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Method and detection kit for identifying lymphoma biomarkers

A technology for biomarkers and lymphoma, applied in the field of medicine, can solve the problems of unsuitability for routine purposes, expensive instruments, large volume of serum samples, etc., and achieve the effects of simplified operation, reasonable price of instruments, and short analysis time.

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
There is currently no literature reporting a pre-column 1-phenyl-5-methylpyrazolone (PMP)-derived HPLC method for the simultaneous detection of free mannose and glucose in the serum of lymphoma patients

Method used

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

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

[0081] (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;

[0082] (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;

[0083] (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;

[0084] (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;

[0085] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, keep the supernatant, repeat 3 times;

[0086] (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 hour 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 ...

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Abstract

The invention provides a method and a detection kit for identifying lymphoma biomarkers. The biomarkers are free glucose and free mannose which 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 pre-column PMP derivatization high-performance liquid chromatography. According to the technical scheme, the method and the detection kit for identifying the lymphoma biomarkers have the advantages that the pre-treatment is simple, 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 lymphoma 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 shows that by means of the analysis method, the free mannose andthe free glucose in the serum of the lymphoma patient can be fast quantified, so that the method and the detection kit for identifying the lymphoma biomarkers have significant meaning in studying a relation between the free mannose and the free glucose in the serum and lymphoma and finding a novel lymphoma 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 a lymphoma biomarker and a detection kit thereof. Background technique [0002] Lymphoma is a malignant tumor that occurs in the lymphatic hematopoietic system. Its clinical manifestations are painless lymphadenopathy, hepatosplenomegaly, and systemic symptoms such as weight loss, night sweats, and itchy skin. [0003] The main clinically relevant auxiliary examinations for lymphoma are blood and bone marrow smears and biopsies, blood biochemical examinations, pathological examinations, and imaging examinations (B-ultrasound, CT, MRI, and PET / CT). However, lymph node biopsy is still the preferred examination method for clinical diagnosis of lymphoma. Lymph node biopsy includes lymph node print, lymph node pathological section and lymph node aspiration smear examination. Both lymph node prints and lymph node pathological sections need to sel...

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 张丽娟韩敏敏王喆薛宏伟徐涛
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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