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Method of biomarker for identifying cervical cancer and detection kit for identifying cervical cancer

A biomarker and cervical cancer technology, applied in the field of medicine, can solve the problems of being unsuitable for routine purposes, cumbersome processing, large volume of serum samples, etc., and achieve the effects of simplified operation, short analysis time and simple operation steps

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 the simultaneous detection of free mannose and glucose in the serum of patients with cervical cancer by a pre-column 1-phenyl-5-methylpyrazolone (PMP)-derived HPLC method

Method used

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  • Method of biomarker for identifying cervical cancer and detection kit for identifying cervical cancer
  • Method of biomarker for identifying cervical cancer and detection kit for identifying cervical cancer
  • Method of biomarker for identifying cervical cancer and detection kit for identifying cervical cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] (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;

[0052] (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;

[0053] (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;

[0054] (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;

[0055] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloro...

Embodiment 2

[0079] (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;

[0080] (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;

[0081] (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;

[0082] (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;

[0083] (5) Extraction: add 500 μL chloroform to each tube, vortex, centrifuge, remove the lower layer of chloroform, keep the supernatant, repeat 3 times;

[0084] (6) Centrifuge the sample at 13000r / min for 10min, take 80μL su...

Embodiment 3

[0106] (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;

[0107] (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;

[0108] (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;

[0109] (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;

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

[0111] (6) Centrifuge the sample at 130...

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Abstract

The invention provides a method of a biomarker for identifying cervical cancer and a detection kit for identifying cervical cancer. The biomarker comprises free mannose and glucose obtained by high performance liquid chromatography derived from pre-column 1-phenyl-5-methyl pyrazolone(PMP) in serum. The detection method is a pre-column PMP derived high performance liquid chromatography method. According to the technical scheme, the method and the detection kit have the advantages that pretreatment is easy, the analysis time is short, the instrument price is reasonable, the method is compliancewith conventional use, operation steps are simple and easy to learn, the accuracy of detection results is high, normal people can be distinguished from patients with cervical cancer only by collectingblood, the amount of required serum is small, and the amount of the collected blood is less than 1mL . The results show that the analytical method can rapidly quantify the free mannose and glucosein the serum of patients with cervical cancer, the method and the detection kit are of great significance for studying the relationship between serum free mannose and glucose and cervical cancer and finding new clinical detection markers for cervical cancer.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for identifying a biomarker of cervical cancer and a detection kit thereof. Background technique [0002] Cervical cancer is the third most common malignancy in women worldwide after breast cancer and colorectal cancer. In 2008, it was estimated that there were 529,800 new cervical cancer cases and 255,100 deaths in the world, and 85% of the new cases were in developing countries. The preoperative diagnostic examination methods for cervical cancer generally include cervical cytology, colposcopy, and endocervical biopsy. Pathological diagnosis is the gold standard for the diagnosis of cervical cancer. The positive diagnostic rate of squamous cell carcinoma antigen (SCC), cytokeratin 21-1 (CYFRA21-1), CA125, CA19-9 and CEA, the most commonly used clinical detection markers in the determination of tumor markers, for cervical adenocarcinoma They were 27-75%, 3...

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 张丽娟曾鹏娇王言奎
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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