CN110412276A - PDE9A autoantibody detection reagent is preparing the purposes in screening lung cancer kit - Google Patents
PDE9A autoantibody detection reagent is preparing the purposes in screening lung cancer kit Download PDFInfo
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- CN110412276A CN110412276A CN201910563584.6A CN201910563584A CN110412276A CN 110412276 A CN110412276 A CN 110412276A CN 201910563584 A CN201910563584 A CN 201910563584A CN 110412276 A CN110412276 A CN 110412276A
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- 102100024227 High affinity cGMP-specific 3',5'-cyclic phosphodiesterase 9A Human genes 0.000 title claims abstract description 47
- 101001117259 Homo sapiens High affinity cGMP-specific 3',5'-cyclic phosphodiesterase 9A Proteins 0.000 title claims abstract description 47
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 47
- 206010058467 Lung neoplasm malignant Diseases 0.000 title claims abstract description 46
- 201000005202 lung cancer Diseases 0.000 title claims abstract description 46
- 208000020816 lung neoplasm Diseases 0.000 title claims abstract description 46
- 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
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 238000004458 analytical method Methods 0.000 claims description 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
- 239000000243 solution Substances 0.000 description 6
- 238000004140 cleaning Methods 0.000 description 5
- 210000002381 plasma Anatomy 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- ZOOGRGPOEVQQDX-UUOKFMHZSA-N 3',5'-cyclic GMP Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=C(NC2=O)N)=C2N=C1 ZOOGRGPOEVQQDX-UUOKFMHZSA-N 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102100025570 Cancer/testis antigen 1 Human genes 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
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 230000008859 change Effects 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
- 230000001419 dependent effect Effects 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
- 238000005516 engineering process Methods 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 210000004072 lung Anatomy 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
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000003147 molecular marker Substances 0.000 description 1
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- 230000032696 parturition Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000008447 perception Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 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
- 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
- 210000001519 tissue Anatomy 0.000 description 1
- 239000000439 tumor marker Substances 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
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Peptides Or Proteins (AREA)
Abstract
The present invention relates to external diagnosis reagent fields, and in particular to PDE9A autoantibody detection reagent is preparing the purposes in screening lung cancer kit.Present invention firstly discovers that the autoantibody of PDE9A albumen is significantly higher than healthy patients in Serum of Patients with Lung Cancer.The reagent that the present invention will test PDE9A 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 PDE9A 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)。
PDE9A (Ensembl:ENSG00000160191MIM:602973) is the cycli phosphate bird of wide expression in the tissue
Glycosides (cGMP) enzyme-specific is important in activation cGMP dependent signals pathway.Have not yet to see PDE9A albumen
The relevant report of autoantibody also has no its prior art relevant to lung 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 PDE9A albumen autoantibody is preparing the purposes in screening lung cancer kit.
The reagent of purposes as the aforementioned, the detection PDE9A albumen autoantibody is enzyme-linked immunosorbent assay reagent
Or linked immune analysis reagent.
The reagent of purposes as the aforementioned, the detection PDE9A albumen autoantibody is western blot reagent.
The reagent of purposes as the aforementioned, the detection PDE9A albumen autoantibody is protein chip detection method reagent.
Purposes as the aforementioned, it is described detection PDE9A albumen autoantibody reagent be detection human serum in PDE9A albumen from
The reagent of body antibody.
A kind of screening lung cancer kit, it includes the reagent for detecting PDE9A albumen autoantibody.
Kit as the aforementioned, the reagent of the detection PDE9A albumen autoantibody are enzyme-linked immunosorbent assay examination
Agent or ELISA reagent.
The reagent of kit as the aforementioned, the detection PDE9A albumen autoantibody is western blot reagent.
Kit as the aforementioned, the reagent of the detection PDE9A albumen autoantibody are the examination of protein chip detection method
Agent.
Kit as the aforementioned, the reagent of the detection PDE9A albumen autoantibody are PDE9A 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.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 " PDE9A autoantibody " refers to " PDE9A albumen autoantibody ".
Detailed description of the invention
Fig. 1: PDE9A autoantibody comparison in patients with lung cancer (LC), normal healthy controls (NC) serum slurry.
Fig. 2: patients with lung cancer (LC) and normal healthy controls (NC) ROC are analyzed.
Specific embodiment
The relationship of PDE9A autoantibody and lung cancer in 1 blood plasma of embodiment
One, clinical data
Patients with lung cancer 30 are chosen, 0 normal healthy controls 30, essential information is as follows:
| Essential information | Patients with lung cancer | Normal healthy controls |
| Number | 30 | 30 |
| Age | 58±10.5 | 42±8.9 |
| Masculinity proportion | 20 (66.7%) | 13 (46.7%) |
Two, testing principle
HuProtTMBe fixed with PDE9A albumen on human protein's custom chip, after increase serum is incubated for, in serum PDE9A 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
The reagent that this part is used is as follows:
Specific steps are 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 PDE9A autoantibody in Plasma of The Patients With Lung Cancer is 39.3SNR (fluorescence signal relative quantification
Ratio), the Average expression level of PDE9A autoantibody is 24.4SNR in normal healthy controls blood plasma.Lung cancer group and healthy control group (p
< 0.01) statistically significant (Fig. 1) is compared.It is 93.1% that the ROC of lung cancer group and normal healthy controls, which analyzes result specificity, quick
Perception is 37.9% (Fig. 2), shows that PDE9A autoantibody can specifically distinguish lung cancer and normal healthy controls.
It can be seen from the above result that the level difference of PDE9A autoantibody is aobvious in patients with lung cancer and the serum of non-lung cancer patient
It writes, by the level of PDE9A 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 PDE9A autoantibody in serum ".
Kit of the invention can be by the level of PDE9A autoantibody in detection serum, can be with screening crowd to be checked
Suffer from the risk of lung cancer: if PDE9A autoantibody is high (for Healthy People), the risk for suffering from lung cancer is high, if PDE9A
Autoantibody is low, then the risk for suffering from lung cancer is low.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 PDE9A 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 PDE9A 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 PDE9A albumen autoantibody reagent be
Western blot reagent.
4. purposes as described in claim 1, which is characterized in that the reagent of the detection PDE9A 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 PDE9A albumen autoantibody
It is the reagent for detecting PDE9A albumen autoantibody in human serum.
6. a kind of screening lung cancer kit, which is characterized in that it includes the reagent for detecting PDE9A albumen autoantibody.
7. kit as claimed in claim 6, which is characterized in that the reagent of the detection PDE9A 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 PDE9A albumen autoantibody reagent be
Western blot reagent.
9. kit as claimed in claim 6, which is characterized in that the reagent of the detection PDE9A 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 PDE9A albumen autoantibody
Reagent is the reagent for detecting PDE9A albumen autoantibody in human serum.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910563584.6A CN110412276A (en) | 2019-06-26 | 2019-06-26 | PDE9A autoantibody detection reagent is preparing the purposes in screening lung cancer kit |
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|---|---|---|---|
| CN201910563584.6A CN110412276A (en) | 2019-06-26 | 2019-06-26 | PDE9A autoantibody detection reagent is preparing the purposes in screening lung cancer kit |
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|---|---|
| CN110412276A true CN110412276A (en) | 2019-11-05 |
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| CN201910563584.6A Pending CN110412276A (en) | 2019-06-26 | 2019-06-26 | PDE9A autoantibody detection reagent is preparing the purposes in screening lung cancer kit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114349857A (en) * | 2022-03-15 | 2022-04-15 | 诺赛联合(北京)生物医学科技有限公司 | Treg cell preparation method and application in aspect of autoimmune diseases |
| CN114605553A (en) * | 2022-03-15 | 2022-06-10 | 诺赛联合(北京)生物医学科技有限公司 | Pharmaceutical composition containing Treg cells and use in autoimmune diseases |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090047689A1 (en) * | 2007-06-20 | 2009-02-19 | John Kolman | Autoantigen biomarkers for early diagnosis of lung adenocarcinoma |
-
2019
- 2019-06-26 CN CN201910563584.6A patent/CN110412276A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090047689A1 (en) * | 2007-06-20 | 2009-02-19 | John Kolman | Autoantigen biomarkers for early diagnosis of lung adenocarcinoma |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114349857A (en) * | 2022-03-15 | 2022-04-15 | 诺赛联合(北京)生物医学科技有限公司 | Treg cell preparation method and application in aspect of autoimmune diseases |
| CN114349857B (en) * | 2022-03-15 | 2022-05-24 | 诺赛联合(北京)生物医学科技有限公司 | Treg cell preparation method and application in aspect of autoimmune diseases |
| CN114605553A (en) * | 2022-03-15 | 2022-06-10 | 诺赛联合(北京)生物医学科技有限公司 | Pharmaceutical composition containing Treg cells and use in autoimmune diseases |
| CN114605553B (en) * | 2022-03-15 | 2022-09-02 | 诺赛联合(北京)生物医学科技有限公司 | Pharmaceutical composition containing Treg cells and use in autoimmune diseases |
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