CN104360079A - PIVKA-II detection kit - Google Patents
PIVKA-II detection kit Download PDFInfo
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- CN104360079A CN104360079A CN201410735809.9A CN201410735809A CN104360079A CN 104360079 A CN104360079 A CN 104360079A CN 201410735809 A CN201410735809 A CN 201410735809A CN 104360079 A CN104360079 A CN 104360079A
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- pivka
- antagonist
- vitamin
- deficiency
- albumen
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- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
Abstract
The invention relates to the field of biological detection and in particular relates to a detection kit used for quantitatively detecting a protein PIVKA-II produced due to deficiency of vitamin K or induction of an antagonist-II as well as a preparation method and application of the detection kit. The detection kit comprises a test paper card, wherein the test paper card comprises a base plate as well as a sample pad, a gold mark pad, a nitrocellulose membrane and a water absorption pad which are sequentially arranged from a sampling end positioned on the surface of the base plate, the gold mark pad contains a protein PIVKA-II antibody produced due to deficiency of vitamin K or induction of the antagonist-II, the nitrocellulose membrane is coated with a detection line and a quality control line, and the protein PIVKA-II antibody produced due to deficiency of vitamin K or the antagonist-II on the gold mark pad is marked by adopting a fluorescent microsphere. The detection kit provided by the invention can be used for detecting PIVKA-II by adopting a fluorescent microsphere immunochromatographic technology initially, has sensitivity and specificity and has the advantages of quickness and simplicity in operation, accurate result, economic applicability and the like.
Description
Technical field
The present invention relates to field of biological detection, particularly relate to protein detection kit of a kind of vitamin K deficiency or antagonist-II induction generation and its production and use.
Background technology
Vitamin K deficiency or antagonist-II induce albumen (the Protein Induced by Vitamin K Absence orAntagonist-II produced, PIVKA-II), be again de--γ-carboxyl factor (Des-gamma-carboxy prothrombin, DCP) or abnormal prothrombin, the mark of primary carcinoma of liver within 1984, is identified as first.In patients with hepatocellular carcinoma serum, PIVKA-II concentration level is far above cirrhosis and metastatic hepatic carcinoma patient, and the dynamic change of the change of its serum-concentration and Patients ' Hepatocytes cancer (operation, treatment and recurrence etc.) is relevant.Research afterwards confirms, PIVKA-II and PIVKA-II detects and can complement one another, and the diagnosis of hepatocellular carcinoma can be brought up to 84% by the joint-detection of the two, and only the patients with hepatocellular carcinoma of about 16% is both negative result.Current PIVKA-II detects and is put in " Japanese liver cancer association liver cancer diagnosis and treatment specification version in 2009 ", for the examination of hepatocellular carcinoma people at highest risk and the auxiliary diagnosis of primary carcinoma of liver; Mark for hepatocellular carcinoma auxiliary diagnosis in " primary carcinoma of liver diagnosis and treatment specification (version in 2011) " that the Ministry of Public Health of China issues also comprises PIVKA-II.
Summary of the invention
The shortcoming of prior art in view of the above, the object of the present invention is to provide vitamin K deficiency or antagonist-II in a kind of Fast Measurement serum or blood plasma to induce the kit of albumen PIVKA-II content produced, for solving the problems of the prior art.
For achieving the above object and other relevant objects, the invention provides vitamin K deficiency or antagonist-II in a kind of Fast Measurement serum or blood plasma and induce the kit of albumen PIVKA-II content produced, comprise test card, described test card comprises base plate, and be positioned at the sample pad be arranged in order from application of sample end of backplate surface, gold mark pad, nitrocellulose filter and adsorptive pads, described gold mark pad comprises albumen PIVKA-II monoclonal antibody of vitamin K deficiency or antagonist-II induction generation, described nitrocellulose filter is coated with detection line and nature controlling line, vitamin K deficiency on described gold mark pad or antagonist-II induce albumen PIVKA-II monoclonal antibody produced to adopt fluorescent microsphere mark.
Preferably, vitamin K deficiency on described gold mark pad or antagonist-II induce albumen PIVKA-II monoclonal antibody produced to adopt 180nm fluorescent microsphere mark, EX (nm)=650/Em (nm)=670, there is signal little by background interference, detection sensitivity is high, the advantage that result is reproducible.
Preferably, described base plate is PVC base plate.
Preferably, on described nitrocellulose filter, detection line is positioned at side close to application of sample end, and nature controlling line is positioned at side away from application of sample end.
Preferably, described detection line is coated with albumen PIVKA-II monoclonal antibody of vitamin K deficiency or antagonist-II induction generation.Vitamin K deficiency on described gold mark pad or antagonist-II induce albumen PIVKA-II monoclonal antibody produced to induce albumen PIVKA-II monoclonal antibody produced can be identical antibody with the vitamin K deficiency on detection line or antagonist-II, also can be different antibody.
Preferably, nature controlling line wraps by sheep anti-mouse antibody.
Preferably, described sample pad adopts damping fluid process, described damping fluid is selected from one or more the combination in PBS damping fluid, Tris-HCl damping fluid, glycine buffer, borate buffer solution and citrate-phosphate salt buffer, and the concentration of damping fluid is 20-200mM.
Preferably, gold mark of the present invention is paid somebody's debt and expected repayment later through pre-service, and the pre-treatment buffer used during pre-service is selected from one or more combination of glycine buffer, Tris-HCl damping fluid, borate buffer solution, and the concentration of damping fluid is 5-50mM.
Preferably, described damping fluid also comprises increased response agent, described increased response agent be selected from PEG4000, PEG6000, PEG8000 and PEG20000 any one, the concentration of described increased response agent is 10 ~ 50g/L.
Preferably, described buffer solution also comprises surfactant, described surfactant is selected from any one or multiple combination in S-19TWEEN 20, S-20TWEEN 80, S-13TRITON X-45, S-14TRITON X-100, S-15TRITON X305, and the concentration of described surfactant is 10 ~ 50g/L.
Preferably, in order to make kit, there is better sensitivity and color developing effect, gold mark of the present invention pads the pre-treatment buffer used when pre-service and comprises following component: stachyose, alum, Fructose Diphosphate, sodium hexametaphosphate and glycocoll, and the total concentration of stachyose, alum, Fructose Diphosphate, sodium hexametaphosphate, glycocoll is 3.5-7.5g/L, the pH value of damping fluid is 7.2-7.6.
Preferably, the concentration of each component in damping fluid is:
The solvent of described pre-treatment buffer is water.
Described pretreated concrete steps are: gold mark pad is soaked 1.5 ~ 2h in pretreatment fluid, take out and are put in 36 ~ 38 DEG C of oven dry.
Described pre-treatment buffer can use the various conventional pH adjusting agent in this area to carry out the adjustment of pH value.
Preferably, described kit also comprises and getting stuck, described getting stuck comprises back of the body card and upper cover, the described back of the body is arranged with test card draw-in groove, described test card is embedded in described test card draw-in groove, be covered with testing window and well on described, matching with the position of described detection line and nature controlling line in the position of described testing window, matches with the position of described sample pad in the position of described well.
Preferred, described in get stuck for plastics get stuck.
Preferably, described detection kit is used for quantitatively detecting the content of the albumen PIVKA-II that vitamin K deficiency or antagonist-II induction produce in serum or blood plasma.
Quantitative detection vitamin K deficiency provided by the present invention or antagonist-II induce the detection kit of the albumen PIVKA-II produced, and can supporting immune quantitative analytical instrument use.Immunoassay instrument, by acquisition testing line (T) and nature controlling line (C) band fluorescence signal, calculates T/C signal value.First various criterion product are added drop-wise on test card before using, analyzing and processing sets up calibration curve (relation of T/C signal value and standard items actual value), again the T/C value that obtains when detecting sample is compared with typical curve, albumen PIVKA-II content detecting vitamin K deficiency in sample or antagonist-II induction generation can be obtained.
Second aspect present invention provides described quantitative detection lipoprotein vitamin K deficiency or antagonist-II to induce the preparation method of the detection kit of the albumen PIVKA-II produced, and comprises the steps:
1) spray golden mark with albumen PIVKA-II antibody-solutions that vitamin K deficiency or the antagonist-II induction of fluorescent microsphere mark produce to pad, the obtained gold mark pad comprising albumen PIVKA-II antibody of vitamin K deficiency or antagonist-II induction generation;
2) on the detection line and nature controlling line of nitrocellulose filter, spray vitamin K deficiency or antagonist-II respectively induce albumen PIVKA-II antibody and sheep anti-mouse antibody that produce, obtained bag by after nitrocellulose filter;
3) by sample pad, step 1) preparation gold mark pad, step 2) nitrocellulose filter, the adsorptive pads prepared be pasted onto on base plate successively, cutting obtains Test paper card; Finally Test paper is snapped fits into the obtained detection kit that gets stuck.
Preferably, gold mark of the present invention is paid somebody's debt and expected repayment later through pre-service, and the pre-treatment buffer used during pre-service is selected from one or more combination of glycine buffer, Tris-HCl damping fluid, borate buffer solution, and the concentration of damping fluid is 5-50mM.
Preferably, in order to make kit, there is better sensitivity and color developing effect, gold mark of the present invention pads the pre-treatment buffer used when pre-service and comprises following component: stachyose, alum, Fructose Diphosphate, sodium hexametaphosphate and glycocoll, and the total concentration of stachyose, alum, Fructose Diphosphate, sodium hexametaphosphate, glycocoll is 3.5-7.5g/L, the pH value of damping fluid is 7.2-7.6.
Preferably, the concentration of each component in damping fluid is:
The solvent of described pre-treatment buffer is water.
Described pretreated concrete steps are: gold mark pad is soaked 1.5 ~ 2h in pretreatment fluid, take out and are put in 36 ~ 38 DEG C of oven dry.
Described pre-treatment buffer can use the various conventional pH adjusting agent in this area to carry out the adjustment of pH value.
Third aspect present invention provides described quantitative detection lipoprotein vitamin K deficiency or antagonist-II to induce the detection kit of the albumen PIVKA-II produced to induce the purposes of albumen PIVKA-II detection field produced at vitamin K deficiency or antagonist-II.
Beneficial effect of the present invention is:
Quantitative detection vitamin K deficiency provided by the present invention or antagonist-II induce the detection kit of the albumen PIVKA-II produced to induce the albumen PIVKA-II produced to be detected by fluorescent micro-ball immune chromatography technology vitamin K deficiency or antagonist-II first, have high sensitivity and high specific concurrently, the content of the albumen PIVKA-II of vitamin K deficiency or antagonist-II induction generation can be detected fast.In addition, described detection kit has the advantages such as operation is fast and convenient, result is accurate, economic and practical, disturb little by the serious blood fat of serum (or blood plasma), haemolysis, as serum (or blood plasma) haemoglobin≤500mg/L, triglyceride≤30mg/dL, variation < 10% is affected on accuracy.
Embodiment
Below by way of specific instantiation, embodiments of the present invention are described, those skilled in the art the content disclosed by this instructions can understand other advantages of the present invention and effect easily.The present invention can also be implemented or be applied by embodiments different in addition, and the every details in this instructions also can based on different viewpoints and application, carries out various modification or change not deviating under spirit of the present invention.
Before further describing the specific embodiment of the invention, should be understood that protection scope of the present invention is not limited to following specific specific embodiments; It is also understood that the term used in the embodiment of the present invention is to describe specific specific embodiments, instead of in order to limit the scope of the invention; In instructions of the present invention and claims, unless explicitly pointed out in addition in literary composition, singulative " ", " one " and " this " comprise plural form.
When embodiment provides numerical range, should be understood that except non-invention is otherwise noted, between two end points of each numerical range and two end points, any one numerical value all can be selected.Unless otherwise defined, all technology used in the present invention are identical with the meaning that those skilled in the art of the present technique understand usually with scientific terminology.Except the concrete grammar used in embodiment, equipment, material, according to those skilled in the art to the grasp of prior art and record of the present invention, any method of prior art that is similar with the method described in the embodiment of the present invention, equipment, material or that be equal to, equipment and material can also be used to realize the present invention.
Unless otherwise indicated, disclosed in the present invention experimental technique, detection method, preparation method all adopt the routine techniques of the molecular biology of the art routine, biological chemistry, chromatin Structure and analysis, analytical chemistry, cell chulture, recombinant DNA technology and association area.These technology are existing in existing document improves explanation, specifically can see the MOLECULAR CLONING:A LABORATORY MANUAL such as Sambrook, Second edition, Cold Spring HarborLaboratory Press, 1989and Third edition, 2001; Ausubel etc., CURRENT PROTOCOLS INMOLECULAR BIOLOGY, John Wiley & Sons, New York, 1987and periodic updates; The seriesMETHODS IN ENZYMOLOGY, Academic Press, San Diego; Wolffe, CHROMATINSTRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; METHODSIN ENZYMOLOGY, Vol.304, Chromatin (P.M.Wassarman and A.P.Wolffe, eds.), AcademicPress, San Diego, 1999; With METHODS IN MOLECULAR BIOLOGY, Vol.119, ChromatinProtocols (P.B.Becker, ed.) Humana Press, Totowa, 1999 etc.
Embodiment 1
The preparation of test card of the present invention:
1) pre-treatment buffer is used to carry out pre-service to gold mark pad, pre-treatment buffer is: stachyose 2g/L, alum 0.5g/L, Fructose Diphosphate 1.5g/L, sodium hexametaphosphate 0.3g/L, the aqueous solution of glycocoll 1.88g/L, pH=7.4, pretreated concrete steps are: gold mark pad is soaked 2h in pretreatment fluid, take out and are put in 37 DEG C of oven dry; Then pad with the pretreated gold mark of albumen PIVKA-II antibody-solutions spraying that vitamin K deficiency or the antagonist-II induction of fluorescent microsphere mark produce, the gold mark pad of albumen PIVKA-II antibody produced induced by obtained bag by vitamin K deficiency or antagonist-II, in solution, the mass ratio of fluorescent microsphere and antibody is 5:1, the concentration of solution is 10mg/ml, and quantity for spray is 4ul/cm;
2) on the detection line and nature controlling line of nitrocellulose filter, spray 1mg/ml respectively vitamin K deficiency or antagonist-II induce albumen PIVKA-II antibody-solutions and sheep anti-mouse antibody solution that produce, quantity for spray is 1ul/cm, obtained bag by after nitrocellulose filter;
3) by sample pad, step 1) preparation gold mark pad, step 2) nitrocellulose filter, the adsorptive pads prepared be pasted onto on PVC base plate successively, cutting obtains the Test paper card of wide 3-5mm; Finally Test paper is snapped fits into the obtained detection kit that gets stuck.Standard lines curve:
Be 0 by concentration respectively, 1.0, 1.5, 2.0, 5.0, 20, 100, 200, 500, 800, the vitamin K deficiency of 1000ng/ml or antagonist-II induce albumen PIVKA-II buffer solution produced to drip in sample pad, each concentration establishes 5 repetitions (testing result gets 5 mean values repeated), after rete is analysed 10 minutes, use immunoassay instrument by acquisition testing line (T) and nature controlling line (C) band fluorescence signal, the sensing range of analyser to fluorescence signal is AD value 0-10000, calculate T/C signal value, set up calibration curve, wherein Y-axis is T/C signal value, X-axis is standard items actual value.
Vitamin K deficiency or antagonist-II induce albumen PIVKA-II anti-interference and specific detection that produce:
Detection sample drop is added in sample pad, 5 repetitions (testing result gets 5 mean values repeated) established by each sample, after rete is analysed 10 minutes, the T/C value obtained when detecting sample is compared with typical curve, vitamin K deficiency in acquisition detection sample or antagonist-II induce the detection data of albumen PIVKA-II content produced, albumen PIVKA-II content data and true vitamin K deficiency produced or antagonist-II the vitamin K deficiency or antagonist-II that detect acquisition is induced to induce albumen PIVKA-II content data produced to contrast again, obtain accuracy and affect deviate.
Sample 1:1.5ng/ml vitamin K deficiency or antagonist-II induce the albumen PIVKA-II, 50mg/L haemoglobin, the 50mg/dL triglyceride that produce;
Sample 2:3ng/ml vitamin K deficiency or antagonist-II induce the albumen PIVKA-II, 500mg/L haemoglobin, the 10mg/dL triglyceride that produce;
Sample 3:6ng/ml vitamin K deficiency or antagonist-II induce the albumen PIVKA-II, 100mg/L haemoglobin, the 20mg/dL triglyceride that produce;
Sample 4:12ng/ml vitamin K deficiency or antagonist-II induce the albumen PIVKA-II, 150mg/L haemoglobin, the 30mg/dL triglyceride that produce;
Sample 5:18ng/ml vitamin K deficiency or antagonist-II induce the albumen PIVKA-II, 200mg/L haemoglobin, the 40mg/dL triglyceride that produce;
Blank: 50mg/L haemoglobin, 50mg/dL triglyceride;
The vitamin K deficiency of the detection that sample 1-5 obtains or antagonist-II induce albumen PIVKA-II content data produced to be respectively 1.49ng/ml, 3.01ng/ml, 5.99ng/ml, 11.95ng/ml, 18.02ng/ml, accuracy affect variation < 10%, do not find when detecting in blank that obvious fluorescence signal changes.
Embodiment 2
The preparation of comparative example test card: adopt 25mM glycine buffer pre-service gold mark pad, other reagent and experimental technique are all with embodiment 1.
1) 25mM glycine buffer pre-service gold mark pad is adopted, then pad with the pretreated gold mark of albumen PIVKA-II antibody-solutions spraying that vitamin K deficiency or the antagonist-II induction of fluorescent microsphere mark produce, the gold mark pad of albumen PIVKA-II antibody produced induced by obtained bag by vitamin K deficiency or antagonist-II, in solution, the mass ratio of fluorescent microsphere and antibody is 5:1, the concentration of solution is 10mg/ml, and quantity for spray is 4ul/cm;
2) on the detection line and nature controlling line of nitrocellulose filter, spray 1mg/ml respectively vitamin K deficiency or antagonist-II induce albumen PIVKA-II antibody-solutions and sheep anti-mouse antibody solution that produce, quantity for spray is 1ul/cm, obtained bag by after nitrocellulose filter;
3) by sample pad, step 1) preparation gold mark pad, step 2) nitrocellulose filter, the adsorptive pads prepared be pasted onto on PVC base plate successively, cutting obtains the Test paper card of wide 3-5mm; Finally Test paper is snapped fits into the obtained detection kit that gets stuck.
Vitamin K deficiency or antagonist-II induce sensitivity and the detectability contrast experiment of the albumen PIVKA-II produced:
Judge with fluorescent instrument, analyser be AD value 0-10000 to the sensing range of fluorescence signal, according to the performance of instrument, CUTOFF value is 50, under certain concentration, more than 90% test example AD value >=50, namely think that kit can be used in the detection under this concentration.
Adopt 5%BSA normal saline solution as dummy, dummy should not contain measured object.The vitamin K deficiency or the antagonist-II that get 0.1 ~ 60ug/L gradient concentration induce albumen PIVKA-II known sample produced to carry out sensitivity detection, and arrange a gradient at interval of 0.2ng/ml, each gradient arranges 300 samples, record testing result.The lowest detection of the kit of result display prepared by embodiment 1 is limited to 0.2ng/ml; And the lowest detectable limit of kit prepared by embodiment 2 is higher than 60ug/L.
In sum, detection kit provided by the present invention has good anti-interference and specificity, and has good sensitivity, and negative background is lower, effectively overcomes various shortcoming of the prior art and tool high industrial utilization.
Above-described embodiment is illustrative principle of the present invention and effect thereof only, but not for limiting the present invention.Any person skilled in the art scholar all without prejudice under spirit of the present invention and category, can modify above-described embodiment or changes.Therefore, such as have in art usually know the knowledgeable do not depart from complete under disclosed spirit and technological thought all equivalence modify or change, must be contained by claim of the present invention.
Claims (10)
1. one kind is quantitatively detected the detection kit of the albumen PIVKA-II of vitamin K deficiency or antagonist-II induction generation, comprise test card, test card comprises base plate, and be positioned at the sample pad be arranged in order from application of sample end of backplate surface, gold mark pad, nitrocellulose filter and adsorptive pads, described gold mark pad comprises albumen PIVKA-II antibody of vitamin K deficiency or antagonist-II induction generation, described nitrocellulose filter is coated with detection line and nature controlling line, vitamin K deficiency on described gold mark pad or antagonist-II induce albumen PIVKA-II antibody produced to adopt fluorescent microsphere mark.
2. the detection kit quantitatively detecting the albumen PIVKA-II of vitamin K deficiency or antagonist-II induction generation as claimed in claim 1, it is characterized in that, on described nitrocellulose filter, detection line is positioned at side close to application of sample end, and nature controlling line is positioned at side away from application of sample end.
3. the detection kit quantitatively detecting the albumen PIVKA-II of vitamin K deficiency or antagonist-II induction generation as claimed in claim 1, it is characterized in that, described detection line is coated with albumen PIVKA-II antibody of vitamin K deficiency or antagonist-II induction generation.
4. the detection kit quantitatively detecting the albumen PIVKA-II of vitamin K deficiency or antagonist-II induction generation as claimed in claim 1, is characterized in that, nature controlling line wraps by sheep anti-mouse antibody.
5. the detection kit quantitatively detecting the albumen PIVKA-II of vitamin K deficiency or antagonist-II induction generation as claimed in claim 1, it is characterized in that, described sample pad adopts damping fluid process, described damping fluid is selected from one or more the combination in PBS damping fluid, Tris-HCl damping fluid, glycine buffer, borate buffer solution and citrate-phosphate salt buffer, and the concentration of damping fluid is 20-200mM.
6. the detection kit quantitatively detecting the albumen PIVKA-II of vitamin K deficiency or antagonist-II induction generation as claimed in claim 1, it is characterized in that, described gold mark pad adopts damping fluid process, described damping fluid is selected from one or more combination of glycine buffer, Tris-HCl damping fluid, borate buffer solution, and the concentration of damping fluid is 5-50mM.
7. quantitative detection vitamin K deficiency according to claim 6 or antagonist-II induce the detection kit of the albumen PIVKA-II produced, it is characterized in that, increased response agent is also comprised in described damping fluid, described increased response agent be selected from PEG4000, PEG6000, PEG8000 and PEG20000 any one, the concentration of described increased response agent is 10 ~ 50g/L.
8. the detection kit quantitatively detecting the albumen PIVKA-II of vitamin K deficiency or antagonist-II induction generation as claimed in claim 1, it is characterized in that, also comprise and getting stuck, described getting stuck comprises back of the body card and upper cover, the described back of the body is arranged with test card draw-in groove, described test card is embedded in described test card draw-in groove, testing window and well is covered with on described, matching with the position of described detection line and nature controlling line in the position of described testing window, matches with the position of described sample pad in the position of described well.
9. the detection kit quantitatively detecting the albumen PIVKA-II of fat vitamin K deficiency or antagonist-II induction generation as claimed in claim 1, it is characterized in that, described detection kit is used for quantitatively detecting vitamin K deficiency or antagonist-II in serum or blood plasma and induces the content of the albumen PIVKA-II produced.
10. the quantitative detection fat vitamin K deficiency according to the arbitrary claim of claim 1 ~ 9 or antagonist-II induce the preparation method of the detection kit of the albumen PIVKA-II produced, and specifically comprise the steps:
1) spray pretreated golden mark with albumen PIVKA-II antibody-solutions that vitamin K deficiency or the antagonist-II induction of fluorescent microsphere mark produce to pad, the obtained gold mark pad comprising albumen PIVKA-II antibody of vitamin K deficiency or antagonist-II induction generation;
2) on the detection line and nature controlling line of nitrocellulose filter, spray vitamin K deficiency or antagonist-II respectively induce albumen PIVKA-II antibody and sheep anti-mouse antibody that produce, obtained bag by after nitrocellulose filter;
3) by sample pad, step 1) preparation gold mark pad, step 2) nitrocellulose filter, the adsorptive pads prepared be pasted onto on base plate successively, cutting obtains Test paper card; Finally Test paper is snapped fits into the obtained detection kit that gets stuck.
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| CN201410735809.9A CN104360079B (en) | 2014-12-05 | 2014-12-05 | A kind of PIVKA-II detection kit |
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| CN201410735809.9A CN104360079B (en) | 2014-12-05 | 2014-12-05 | A kind of PIVKA-II detection kit |
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| CN104360079B CN104360079B (en) | 2016-01-20 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109444422A (en) * | 2018-12-17 | 2019-03-08 | 福建省立医院 | A kind of detection card and its construction method and application based on UPT-LF quantitative determination blood-serum P IVKA-II |
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| US6893831B1 (en) * | 1999-12-14 | 2005-05-17 | Sanko Junyaku Co., Ltd. | Immunoassay of PIVKA-II |
| CN101377505A (en) * | 2008-04-16 | 2009-03-04 | 北京科美东雅生物技术有限公司 | Des-gamma-carboxy-pro-thrombin microplate chemiluminescence immune analysis determination reagent kit and preparing method thereof |
| CN102226779A (en) * | 2011-03-28 | 2011-10-26 | 中国人民解放军第三军医大学第三附属医院 | Electrochemical immunodetection method |
| CN102879559A (en) * | 2011-07-12 | 2013-01-16 | 上海执诚生物科技股份有限公司 | Real-time quantitative detection reagent and method of time-resolved fluorescence immune chromatography |
| CN103597351A (en) * | 2011-05-23 | 2014-02-19 | 积水医疗株式会社 | Method for inhibiting non-specific reaction in PIVKA-II measurement reagent |
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2014
- 2014-12-05 CN CN201410735809.9A patent/CN104360079B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6893831B1 (en) * | 1999-12-14 | 2005-05-17 | Sanko Junyaku Co., Ltd. | Immunoassay of PIVKA-II |
| CN101377505A (en) * | 2008-04-16 | 2009-03-04 | 北京科美东雅生物技术有限公司 | Des-gamma-carboxy-pro-thrombin microplate chemiluminescence immune analysis determination reagent kit and preparing method thereof |
| CN102226779A (en) * | 2011-03-28 | 2011-10-26 | 中国人民解放军第三军医大学第三附属医院 | Electrochemical immunodetection method |
| CN103597351A (en) * | 2011-05-23 | 2014-02-19 | 积水医疗株式会社 | Method for inhibiting non-specific reaction in PIVKA-II measurement reagent |
| CN102879559A (en) * | 2011-07-12 | 2013-01-16 | 上海执诚生物科技股份有限公司 | Real-time quantitative detection reagent and method of time-resolved fluorescence immune chromatography |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109444422A (en) * | 2018-12-17 | 2019-03-08 | 福建省立医院 | A kind of detection card and its construction method and application based on UPT-LF quantitative determination blood-serum P IVKA-II |
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| CN104360079B (en) | 2016-01-20 |
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