CN104483479B - A kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile - Google Patents
A kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile Download PDFInfo
- Publication number
- CN104483479B CN104483479B CN201410845993.2A CN201410845993A CN104483479B CN 104483479 B CN104483479 B CN 104483479B CN 201410845993 A CN201410845993 A CN 201410845993A CN 104483479 B CN104483479 B CN 104483479B
- Authority
- CN
- China
- Prior art keywords
- film
- solution
- iii
- aperture
- solution penetrates
- 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.)
- Expired - Fee Related
Links
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 title claims abstract description 15
- 239000010931 gold Substances 0.000 title claims abstract description 15
- 229910052737 gold Inorganic materials 0.000 title claims abstract description 15
- 239000004753 textile Substances 0.000 title claims abstract description 10
- 238000003556 assay Methods 0.000 title claims abstract description 8
- 238000001914 filtration Methods 0.000 title claims abstract description 8
- 238000000034 method Methods 0.000 title claims abstract description 8
- 239000000084 colloidal system Substances 0.000 claims abstract description 7
- 239000000243 solution Substances 0.000 claims description 50
- 229910019142 PO4 Inorganic materials 0.000 claims description 20
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 20
- 239000010452 phosphate Substances 0.000 claims description 20
- 108010022355 Fibroins Proteins 0.000 claims description 10
- 230000000274 adsorptive effect Effects 0.000 claims description 10
- 239000008366 buffered solution Substances 0.000 claims description 10
- 229920003023 plastic Polymers 0.000 claims description 10
- 239000004033 plastic Substances 0.000 claims description 10
- 239000000020 Nitrocellulose Substances 0.000 claims description 5
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 5
- 239000012888 bovine serum Substances 0.000 claims description 5
- 239000013642 negative control Substances 0.000 claims description 5
- 229920001220 nitrocellulos Polymers 0.000 claims description 5
- 239000013641 positive control Substances 0.000 claims description 5
- 239000012460 protein solution Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000000427 antigen Substances 0.000 abstract description 4
- 102000036639 antigens Human genes 0.000 abstract description 4
- 108091007433 antigens Proteins 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 238000004451 qualitative analysis Methods 0.000 abstract description 2
- 230000035484 reaction time Effects 0.000 abstract description 2
- 238000001514 detection method Methods 0.000 description 3
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011368 organic material Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention discloses a kind of dot immuno gold filtration assay and measure the method for Ancient Silk Textile, adopt miillpore filter as carrier, by antigen or antibody point on film, add sample to be tested after closing, wash the corresponding antigen of antibody test of rear colloid gold label; Can according to whether carrier occurring redness carries out qualitative analysis.The invention has the beneficial effects as follows: adopt the method for dot immuno gold filtration assay to detect Ancient Silk Textile, on the one hand, highly sensitive, simple to operate.On the other hand, cost is low, and the reaction time is short.
Description
Technical field
The invention belongs to the detection of Ancient Silk Textile, particularly relate to a kind of method that dot immuno gold filtration assay measures Ancient Silk Textile.
Background technology
Silk goods are woven by silk and form, it is Chinese nation's crystallization of wisdom, the precious legacy of China, but silk is as a kind of high-molecular organic material, bury in the environment of grave for a long time, will inevitably be subject to the impact of the factors such as oxygen, water, microorganism and degrade, to such an extent as to outward appearance is seriously damaged and rotten, the basic None-identified of naked eyes.Conventional detection method has Infrared spectroscopy, amino acid analysis, liquid-phase chromatographic analysis etc., but because grain rotten silk goods impurity content is many, spectrum elucidation difficulty, length consuming time, sensitivity is low, and can not get rid of the interference of other oroteins, brings certain difficulty to research worker.
Summary of the invention
The technical problem to be solved in the present invention is for this reason: provide a kind of quick, directly perceived, easy Ancient Silk Textile detection method for above-mentioned existing problem.
Adopt following technical scheme: a kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile for this reason, it is characterized in that the step adopted is as follows:
A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, and making a call to a diameter in the central authorities of lid is respectively the aperture of 0.3-0.8cm; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 1-2ml0.5-3mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100mlpH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 80-100 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;
B), on the NC film below I, II, III aperture, the mass concentration dripping 80-100 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;
C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit that 80-100 μ L50-100 doubly dilutes, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that pH is 7.4, and solution penetrates into cassette interior;
D), on the NC film below I, III aperture, the pH dripping 80-100 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;
E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label that 80-100 μ L100-400 doubly dilutes, solution penetrates into cassette interior;
F), on the NC film below I, II, III, the pH dripping 80-100 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;
G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.
The present invention adopts miillpore filter as carrier, by antigen or antibody point on film, adds sample to be tested, wash the corresponding antigen of antibody test of rear colloid gold label after closing; Can according to whether carrier occurring redness carries out qualitative analysis.
The invention has the beneficial effects as follows: adopt the method for dot immuno gold filtration assay to detect Ancient Silk Textile, on the one hand, highly sensitive, simple to operate.On the other hand, cost is low, and the reaction time is short.
Embodiment
The step that embodiment 1 adopts is as follows:
A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, and making a call to a diameter in the central authorities of lid is respectively the aperture of 0.3cm; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 1ml0.5mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100mlpH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 80 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;
B), on the NC film below I, II, III aperture, the mass concentration dripping 80 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;
C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit that 80 μ L50 doubly dilute, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that pH is 7.4, and solution penetrates into cassette interior;
D) on the NC film below I, III aperture, the pH dripping 80 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;
E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label that 80 μ L100 doubly dilute, solution penetrates into cassette interior;
F) on the NC film below I, II, III, the pH dripping 80 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;
G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.
The step that embodiment 2 adopts is as follows:
A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, and making a call to a diameter in the central authorities of lid is respectively the aperture of 0.6cm; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 1.5ml2mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100mlpH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 90 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;
B), on the NC film below I, II, III aperture, the mass concentration dripping 90 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;
C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit that 90 μ L700 doubly dilute, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that pH is 7.4, and solution penetrates into cassette interior;
D) on the NC film below I, III aperture, the pH dripping 90 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;
E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label that 90 μ L300 doubly dilute, solution penetrates into cassette interior;
F) on the NC film below I, II, III, the pH dripping 90 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;
G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.
The step that embodiment 3 adopts is as follows:
A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, and making a call to a diameter in the central authorities of lid is respectively the aperture of 0.8cm; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 2ml3mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100mlpH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 100 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;
B), on the NC film below I, II, III aperture, the mass concentration dripping 100 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;
C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit that 100 μ L100 doubly dilute, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that pH is 7.4, and solution penetrates into cassette interior;
D) on the NC film below I, III aperture, the pH dripping 100 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;
E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label that 100 μ L400 doubly dilute, solution penetrates into cassette interior;
F) on the NC film below I, II, III, the pH dripping 100 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;
G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.
Claims (1)
1. dot immuno gold filtration assay measures a method for Ancient Silk Textile, it is characterized in that the step adopted is as follows:
A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, and making a call to a diameter in the central authorities of lid is respectively the aperture of 0.3-0.8cm; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 1-2ml0.5-3mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100mlpH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 80-100 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;
B), on the NC film below I, II, III aperture, the mass concentration dripping 80-100 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;
C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit that 80-100 μ L50-100 doubly dilutes, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that pH is 7.4, and solution penetrates into cassette interior;
D), on the NC film below I, III aperture, the pH dripping 80-100 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;
E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label that 80-100 μ L100-400 doubly dilutes, solution penetrates into cassette interior;
F), on the NC film below I, II, III, the pH dripping 80-100 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;
G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410845993.2A CN104483479B (en) | 2014-12-31 | 2014-12-31 | A kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410845993.2A CN104483479B (en) | 2014-12-31 | 2014-12-31 | A kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104483479A CN104483479A (en) | 2015-04-01 |
| CN104483479B true CN104483479B (en) | 2016-02-17 |
Family
ID=52758046
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410845993.2A Expired - Fee Related CN104483479B (en) | 2014-12-31 | 2014-12-31 | A kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104483479B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105628933A (en) * | 2016-02-04 | 2016-06-01 | 中国丝绸博物馆 | Detection method for determining ancient wool fabrics by dot fluorescent immunofiltration |
| CN106290927B (en) * | 2016-07-26 | 2018-12-14 | 鲁东大学 | Fortimicin quick detection kit and its preparation, application method |
| CN109187699B (en) * | 2018-09-05 | 2020-06-23 | 浙江理工大学 | Method for detecting ancient silk fabric |
| CN109187935B (en) * | 2018-09-11 | 2020-12-18 | 浙江理工大学 | Method for detecting ancient silk fabric based on microwave |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0543600A (en) * | 1991-08-08 | 1993-02-23 | Kanebo Ltd | Antibody-or antigen-immobilized silk fibroin membrane and sensor for measuring immune |
| JPH0572205A (en) * | 1991-09-14 | 1993-03-23 | Kanebo Ltd | Immunoassay and apparatus therefor |
| CN1866014A (en) * | 2006-05-19 | 2006-11-22 | 浙江省农业科学院 | Dot immunogold filtration assay kit for diagnosing cysticercosis cellulosae, preparation and application thereof |
| CN103509108A (en) * | 2013-08-07 | 2014-01-15 | 浙江大学 | Method of preparing bombyx mori silk fibroin specific antibody by utilizing characteristic dodecapeptide |
| CN103509107A (en) * | 2013-08-07 | 2014-01-15 | 浙江大学 | Method of preparing bombyx mori silk fibroin specific antibody by utilizing characteristic polypeptide |
| CN104059131A (en) * | 2014-07-04 | 2014-09-24 | 丝科普乐（北京）生物科技有限公司 | Anti-silk fibroin polyclonal antibody and preparation method thereof |
-
2014
- 2014-12-31 CN CN201410845993.2A patent/CN104483479B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0543600A (en) * | 1991-08-08 | 1993-02-23 | Kanebo Ltd | Antibody-or antigen-immobilized silk fibroin membrane and sensor for measuring immune |
| JPH0572205A (en) * | 1991-09-14 | 1993-03-23 | Kanebo Ltd | Immunoassay and apparatus therefor |
| CN1866014A (en) * | 2006-05-19 | 2006-11-22 | 浙江省农业科学院 | Dot immunogold filtration assay kit for diagnosing cysticercosis cellulosae, preparation and application thereof |
| CN103509108A (en) * | 2013-08-07 | 2014-01-15 | 浙江大学 | Method of preparing bombyx mori silk fibroin specific antibody by utilizing characteristic dodecapeptide |
| CN103509107A (en) * | 2013-08-07 | 2014-01-15 | 浙江大学 | Method of preparing bombyx mori silk fibroin specific antibody by utilizing characteristic polypeptide |
| CN104059131A (en) * | 2014-07-04 | 2014-09-24 | 丝科普乐（北京）生物科技有限公司 | Anti-silk fibroin polyclonal antibody and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 利用丝素蛋白抗体鉴定古代丝织品;郑秦等;《蚕业科学》;20140615;第40卷(第3期);520-526 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104483479A (en) | 2015-04-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104483479B (en) | A kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile | |
| CN103852460B (en) | Based on the method that how residual the magnetic nano fluorescent sensor detection of antibiotics of aptamers is | |
| CN108080042A (en) | Micro-fluidic chip of binding time resolved fluorometric technology and its preparation method and application | |
| CN101464464A (en) | Fluorescent microsphere immunity chromatography test paper for detecting food-borne pathogenic microbe | |
| CN203405465U (en) | Test strip for detecting propepsin II and reagent card with same | |
| CN102338801B (en) | High-sensitivity immunochip detection system and application method thereof | |
| CN103439497A (en) | Salmonella enrichment and rapid detection method | |
| CN101566632B (en) | Method for ELISA rapid detection of thermoduric bacteria | |
| CN109374907B (en) | Colistin colloidal gold detection kit and application thereof | |
| CN110308187A (en) | A kind of preparation method and application of porous nano pucherite/bismuth sulfide optical electro-chemistry aptamer sensor based on zinc and cobalt doped | |
| CN106662529A (en) | Assay devices and method of detecting a target analyte | |
| CN106248894A (en) | A kind of paper chip detecting Residue of Antibiotics in Milk | |
| CN202794177U (en) | Kit for enzyme-linked immune-chromatography | |
| CN104459163A (en) | Method for detecting antique silk fabric | |
| CN105651991B (en) | A kind of method of rapid detection of enterobacter sakazakii | |
| CN104977407A (en) | Colloidal gold immunochromatography test paper strip for detecting tetracycline drugs, and preparation method thereof | |
| CN203965377U (en) | A kind of threonine phosphoric acid lyase double-layer nanometer gold membrane electrochemical immunosensor | |
| Yi et al. | A portable toolbox based on time-resolved fluoroimmunoassay and immunomagnetic separation for Cronobacter sakazakii on-site detection in dairy | |
| CN205786331U (en) | The detection device of chemically based luminescence method | |
| CN102331500A (en) | Method and enzyme linked immunosorbent assay kit for detecting lemon yellow | |
| CN104089997A (en) | Electrochemical immunosensor, and preparation method and application thereof | |
| CN103383354B (en) | Detection method of magnetic particle chemiluminescence kit for detecting enterotoxin SEA | |
| CN106053805A (en) | Sensitivity enhanced type nucleic acid aptamer test strip for detecting salmonella and method for preparing sensitivity enhanced type nucleic acid aptamer test strip | |
| CN103115934B (en) | A kind of based on Fe 3o 4the NMR food-borne pathogen rapid detection of@Au composite nanoparticle | |
| CN203275412U (en) | Kit for chemical luminescence immunoassay of CA242 (tumor marker) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160217 Termination date: 20161231 |