CN110412270A - SSNA1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit - Google Patents

SSNA1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit Download PDF

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Publication number
CN110412270A
CN110412270A CN201910562539.9A CN201910562539A CN110412270A CN 110412270 A CN110412270 A CN 110412270A CN 201910562539 A CN201910562539 A CN 201910562539A CN 110412270 A CN110412270 A CN 110412270A
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CN
China
Prior art keywords
reagent
ssna1
lung cancer
autoantibody
detection
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201910562539.9A
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Chinese (zh)
Inventor
杨桢
张立
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
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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Publication date
Application filed by West China Precision Medicine Industrial Technology Institute, West China Hospital of Sichuan University filed Critical West China Precision Medicine Industrial Technology Institute
Priority to CN201910562539.9A priority Critical patent/CN110412270A/en
Publication of CN110412270A publication Critical patent/CN110412270A/en
Pending legal-status Critical Current

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Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407Specifically defined cancers
    • G01N33/57423Specifically defined cancers of lung

Abstract

The present invention relates to external diagnosis reagent fields, and in particular to SSNA1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit.Present invention firstly discovers that the autoantibody of SSNA1 albumen is significantly higher than healthy patients in Serum of Patients with Lung Cancer.The reagent that the present invention will test SSNA1 albumen autoantibody is used to prepare screening lung cancer kit, can be realized effective screening of lung cancer.

Description

SSNA1 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 SSNA1 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)。
SSNA1 (Ensembl:ENSG00000176101) also known as NA14, Quan Mingwei SS core autoantigen 1, in testis, kidney Generally existing expression in dirty equal tissues.But the accurate function of SSNA1 and clinical meaning are substantially unknown.It has not yet to see The relevant report of SSNA1 albumen 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 SSNA1 albumen autoantibody is preparing the purposes in screening lung cancer kit.
The reagent of purposes as the aforementioned, the detection SSNA1 albumen autoantibody is enzyme-linked immunosorbent assay reagent Or linked immune analysis reagent.
The reagent of purposes as the aforementioned, the detection SSNA1 albumen autoantibody is western blot reagent.
The reagent of purposes as the aforementioned, the detection SSNA1 albumen autoantibody is protein chip detection method reagent.
Purposes as the aforementioned, it is described detection SSNA1 albumen autoantibody reagent be detection human serum in SSNA1 albumen from The reagent of body antibody.
A kind of screening lung cancer kit, it includes the reagent for detecting SSNA1 albumen autoantibody.
Kit as the aforementioned, the reagent of the detection SSNA1 albumen autoantibody are enzyme-linked immunosorbent assay examination Agent or ELISA reagent.
The reagent of kit as the aforementioned, the detection SSNA1 albumen autoantibody is western blot reagent.
Kit as the aforementioned, the reagent of the detection SSNA1 albumen autoantibody are the examination of protein chip detection method Agent.
Kit as the aforementioned, the reagent of the detection SSNA1 albumen autoantibody are SSNA1 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.
" SSNA1 autoantibody " herein refers to " SSNA1 albumen autoantibody ".
Detailed description of the invention
Fig. 1: SSNA1 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 SSNA1 autoantibody and lung cancer in 1 blood plasma of embodiment
One, clinical data
Patients with lung cancer 30 are chosen, 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 SSNA1 albumen on human protein's custom chip, after increase serum is incubated for, in serum SSNA1 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 SSNA1 autoantibody in Plasma of The Patients With Lung Cancer is 23.3SNR (fluorescence signal relative quantification Ratio), the Average expression level of SSNA1 autoantibody is 31.9SNR in normal healthy controls blood plasma.Lung cancer group and healthy control group (p < 0.01) statistically significant (Fig. 1) is compared.It is 86.7% that the ROC of lung cancer group and normal healthy controls, which analyzes result specificity, quick Perception is 26.7% (Fig. 2), shows that SSNA1 autoantibody can specifically distinguish lung cancer and normal healthy controls.
It can be seen from the above result that the level difference of SSNA1 autoantibody is aobvious in patients with lung cancer and the serum of non-lung cancer patient It writes, by the level of SSNA1 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 SSNA1 autoantibody in serum ".
Kit of the invention can be by the level of SSNA1 autoantibody in detection serum, can be with screening crowd to be checked Suffer from the risk of lung cancer: if SSNA1 autoantibody is low (for Healthy People), the risk for suffering from lung cancer is high, if SSNA1 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 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 SSNA1 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 SSNA1 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 SSNA1 albumen autoantibody reagent be Western blot reagent.
4. purposes as described in claim 1, which is characterized in that the reagent of the detection SSNA1 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 SSNA1 albumen autoantibody It is the reagent for detecting SSNA1 albumen autoantibody in human serum.
6. a kind of screening lung cancer kit, which is characterized in that it includes the reagent for detecting SSNA1 albumen autoantibody.
7. kit as claimed in claim 6, which is characterized in that the reagent of the detection SSNA1 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 SSNA1 albumen autoantibody reagent be Western blot reagent.
9. kit as claimed in claim 6, which is characterized in that the reagent of the detection SSNA1 albumen autoantibody is egg White chip detecting method reagent.
10. the kit as described in claim 6~9 is any, which is characterized in that the detection SSNA1 albumen autoantibody Reagent is the reagent for detecting SSNA1 albumen autoantibody in human serum.
CN201910562539.9A 2019-06-26 2019-06-26 SSNA1 autoantibody detection reagent is preparing the purposes in screening lung cancer kit Pending CN110412270A (en)

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WO2018217953A1 (en) * 2017-05-23 2018-11-29 Essenlix Corporation Rapid sample temperature changing for assaying
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Publication number Priority date Publication date Assignee Title
CN1852974A (en) * 2003-06-09 2006-10-25 密歇根大学董事会 Compositions and methods for treating and diagnosing cancer
US20110207613A1 (en) * 2008-06-11 2011-08-25 Sense Proteomic Limited Biomarkers for lupus
US20180356405A1 (en) * 2015-09-29 2018-12-13 Essenlix Corp. Method of Detecting an Analyte in a Sample
US20190079083A1 (en) * 2016-03-30 2019-03-14 Macroarray Diagnostics Gmbh Antigen array
WO2017180587A2 (en) * 2016-04-11 2017-10-19 Obsidian Therapeutics, Inc. Regulated biocircuit systems
WO2018148342A1 (en) * 2017-02-07 2018-08-16 Essenlix Corporation Compressed open flow assay and use
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Application publication date: 20191105