CN103389375A - Liquid chip kit for diagnosing lung cancer - Google Patents
Liquid chip kit for diagnosing lung cancer Download PDFInfo
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- CN103389375A CN103389375A CN2013102900791A CN201310290079A CN103389375A CN 103389375 A CN103389375 A CN 103389375A CN 2013102900791 A CN2013102900791 A CN 2013102900791A CN 201310290079 A CN201310290079 A CN 201310290079A CN 103389375 A CN103389375 A CN 103389375A
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Abstract
The invention discloses a liquid chip kit for diagnosing lung cancer. The kit comprises microballs coated with capturing antibodies, detecting antibodies labeled by biotin, microballs coupled with Halotag Amine (O2) ligand, antigen proteins, streptavidin-phycoerythrin and anti-human immune globulin G labeled by biotin; wherein the capturing antibodies contain at least one component, which is selected from the following capturing antibodies: CRP, Prolactin, CEA, NSE, CK19, Axl and ADAM8; the detecting antibodies contain at least one component, which is selected from the following detecting antibodies: CRP, Prolactin, CEA, NSE, CK19, Axl and ADAM8, and are corresponding to the capturing antibodies; the antigen proteins contain at least one component, which is selected from following components: p62, CTAG, p53 and CAGE. Microballs with different antibodies or antigen proteins have different color codes. The liquid chip kit carries out detections of antigen and autoantibody at the same time, and the precision rate is high.
Description
Technical field
The present invention relates to the medicine bioengineering field, relate in particular to a kind of liquid phase chip reagent box for pulmonary cancer diagnosis.
Background technology
Lung cancer is the incidence of disease and the highest malignant tumour of mortality ratio in global range at present.Over nearly 20 years, owing to carrying out energetically smoking cessation, the incidence of disease of western countries' male lung cancers such as Europe and the U.S. has started to descend.But the incidence of disease of female lung cancer continues to rise.China is cigarette production and selling big country, no matter the male sex or women.The incidence of disease of lung cancer all is lasting ascendant trend, especially with women's incidence of disease, rises faster.Clinical research shows, the carcinoma in situ cure rate is near 100%.5 years survival rates of I phase patients with lung cancer reach 60%～90%, and 5 years survival rates of IIIb and IV patient only 5%～20%.The prompting early diagnosis is the key of improving lung cancer for prognosis.According to the statistical data demonstration of national cancer institute, if can make diagnosis in the lung cancer I phase, its five year survival rate can reach more than 90%; And in II after the phase, five year survival rate sharply is down to below 20%; Develop III, IV phase, perform the operation not only unhelpful, but also likely impel tumour to accelerate diffusion, accelerate dead.Therefore, how to improve lung cancer early diagnosis level and become the serious and urgent task that lung cancer preventing and controlling person faces.
Detection means to lung cancer now depends on the image detecting instruments such as CT, MR, PET, but these image means generally can only be found the tumour that 1-2cm is above, and tumour is from pathology to growing into 1-2cm, generally will pass through the even longer time of 2-5, so the early stage rate of missed diagnosis of cancer is up to 80-90%.
At present, the early diagnosis cancer is normally found tumor markers by blood count, particularly wherein albumen sign.The assay method of blood serum tumor markers mainly contains radiommunoassay, EIA enzyme immunoassay, chemiluminescence immune assay, electrochemiluminescence immunoassay etc.Yet these methods are all the detection methods of single index: and it is not high that the detection that tumour is carried out the unique identification thing is existed the deficiencies, the particularly recall rate to infantile tumour such as specificity is strong, sensitivity is low all the time, therefore may cause some patients were to fail to pinpoint a disease in diagnosis.
The marks such as the tumour antigen in serum can induce body to produce autoantibody, tumour occur also to fail by the clinical examination means detect early stage, body immune system just can monitor the existence of the tumour antigen of low expression level, and initiation immune response, produce a large amount of antibody, play effective bio signal amplification.
Contrast clinical tumor marker albumen commonly used, the detection of autoantibody accounts for great advantage in diagnosing tumor.As, can carry out early diagnosis to tumour, be convenient to early treatment, improve cure rate.Multinomial research shows the existence that autoantibody can be detected before imaging examination is made a definite diagnosis solid carcinoma several months to the several years, even 2-10 just can detect in serum the antibody that has for tumour antigen before tumour is made a definite diagnosis, and autoantibody is higher than the titre of corresponding tumour antigen simultaneously.
However, the detection of autoantibody also has its limitation, and such as in the ordinary course of things, tumour of the same race can produce one or more tumour autoantibodies singularly, and the histological types of different tumours or tumour of the same race both can detect common tumour antibody, also can detect different tumour antibodies.
Therefore, how the detection of autoantibody and tumour antigen is joined together, filter out suitable tumor marker combination, susceptibility and the accuracy that improves diagnosing tumor had great importance.
Summary of the invention
The invention provides a kind of liquid phase chip reagent box for pulmonary cancer diagnosis,, to the antigen in serum and antibody parallel detection, do not have cross reaction, accuracy rate is high, but high flux, realizes fast the diagnosis of the early stage of lung cancer.
A kind of liquid phase chip reagent box for pulmonary cancer diagnosis comprises: microballoon, the biotin labeled detection antibody of coated capture antibody, be coupled Halotag Amine (O
2) microballoon, antigen protein, SA-PE and the biotin labeled anti-human immunoglobulin G of ligand; Wherein,
Capture antibody is at least a in CRP capture antibody, Prolactin capture antibody, CEA capture antibody, NSE capture antibody, CK19 capture antibody, Axl capture antibody and ADAM8 capture antibody;
Detect antibody and be CRP and detect antibody, Prolactin and detect antibody, CEA and detect antibody, NSE and detect antibody, CK19 and detect antibody, Axl and detect antibody and ADAM8 and detect at least a in antibody, and corresponding with capture antibody;
Antigen protein is at least a in p62, CTAG, p53 and CAGE;
Microballoon with different capture antibodies or antigen protein has different color codings.
NSE: nerve specificity olefinic alcohol enzyme, enolase are a kind of glycolytic ferments, are prevalent in mammiferous tissue, with the dimeric form of serial isodynamic enzyme, have (α α, α β, β β and γ γ).
CK19:Cyfra21-1, cytokeratin 19 fragment, be approximately one of the keratic soluble component of epithelial cell of 30kDa of molecular weight, is a kind of mark of extraordinary discriminating lung benign and malignant diseases, especially to the detection better effects if of lung squamous cancer.
CEA: carcinomebryonic antigen is a kind of tumour antigen that is present in kinds of tumor cells, and lung carcinoma cell can directly produce CEA.
The CRP:C reactive protein, as a kind of acute phase protein, affected by the factors such as age, immune state, medicine less, it is to clinical various acute and chronic infection, and the meaning of the diagnosis of the diseases such as tissue damage, observation of curative effect and prognosis judgement is familiar with by people.
Axl: the UFO that is otherwise known as, ARK, or Tyro7, be found as a transformed gene in leukaemia the earliest.This albumen is comprised of 894 amino acid, and molecular weight is 140KD, roughly is evenly distributed on the both sides of cell membrane.
ADAM8: disintegrin-metalloproteinases (a disintegrin and metalloproteases, ADAMs) be that a class of discovered in recent years is combined glycoprotein family with cell membrane, participate in the processes such as the degraded of glutinous company, Fusion of Cells, extracellular matrix of glutinous company, the cell-matrix of cell-cell and signal conduction.
Prolactin: prolactin(PRL, a kind of polypeptide protein hormone for the anterior pituitary secretion, also be prolactin (PRL).
CAGE: tumor-related gene, belong to cancer-testis (cancer-testis CT) antigen family member, that the people such as Cho in 2002 adopt recombinant clone to express the serological analysis technology of antigen, a new Cancer-testis antigen encoding gene of finding during examination testis tissue cDNA library.
P62: nucleoporin is that relative molecular mass is about 62 * 10
31 tumor associated antigen, belong to IMA-IGF2BP3-001 (IGF2) mRNA a member in conjunction with protein family, claim again MP2/IGF2BP2.
CTAG:NY-ESO-1, that chen in 1997 etc. utilize the SEREX technology to screen the tumour antigen that obtains from cancer of the esophagus cDNA library, belong to CTA (cancer-testis antigen) family, this family member only expresses in testis tissue and some tumor tissues.
The present invention is used autoantibody that seven kinds of autoantigens and four kinds of antigen produces as detected object, in liquid phase chip reagent box of the present invention, capture antibody and detect that antibody all can be special with corresponding Sera of Lung Cancer mark (being CRP, Prolactin, CEA, NSE, CK19, Axl or ADAM8) combination, and SA-PE can with the combination of biotin high degree of specificity, therefore can form " microballoon-capture antibody+blood serum designated object+detection antibody+SA-PE " compound for the Sera of Lung Cancer mark; In addition, antigen protein and anti-human immunoglobulin G all can be special with corresponding Sera of Lung Cancer in antibody (p62 antibody, NY-ESO-1 antibody, p53 antibody or CAGE antibody) combination, and anti-human immunoglobulin G is by the phycoerythrin mark, therefore can form " microballoon-antigen protein+serum antibody+anti-human immunoglobulin G+SA-PE " compound for Sera of Lung Cancer antibody
Detect by instrument, definite reaction types different from the microballoon color, excite phycoerythrin with green laser, measures the quantity of the report fluorescence molecule of combination on microballoon, is used for indirectly determining the Sera of Lung Cancer mark of combination on microballoon and the content of serum antibody.
The microballoon that described microballoon can adopt conventional liquid-phase chip to use gets final product.
Described capture antibody is at least two kinds in CRP capture antibody, Prolactin capture antibody, CEA capture antibody, NSE capture antibody, CK19 capture antibody, Axl capture antibody and ADAM8 capture antibody.
Described capture antibody is CRP capture antibody and Prolactin capture antibody.
During the coated microballoon of preparation, the addition of capture antibody is 20～30 μ g/6.25 * 10
6Individual microballoon.
Preferably, described antigen protein is CTAG.
The concentration of the microballoon of every kind of coated capture antibody is 1.2～1.8 * 10
4Individual/μ l, be preferably 1.2 * 10
4Individual/μ l.
Be coupled Halotag Amine (O
2) concentration of microballoon of ligand is 1.2～1.8 * 10
4Individual/μ l, be preferably 1.2 * 10
4Individual/μ l.
The concentration of every kind of biotin labeled detection antibody is 0.8～1.2 μ g/ml, is preferably 1 μ g/ml.
The concentration of biotin labeled anti-human immunoglobulin G is 0.8～1.2 μ g/ml, is preferably 1 μ g/ml.
Compared with prior art, beneficial effect of the present invention is:
The present invention is used for the liquid phase chip reagent box of pulmonary cancer diagnosis based on the liquid-phase chip technology platform, can be to carrying out joint-detection in CRP, Prolactin, CEA, NSE, CK19, Axl and seven kinds of blood serum designated objects of ADAM8 and p62 antibody, CTAG antibody, p53 antibody and four kinds of serum antibodies of CAGE antibody in serum, by CTAG antibody, CRP and Prolactin are carried out joint-detection, the accuracy rate of pulmonary cancer diagnosis is up to 91%, reaction false positive≤2% is high to the accuracy rate of diagnosis of lung cancer.
The liquid phase chip reagent box that the present invention is used for pulmonary cancer diagnosis has and has no side effect, high flux, and susceptibility is high, and good reproducibility detects fast, accurately, the advantages such as cost effective.
Description of drawings
Fig. 1 is for detecting the typical curve schematic diagram of CRP;
Fig. 2 is for detecting the typical curve schematic diagram of Prolactin;
Fig. 3 is for detecting the typical curve schematic diagram of CTAG autoantibody.
Embodiment
In order to understand better technical scheme of the present invention, be further described below in conjunction with specific embodiment, but those of ordinary skill in the art will be appreciated that, the present invention is not limited to these embodiment.
Embodiment 1 is used for the preparation of Sera of Lung Cancer sample CTAG antibody, CRP and Prolactin marker detection kit
1, reagent and solution
(1) 0.1M NaH
2PO
4, pH6.2: weighing 3.5814g NaH
2PO
4In 90mL ddH
2In O, NaOH is settled to 100mL, the 0.22um membrane filtration after regulating pH to 6.2;
(2) 0.05M MES, pH5.0: weighing 0.976g MES is in 90mL ddH
2In O, NaOH is settled to 100mL, the 0.22um membrane filtration after regulating pH to 5.0;
(3) 0.1M MES, pH6.0: weighing 1.952g MES is in 90mL ddH
2In O, NaOH is settled to 100mL, the 0.22um membrane filtration after regulating pH to 6.0;
(4) 0.1M MES, pH4.5: weighing 1.952g MES is in 90mL ddH
2In O, NaOH is settled to 100mL, the 0.22um membrane filtration after regulating pH to 4.5;
(5) PBS:NaCl, 137mM; KCl, 2.7mM; Na
2HPO
4, 8.1mM; KH
2PO
4, 1.5mM; PH7.2-7.4, the 0.22um membrane filtration;
(6) contain 0.02%Proclin300 in 1%PBSB:0.1%PBS, the 0.22um membrane filtration;
(7) contain 0.1%tween-20 in 0.1%PBST:0.1%PBS, the 0.22um membrane filtration;
(8) EDC (1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate);
(9) S-NHS (N-hydroxy thiosuccinimide).
2, be used for the liquid phase chip reagent box of lung cancer early diagnosis, include:
(1) the coated microballoon of 2-plex: contain the 34# microballoon of coated CRP capture antibody, the 45# microballoon of coated Prolactin capture antibody; The concentration of coated microballoon of the same race is not 1.2 * 10
4Individual/μ l;
(2) be coupled HaloTag (O
2) 55# and the 47# microballoon of amine ligand, be coupled goat anti-human igg's 42# microballoon;
(3) the 2-plex biotin labeling detects antibody: the mixed liquor that detects respectively antibody and Prolactin detection antibody with biotin labeled CRP;
(4) CTAG and Halo antigen protein;
(5) SA-PE;
(6) goat anti-human immunoglobulin G-biotin and immunoglobulin G while;
(7) reaction buffer: 1%PBSB;
(8) dilution buffer liquid: 1%PBSB;
(9) sealed membrane;
(10) 96 orifice plates;
(11) control liquid (CRP standard protein, prolactin standard protein).
3, prepare above-mentioned liquid phase chip reagent box, comprise the steps:
3.1 by the composition of mentioned reagent box, every kind of coated corresponding microballoon of capture antibody, the preparation method is identical:
(1) (after vortex and ultrasonic 20s, each draws 200ul (6.25 * 10 respectively for magnetic microsphere, bio-rad) suspension to get 45#, 34#, 42# microballoon
6Individual microballoon) to centrifuge tube;
(2) 14000g, centrifugal 4min, suck supernatant;
(3) add 100ul ddH
2O, vortex and ultrasonic 20s respectively;
(4) 14000g, centrifugal 4min, suck supernatant, adds 160ul0.1M, pH6.2NaH
2PO
4, difference vortex and ultrasonic 20s;
(5) add rapidly the S-NHS of 20ul50mg/mL, vortex mixes;
(6) add rapidly the EDC of 20ul50mg/mL, respectively vortex and ultrasonic approximately 20s;
(7) 20min is hatched in the lucifuge rotation, can mix once every the 10min vortex;
(8) 14000g, centrifugal 4min, carefully suck supernatant, adds 250 μ L0.05M, the MES of pH5.0, vortex and ultrasonic approximately 20s respectively;
(9) 14000g, centrifugal 4min, carefully suck supernatant, repeats above-mentioned washing step once;
(10) carefully suck supernatant, use 50ul0.05M, the resuspended microballoon of the MES of pH5.0, vortex and ultrasonic approximately 20s respectively;
(11) add 25ug CRP capture antibody (R﹠amp; D Systems, production code member are KXL0211061), Prolactin capture antibody (R﹠amp; D Systems, production code member is DJJ0111061) and anti-human immunoglobulin G (the Jackson ImmunoResearch Laboratories in goat source, article No.: 109-005-003) in corresponding model microballoon, add 0.05M, the MES of pH5.0 is 500ul to final volume, difference vortex and ultrasonic approximately 20s;
(12) 2h is hatched in the lucifuge rotation;
(13) 14000g, centrifugal 4min, carefully suck supernatant, adds 1mL0.01%PBST, respectively vortex and ultrasonic approximately 20s;
(13) 14000g, centrifugal 4min, carefully suck supernatant, repeats above-mentioned washing step once;
(14) add 1mL1%PBSB, respectively vortex and ultrasonic approximately 20s;
(15) 30min is hatched in the lucifuge rotation, the activated carboxyl site of sealing microsphere surface remnants;
(16) 14000g, centrifugal 4min, carefully suck supernatant, adds 1mL1%PBSB, respectively vortex and ultrasonic approximately 20s;
(17) carefully suck supernatant, adding 1%PBSB is 200ul to final volume, difference vortex and ultrasonic approximately 20s;
(18) get 2ul in 38ul1%PBSB, after vortex and ultrasonic approximately 20s, blood counting chamber is counted respectively;
(19) on centrifuge tube mark microballoon concentration, be coupled the date, then place 4 the degree keep in Dark Place;
3.2 by the composition of mentioned reagent box, Halo Tag (O
2) Amine ligand is coupled corresponding microballoon, the preparation method is identical:
1. take out 55#, 47# microballoon stoste, respectively vortex and ultrasonic approximately 20s, get 200ul (approximately 6.25 * 10
6Individual) in centrifuge tube, 14000g, centrifugal 4min;
2. suck gently supernatant, add 1ml0.1M, the MES of pH6.0 is vortex and ultrasonic approximately 20s respectively;
3. 14000g, centrifugal 4min, remove supernatant;
4. add 40ul5mg/mLHalo Tag (O
2) Amine ligand, supplementing 0.1M, the MES of pH6.0 is to final volume 450ul, and vortex mixes;
5. add 50ul50mg/mL EDC (now with the current, 0.1M, the MES dissolving of pH6.0), respectively vortex and ultrasonic approximately 20s;
6. 2h is hatched in the lucifuge rotation;
7. add 500ul0.1M, the MES of pH4.5, vortex 20s, 14000g, centrifugal 4min;
8. suck gently supernatant, use 500ul0.1M, the MES of pH4.5 washes twice;
9. 14000g, centrifugal 4min, after carefully sucking supernatant, add 0.1M, and the MES of pH4.5 is to final volume 200ul, respectively vortex and ultrasonic approximately 20s;
10. get 2ul in 38ul1%PBSB, after vortex and ultrasonic approximately 20s, blood counting chamber is counted respectively;
(1) first take out all reagent before the use, place balance to room temperature;
(2) get 2-plex Halo Tag (O
2) Amine ligand is coupled microballoon, vortex mixes 20s, and ultrasonic 20s gets in right amount in 500ul1%PBSB by 2500, every hole microballoon;
(3) lucifuge sealing 1h;
(4) 14000g, centrifugal 4min, add corresponding albumen after drawing supernatant, wherein, CTAG (Creative Biomart, article No. CTAG1B-1512H), each 0.2ug/ hole of Halo, supplement 1%PBSB to final volume be 500ul;
(5) the room temperature lucifuge is hatched 1h;
(6), with after above-mentioned microballoon vortex and ultrasonic approximately 20s, divide and install in 96 orifice plates, the 10ul/ hole;
(7) prepare CRP (R﹠amp; D Systems, production code member are 1229006), Prolactin (R﹠amp; DSystems, production code member are 1175435), the human IgG standard protein, get 8 centrifuge tubes S1, S2, S3, S4, S5, S6, S7, S8 on marks respectively, 2 times of gradient series dilutions, specifically as table 1:
Table 1
Wherein as dilution, human IgG uses the serum matrix that does not contain IgG as dilution to standard protein, and is 4 times of dilutions with 1%PBSB.
(9)---Prime----rinse (channel2)---Prime---places 96 the orifice plates----Run5:MAGX3 that opens the plate machine of washing;
(10) get the coated microballoon of 2-plex capture antibody, vortex also mixes 20s, by 2500, every hole microballoon, joins in 1%PBSB;
(11) add immediately standard items after dilution, each 30ul/ hole;
(12) wrap flat board with aluminium-foil paper, concussion incubation reaction 60min;
(13) 0.1%PBST washes three times, Run5:MAGX3;
(14) (biotin labeled CRP detects antibody, and production code member is DY2648, R﹠amp to dilute 2-plex biotin labeling detection antibody with the ratio of 1:180 with 1%PBSB; D system, biotin labeled Prolactin detects antibody, and production code member is DY682, R﹠amp; D system), each concentration that detects antibody is 1ug/ml, is distributed into the 50ul/ hole;
(15) wrap flat board with aluminium-foil paper, concussion incubation reaction 60min;
(16) 0.1%PBST washes three times, Run5:MAGX3;
(17) 1%PBSB dilution anti-human IgG-biotin (concentration is 1ug/ml) and SAPE (concentration is 1ug/ml), be distributed into the 50ul/ hole;
(18) wrap flat board with aluminium-foil paper, concussion incubation reaction 60min;
(19) 0.1%PBST washes three times, Run5:MAGX3;
(20) the resuspended microballoon of 100ul/well1%PBSB, RT, concussion 3-5min;
(21) reading result on the liquid-phase chip analyser, instrument is the drawing standard curve automatically, and calculates the measured value of sample to be tested.
3.4 interpretation of result
During the production standard curve, the prediction concentrations of CRP, Prolactin and human IgG reference material, measured concentration and MFI value are referring to table 2, and the detection curve of CRP, Prolactin and autoantibody as shown in Figures 1 to 3.
Calculate,
The typical curve of CRP is: FI=99.8135+ (4893.65-99.8135)/((1+ (Conc/13.9421) ^-0.878012)) ^1.02009;
The typical curve of Prolactin is: FI=7.03791+ (852.475-7.03791)/((1+ (Conc/25.6551) ^-1.09938)) ^0.920591;
The typical curve of autoantibody is: FI=15.1731+ (12382-15.1731)/((1+ (Conc/517.673) ^-1.2711)) ^0.857029.
The making of the typical curve of table 2 reference material
Can find out from accompanying drawing and above-mentioned table, CRP, the linearity of each index typical curve of Prolactin and CTAG is R in the concentration range that detects
2〉=0.99, with kit measurement external quality-control product, relative error≤± 5%, the accuracy rate that four index associatings can make pulmonary cancer diagnosis is up to 91%, reaction false positive≤ 2%.Detect required serum amount few (2 microlitre), and can complete in three hours.
Claims (8)
1. liquid phase chip reagent box that is used for pulmonary cancer diagnosis comprises: microballoon, the biotin labeled detection antibody of coated capture antibody, be coupled Halotag Amine (O
2) microballoon, antigen protein, SA-PE and the biotin labeled anti-human immunoglobulin G of ligand; Wherein,
Capture antibody is at least a in CRP capture antibody, Prolactin capture antibody, CEA capture antibody, NSE capture antibody, CK19 capture antibody, Axl capture antibody and ADAM8 capture antibody;
Detect antibody and be CRP and detect antibody, Prolactin and detect antibody, CEA and detect antibody, NSE and detect antibody, CK19 and detect antibody, Axl and detect antibody and ADAM8 and detect at least a in antibody, and corresponding with capture antibody;
Antigen protein is at least a in p62, CTAG, p53 and CAGE;
Microballoon with different capture antibodies or antigen protein has different color codings.
2. liquid phase chip reagent box as claimed in claim 1, it is characterized in that, described capture antibody is at least two kinds in CRP capture antibody, Prolactin capture antibody, CEA capture antibody, NSE capture antibody, CK19 capture antibody, Axl capture antibody and ADAM8 capture antibody.
3. liquid phase chip reagent box as claimed in claim 2, is characterized in that, described capture antibody is CRP capture antibody and Prolactin capture antibody.
4. liquid phase chip reagent box as claimed in claim 1, is characterized in that, during the coated microballoon of preparation, the addition of capture antibody is 20～30 μ g/6.25 * 10
6Individual microballoon.
5. liquid phase chip reagent box as claimed in claim 1, is characterized in that, described antigen protein is CTAG.
6. liquid phase chip reagent box as claimed in claim 1, is characterized in that, the concentration of the microballoon of every kind of coated capture antibody is (1.2～1.8) * 10
4Individual/μ l.
7. liquid phase chip reagent box as claimed in claim 1, is characterized in that, is coupled Halotag Amine (O
2) concentration of microballoon of ligand is (1.2～1.8) * 10
4Individual/μ l.
8. liquid-phase chip as claimed in claim 1, is characterized in that, the concentration of every kind of biotin labeled detection antibody is 0.8～1.2 μ g/ml.
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| CN103869086A (en) * | 2014-04-14 | 2014-06-18 | 杭州凯保罗生物科技有限公司 | Serum autoantibody detection kit |
| CN105510586A (en) * | 2015-12-22 | 2016-04-20 | 湖北鹊景生物医学有限公司 | Kit for lung cancer diagnosis and use method of kit |
| CN105548547A (en) * | 2016-02-18 | 2016-05-04 | 山东信力科生物科技有限公司 | Flow type array immunoassay kit for detecting lung cancer markers based on flow cytometry |
| CN106680515A (en) * | 2016-10-21 | 2017-05-17 | 杭州金式麦生物科技有限公司 | Polymolecular marker composition used for lung cancer diagnosis |
| CN109682969A (en) * | 2019-02-18 | 2019-04-26 | 四川大学华西医院 | A kind of liquid phase chip reagent box detecting intractable epilepsy disease |
| CN110286220A (en) * | 2019-06-04 | 2019-09-27 | 郑州大学第一附属医院 | A kind of application of the molecular marker group and its capture albumen of the early stage cancer of the esophagus in kit |
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