CN203490226U - Immunofluorescence chromatography test strip for detection of Miltenberger antibody - Google Patents
Immunofluorescence chromatography test strip for detection of Miltenberger antibody Download PDFInfo
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- CN203490226U CN203490226U CN201320431392.8U CN201320431392U CN203490226U CN 203490226 U CN203490226 U CN 203490226U CN 201320431392 U CN201320431392 U CN 201320431392U CN 203490226 U CN203490226 U CN 203490226U
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Abstract
The utility model discloses an immunofluorescence chromatography test strip for detection of Miltenberger antibody. The test strip comprises a sample pad, a reaction membrane and an absorbent pad which are sequentially lapped on a lining plate, wherein two ends of the sample pad are respectively provided with a first sample adding point and a second sample adding point, an inner quality control belt and at least an antibody detection belt are fixed on the reaction membrane. By adopting the immunofluorescence chromatography test strip, the problem that simple and practical conventional clinical Miltenberger blood group antibody detection technologies and reagents are lack in China or even in the world is solved, the test strip can be used for assisting in diagnosing diseases such as hemolytic disease of the newborn and adverse effect of blood transfusion caused by Miltenberger blood group antibody in clinic, and guarantees safe, effective and scientific blood transfusion in clinic.
Description
Technical field
The utility model relates to medical science detection field, particularly relates to a kind of Miltenberger antibody test immunofluorescence chromatograph test strip.
Background technology
MNS blood group system is second blood group system finding after ABO, and the polymorphism number of its antigen is only second to Rh blood group system.In MNS blood group system, have the sub-blood group system of the special Miltenberger of a class, its blood group antigens are red blood cell phenotypes of a series of low frequency antigens combinations of being identified with human serum immune antiboidy.In this subsystem, the occurrence frequency of several antigens such as Mur, Mil, lower than per mille in European white man, and is 3% ~ 10% in Chinese and other asian populations.Investigation for the 844Ming Guangxi crowd of a Dong nationality demonstration, Mur rare blood group antigen positive rate is 15.4%.And the people such as Lin Mali are in nineteen ninety, the investigation for each race of Taiwan shows, the Mur blood group positive rate of high mountain aborigines' AMIS is up to 88.4%.
Anti-Mur incidence is very high in south east asia, and particularly in China Taiwan, Hong Kong crowd, the occurrence frequency of anti-Mur is even only second to the anti-B of anti-A/.Clinical report confirms that anti-Mur antibody can cause alloimmunity disease, as blood transfusion bad reaction, neonatal hemolytic disease etc.Follow globalization process, each race and various countries' population, in global flow faster, make Mur antigen become more and more important at the clinical meaning of countries in the world.Haematogenic immunity theory and clinical practice all clearly propose, and in the spectrum cell of identifying, should have Mur antigen positive red blood cell at blood group antibody examination red blood cell group and the blood group antibody of countries in Asia's application.Owing to lacking related reagent, in the Antibody screening cell that most medical institutions are used, do not comprise Miltenberger blood group antigens, the specificity of the fubaritic antibody of conventional sense, causes existing the patient of this antibody undetected.Due to fresh red blood cell holding time very short (maximum 3 months), Miltenberger antigen positive red blood cell source is very limited, and the detection of the examination of Miltenberger blood group and antibody thereof is greatly limited.Several subject matters that Miltenberger blood group antibody faces in detecting are at present: 1), owing to not using corresponding screening red blood cell in clinical detection spectrum cell, irregular antibody is often undetected; 2) the erythrocytic stable supply of screening that contains private antigen can not get long-term assurance; 3) the fresh red blood cell holding time is extremely short.
Due to so far also without the monoclonal antibody of the anti-Miltenberger blood group antigens of commercialized supply, the inherent defect of people source blood grouping reagent (human red blood cell reagent and Serum Antibodies reagent) and there is no suitable test method, Miltenbergerr blood group compatibility test can not routine be carried out at present clinically in countries in the world, the check work of only in large-scale Blood Center and the blood group reference test chamber of minority, some difficult blood group incompatibility disease patients being correlated with.So far, concrete obstacle and the problem that can not conventional carry out clinically the experiment of Miltenberger blood group compatibility are: Miltenberger antigen reagent and antibody reagent be shortage all, the inherent defect of red blood cell blood cell reagent and Serum Antibodies reagent, and the shortcoming of the immunoserology experimental technique method of applying at present etc.China is clinical for the importance of Miltenberger blood group antigens antibody and the correlative study of detection method also seldom, reports at present only for case.
Australia CSL company utilizes Kode
tMtechnology, is connected to synthetic polypeptide antigen on red blood cell, prepares the red blood cell of the anti-Mur antibody of screening and anti-MUT antibody.This technology is by red blood cell and contains function (epi-position) group-connection chain-fat group (function-epitope-spacer lipid, FSL) liquid mixes a period of time, the FSL that carries special epitope can simulate glycolipid structure and be incorporated into erythrocyte surface, thereby changes erythrocytic blood group.But this technology can only increase antigen, and can not seal original red cell antigens.This examination red blood cell has been used to carry out large-scale antibody screening.DanCSL company utilizes Kode
tMtechnology is prepared remains based on the erythrocytic blood group antigens reagent of survival, and above-mentioned several root problems of mentioning are not resolved, and is present in the blood group antigens on the erythrocyte membrane of survival, and due to the easy haemolysis of red blood cell, antigenicity is also withered away thereupon.
Utility model content
The technical matters that the utility model mainly solves is to provide a kind of Miltenberger antibody test immunofluorescence chromatograph test strip, and the operation of immunofluorescence chromatographic technique is fast and convenient, in conjunction with detecting instrument, can realize sxemiquantitative or quantitatively detect; This technology can solve China and even lack at present in the world simple and practical routine clinical Miltenberger blood group antibody detection technique and a difficult problem for reagent, the diagnosis of the diseases such as the auxiliary clinical neonatal hemolytic disease causing because of Miltenberger blood group antibody, blood transfusion bad reaction, and compatibility experiment detects before transfusing blood, ensure clinical safety, effective, science blood transfusion.
For solving the problems of the technologies described above, the technical scheme that the utility model adopts is: a kind of Miltenberger antibody test immunofluorescence chromatograph test strip is provided, comprise reaction film, sample pad, absorption pad and liner plate, described reaction film is positioned on described base plate, on described reaction film, be fixed with and detect band and quality control band, described sample pad and described absorption pad are overlapping with the two side portions of described reaction film respectively, and be positioned on described reaction film and described base plate, in described sample pad, comprise that first adds sampling point and second and add sampling point, described first adds sampling point is arranged on away from described sample pad and the overlapping side of described reaction film, described second adds sampling point is arranged on and approaches described sample pad and the overlapping side of described reaction film.
In preferred embodiment of the utility model, the length of the both sides lap of described sample pad or described absorption pad and described reaction film is 0.5 mm.
In preferred embodiment of the utility model, the material of described reaction film is a kind of in nitrocellulose filter, cellulose acetate film, nylon membrane and poly tetrafluoroethylene.
In preferred embodiment of the utility model, on described quality control band, be fixed with human IgG and people IgM, described detection band is at least one, described detection is with and is fixed with Streptavidin.
The beneficial effects of the utility model are: the operation of immunofluorescence chromatographic technique is fast and convenient, in conjunction with detecting instrument, can realize sxemiquantitative or quantitatively detect; This technology can solve China and even lack at present in the world simple and practical routine clinical Miltenberger blood group antibody detection technique and a difficult problem for reagent, the diagnosis of the diseases such as the auxiliary clinical neonatal hemolytic disease causing because of Miltenberger blood group antibody, blood transfusion bad reaction, and compatibility experiment detects before transfusing blood, ensure clinical safety, effective, science blood transfusion.
Accompanying drawing explanation
In order to be illustrated more clearly in the technical scheme in the utility model embodiment, below the accompanying drawing of required use during embodiment is described is briefly described, apparently, accompanying drawing in the following describes is only embodiment more of the present utility model, for those of ordinary skills, do not paying under the prerequisite of creative work, can also according to these accompanying drawings, obtain other accompanying drawing, wherein:
Fig. 1 is the structural representation of the utility model Miltenberger antibody test immunofluorescence chromatograph test strip one preferred embodiment;
In accompanying drawing, the mark of each parts is as follows: 1, first adds sampling point, and 2, sample pad, 3, second adds sampling point, and 4, detect band, 5, reaction film, 6, quality control band, 7, absorption pad, 8, liner plate.
Embodiment
Below the technical scheme in the utility model embodiment is clearly and completely described, obviously, described embodiment is only a part of embodiment of the present utility model, rather than whole embodiment.Embodiment based in the utility model, those of ordinary skills are not making all other embodiment that obtain under creative work prerequisite, all belong to the scope of the utility model protection.
Embodiment mono-:
Refer to Fig. 1, a kind of Miltenberger antibody test immunofluorescence chromatograph test strip, comprise reaction film 5, sample pad 2, absorption pad 7 and liner plate 8, described reaction film 5 is positioned on described liner plate 8, on described reaction film 5, be fixed with to detect and be with 4 and quality control band 6, described sample pad 2 and described absorption pad 7 are overlapping with the two side portions of described reaction film 5 respectively, and be positioned on described reaction film 5 and described liner plate 8, in described sample pad 2, comprise that first adds sampling point 1 and second and add sampling point 3, described first adds sampling point 1 is arranged on away from described sample pad 2 side overlapping with described reaction film 5, described second adds sampling point 3 is arranged on and approaches described sample pad 2 side overlapping with described reaction film 5.
Preferably, the material of described reaction film 5 is a kind of in nitrocellulose filter, cellulose acetate film, nylon membrane and poly tetrafluoroethylene.On described quality control band 6, be fixed with human IgG and people IgM.Described detection is with 4 to be at least one, and described detection is with and on 4, is fixed with Streptavidin.
Embodiment bis-:
Provide described in a kind of embodiment mono-test strips for the detection method of Miltenberger blood group antibody screening:
(1) preparation of test strips:
1) choose Millipore HF 180 tape backing films as reaction film, be cut into 30 * 1.8 cm sizes standby, using PBS that the pH value of 0.01 M is 7.2 to regulate its concentration human IgG and people IgM is 0.5 mg/mL, adding volume fraction is 1% Tween-20, use a stroke film instrument to be sprayed on NC film surface as quality control band, drawing a film amount is 0.5 μ L/cm.Streptavidin is applied to PBS that the pH value of 0.15 M is 7.2, and to be adjusted to concentration be 0.1 mg/mL, use a stroke film instrument to be sprayed on NC film surface as detecting band, every line is spaced apart 5 mm, draws after film finishes and puts into immediately 37 ℃ of baking oven dried overnight, and room temperature preservation is standby;
2) assembling of test strips
Choose domestic high-quality glass fibre membrane BT100 as the material of sample pad, be cut into 30 * 1.8 cm sizes standby, choose domestic high-quality thieving paper CH37K as the material of absorption pad, and it is standby to be cut into 30 * 1.8 cm sizes, in specification, be first to paste reaction film on the PVC liner plate of 30 * 6 cm, then take the method for overlap joint to stick on liner plate sample pad and absorption pad, the length of overlap joint is 0.5 mm, so that the diffusion of reactant liquor, after assembling, be cut into the wide test strips of 4 mm, and be packaged in plastic casing, be put in aluminium foil bag, 4 ℃ of sealings are preserved.
3) preparation of fluorescence reaction liquid
It is 2 mg/mL that anti-human IgG or anti-human IgM antibody-solutions phosphate buffer are adjusted to concentration, pH is 7.5 to 8.0, get 1 mL antibody-solutions, slowly adding 100 μ L concentration is the Cy5 fluorescent dye of 1 mg/mL, 4 ℃ slowly stir 3 h after at room temperature reaction 1 h, with the phosphate buffer that pH is 7.4, dialyse again, remove unreacted Cy5 dyestuff.The good fluorescence antibody of recovery coupling after dislysate completes, measures after coupling efficiency and antibody concentration, at 4 ℃, keeps in Dark Place.
(2) detect the preparation of sample
1) patients serum to be checked
Extract Venous Blood separation of serum to be checked;
2) serum to be checked and polypeptide antigen are hatched
By Mia (MNS7), Mur (MNS10), Hil(MNS20 in the sub-blood group system of corresponding Miltenberger) and the polypeptide of MUT (MNS35) blood group antigens amino acid sequence with 0.15 M pH 7.2 PBS, be dissolved to 10 mg/mL, after getting 50 μ L polypeptide antigens and 50 μ L serum to be checked and mixing, 1 h is standby for incubated at room.
(3) detecting step
1) by test strips and reactant liquor balance to room temperature (18-25 ℃);
2) add the mixed liquor of 10 μ L serum to be checked and polypeptide antigen to sample pad;
3) adding 10 μ L to contain volume fraction is that the pH of 0.5% the Tween-20 0.15M PBS that is 7.2 is to sample pad;
4) repeating step 3) 3 times;
5) add the 5 anti-human IgG of μ L Cy5-and/or anti-human IgM to sample pad;
6) repeating step 3) 3 times;
7) test strips is placed in to fluorescence signal and reads instrument, read fluorescence signal and measure.
Embodiment tri-:
Provide described in a kind of embodiment mono-test strips for the detection method of Miltenberger blood group antibody specificity identification:
(1) preparation of test strips:
1) choose Millipore HF 180 tape backing films as reaction film, be cut into 30 * 1.8 cm sizes standby, using PBS that the pH value of 0.01 M is 7.2 to regulate its concentration human IgG and people IgM is 0.5 mg/mL, adding volume fraction is 1% Tween-20, use a stroke film instrument to be sprayed on NC film surface as quality control band 6, drawing a film amount is 0.5 μ L/cm.Streptavidin is applied to PBS that the pH value of 0.15 M is 7.2, and to be adjusted to concentration be 0.1 mg/mL, use a stroke film instrument to be sprayed on NC film surface as detecting band, every line is spaced apart 5 mm, draws after film finishes and puts into immediately 37 ℃ of baking oven dried overnight, and room temperature preservation is standby;
2) assembling of test strips
Choose domestic high-quality glass fibre membrane BT100 as the material of sample pad, be cut into 30 * 1.8 cm sizes standby, choose domestic high-quality thieving paper CH37K as the material of absorption pad, and it is standby to be cut into 30 * 1.8 cm sizes, in specification, be first to paste reaction film on the PVC liner plate of 30 * 6 cm, then take the method for overlap joint to stick on liner plate sample pad and absorption pad, the length of overlap joint is 0.5 mm, so that the diffusion of reactant liquor, after assembling, be cut into the wide test strips of 4 mm, and be packaged in plastic casing, be put in aluminium foil bag, 4 ℃ of sealings are preserved.
3) preparation of fluorescence reaction liquid
It is 2 mg/mL that anti-human IgG or anti-human IgM antibody-solutions phosphate buffer are adjusted to concentration, pH is 7.5 to 8.0, get 1 mL antibody-solutions, slowly adding 100 μ L concentration is the Cy5 fluorescent dye of 1 mg/mL, 4 ℃ slowly stir 3 h after at room temperature reaction 1 h, with the phosphate buffer that pH is 7.4, dialyse again, remove unreacted Cy5 dyestuff.The good fluorescence antibody of recovery coupling after dislysate completes, measures after coupling efficiency and antibody concentration, at 4 ℃, keeps in Dark Place.
(2) detect the preparation of sample
1) patients serum to be checked
Extract Venous Blood separation of serum to be checked;
2) serum to be checked and polypeptide antigen are hatched
By Mia (MNS7), Mur (MNS10), Hil(MNS20 in the sub-blood group system of corresponding Miltenberger) and the polypeptide of MUT (MNS35) blood group antigens amino acid sequence with 0.15 M pH 7.2 PBS, be dissolved to 10 mg/mL respectively, after getting 50 μ L polypeptide antigens and 50 μ L serum to be checked and mixing, 1 h is standby for incubated at room.
(3) detecting step
1) by test strips and reactant liquor balance to room temperature (18-25 ℃);
2) add the mixed liquor of 10 μ L serum to be checked and polypeptide antigen to sample pad;
3) adding 10 μ L to contain volume fraction is that the pH of 0.5% the Tween-20 0.15M PBS that is 7.2 is to sample pad;
4) repeating step 3) 3 times;
5) add the 5 anti-human IgG of μ L Cy5-and/or anti-human IgM to sample pad;
6) repeating step 3) 3 times;
7) test strips is placed in to fluorescence signal and reads instrument, read fluorescence signal and measure.
The beneficial effect that the utility model Miltenberger blood group antibody detects immunofluorescence chromatograph test strip is: the operation of immunofluorescence chromatographic technique is fast and convenient, in conjunction with detecting instrument, can realize sxemiquantitative or quantitatively detect; This technology can solve China and even lack at present in the world simple and practical routine clinical Miltenberger blood group antibody detection technique and a difficult problem for reagent, the diagnosis of the diseases such as the auxiliary clinical neonatal hemolytic disease causing because of Miltenberger blood group antibody, blood transfusion bad reaction, and compatibility experiment detects before transfusing blood, ensure clinical safety, effective, science blood transfusion.
The foregoing is only embodiment of the present utility model; not thereby limit the scope of the claims of the present utility model; every equivalent structure or conversion of equivalent flow process that utilizes the utility model description to do; or be directly or indirectly used in other relevant technical field, be all in like manner included in scope of patent protection of the present utility model.
Claims (4)
1. a Miltenberger antibody test immunofluorescence chromatograph test strip, it is characterized in that, comprise reaction film, sample pad, absorption pad and liner plate, described reaction film is positioned on described base plate, on described reaction film, be fixed with and detect band and quality control band, described sample pad and described absorption pad are overlapping with the two side portions of described reaction film respectively, and be positioned on described reaction film and described base plate, in described sample pad, comprise that first adds sampling point and second and add sampling point, described first adds sampling point is arranged on away from described sample pad and the overlapping side of described reaction film, described second adds sampling point is arranged on and approaches described sample pad and the overlapping side of described reaction film.
2. test strips according to claim 1, is characterized in that, the length of the both sides lap of described sample pad or described absorption pad and described reaction film is 0.5 mm.
3. test strips according to claim 1, is characterized in that, the material of described reaction film is a kind of in nitrocellulose filter, cellulose acetate film, nylon membrane and poly tetrafluoroethylene.
4. test strips according to claim 1, is characterized in that, is fixed with human IgG and people IgM on described quality control band, and described detection band is at least one, and described detection is with and is fixed with Streptavidin.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201320431392.8U CN203490226U (en) | 2013-07-19 | 2013-07-19 | Immunofluorescence chromatography test strip for detection of Miltenberger antibody |
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| CN201320431392.8U CN203490226U (en) | 2013-07-19 | 2013-07-19 | Immunofluorescence chromatography test strip for detection of Miltenberger antibody |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104360087A (en) * | 2014-12-10 | 2015-02-18 | 中国科学院苏州生物医学工程技术研究所 | Method for detecting Miltenberger blood group antibody |
| CN111892657A (en) * | 2019-05-06 | 2020-11-06 | 医疗财团法人台湾血液基金会 | Antibody and fragment thereof, kit and method for detecting blood group antigen of Mitigo |
-
2013
- 2013-07-19 CN CN201320431392.8U patent/CN203490226U/en not_active Expired - Lifetime
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104360087A (en) * | 2014-12-10 | 2015-02-18 | 中国科学院苏州生物医学工程技术研究所 | Method for detecting Miltenberger blood group antibody |
| CN111892657A (en) * | 2019-05-06 | 2020-11-06 | 医疗财团法人台湾血液基金会 | Antibody and fragment thereof, kit and method for detecting blood group antigen of Mitigo |
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Effective date of registration: 20160803 Address after: Science and Technology City kolding road high tech Zone of Suzhou City, Jiangsu Province, No. 88 215163 Patentee after: SUZHOU GUOKE MEDICAL TECHNOLOGY DEVELOPMENT Co.,Ltd. Address before: Science and Technology City kolding road high tech Zone of Suzhou City, Jiangsu Province, No. 88 215163 Patentee before: Suzhou Institute of Biomedical Engineering and Technology Chinese Academy of Sciences |
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Effective date of registration: 20160912 Address after: 215163 Suzhou high tech Zone, Jinfeng Road, No. 8, Jiangsu Patentee after: SUZHOU GUOKE SIBEIDA BIOTECHNOLOGY CO.,LTD. Address before: Science and Technology City kolding road high tech Zone of Suzhou City, Jiangsu Province, No. 88 215163 Patentee before: SUZHOU GUOKE MEDICAL TECHNOLOGY DEVELOPMENT Co.,Ltd. |
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Effective date of registration: 20230506 Address after: 215163 Suzhou 88 high tech Zone, Jiangsu science and Technology City Patentee after: Suzhou Guoke medical technology development (Group) Co.,Ltd. Address before: 215163 Kam Peak Road, Suzhou high tech Zone, Suzhou, Jiangsu Patentee before: SUZHOU GUOKE SIBEIDA BIOTECHNOLOGY CO.,LTD. |
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