CN104515768A - Tumor specific growth factor detection kit - Google Patents
Tumor specific growth factor detection kit Download PDFInfo
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- CN104515768A CN104515768A CN201410435931.4A CN201410435931A CN104515768A CN 104515768 A CN104515768 A CN 104515768A CN 201410435931 A CN201410435931 A CN 201410435931A CN 104515768 A CN104515768 A CN 104515768A
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Abstract
The invention relates to a tumor specific growth factor detection kit. The kit comprises an R1 reagent, an R2 reagent and a standard substance solution, the R1 reagent comprises phosphate, disodium ethylene diamine tetraacetate, Kathon and 4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid, and the solvent of the R1 reagent is purified water; the R2 reagent comprises a developer, Kathon, sodium citrate and glycerin, and the solvent of the R2 reagent is purified water; and the standard substance solution comprises a solution A and a solution B, the solution A is purified water, and the solution B is a sialic acid, homocysteine and fructosamine mixed solution. The kit has the advantages of substantial simplification of detection steps, cost saving, simple practical operation, no need of a precious device, and extremely easy popularization.
Description
Technical field
The present invention relates to kit, be specifically related to a kind of TSGF detection kit.
Background technology
Be distributed widely in intraor extracellular and various body fluid because glucide can form glycolipid, glycoprotein, oligosaccharide etc., when cell generation canceration, its metabolic disorder can cause the content in body fluid to raise, and is internationally recognized tumor markers; Amino acid and metabolic product thereof are also applicable to generaI investigation screening because its knurl species specificity is little.Through studying for a long period of time, in this several micromolecular tumor markers, selected several set of landmarks is combined and is collectively referred to as TSGF(Tumor Specific Growth of Factor, TSGF).When tumour produces, in blood, TSGF content raises, and is the important clue of the early detection of tumour, postoperative and Radiotherapy chemotherapy, transfer, recurrence.Clinical testing proves, measures the content of serum tumor specificity growth factor, and will be the early detection of tumour, examination, the dynamic monitoring that the discriminating of innocent and malignant tumour and conditions of patients change provides effective reference frame.
Several relevant tumor markers of joint-detection can improve the extensive common recognition that nodule detection sensitivity has become clinical.Fujian Newland Life Technologies (Holdings) Co., Ltd. successfully develops TSGF detection kit at the beginning of 2000, carry out the clinical verification of millions of example at the situation of all-level hospitals in China tens big or middle cities, because of its specificity and sensitivity higher and be widely used, delivered nearly 500 sections of correlative theses.Along with improving constantly of market polymeric monomer inspection use amount, expose the shortcoming that this kit uses operation gradually: 1) reagent has hydrochloric acid and acetic acid composition, has certain pungency, has impact to environment and operator; 2) bath temperature accuracy requirement is high, 100 DEG C ± 1 DEG C, and the difference of altitude change and well heater all can cause the fluctuation of temperature, and fluctuating range, very likely more than 1 DEG C, causes test findings unreliable; 3) manual application of sample, manual colorimetric, not only complex operation, and also stochastic error is larger.
Summary of the invention
The object of the invention is to the defect overcoming prior art, a kind of TSGF detection kit changed accurately, is fast and automatically provided.
For achieving the above object, the present invention adopts following technical scheme: a kind of TSGF detection kit, comprises R1 reagent, R2 reagent and standard solution;
The each component of described R1 reagent and the mass volume ratio in R1 reagent as follows:
Phosphate 3.00 ~ 3.20g/100mL
Disodium ethylene diamine tetraacetate 0.59 ~ 0.61g/100mL
KF88 0.10 ~ 0.50g/100mL
4-hydroxyethyl piperazine propane sulfonic acid 0.02 ~ 0.10 g/100mL
Its solvent is purified water;
The each component of described R2 reagent and the mass volume ratio in R2 reagent as follows:
Developer 0.08 ~ 0.25g/100mL
KF88 0.10 ~ 0.50g/100mL
Sodium citrate 1.50 ~ 3.50g/100mL
Glycerine 1.00 ~ 8.00g/100mL
Its solvent is purified water;
Described standard solution comprises A liquid and B liquid, and described A liquid is purified water, each component of described B liquid and the mass volume ratio in B liquid as follows:
Sialic acid 0.10 ~ 0.20g/100mL
Homocysteine 0.01 ~ 0.03g/100mL
Fructosamine 0.05 ~ 0.08/g/100mL
Its solvent is purified water.
Preferably, each component of described R1 reagent and the mass volume ratio in R1 reagent as follows:
Phosphate 3.10g/100mL
Disodium ethylene diamine tetraacetate 0.60g/100mL
KF88 0.26g/100mL
4-hydroxyethyl piperazine propane sulfonic acid 0.05 g/100mL
Its solvent is purified water;
The each component of described R2 reagent and the mass volume ratio in R2 reagent as follows:
Developer 0.15g/100mL
KF88 0.26g/100mL
Sodium citrate 2.10g/100mL
Glycerine 5.00g/100mL
Its solvent is purified water;
The each component of B liquid in described standard solution and the mass volume ratio in B liquid as follows:
Sialic acid 0.145g/100mL
Homocysteine 0.023g/100mL
Fructosamine 0.068g/100mL
Its solvent is purified water.
Described phosphate is sodium dihydrogen phosphate.
Described developer is triketohydrindene hydrate.
The principle that this kit detects TSGF is, utilize mixed liquor in kit under 37 DEG C of testing conditions with the TSGF (glycolipid of certain band amino group in testing sample, the materials such as the metabolic product of glycoprotein and free a-amino acid) carry out superposition coupling reaction, reaction product carries out chromogenic reaction with the developer in kit again under same reaction conditions, form blue product, the absorbance of this color product is measured at wavelength 570nm place, and the TSGF content conversed by standard concentration curve and formula (TSGF clinical concentration=100-0.18TSGF N) in testing sample.
The present invention can use on automatic clinical chemistry analyzer, achieve automatic sample, automatic colorimetric, improve precision and the efficiency of detection, comply with clinical examination THE TRENDS OF DEVELOPMENT IN AUTOMATION, there is the advantage that automaticity is high, the test duration is short, reproducible, error is little, the demand of large sample health check-up can be met.
The present invention adopts above technical scheme, under same detection system, adopt kit, to glycolipid, the disposable common colour developing of material such as glycoprotein metabolism product and free a-amino acid of the certain band amino group that several malignant growths in serum are correlated with, sensitivity and the specificity of detection is improve by joint-detection, can detect on automatic clinical chemistry analyzer, achieve and turn to robotization accurately, fast and easily, standardized operation by manual operations, detection efficiency greatly improves, and has complied with the trend of current product automation.Compared with the detection of other TSGFs, the present invention enormously simplify the step of detection, not only cost saving, and practical operation is easy, without the need to expensive instrument, very easily promotes.
Embodiment
A kind of TSGF detection kit, comprises R1 reagent, R2 reagent and standard solution;
For achieving the above object, the present invention adopts following technical scheme: a kind of TSGF detection kit, comprises R1 reagent, R2 reagent and standard solution;
The each component of described R1 reagent and the mass volume ratio in R1 reagent as follows:
Phosphate 3.00 ~ 3.20g/100mL
Disodium ethylene diamine tetraacetate 0.59 ~ 0.61g/100mL
KF88 0.10 ~ 0.50g/100mL
4-hydroxyethyl piperazine propane sulfonic acid 0.02 ~ 0.10 g/100mL
Its solvent is purified water;
The each component of described R2 reagent and the mass volume ratio in R2 reagent as follows:
Developer 0.08 ~ 0.25g/100mL
KF88 0.10 ~ 0.50g/100mL
Sodium citrate 1.50 ~ 3.50g/100mL
Glycerine 1.00 ~ 8.00g/100mL
Its solvent is purified water;
Described standard solution comprises A liquid and B liquid, and described A liquid is purified water, each component of described B liquid and the mass volume ratio in B liquid as follows:
Sialic acid 0.10 ~ 0.20g/100mL
Homocysteine 0.01 ~ 0.03g/100mL
Fructosamine 0.05 ~ 0.08/g/100mL
Its solvent is purified water.
Preferably, each component of described R1 reagent and the mass volume ratio in R1 reagent as follows:
Phosphate 3.10g/100mL
Disodium ethylene diamine tetraacetate 0.60g/100mL
KF88 0.26g/100mL
4-hydroxyethyl piperazine propane sulfonic acid 0.05 g/100mL
Its solvent is purified water;
The each component of described R2 reagent and the mass volume ratio in R2 reagent as follows:
Developer 0.15g/100mL
KF88 0.26g/100mL
Sodium citrate 2.10g/100mL
Glycerine 5.00g/100mL
Its solvent is purified water;
The each component of B liquid in described standard solution and the mass volume ratio in B liquid as follows:
Sialic acid 0.145g/100mL
Homocysteine 0.023g/100mL
Fructosamine 0.068g/100mL
Its solvent is purified water.
Described phosphate is sodium dihydrogen phosphate.
Described developer is triketohydrindene hydrate.
When using this kit to detect sample, the volume ratio of R1 reagent and R2 reagent is 1:1.
In described standard solution, the N of A liquid and B liquid is respectively 0U/mL and 200U/mL, and absorbance corresponding to described 1U/mL N is 0.00011, and namely after the reaction of per unit standard items, standard absorbance value is 0.00011.
Embodiment 1
The preparation of R1 reagent: take following composition
Sodium dihydrogen phosphate 3.00g
Disodium ethylene diamine tetraacetate 0.59g
KF88 0.30g
4-hydroxyethyl piperazine propane sulfonic acid 0.10g
Dissolve by purified water, be mixed with the solution of 100mL, be R1 reagent;
The preparation of R2 reagent: take following composition
Triketohydrindene hydrate 0.08g
KF88 0.50g
Sodium citrate 2.50g
Glycerine 1.00g
Dissolve by purified water, be mixed with the solution of 100mL, be R2 reagent;
The preparation of standard solution:
The preparation of A liquid (N is the standard solution of 0U/mL): i.e. purified water;
The preparation of B liquid (N is the standard solution of 200U/mL): take following composition
Sialic acid 0.10g
Homocysteine 0.03g
Fructosamine 0.065g
Dissolve by purified water, be mixed with the solution of 100mL, be B liquid.
Embodiment 2
The preparation of R1 reagent: take following composition
Sodium dihydrogen phosphate 3.10g
Disodium ethylene diamine tetraacetate 0.61g
KF88 0.50g
4-hydroxyethyl piperazine propane sulfonic acid 0.02g
Dissolve by purified water, be mixed with the solution of 100mL, be R1 reagent;
The preparation of R2 reagent: take following composition
Triketohydrindene hydrate 0.25g
KF88 0.30g
Sodium citrate 3.50g
Glycerine 4.50g
Dissolve by purified water, be mixed with the solution of 100mL, be R2 reagent;
The preparation of standard solution:
The preparation of A liquid (N is the standard solution of 0U/mL): i.e. purified water;
The preparation of B liquid (N is the standard solution of 200U/mL): take following composition
Sialic acid 0.15g
Homocysteine 0.01g
Fructosamine 0.08g
Dissolve by purified water, be mixed with the solution of 100mL, be B liquid.
Embodiment 3
The preparation of R1 reagent: take following composition
Sodium dihydrogen phosphate 3.20g
Disodium ethylene diamine tetraacetate 0.60g
KF88 0.10g
4-hydroxyethyl piperazine propane sulfonic acid 0.06g
Dissolve by purified water, be mixed with the solution of 100mL, be R1 reagent;
The preparation of R2 reagent: take following composition
Triketohydrindene hydrate 0.16g
KF88 0.10g
Sodium citrate 1.50g
Glycerine 8.00g
Dissolve by purified water, be mixed with the solution of 100mL, be R2 reagent;
The preparation of standard solution:
The preparation of A liquid (N is the standard solution of 0U/mL): i.e. purified water;
The preparation of B liquid (N is the standard solution of 200U/mL): take following composition
Sialic acid 0.20g
Homocysteine 0.02g
Fructosamine 0.05g
Dissolve by purified water, be mixed with the solution of 100mL, be B liquid.
Embodiment 4
The preparation of R1 reagent: take following composition
Sodium dihydrogen phosphate 3.10g
Disodium ethylene diamine tetraacetate 0.60g
KF88 0.26g
4-hydroxyethyl piperazine propane sulfonic acid 0.05g
Dissolve by purified water, be mixed with the solution of 100mL, be R1 reagent;
The preparation of R2 reagent:
Triketohydrindene hydrate 0.15g
KF88 0.26g
Sodium citrate 2.10g
Glycerine 5.00g
Dissolve by purified water, be mixed with the solution of 100mL, be R2 reagent;
The preparation of standard solution:
The preparation of A liquid (N is the standard solution of 0U/mL): i.e. purified water;
The preparation of B liquid (N is the standard solution of 200U/mL): take following composition
Sialic acid 0145g
Homocysteine 0.023g
Fructosamine 0.068g
Dissolve by purified water, be mixed with the solution of 100mL, be B liquid.
Selecting parameter when adopting automatic clinical chemistry analyzer to detect is as follows:
Predominant wavelength: 570nm(also can select the wavelength within the scope of 560nm ~ 580nm)
Commplementary wave length: do not set (if instrument requirements must set, then setting within the scope of 700nm ~ 800nm)
Reaction method: rate method the Direction of Reaction: upwards temperature: 37 DEG C
The ratio of testing sample volume and R1, R2 reagent cumulative volume is selected according to the automatic clinical chemistry analyzer of different model.
Time delay/minute: take the circumstances into consideration selected according to the linear section added after reagent R2 in example reaction curve.
Typical curve adopts two point Linear calibrations.
Adopt the detection of the kit of above embodiment 1 to the TSGF content in sample as follows: the operation steps on automatic clinical chemistry analyzer is as follows:
(1) R1 reagent and R2 reagent are put into corresponding reagent position, the volume ratio of R1 reagent and R2 reagent is 1:1;
(2) set factors is set according to type parameter list;
(3) take out N and be respectively 0U/mL(A liquid) and 200U/mL(B liquid) standard solution (corresponding clinical concentration is respectively 100U/mL and 64U/mL) put on standard items (calibration object) fixed mount, the calibration of upper machine;
(4) sample is put on sample rack, upper machine testing.
Three kinds of situations of upper machine testing time-division:
(1) if automatic clinical chemistry analyzer typical curve arranges when allowing negative slope calibration (as Hitachi is serial, Olympus is serial), first use formula: TSGF clinical concentration=100-0.18TSGF N, after standard items are converted into clinical concentration (100U/ml and 64U/ml) by N (0U/ml and 200U/ml), carry out two point Linear calibrations again, institute's measured value is TSGF clinical concentration value.
(2) if when the setting of automatic clinical chemistry analyzer typical curve does not allow negative slope to calibrate, if instrument allows input formula, after then inputting formula (TSGF clinical concentration=100-0.18TSGF N), carry out two point Linear calibrations, institute's measured value is also TSGF clinical concentration value.
(3) if when the setting of automatic clinical chemistry analyzer typical curve does not allow negative slope to calibrate, and instrument does not allow to input formula, then institute's measured value is TSGF N value, and N value is converted into clinical concentration value by formula (TSGF clinical concentration=100-0.18TSGF N).
Detect the measured object content of 543 routine normal human serums, testing result is normal distribution, mean value X=49.30U/ml, standard deviation S D=7.47U/ml, X+1.96S is adopted to determine lower limit, draw reference range: TSGF clinical concentration >=64U/ml is for positive, and TSGF clinical concentration < 64U/ml be feminine gender, oneself term of reference can be set up in each laboratory.
Adopt the detecting step of the kit of above embodiment 2 to 4 to the TSGF content in sample similar with embodiment 1, the present invention is no longer described in detail.
Claims (4)
1. a TSGF detection kit, is characterized in that: it comprises R1 reagent, R2 reagent and standard solution;
The each component of described R1 reagent and the mass volume ratio in R1 reagent as follows:
Phosphate 3.00 ~ 3.20g/100mL
Disodium ethylene diamine tetraacetate 0.59 ~ 0.61g/100mL
KF88 0.10 ~ 0.50g/100mL
4-hydroxyethyl piperazine propane sulfonic acid 0.02 ~ 0.10 g/100mL
Its solvent is purified water;
The each component of described R2 reagent and the mass volume ratio in R2 reagent as follows:
Developer 0.08 ~ 0.25g/100mL
KF88 0.10 ~ 0.50g/100mL
Sodium citrate 1.50 ~ 3.50g/100mL
Glycerine 1.00 ~ 8.00g/100mL
Its solvent is purified water;
Described standard solution comprises A liquid and B liquid, and described A liquid is purified water, each component of described B liquid and the mass volume ratio in B liquid as follows:
Sialic acid 0.10 ~ 0.20g/100mL
Homocysteine 0.01 ~ 0.03g/100mL
Fructosamine 0.05 ~ 0.08/g/100mL
Its solvent is purified water.
2. a kind of TSGF detection kit according to claim 1, is characterized in that: each component of described R1 reagent and the mass volume ratio in R1 reagent as follows:
Phosphate 3.10g/100mL
Disodium ethylene diamine tetraacetate 0.60g/100mL
KF88 0.26g/100mL
4-hydroxyethyl piperazine propane sulfonic acid 0.05 g/100mL
Its solvent is purified water;
The each component of described R2 reagent and the mass volume ratio in R2 reagent as follows:
Developer 0.15g/100mL
KF88 0.26g/100mL
Sodium citrate 2.10g/100mL
Glycerine 5.00g/100mL
Its solvent is purified water;
The each component of B liquid in described standard solution and the mass volume ratio in B liquid as follows:
Sialic acid 0.145g/100mL
Homocysteine 0.023g/100mL
Fructosamine 0.068g/100mL
Its solvent is purified water.
3. a kind of TSGF detection kit according to claim 1 and 2, is characterized in that: described phosphate is sodium dihydrogen phosphate.
4. a kind of TSGF detection kit according to claim 1 and 2, is characterized in that: described developer is triketohydrindene hydrate.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106053444A (en) * | 2016-04-28 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | A kit for measuring a specific growth factor and a preparing method thereof |
| CN109425580A (en) * | 2017-08-28 | 2019-03-05 | 江苏福隆医疗器械有限公司 | A kind of tumor specific growth factor detection kit and its application method |
| CN109799331A (en) * | 2019-03-18 | 2019-05-24 | 湖南海源医疗科技股份有限公司 | A kind of Tumor specific factor TSGF analysis strip |
| CN111721934A (en) * | 2020-06-24 | 2020-09-29 | 湖南新大陆生物技术有限公司 | Improved specific growth factor detection kit and application thereof |
| CN113740539A (en) * | 2021-08-11 | 2021-12-03 | 重庆中元汇吉生物技术有限公司 | Kit for determining specific growth factor |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1139811C (en) * | 1999-11-09 | 2004-02-25 | 赵永同 | Tumor early detection and evaluation kit and preparation process thereof |
| KR20030021159A (en) * | 2000-05-17 | 2003-03-12 | 루드빅 인스티튜트 포 캔서 리서치 | Methods for detecting for the presence of tumor cells and for screening for anti-tumor agents |
| CN100475270C (en) * | 2006-01-20 | 2009-04-08 | 清华大学 | A kind of medicine for treating tumor and application thereof |
| CN101097217A (en) * | 2006-06-29 | 2008-01-02 | 王珊珊 | Technology for enhancing blood serum specificity growth factor immunologic assay sensitivity |
| GB201212334D0 (en) * | 2012-07-11 | 2012-08-22 | Warwick The | Therapeutic targets for alzheimers disease |
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2014
- 2014-08-29 CN CN201410435931.4A patent/CN104515768B/en active Active
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106053444A (en) * | 2016-04-28 | 2016-10-26 | 安徽伊普诺康生物技术股份有限公司 | A kit for measuring a specific growth factor and a preparing method thereof |
| CN109425580A (en) * | 2017-08-28 | 2019-03-05 | 江苏福隆医疗器械有限公司 | A kind of tumor specific growth factor detection kit and its application method |
| CN109799331A (en) * | 2019-03-18 | 2019-05-24 | 湖南海源医疗科技股份有限公司 | A kind of Tumor specific factor TSGF analysis strip |
| CN111721934A (en) * | 2020-06-24 | 2020-09-29 | 湖南新大陆生物技术有限公司 | Improved specific growth factor detection kit and application thereof |
| CN113740539A (en) * | 2021-08-11 | 2021-12-03 | 重庆中元汇吉生物技术有限公司 | Kit for determining specific growth factor |
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