CN110456060A - EFCAB13 autoantibody detection reagent is preparing the purposes in screening lung cancer kit - Google Patents

EFCAB13 autoantibody detection reagent is preparing the purposes in screening lung cancer kit Download PDF

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Publication number
CN110456060A
CN110456060A CN201910702648.6A CN201910702648A CN110456060A CN 110456060 A CN110456060 A CN 110456060A CN 201910702648 A CN201910702648 A CN 201910702648A CN 110456060 A CN110456060 A CN 110456060A
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reagent
efcab13
lung cancer
autoantibody
detection
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CN201910702648.6A
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CN110456060B (en
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刘丹
李为民
张立
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West China Precision Medicine Industrial Technology Institute
West China Hospital of Sichuan University
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West China Precision Medicine Industrial Technology Institute
West China Hospital of Sichuan University
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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
    • 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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  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Urology & Nephrology (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
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Abstract

The present invention relates to external diagnosis reagent fields, and in particular to EFCAB13 autoantibody detection reagent is preparing the purposes in screening lung cancer kit.Present invention firstly discovers that the autoantibody of EFCAB13 albumen is significantly higher than healthy patients in Serum of Patients with Lung Cancer.The reagent that the present invention will test EFCAB13 albumen autoantibody is used to prepare screening lung cancer kit, can be realized effective screening of lung cancer.

Description

EFCAB13 autoantibody detection reagent is preparing the purposes in screening lung cancer kit
Technical field
The present invention relates to external diagnosis reagent fields, and in particular to EFCAB13 autoantibody detection reagent is in preparation lung cancer Purposes in kit for screening.
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)。
The SNP Id of EFCAB13 is rs118004742, is located on 17q21.3 chromosome, and pertinent literature discovery is consulted: though It is related with prostate cancer that so evidence suggests EFCAB13, but some researches show that EFCAB13 genes in prostate cancer patients There are significant differences with normal person for Level of Expression of Retinoic Acid.Up to now there is not yet the related report of EFCAB13 albumen autoantibody Road 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 EFCAB13 albumen autoantibody is preparing the purposes in screening lung cancer kit.
Purposes as the aforementioned, the reagent of the detection EFCAB13 albumen autoantibody are enzyme-linked immunosorbent assay examination Agent or linked immune analysis reagent.
The reagent of purposes as the aforementioned, the detection EFCAB13 albumen autoantibody is western blot reagent.
Purposes as the aforementioned, the reagent of the detection EFCAB13 albumen autoantibody are the examination of protein chip detection method Agent.
Purposes as the aforementioned, the reagent of the detection EFCAB13 albumen autoantibody are EFCAB13 eggs in detection human serum The reagent of white autoantibody.
A kind of screening lung cancer kit, it includes the reagent for detecting EFCAB13 albumen autoantibody.
Kit as the aforementioned, the reagent of the detection EFCAB13 albumen autoantibody are enzyme-linked immunosorbent assay use Reagent or ELISA reagent.
The reagent of kit as the aforementioned, the detection EFCAB13 albumen autoantibody is western blot reagent.
Kit as the aforementioned, the reagent of the detection EFCAB13 albumen autoantibody are protein chip detection method use Reagent.
Kit as the aforementioned, the reagent of the detection EFCAB13 albumen autoantibody are EFCAB13 in detection human serum The reagent of albumen 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 " EFCAB13 autoantibody " refers to " EFCAB13 albumen autoantibody ".
Detailed description of the invention
Fig. 1: EFCAB13 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 EFCAB13 autoantibody and lung cancer in 1 blood plasma of embodiment
One, clinical data
Patients with lung cancer 59 are chosen, normal healthy controls 30, essential information is as follows:
Essential information Patients with lung cancer Normal healthy controls
Number 59 30
Age 54.5±7.6 42±8.9
Masculinity proportion 37 (62.7%) 13 (46.7%)
Two, testing principle
HuProtTMEFCAB13 albumen is fixed on human protein's custom chip, after increase serum is incubated for, in serum EFCAB13 autoantibody (including mainly IgG, IgM type antibody, there are also other types of antibody) can be cleaned in conjunction with up Removal unbonded antibody and other oroteins, then with anti-human IgM fluorescent marker secondary antibody (cy5 label, red is presented) and anti-human IgG fluorescence secondary antibody (cy3 label, green is presented) detection, reads signal, the power of signal and the parent of antibody by Fluorescence Scanner It is positively correlated with power and quantity.
Three, method
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 EFCAB13 autoantibody in Plasma of The Patients With Lung Cancer is that (fluorescence signal is relatively fixed by 14.1SNR Measure ratio), the Average expression level of EFCAB13 autoantibody is 23.5SNR in normal healthy controls blood plasma.Lung cancer group with and health it is right Statistically significant (Fig. 1) is compared according to group (p < 0.05).The ROC of lung cancer group and normal healthy controls analyzes result specificity 96.6%, sensibility is 20.3% (Fig. 2), shows that EFCAB13 autoantibody can specifically distinguish lung cancer and normal healthy controls.
It can be seen from the above result that the level difference of patients with lung cancer and EFCAB13 autoantibody in the serum of non-lung cancer patient Significantly, by the level of EFCAB13 autoantibody in detection serum, it 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 EFCAB13 autoantibody in serum ".
Kit of the invention can be by the level of EFCAB13 autoantibody in detection serum, can be with screening people to be checked Group suffers from the risk of lung cancer: if EFCAB13 autoantibody is low (for Healthy People), the risk for suffering from lung cancer is high, if EFCAB13 autoantibody is high, then the risk for suffering from lung cancer is low.It can be used for the auxiliary diagnosis of clinical lung cancer, take phase for patient The remedy measures or decision of pass provide effective foundation, and potential applicability in clinical practice is good.

Claims (10)

1. the reagent of detection EFCAB13 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 EFCAB13 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 EFCAB13 albumen autoantibody reagent be Western blot reagent.
4. purposes as described in claim 1, which is characterized in that the reagent of the detection EFCAB13 albumen autoantibody is egg White chip detecting method reagent.
5. the purposes as described in Claims 1 to 4 is any, which is characterized in that the examination of the detection EFCAB13 albumen autoantibody Agent is the reagent for detecting EFCAB13 albumen autoantibody in human serum.
6. a kind of screening lung cancer kit, which is characterized in that it includes the reagent for detecting EFCAB13 albumen autoantibody.
7. kit as claimed in claim 6, which is characterized in that it is described detection EFCAB13 albumen autoantibody reagent be Enzyme-linked immunosorbent assay reagent or ELISA reagent.
8. kit as claimed in claim 6, which is characterized in that it is described detection EFCAB13 albumen autoantibody reagent be Western blot reagent.
9. kit as claimed in claim 6, which is characterized in that it is described detection EFCAB13 albumen autoantibody reagent be Protein chip detection method reagent.
10. the kit as described in claim 5~9 is any, which is characterized in that the detection EFCAB13 albumen autoantibody Reagent be detect human serum in EFCAB13 albumen autoantibody reagent.
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140329704A1 (en) * 2013-03-28 2014-11-06 President And Fellows Of Harvard College Markers for mature beta-cells and methods of using the same
US20160376651A1 (en) * 2013-07-24 2016-12-29 The Chinese University Of Hong Kong Biomarkers for premature birth
US20180282809A1 (en) * 2015-09-29 2018-10-04 Max-Delbrück-Centrum Für Molekulare Medizin In Der Helmholtz-Gemeinschaft A METHOD FOR DIAGNOSING A DISEASE BY DETECTION OF circRNA IN BODILY FLUIDS
US20180305689A1 (en) * 2015-04-22 2018-10-25 Mina Therapeutics Limited Sarna compositions and methods of use
WO2019008415A1 (en) * 2017-07-05 2019-01-10 Datar Rajan Exosome and pbmc based gene expression analysis for cancer management
WO2019008414A1 (en) * 2017-07-05 2019-01-10 Datar Rajan Exosome based gene expression analysis for cancer management

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140329704A1 (en) * 2013-03-28 2014-11-06 President And Fellows Of Harvard College Markers for mature beta-cells and methods of using the same
US20160376651A1 (en) * 2013-07-24 2016-12-29 The Chinese University Of Hong Kong Biomarkers for premature birth
US20180305689A1 (en) * 2015-04-22 2018-10-25 Mina Therapeutics Limited Sarna compositions and methods of use
US20180282809A1 (en) * 2015-09-29 2018-10-04 Max-Delbrück-Centrum Für Molekulare Medizin In Der Helmholtz-Gemeinschaft A METHOD FOR DIAGNOSING A DISEASE BY DETECTION OF circRNA IN BODILY FLUIDS
WO2019008415A1 (en) * 2017-07-05 2019-01-10 Datar Rajan Exosome and pbmc based gene expression analysis for cancer management
WO2019008414A1 (en) * 2017-07-05 2019-01-10 Datar Rajan Exosome based gene expression analysis for cancer management

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CDI-LAB.COM: "HuProt v3 block 20", 《HTTPS://COLLECTION.CDI-LAB.COM/PUBLIC/GAL/3/20?PAGE=1》 *
YANINA NATANZON，ET AL.: "Genomic Analysis Using Regularized Regression in High-Grade Serous Ovarian Cancer", 《CANCER INFORM》 *
夏野峰: "肺癌CIRCRNA生物信息学分析与数据库开发", 《中国优秀博硕士学位论文全文数据库(硕士)基础科学辑》 *

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