CN110488019A - REPS1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit - Google Patents
REPS1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit Download PDFInfo
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- CN110488019A CN110488019A CN201910706018.6A CN201910706018A CN110488019A CN 110488019 A CN110488019 A CN 110488019A CN 201910706018 A CN201910706018 A CN 201910706018A CN 110488019 A CN110488019 A CN 110488019A
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- 206010058467 Lung neoplasm malignant Diseases 0.000 title claims abstract description 49
- 201000005202 lung cancer Diseases 0.000 title claims abstract description 49
- 208000020816 lung neoplasm Diseases 0.000 title claims abstract description 49
- 101001095989 Homo sapiens RalBP1-associated Eps domain-containing protein 1 Proteins 0.000 title claims abstract description 47
- 102100037881 RalBP1-associated Eps domain-containing protein 1 Human genes 0.000 title claims abstract description 47
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 47
- 238000001514 detection method Methods 0.000 title claims abstract description 33
- 238000012216 screening Methods 0.000 title claims abstract description 17
- 210000002966 serum Anatomy 0.000 claims abstract description 21
- 238000012360 testing method Methods 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 6
- 238000002965 ELISA Methods 0.000 claims description 4
- 238000001262 western blot Methods 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims 2
- 108090000790 Enzymes Proteins 0.000 claims 2
- 239000003547 immunosorbent Substances 0.000 claims 2
- 238000001179 sorption measurement Methods 0.000 claims 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical group CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims 1
- 102000002322 Egg Proteins Human genes 0.000 claims 1
- 108010000912 Egg Proteins Proteins 0.000 claims 1
- 235000014103 egg white Nutrition 0.000 claims 1
- 210000000969 egg white Anatomy 0.000 claims 1
- 238000003745 diagnosis Methods 0.000 abstract description 3
- 201000010099 disease Diseases 0.000 description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 12
- 210000004072 lung Anatomy 0.000 description 9
- 239000000243 solution Substances 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 5
- 238000004140 cleaning Methods 0.000 description 5
- 210000002381 plasma Anatomy 0.000 description 5
- 206010028980 Neoplasm Diseases 0.000 description 4
- 239000007788 liquid Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 101100301555 Homo sapiens REPS1 gene Proteins 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 102100025570 Cancer/testis antigen 1 Human genes 0.000 description 1
- 208000002927 Hamartoma Diseases 0.000 description 1
- 101000856237 Homo sapiens Cancer/testis antigen 1 Proteins 0.000 description 1
- 108090000144 Human Proteins Proteins 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 238000012864 cross contamination Methods 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 239000000104 diagnostic biomarker Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000010197 meta-analysis Methods 0.000 description 1
- 238000002493 microarray Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003147 molecular marker Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000032696 parturition Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000006748 scratching Methods 0.000 description 1
- 230000002393 scratching effect Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 239000000439 tumor marker Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57423—Specifically defined cancers of lung
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6884—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from lung
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- Urology & Nephrology (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
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Abstract
The present invention relates to external diagnosis reagent fields, and in particular to REPS1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit.Present invention firstly discovers that the autoantibody of REPS1 albumen is significantly higher than healthy patients in Serum of Patients with Lung Cancer.The reagent that the present invention will test REPS1 albumen autoantibody is used to prepare screening lung cancer kit, can be realized effective screening of lung cancer.
Description
Technical field
The present invention relates to external diagnosis reagent fields, and in particular to REPS1 autoantibody detection reagent is in preparation lung cancer sieve
Look into the purposes in kit.
Background technique
Lung cancer is that one of most common malignant tumour, morbidity and mortality are in rise year by year trend in the world, at present
Disease incidence occupies first place in the world, and seriously threatens human health and life.
Lung cancer is a kind of disease for being good at concealment, often develops to advanced stage in disease and just shows clinical symptoms, 70~
80% patients with lung cancer has been middle and advanced stage when being diagnosed to be with Lung Cancer Symptoms, and cancer cell has been spread, and misses best healing
Opportunity, five year survival rate are low.For the patients with lung cancer of early stage, giving birth to for 5 years or more for patient is greatly improved by treating in time
Deposit rate and life quality.Therefore the early diagnosis and the effective screening of progress of lung cancer are most important.
The screening of lung cancer, referring to does not have the related indication crowd of lung cancer to carry out routine physical examination those, before there is symptom
Discovery lung cancer in time.If the lung cancer molecular marker inside blood plasma can be found, for prompting clinician's early stage to patient
Relevant remedy measures or decision are taken to have great importance.
Autoantibody refers to the antibody that body generates itself organ, cell or cell component.Currently, certain albumen from
Body antibody has become the marker of lung cancer, such as: p53, NY-ESO-1, CYFRA (Tang Z-M, Ling Z-G, Wang
C-M, Wu Y-B, Kong J-L (2017) Serum tumor-associated autoantibodies as diagnostic
Biomarkers for lung cancer:A systematic review and meta-analysis.PLoS ONE 12
(7): e0182117).
REPS1 gene (Ensembl:ENSG00000135597) coding has the signal transducer there are two EH structural domain,
It interacts with signal transduction, the protein that endocytosis and cytoskeleton change is participated in.
Have not yet to see REPS1 gene, albumen or REPS1 autoantibody report relevant to cancer.
Summary of the invention
The purpose of the present invention is to provide the detections of the lung cancer marker and the marker of a kind of new autoantibody class
Reagent is preparing the purposes in screening lung cancer kit.
Technical solution of the present invention includes:
The reagent of detection REPS1 albumen autoantibody is preparing the purposes in screening lung cancer kit.
The reagent of purposes as the aforementioned, the detection REPS1 albumen autoantibody is enzyme-linked immunosorbent assay reagent
Or linked immune analysis reagent.
The reagent of purposes as the aforementioned, the detection REPS1 albumen autoantibody is western blot reagent.
The reagent of purposes as the aforementioned, the detection REPS1 albumen autoantibody is protein chip detection method reagent.
Purposes as the aforementioned, it is described detection REPS1 albumen autoantibody reagent be detection human serum in REPS1 albumen from
The reagent of body antibody.
A kind of screening lung cancer kit, it includes the reagent for detecting REPS1 albumen autoantibody.
Kit as the aforementioned, the reagent of the detection REPS1 albumen autoantibody are enzyme-linked immunosorbent assay examination
Agent or ELISA reagent.
The reagent of kit as the aforementioned, the detection REPS1 albumen autoantibody is western blot reagent.
Kit as the aforementioned, the reagent of the detection REPS1 albumen autoantibody are the examination of protein chip detection method
Agent.
Kit as the aforementioned, the reagent of the detection REPS1 albumen autoantibody are REPS1 albumen in detection human serum
The reagent of autoantibody.
The present invention provides a kind of new screening lung cancer markers and a kind of new screening lung cancer kit, can be realized lung
Effective screening of cancer；And patient can be injured very low using serum as test sample.
Kit of the invention can also distinguish lung cancer and lung's benign disease.
The present invention has good application prospect.
Obviously, above content according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field
Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.
Above content of the invention is described in further detail again below by way of specific embodiment.But it should not be by this
The range for being interpreted as the above-mentioned theme of the present invention is only limitted to example below.All technologies realized based on above content of the present invention are equal
Belong to the scope of the present invention.
Hereinafter " REPS1 autoantibody " refers to " REPS1 albumen autoantibody ".
Detailed description of the invention
Fig. 1: patients with lung cancer (LC), lung's benign disease (DC), normal healthy controls (NC) serum slurry in REPS1 autoantibody water
Flat comparison.
Fig. 2: patients with lung cancer (LC) and lung's benign disease (DC) ROC are analyzed.
Fig. 3: patients with lung cancer (LC) and normal healthy controls (NC) ROC are analyzed.
Specific embodiment
The relationship of REPS1 autoantibody and lung cancer in 1 blood plasma of embodiment
One, clinical data
Selection patients with lung cancer 59, lung's benign disease (non-malignant tumors such as tuberculosis, hamartoma) 29, normal healthy controls 30
Example, essential information are as follows:
| Essential information | Patients with lung cancer | Lung's benign disease | Normal healthy controls |
| Number | 59 | 29 | 30 |
| Age | 54.5±7.6 | 46.5±10.0 | 42±8.9 |
| Masculinity proportion | 37 (62.7%) | 12 (41.4%) | 13 (46.7%) |
Two, testing principle
HuProtTMBe fixed with REPS1 albumen on human protein's custom chip, after increase serum is incubated for, in serum REPS1 from
Body antibody (including mainly IgG, IgM type antibody, there are also other types of antibody) can be in conjunction with up, and cleaning removal is unbonded
Antibody and other oroteins, then with anti-human IgM fluorescent marker secondary antibody (cy5 mark, present red) and anti-human igg fluorescence secondary antibody
(cy3 label, green is presented) detection reads signal, the power and the affinity and quantity of antibody of signal by Fluorescence Scanner
It is positively correlated.
Three, method
Reagent used in this part is as follows:
Specific step is as follows:
1) rewarming: chip is taken out from -80 DEG C of refrigerators, 4 DEG C of refrigerator rewarming half an hour is placed in, is subsequently placed in room temperature rewarming
15min；
2) close: the chip after rewarming fixes 14 blocks fences, and after fixing, envelope is added into each block
Liquid is closed, and is placed on side-sway shaking table, room temperature closes 3hr；
3) serum sample is incubated for: after the completion of closing, then confining liquid to the greatest extent is rapidly added preprepared serum and is incubated for
Liquid, every chip can be incubated for 14 serum samples, and the loading volume of each serum sample is 200 μ L, side-sway shaking table 20rpm, and 4 DEG C
It is incubated overnight (serum sample is firstly placed on 4 DEG C of chromatography cabinet freeze thawing, adds Incubating Solution with 1: 50 dilution proportion, obtains serum Incubating Solution)；
4) it cleans: chip and chip fence being taken out together, suck sample, is then rapidly added isometric PBST, such as
This circulation for several times, guarantees when removing chip fence no cross contamination between serum sample.After removing chip fence, chip is placed in
Added with the chip cleaning box of cleaning solution, horizontal shaker, room temperature 80rpm is cleaned 3 times, each 10min；
5) secondary antibody is incubated for: chip being transferred in the incubation box that joined 3mL secondary antibody Incubating Solution, side-sway shaking table 40rpm is kept away
Light, room temperature 1hr；
6) it cleans: chip being taken out into (upper surface for paying attention to touch or scratching chip), is placed in the core added with cleaning solution
Piece cleaning box, horizontal shaker, room temperature 80rpm are cleaned 3 times, each 10min.It is cleaned 2 times with ddH2O after the completion, each 10min；
7) dry；
8) it scans: being scanned using brilliant core LuxScan 10K micro-array chip scanner；
9) data are extracted: corresponding GAL file (having recorded albumen position in chip) is opened, by chip image and
The each array of GAL file is integrally aligned, and presses automatic aligning button, is extracted data and is saved.
Four, result
The Average expression level of REPS1 autoantibody in Plasma of The Patients With Lung Cancer is 41.6SNR (fluorescence signal relative quantification
Ratio), the Average expression level of REPS1 autoantibody is 61.6SNR in lung's benign disease blood plasma, in normal healthy controls blood plasma
The Average expression level of REPS1 autoantibody is 56.9SNR.Lung cancer group and lung's benign disease group (p < 0.001) and health are right
Statistically significant (Fig. 1) is compared according to group (p < 0.01).The ROC of lung cancer group and benign disease analyzes result specificity
92.9%, sensibility is 17.0% (Fig. 2)；It is 93.1% that the ROC of lung cancer group and normal healthy controls, which analyzes result specificity, sensibility
For 13.6% (Fig. 3), show that REPS1 autoantibody can specifically distinguish lung cancer and lung's benign disease and normal healthy controls.
It can be seen from the above result that the level difference of REPS1 autoantibody is aobvious in patients with lung cancer and the serum of non-lung cancer patient
It writes, by the level of REPS1 autoantibody in detection serum, can achieve the purpose that screening lung cancer.
The composition and its application method of the detection kit of the invention of embodiment 2
One, kit forms
Detection kit (14 person-portion):
Two, kit application method
With 1 Part III of embodiment --- " detection of REPS1 autoantibody in serum ".
Kit of the invention can be by the level of REPS1 autoantibody in detection serum, can be with screening crowd to be checked
Suffer from the risk of lung cancer: if REPS1 autoantibody is high (for Healthy People), the risk for suffering from lung cancer is low, if REPS1
Autoantibody is low, then the risk for suffering from lung cancer is high.It can be used for the auxiliary diagnosis of clinical lung cancer, take relevant treatment for patient
Measure or decision provide effective foundation, and potential applicability in clinical practice is good.
Claims (10)
1. the reagent of detection REPS1 albumen autoantibody is preparing the purposes in screening lung cancer kit.
2. purposes as described in claim 1, which is characterized in that the reagent of the detection REPS1 albumen autoantibody is enzyme-linked
Immunosorbent adsorption test reagent or linked immune analysis reagent.
3. purposes as described in claim 1, which is characterized in that it is described detection REPS1 albumen autoantibody reagent be
Western blot reagent.
4. purposes as described in claim 1, which is characterized in that the reagent of the detection REPS1 albumen autoantibody is albumen
Chip detecting method reagent.
5. the purposes as described in Claims 1 to 4 is any, which is characterized in that the reagent of the detection REPS1 albumen autoantibody
It is the reagent for detecting REPS1 albumen autoantibody in human serum.
6. a kind of screening lung cancer kit, which is characterized in that it includes the reagent for detecting REPS1 albumen autoantibody.
7. kit as claimed in claim 6, which is characterized in that the reagent of the detection REPS1 albumen autoantibody is enzyme
Linked immunosorbent adsorption test reagent or ELISA reagent.
8. kit as claimed in claim 6, which is characterized in that it is described detection REPS1 albumen autoantibody reagent be
Western blot reagent.
9. kit as claimed in claim 6, which is characterized in that the reagent of the detection REPS1 albumen autoantibody is egg
White chip detecting method reagent.
10. the kit as described in claim 5~9 is any, which is characterized in that the detection REPS1 albumen autoantibody
Reagent is the reagent for detecting REPS1 albumen autoantibody in human serum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910706018.6A CN110488019A (en) | 2019-07-31 | 2019-07-31 | REPS1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910706018.6A CN110488019A (en) | 2019-07-31 | 2019-07-31 | REPS1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit |
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| Publication Number | Publication Date |
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| CN110488019A true CN110488019A (en) | 2019-11-22 |
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| CN201910706018.6A Pending CN110488019A (en) | 2019-07-31 | 2019-07-31 | REPS1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140309123A1 (en) * | 2011-03-28 | 2014-10-16 | Rosetta Genomics Ltd. | Methods for lung cancer classification |
| US20150099643A1 (en) * | 2011-05-02 | 2015-04-09 | Rheinische Friedrich-Wilhelms-Universitat Bonn | Blood-based gene expression signatures in lung cancer |
-
2019
- 2019-07-31 CN CN201910706018.6A patent/CN110488019A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140309123A1 (en) * | 2011-03-28 | 2014-10-16 | Rosetta Genomics Ltd. | Methods for lung cancer classification |
| US20150099643A1 (en) * | 2011-05-02 | 2015-04-09 | Rheinische Friedrich-Wilhelms-Universitat Bonn | Blood-based gene expression signatures in lung cancer |
Non-Patent Citations (1)
| Title |
|---|
| 裴露: "基于SEREX技术及Oncomine数据库对肺癌相关抗原的筛选与鉴定", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
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Application publication date: 20191122 |