CN106769349B - Method for detecting abnormal glycosylated protein cells in blood - Google Patents
Method for detecting abnormal glycosylated protein cells in blood Download PDFInfo
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- CN106769349B CN106769349B CN201710009376.2A CN201710009376A CN106769349B CN 106769349 B CN106769349 B CN 106769349B CN 201710009376 A CN201710009376 A CN 201710009376A CN 106769349 B CN106769349 B CN 106769349B
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- blood
- smear
- glycosylated protein
- automatic
- protein cells
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- 210000004369 blood Anatomy 0.000 title claims abstract description 39
- 239000008280 blood Substances 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 33
- 102000035122 glycosylated proteins Human genes 0.000 title claims abstract description 18
- 108091005608 glycosylated proteins Proteins 0.000 title claims abstract description 18
- 230000002159 abnormal effect Effects 0.000 title abstract description 15
- 238000004043 dyeing Methods 0.000 claims abstract description 13
- 210000005259 peripheral blood Anatomy 0.000 claims abstract description 7
- 239000011886 peripheral blood Substances 0.000 claims abstract description 7
- 238000006243 chemical reaction Methods 0.000 claims abstract description 4
- 238000012937 correction Methods 0.000 claims abstract description 4
- 238000001035 drying Methods 0.000 claims abstract description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 239000001045 blue dye Substances 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000012128 staining reagent Substances 0.000 claims description 5
- 239000011521 glass Substances 0.000 claims description 3
- 238000012790 confirmation Methods 0.000 claims description 2
- 206010028980 Neoplasm Diseases 0.000 abstract description 16
- 238000003745 diagnosis Methods 0.000 abstract description 9
- 201000011510 cancer Diseases 0.000 abstract description 7
- 238000012216 screening Methods 0.000 abstract description 6
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 5
- 238000002360 preparation method Methods 0.000 abstract description 3
- 239000003086 colorant Substances 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 20
- 238000001514 detection method Methods 0.000 description 8
- 239000000975 dye Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000012670 alkaline solution Substances 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000013595 glycosylation Effects 0.000 description 2
- 238000006206 glycosylation reaction Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000013529 alpha-Fetoproteins Human genes 0.000 description 1
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a method for detecting abnormal glycosylated protein cells in blood, which comprises the following steps: (1) making a blood smear by using fasting peripheral blood or fasting venous whole blood of a patient; (2) adding related coloring agents, dripping a plurality of parts of the same smear, and placing the smear in related equipment for drying reaction; (3) the dried smear is processed by automatic film shooting, automatic calculation and analysis, manual correction and report printing. The method for detecting the abnormal glycosylated protein cells in the blood provided by the invention determines the content of the abnormal glycosylated protein cells in the blood by a dyeing method, and has the advantages of unique method, easy preparation of reagents and higher accuracy. The method is suitable for early pre-diagnosis, screening and cancer auxiliary diagnosis of various tumors, and is simple, effective and convenient.
Description
Technical Field
The invention belongs to the technical field of blood detection, relates to a blood detection method, and particularly relates to a detection method of abnormal glycosylated protein cells in blood.
Background
Many researches have found that the glycosylation of protein cells in cancer cells and tumor tissues of individual cancer patients is abnormal, the principle is that after the protein cells are invaded by cancer cells, an enzyme is generated to modify the cell surface, so that the glycosylation of the protein cells and the variation of surface sugar chains are abundant, such variant glycosylated cells also exist in human blood, and by detecting the variation of the content of the variant sugar chain glycosylated protein cells, the company finds a novel marker method for detecting the variant sugar chain glycosylated protein cells, and the detection can be used for various tumor prediction and cancer auxiliary diagnosis.
At present, the detection methods for tumors are more and more single, such as various sugar chain antigens, alpha fetoprotein, polypeptide antigens and the like, and the existing methods are all used for screening and diagnosing single tumors.
In view of the above, there is an urgent need to design a new detection method to overcome the above-mentioned defects of the existing detection methods.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: the method for detecting the abnormal glycosylated protein cells in the blood is provided, the content of the abnormal glycosylated protein cells in the blood is determined by a dyeing method, and the method is unique, the reagent is easy to prepare, and the accuracy rate is higher; the method is suitable for early pre-diagnosis and screening of various tumors, and is simple, effective and convenient.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for detecting abnormally glycosylated protein cells in blood, the method comprising:
(1) making a blood smear by using fasting peripheral blood or fasting venous whole blood of a patient;
(2) adding related dyeing materials, namely water, sodium hydroxide, dye and disperse blue dye, wherein the dye is alkaline solution with the Ph value of 8-11 and the disperse blue dye with the mass percentage of 0.01-20; dripping a plurality of parts of the same smear, and placing the smear in related equipment for drying reaction; the dyeing material formula comprises: 0.5-2% of disperse blue dye and 98-99.5% of water, and adjusting the pH value to 8-11 by using sodium hydroxide;
(3) the dried smear is subjected to automatic film shooting, automatic calculation and analysis, manual correction and report printing; three drops of peripheral blood or whole blood of a person to be detected are taken and put on a glass slide, a staining reagent is added into each drop of blood, the blood is naturally dried at constant temperature and humidity, and automatic shooting, automatic analysis, manual calibration confirmation and report printing are carried out on related automatic equipment.
As a preferred scheme of the invention, the content of abnormal glycosylated protein cells in blood is determined by a dyeing method, and the method is unique, the reagent is easy to prepare, and the accuracy rate is higher.
As a preferred scheme of the invention, the method is suitable for early pre-diagnosis, screening and cancer auxiliary diagnosis of various tumors, and is simple, effective and convenient.
As a preferred embodiment of the present invention, 50. mu.l of the staining reagent is added to each drop of blood.
The invention has the beneficial effects that: the method for detecting the abnormal glycosylated protein cells in the blood provided by the invention determines the content of the abnormal glycosylated protein cells in the blood by a dyeing method, and has the advantages of unique method, easy preparation of reagents and higher accuracy. The method is suitable for early pre-diagnosis, screening and cancer auxiliary diagnosis of various tumors, and is simple, effective and convenient.
Drawings
FIG. 1 is a flow chart of the detection process of the present invention.
Detailed Description
Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
Example one
Referring to FIG. 1, the present invention discloses a method for detecting abnormal glycosylated protein cells in blood, the method comprising:
(1) making a blood smear by using fasting peripheral blood or fasting venous whole blood of a patient;
(2) adding related dyeing materials, namely water, sodium hydroxide, dye and disperse blue dye, wherein the dye is alkaline solution with the Ph value of 8-11 and the disperse blue dye with the mass percentage of 0.01-20; dripping a plurality of parts of the same smear, and placing the smear in related equipment for drying reaction; the dyeing material formula comprises: 0.5-2% of disperse blue dye and 98-99.5% of water, and adjusting the pH value to 8-11 by using sodium hydroxide;
(3) the dried smear is subjected to automatic film shooting, automatic calculation and analysis, manual correction and report printing; three drops of peripheral blood or whole blood of a person to be detected are taken and put on a glass slide, 50 microliter of staining reagent is added into each drop of blood, the blood is naturally dried at constant temperature and humidity, and the blood is automatically photographed, automatically analyzed, manually calibrated and confirmed and printed on related automatic equipment.
In conclusion, the method for detecting the abnormal glycosylated protein cells in the blood provided by the invention determines the content of the abnormal glycosylated protein cells in the blood by a dyeing method, and has the advantages of unique method, easy preparation of reagents and higher accuracy. The method is suitable for early pre-diagnosis and screening of various tumors.
The description and applications of the invention herein are illustrative and are not intended to limit the scope of the invention to the embodiments described above. Variations and modifications of the embodiments disclosed herein are possible, and alternative and equivalent various components of the embodiments will be apparent to those skilled in the art. It will be clear to those skilled in the art that the present invention may be embodied in other forms, structures, arrangements, proportions, and with other components, materials, and parts, without departing from the spirit or essential characteristics thereof. Other variations and modifications of the embodiments disclosed herein may be made without departing from the scope and spirit of the invention.
Claims (2)
1. A method for detecting abnormally glycosylated protein cells in blood, the method comprising:
(1) making a blood smear by using fasting peripheral blood or fasting venous whole blood of a patient;
(2) adding related dyeing materials, namely water, sodium hydroxide and disperse blue dye, wherein the mass percentage of the disperse blue dye in the dyeing materials is 0.01-20%, and the Ph value of the dyeing materials is 8-11; dripping a plurality of parts of the same smear, and placing the smear in related equipment for drying reaction; the dyeing material formula comprises: the mass percent of the disperse blue dye is 0.5-2%, the mass percent of the water is 98-99.5%, and the Ph value is adjusted to be 8-11 by sodium hydroxide;
(3) the dried smear is subjected to automatic film shooting, automatic calculation and analysis, manual correction and report printing; three drops of peripheral blood or whole blood of a person to be detected are taken and put on a glass slide, a staining reagent is added into each drop of blood, the blood is naturally dried at constant temperature and humidity, and automatic shooting, automatic analysis, manual calibration confirmation and report printing are carried out on related automatic equipment.
2. The method according to claim 1, wherein the method comprises the steps of:
add 50. mu.l of staining reagent to each drop of blood.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710009376.2A CN106769349B (en) | 2017-01-06 | 2017-01-06 | Method for detecting abnormal glycosylated protein cells in blood |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710009376.2A CN106769349B (en) | 2017-01-06 | 2017-01-06 | Method for detecting abnormal glycosylated protein cells in blood |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106769349A CN106769349A (en) | 2017-05-31 |
| CN106769349B true CN106769349B (en) | 2020-01-21 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710009376.2A Ceased CN106769349B (en) | 2017-01-06 | 2017-01-06 | Method for detecting abnormal glycosylated protein cells in blood |
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| Country | Link |
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| CN (1) | CN106769349B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111751557A (en) * | 2020-07-16 | 2020-10-09 | 上海君联医疗设备有限公司 | Sugar chain protein and calcium-histone detection reagent composition and use thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1079825A (en) * | 1991-12-20 | 1993-12-22 | 洛克菲勒大学 | The immunochemistry of advanced glycosylation end product detects in the body |
| CN1211321A (en) * | 1995-12-26 | 1999-03-17 | 皮考瓦医疗研究院 | Methods for measurment and treatment predicated on presence of advanced glycosylation endproducts in tobacco and its combustion byproducts |
| CN101351706A (en) * | 2005-11-04 | 2009-01-21 | 福罗若技术有限公司 | Method for monitoring hydrolytic activity |
| CN104854458A (en) * | 2012-10-25 | 2015-08-19 | 抗癌治疗研究协会 | Methylglyoxal as a marker of cancer |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2711016A1 (en) * | 2012-09-21 | 2014-03-26 | Covagen AG | Novel IL-17A binding molecules and medical uses thereof |
-
2017
- 2017-01-06 CN CN201710009376.2A patent/CN106769349B/en not_active Ceased
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1079825A (en) * | 1991-12-20 | 1993-12-22 | 洛克菲勒大学 | The immunochemistry of advanced glycosylation end product detects in the body |
| CN1211321A (en) * | 1995-12-26 | 1999-03-17 | 皮考瓦医疗研究院 | Methods for measurment and treatment predicated on presence of advanced glycosylation endproducts in tobacco and its combustion byproducts |
| CN101351706A (en) * | 2005-11-04 | 2009-01-21 | 福罗若技术有限公司 | Method for monitoring hydrolytic activity |
| CN104854458A (en) * | 2012-10-25 | 2015-08-19 | 抗癌治疗研究协会 | Methylglyoxal as a marker of cancer |
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| CN106769349A (en) | 2017-05-31 |
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