CN211148670U - Novel rapid detection kit for coronavirus 2019-nCoV antigen - Google Patents
Novel rapid detection kit for coronavirus 2019-nCoV antigen Download PDFInfo
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- CN211148670U CN211148670U CN202020151387.1U CN202020151387U CN211148670U CN 211148670 U CN211148670 U CN 211148670U CN 202020151387 U CN202020151387 U CN 202020151387U CN 211148670 U CN211148670 U CN 211148670U
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Abstract
The utility model discloses a novel coronavirus 2019-nCoV antigen rapid detection kit belongs to clinical medicine detection area. The test strip is composed of a detection strip and a box body: the box body comprises a box cover and a box bottom, and a detection strip is arranged in the box body. The inspection window and the sampling hole can be seen on the front surface of the box body, and the information recording column, the result judgment description and the sampling hole label are printed on the front surface of the box cover. The utility model discloses monoclonal antibody or many anti based on novel coronavirus specificity antigen S protein adopts "two antibody sandwich" immunochromatography technique to exempt from and has prepared a novel coronavirus antigen short-term test kit, and it is simple to have a detection method, and the result shows accuracy, quick, need not the advantage of special instrument and equipment.
Description
Technical Field
The utility model belongs to clinical medicine detects the field, relates to monoclonal antibody, colloidal gold immunochromatography detection technique, concretely relates to novel coronavirus 2019-nCoV short-term test kit.
Background
The novel coronavirus is a novel coronavirus which can cause pneumonia/pulmonary infection of human, belongs to a coronavirus family with common cold, Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS), and is named as 2019 novel coronavirus, namely 2019-nCoV, by the World Health Organization (WHO).
2019-nCoV is a new strain of coronavirus which has never been found in human body before, and can cause respiratory symptoms such as fever, cough, dyspnea, and the like after infecting the human body, and in more serious cases, infection can cause pneumonia, severe acute respiratory syndrome, renal failure, and even death. At present, no specific treatment medicine for diseases caused by the novel coronavirus infection exists. 2019-nCoV has been discovered to spread at a fast speed in the crowd, and the spreading speed is extremely striking. The central authorities of the prevention and control party and governments at all levels pay high attention to the novel coronavirus, 31 provincial and municipal governments in China start major emergent public health event first-level responses, although a strict joint defense joint control system has been formed in China, rapid propagation of the virus still cannot be blocked, and people have little knowledge about the novel coronavirus and have many unknown conditions. From the current conditions of patients confirmed and treated by clinical detection, most patients have light symptoms, the proportion of severe patients and critical patients is small, the mortality rate is not high, even some patients are detected to be positive by closely contacting the virus nucleic acid, but no clinical symptoms exist, the phenomenon indicates that a plurality of latent virus infectors possibly exist in the population, the latent infectors are possibly a group of more dangerous infectors, and new challenges are brought to virus prevention and control. In addition, the microbial system of hong Kong university teaches Yuan nationally, and in an international authoritative medical journal 'lance' published paper at 24/1 month, researches on the epidemiology and clinical symptoms of the novel coronavirus aggregated infection of a family, and the research result indicates that the novel coronavirus pneumonia symptoms are not 'specific'. In the study cases, the lung of a 10 year old child is infected with virus, but no clinical symptoms appear, which is "rare" in the SARS epidemic of the year. Parents do not have fever symptoms but have diarrhea symptoms immediately after admission. Thus, the authors of this paper believe that in addition to isolating the heat early, the "contacter" should be tracked and isolated early for this new coronavirus pneumonia epidemic, since it is possible that patients with lung infection lesions without symptoms may be a source of spread of the epidemic.
Since the emergence of new coronavirus pneumonia, the whole society has high attention, and a professor group of the perpetual vibration of the university of compound denier strives to analyze gene data of the new coronavirus in the first time of seconds and shares the data on the national center for biotechnology information. A plurality of laboratories and companies in China develop a fluorescent PCR (polymerase chain reaction) novel coronavirus nucleic acid detection kit by means of published novel coronavirus gene data design, and the kit is used for virus infection diagnosis and curative effect evaluation in epidemic situation prevention and control at this time and provides an effective technical means for epidemic situation prevention and control.
The novel coronavirus causing epidemic situation does not appear before, and human cognition is quite limited, so that the difference between the novel coronavirus pneumonia and the SARS epidemic situation in the year is recognized in the epidemic situation prevention and control, a more complete prevention and control strategy is formulated according to the clinical manifestation characteristics of the novel coronavirus pneumonia on the basis of SARS prevention and control experience in the year, and epidemic situation diffusion is controlled more effectively. On the basis of body temperature detection and viral nucleic acid detection, much attention should be paid to tracking, detection and isolation measures of "contacters". Because of the limitation of detection conditions and technologies, the fluorescence PCR virus nucleic acid detection is currently limited to the detection and diagnosis of suspected infected persons with obvious clinical symptoms such as fever and the like and the evaluation of clinical curative effect, and is not suitable for the tracking detection of 'contacters'. Therefore, a novel coronavirus infection detection and diagnosis technology and a reagent product which are rapid, simple and convenient and can be used on site and suitable for screening of 'contacters' and detection and diagnosis of virus infectors without clinical symptoms are urgently needed for epidemic situation prevention and control.
Disclosure of Invention
The utility model aims at providing a novel coronavirus 2019-nCoV antigen rapid detection kit based on gene engineering technology prokaryotic expression novel coronavirus specific antigen S protein, 2019-nCoV S protein specificity monoclonal antibody and gold mark immunochromatography technique, applicable in the suspected infector throat cotton sample novel coronavirus antigen detection. The detection process is simple, the detection can be completed on site or at home without any facility equipment, the detection result is obtained in 3-5 minutes, and the detection result can be directly read manually. The technology is a supplement to the current virus nucleic acid detection technology, and can effectively improve the epidemic situation prevention and control effect. The kit can be suitable for 'screening contacters' and detecting and diagnosing virus infectors without clinical symptoms.
The utility model discloses a following technical scheme realizes: a novel coronavirus 2019-nCoV antigen rapid detection kit based on an S antigen monoclonal antibody comprises a card box and a test strip.
The card box comprises a box cover and a box bottom, the material of the box cover is ABS resin or plastic, the box cover and the box bottom are connected in an inserting mode through columns and column grooves, and the box cover and the box bottom are both in a round-corner rectangular shape; the front surface of the card box can be seen with a detection window and a sample adding hole, the detection window is oblong, and the sample adding hole is oval; the position of the inspection window is opposite to the reaction membrane of the test strip, and the position of the sample adding hole is opposite to the sample pad of the test strip; the front of the box cover is provided with a result mark, a sampling hole mark, a product name mark and an information recording column.
The test strip is composed of a substrate, a sample pad, a mark antibody pad, a reaction film and a water absorption pad which are sequentially adhered on the substrate by double faced adhesive tapes.
The cartridge material may be composed of ABS resin (acrylonitrile-butadiene-styrene copolymer) or plastic (polystyrene, polyethylene, polyvinyl chloride, etc.).
The substrate material can be made of PVC plastic (polyvinyl chloride) or PS plates.
The sample pad may be composed of glass fibers or non-woven fabric.
The marked antibody pad is formed by solidifying a monoclonal antibody or a polyclonal antibody against the novel coronavirus 2019-nCoV S protein marked by a marker on glass fiber or non-woven fabric, wherein the marker can be selected from colloidal gold particles, colored latex particles or fluorescent microspheres.
The reaction membrane is composed of a nitrocellulose membrane, a detection line (T line) and a quality control line (C line) are arranged on the nitrocellulose membrane, the detection line is composed of an anti-novel coronavirus 2019-nCoV S protein monoclonal antibody or a polyclonal antibody coating, and the quality control line is composed of a goat or rabbit anti-mouse IgG polyclonal antibody or a secondary antibody coating aiming at the detection line coating polyclonal antibody.
The water absorption pad is composed of water absorption filter paper or oil filter paper.
The utility model discloses an innovative effect is mainly embodied in that novel coronavirus 2019-nCoV surface specificity antigen S protein is used as an antigen target spot, virus antigen in a sample is detected by adopting an immunochromatography detection technology, and diagnosis reference is made to virus infection, the scheme can be realized by the following process that firstly, published novel coronavirus 2019-nCoV surface specificity antigen S protein gene sequence data are utilized, 2019-nCoV S protein gene is optimized and synthesized, a 2019-nCoV S protein prokaryotic expression vector is constructed, the vector is transferred into escherichia coli E.Coli B L21 (DE3), IPTG induced expression 2019-nCoV S protein is utilized, a purified recombinant 2019-nCoV S protein immune Balb/c mouse is utilized, an anti-2019-nCoV S protein monoclonal antibody is prepared by a monoclonal antibody technology, or 2019-nCoV S protein multiple antibodies are prepared by animals such as immunized sheep and rabbits, a novel coronavirus 2019-nCoV antigen rapid detection kit is prepared by adopting a double-antibody sandwich immunochromatography technology, and a suspected virus infection person can make a respiratory tract rapid diagnosis through detecting a secretion.
The utility model discloses a solid-phase immunodetection technique, antibody albumen are solidified on NC membrane and glass fiber or non-woven fabrics, and antibody albumen is more stable under this kind of solidification dry state, and is not high to the ambient temperature requirement, and the kit can be preserved under normal atmospheric temperature (4 ℃ -30 ℃), and the transportation is simple, does not need the cold chain, simultaneously, the utility model discloses still have easy operation, need not special equipment, direct reading is visualized to the result. The kit is applicable to detection of respiratory tract secretion (throat test specimen), sputum, oral saliva and tracheal lavage fluid samples.
Drawings
Fig. 1 is a front schematic view of the present invention.
Fig. 2 is a schematic view of the back of the box cover of the present invention.
Fig. 3 is a front view of the case bottom of the present invention.
Fig. 4 is a front schematic view of the test strip of the present invention.
Fig. 5 is a schematic side view of the test strip of the present invention.
The figures in the drawings represent the following: 1. a sample application hole; 2. a viewing window; 3. an information recording field; 4. a box cover; 5. a well mark (S); 6. identifying a result; 7. a box bottom; 8. a column; 9. a column groove; 10. a test strip groove; 11. a sample pad; 12. single-sided glue; 13. a reaction film; 14. detecting a line T; 15. a quality control line C; 16. a water absorbent pad; 17. a substrate; 18. double-sided adhesive tape; 19. the antibody pad is labeled.
Detailed Description
The following examples are provided for further understanding of the present invention, and are not intended to be limiting to the preferred embodiments, and are not intended to limit the scope and content of the present invention, and any product identical or similar to the present invention, which is derived by anyone under the teaching of the present invention or by combining the features of the present invention with other prior art, is intended to fall within the scope of the present invention.
Example 1 novel coronavirus 2019-nCoV antigen gold-labeled rapid detection kit
A novel gold-labeled rapid detection kit for coronavirus 2019-nCoV antigens comprises a box cover (4), a box bottom (7) and a test strip arranged in a test strip groove at the bottom of the box. The box cover (4) is provided with a viewing window (2), a result mark (6), a sampling hole mark (5), a sampling hole (1) and an information recording column (3). The sampling hole (1) corresponds to a sample pad (11) of the test strip, the inspection window (2) corresponds to a reaction film (13) of the test strip, the result mark C corresponds to a quality control line (15) on the reaction film, and the T corresponds to a detection line (14) on the reaction film.
The bottom of the card box is provided with a test strip groove (13), and the specification of the test strip groove is matched with the test strip.
The test strip consists of a sample pad (11), a mark antibody pad (19), a reaction film (13) and a water absorption pad (16) which are sequentially bonded by a double-sided adhesive tape (18) on a substrate, wherein the sample pad, the mark antibody pad and the reaction film are staggered by about 15mm and bonded by a single-sided adhesive tape (12), and the reaction film and the water absorption pad are staggered by about 15 mm. The substrate is composed of a PVC plate, the sample pad is composed of non-woven fabric, the reaction membrane is composed of a nitrocellulose membrane (NC) membrane, and the water absorption pad is composed of oil filter paper. The labeled antibody pad (19) is prepared by immersing a colloidal gold labeled anti-novel coronavirus 2019-nCoV monoclonal antibody a into non-woven fabric, drying and curing at 25 ℃, and a detection line T (14) on the reaction film is coated by an anti-novel coronavirus 2019-nCoV monoclonal antibody b and dried at 45 ℃ for 1 hour. The quality control line C (15) on the reaction membrane (13) is prepared by streaking and coating a goat or rabbit anti-mouse IgG polyclonal antibody and drying for 1 hour at 45 ℃. The test strip is placed in a test strip groove (10) of the box bottom (7); the column (8) on the box cover (4) is aligned with the column groove (9) on the box bottom and closed in a plugging way.
The detection method comprises the steps of adding 300 mu L of sample diluent (0.01M PBS (phosphate buffer solution) with pH 7.2) into a special sample tube with a filtering function, immersing a cotton test specimen or a sputum test specimen collected from the throat into the diluent of the sample tube, continuously stirring and extruding along the tube wall to enable virus antigens in the test specimen to be contained in the diluent as much as possible, taking out the cotton swab, adding a filter cover to the sample tube, inverting the sample tube, dripping 2 drops of 100 mu L into the sample adding hole, filling a sample number into an information recording column on the front face of a box cover, starting observation of a result, observing that a quality control line C and a detection line T at a detection window are in a color development state and are effective within 20 minutes, if the quality control line C and the detection line T are both in a mauve color development state, judging that the result is positive, if only the quality control line C and the detection line T are both in a mauve color development state, and judging that the result is negative if the quality control line C and the detection line T are both in a non-effect, directly adding L of the sample tube, and.
The kit of the embodiment is used for detecting throat testees, sputum, oral saliva and tracheal lavage fluid samples.
Example 2 a novel kit for rapid detection of coronavirus 2019-nCoV antigen by latex immunochromatography, the labeled antibody pad (19) constituting the structure of the kit was replaced with a colored latex pad, and the conditions were the same as in example 1, except that the labeled substance used in the colored latex pad was colored latex, and the preparation method was the same as that of the gold-labeled pad. The sample, sample preparation method and sample addition amount were applied, and the result was determined in the same manner as in example 1.
Example 3 a novel coronavirus 2019-nCoV antigen fluorescent microsphere immunochromatography rapid detection kit the constitution structure of the labeled antibody pad (19) of the kit was replaced by a fluorescent microsphere pad, and the other conditions were the same as in example 1, except that the label used in the fluorescent microsphere pad was a fluorescent microsphere, and the preparation method thereof was the same as that of the gold-labeled pad. The sample preparation method and the sample addition amount were the same as those in example 1. And (4) interpretation of results: the reaction film in the inspection window is observed by irradiating the handheld fluorescent lamp, and the quality control line C and the detection line T at the inspection window display the fluorescent reaction condition and are effective within 20 minutes. If the quality control line C and the detection line T both show purple fluorescence, the result is positive; if only the quality control line C shows purple fluorescence and the detection line T does not show color, the result is negative; if the quality control line C and the detection line T do not develop color, the result is invalid.
Claims (1)
1. A novel coronavirus 2019-nCoV antigen rapid detection kit is characterized in that: the test strip is composed of a card box and a test strip; the card box comprises a box cover and a box bottom, the material of the box cover is ABS resin or plastic, the box cover and the box bottom are connected in an inserting mode through columns and column grooves, and the box cover and the box bottom are both in a round-corner rectangular shape; the front surface of the card box can be seen with a detection window and a sample adding hole, the detection window is oblong, and the sample adding hole is oval; the position of the inspection window is opposite to the reaction membrane of the test strip, and the position of the sample adding hole is opposite to the sample pad of the test strip; the front surface of the box cover is provided with a result mark, a sampling hole mark, a product name mark and an information recording column; the test strip is composed of a substrate, a sample pad, a mark antibody pad, a reaction film and a water absorption pad which are sequentially adhered on the substrate by double faced adhesive tapes.
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| CN111929442A (en) * | 2020-08-13 | 2020-11-13 | 北京测尔康生物技术有限公司 | Novel coronavirus antigen and antibody detection kit |
| CN112067712A (en) * | 2020-08-18 | 2020-12-11 | 上海纳米技术及应用国家工程研究中心有限公司 | Volatile marker for diagnosing novel coronavirus and application thereof |
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| CN112415201A (en) * | 2020-08-21 | 2021-02-26 | 北京现代高达生物技术有限责任公司 | Novel coronavirus S protein and N protein joint detection colloidal gold test strip and preparation method and application thereof |
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| CN111929442A (en) * | 2020-08-13 | 2020-11-13 | 北京测尔康生物技术有限公司 | Novel coronavirus antigen and antibody detection kit |
| CN112067712A (en) * | 2020-08-18 | 2020-12-11 | 上海纳米技术及应用国家工程研究中心有限公司 | Volatile marker for diagnosing novel coronavirus and application thereof |
| CN112415201A (en) * | 2020-08-21 | 2021-02-26 | 北京现代高达生物技术有限责任公司 | Novel coronavirus S protein and N protein joint detection colloidal gold test strip and preparation method and application thereof |
| WO2022037010A1 (en) * | 2020-08-21 | 2022-02-24 | 北京现代高达生物技术有限责任公司 | Colloidal gold test strip for joint detection of s protein and n protein of sars-cov-2 |
| CN112213290A (en) * | 2020-09-15 | 2021-01-12 | 深圳技术大学 | Novel coronavirus detection method and system based on luminescent quantum dot material |
| CN112162092A (en) * | 2020-09-30 | 2021-01-01 | 北京金沃夫生物工程科技有限公司 | Novel coronavirus detection kit |
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| AU2021351793B2 (en) * | 2020-09-30 | 2023-03-16 | Beijing Jinwofu Bioengineering Technology Co., Ltd | Detection kit and preparation method thereof and a detection method for novel coronavirus |
| EP4063865A4 (en) * | 2020-09-30 | 2023-07-19 | Beijing Jinwofu Bioengineering Technology Co., Ltd | Kit for detecting novel coronavirus, and preparation method therefor and detection method therefor |
| CN112162092B (en) * | 2020-09-30 | 2024-04-02 | 北京金沃夫生物工程科技有限公司 | Novel coronavirus detection kit |
| CN112557653A (en) * | 2020-11-19 | 2021-03-26 | 安徽瀚海博兴生物技术有限公司 | Colloidal gold kit for antigen-antibody mixed detection of novel coronavirus and preparation method thereof |
| CN112782401A (en) * | 2021-02-08 | 2021-05-11 | 聊城大学 | Method for rapidly detecting novel coronavirus in vitro and application |
| CN113219174A (en) * | 2021-04-14 | 2021-08-06 | 中国科学院西北高原生物研究所 | Novel coronavirus S antigen detection kit and use method thereof |
| WO2022236891A1 (en) * | 2021-05-14 | 2022-11-17 | 沙滨 | Non-invasive detection kit for detecting novel coronavirus antigen, and detection method thereof |
| CN113552348A (en) * | 2021-07-28 | 2021-10-26 | 四川省医学科学院·四川省人民医院 | Nanoparticle solution for detecting SARS-CoV-2 coronavirus S protein, preparation method, kit and application |
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