained through pre-column 1-methyl-5-methyl pyrazolone (PMP)-derived high-performance liquid chromatography. The detection method in the invention is pre-column PMP-derived high-performance liquid chromatography. The technical scheme provided by the invention has the advantages of being simple in preprocessing, short in analysis time, and reasonable in instrument price, the conventional use is conformed to, the operationsteps are simple and easy to learn, the accuracy of the detection result is high, an average person and an esophagus cancer patient can be distinguished only through blood sampling, the number of needed serum is extremely small, the blood collection amount is smaller than 1 mL, and the like. The result shows that the analysis method can quickly quantify free mannose and glucose in serum of esophageal cancer patients and has very important significance in studying the relation between the free glucose and mannose in the serum and the esophagus cancer and finding a novel esophagus cancer clinical detection marker.

Description

Technical field [0001] The invention belongs to the technical field of medicine, and specifically relates to a method for identifying biomarkers of esophageal cancer and a detection kit thereof. Background technique [0002] The esophagus is an organ of the digestive system. It connects the throat to the stomach, closes to the ventral side of the spine, and helps transport food into the stomach. It is the key to food entering the digestive system. Therefore, diseases in the esophagus will have a serious impact on the body. The diagnosis of esophageal cancer generally requires imaging examinations, esophagoscopy and blood tests, combined with clinical manifestations for diagnosis. These diagnostic methods will bring certain drawbacks. Therefore, it is necessary to find a convenient and non-invasive detection method. Blood test has become an ideal test method because of its simple operation and non-invasive characteristics. The most commonly used clinical test markers in blood te...

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 张丽娟杨丹丹尚伟
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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