CN104360082A - AFP and GP73 combined detection kit - Google Patents
AFP and GP73 combined detection kit Download PDFInfo
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- CN104360082A CN104360082A CN201410736170.6A CN201410736170A CN104360082A CN 104360082 A CN104360082 A CN 104360082A CN 201410736170 A CN201410736170 A CN 201410736170A CN 104360082 A CN104360082 A CN 104360082A
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Abstract
The invention relates to the field of biological detection, and particularly relates to a detection kit for quantitatively detecting alpha fetoprotein (AFP) and Golgi protein-73 (GP-73), a preparation method and application thereof. The kit comprises an AFP detection test paper card and a GP-73 detection test paper card which are independent from each other, wherein each detection test paper card comprises a bottom plate, as well as a sample pad, a gold mark pad, a nitrocellulose film and a water absorption pad which are arranged on the surface of the bottom plate and are sequentially arranged from a feeding end. The kit detects AFP and GP73 by using a fluorescent microspherev immunochromatography technology for the first time, has sensitivity and specificity, has the advantages of quick and simple operation, accurate result and the like, and is economical and practical.
Description
Technical field
The present invention relates to field of biological detection, particularly relate to a kind of AFP, GP73 combined detection kit and its production and use.
Background technology
Alpha-fetoprotein (alpha fetoprotein, AFP) be a kind of glycoprotein that development of fetus is synthesized by liver and yolk bag in early days, along with the growth at age after development of fetus and birth, in blood, AFP level declines gradually, and the reference interval of HAS AFP concentration is 1-20 μ g/L.AFP is the special mark of the sensitivity of hepatocellular carcinoma (hepatocellular carcinoma, HCC), serum concentration of AFP more than lasting more than 5 weeks of lasting 4 weeks of 400 μ g/L or 200-400 μ g/L, strong suspicion hepatocellular carcinoma; Serum afp also has the size judgement of available liver cancer, the curative effect evaluation of liver cancer and Index for diagnosis simultaneously.But according to statistics, the patients with hepatocellular carcinoma serum afp of about 20%-30% is not high; The optimum hepatopathy of part, comprises oxyhepatitis, chronic hepatitis and large-area hepatonecrosis, also has the rising of Serum AFP.Therefore, there is false positive to a certain degree and false negative by Serum AFP as the mark of hepatocellular carcinoma.
Golgiosome glycoprotein-73 (GolgiProtein 73, GP73) also known as II type golgiosome memebrane protein (Golgiphosphoprotein2, GOLPH 2), its concentration in the patients serum such as chronic hepatitis, cirrhosis raises, and raise more obvious in hepatocellular carcinoma patients serum, with in significant difference between normal person or optimum hepatopathy patients.There are some researches show, serum GP73 is used for the diagnosis of liver cancer, is better than Serum AFP and detects.According to the literature, the sensitivity that GP73 detects liver cancer is 69%, and specificity is 75%.For the diagnosis of early liver cancer, the susceptibility of GP73 is 62%, and AFP susceptibility is only 25%.In the liver cancer patient of AFP level lower than 20 μ g/L, the crowd GP73 level of 57% is had significantly to raise.This shows, the application of GP73 can improve the recall rate of the liver cancer patient to AFP negative greatly.
To sum up, AFP is the biochemical marker of classical hepatocellular carcinoma, the Typical Representative of GP73 novel liver cancer marker.Comparatively speaking, the susceptibility of GP73 is higher, false negative to a certain degree and false positive is all there is in two kinds of indexs when applying separately, if by appropriate mode and means, joint-detection two indexs, complement one another, will contribute to the false negative and the false positive that reduce hepatocellular carcinoma clinical diagnosis further simultaneously, improving the diagnosis performance of these marks to hepatocellular carcinoma, is clinical and patient service better.
Summary of the invention
The shortcoming of prior art in view of the above, the object of the present invention is to provide detection kit of a kind of quantitative detection AFP, golgiosome glycoprotein-73GP73 and its production and use, for solving the problems of the prior art.
For achieving the above object and other relevant objects, the invention provides one and quantitatively detect AFP, the detection kit of golgiosome glycoprotein-73GP73, comprise mutual independently AFP Test paper card, golgiosome glycoprotein-73GP73 Test paper card, described Test paper card comprises base plate all separately, 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, the gold mark pad of described AFP Test paper card comprises AFP antibody, the gold mark pad of described golgiosome glycoprotein-73GP73 Test paper card comprises golgiosome glycoprotein-73GP73 antibody, described each nitrocellulose filter is coated with detection line and nature controlling line, AFP antibody on described gold mark pad, golgiosome glycoprotein-73GP73 antibody adopts fluorescent microsphere mark.
Preferably, the antibody on each gold mark pad adopts 180nm fluorescent microsphere mark, EX (nm)=650/Em (nm)=670, and have 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 each 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, the detection line of described AFP Test paper card is coated with AFP antibody; The detection line of described golgiosome glycoprotein-73GP73 Test paper card is coated with golgiosome glycoprotein-73GP73 antibody.
Antibody in each test card on gold mark pad can be identical antibody with the antibody on detection line, also can be different antibody.
Preferably, each nature controlling line wraps by sheep anti-mouse antibody.
Preferably, described each 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, each 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, each 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: each gold mark pad is soaked 1.5 ~ 2h in pretreatment fluid, takes out and be 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 two parallel test card draw-in grooves, described test card is embedded in described test card draw-in groove, be covered with two testing windows and two wells on described, matching with the detection line of two test cards and the position of nature controlling line respectively in the position of described two testing windows, matches with the position of the sample pad of two test cards in the position of described two wells.
Preferred, described in get stuck for plastics get stuck.
Preferred, be also provided with the translot of connection two wells between described two wells.
Preferably, described detection kit is used for the content quantitatively detecting AFP, golgiosome glycoprotein-73GP73 in serum or blood plasma.
The detection kit of quantitative detection AFP provided by the present invention, golgiosome glycoprotein-73GP73, 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 obtained when detecting sample is compared with typical curve, the AFP, the golgiosome glycoprotein-73GP73 content that detect in sample can be obtained.
Second aspect present invention provides the preparation method of the detection kit of described quantitative detection lipoprotein AFP, golgiosome glycoprotein-73GP73, comprises the steps:
1) each self-corresponding gold mark pad is sprayed respectively, the obtained gold mark pad comprising AFP antibody, golgiosome glycoprotein-73GP73 antibody respectively with AFP antibody, the golgiosome glycoprotein-73GP73 antibody-solutions of fluorescent microsphere mark;
2) on the detection line and nature controlling line of the nitrocellulose filter of AFP Test paper card, AFP antibody and sheep anti-mouse antibody is sprayed respectively; The detection line and nature controlling line of the nitrocellulose filter of golgiosome glycoprotein-73GP73 Test paper card spray golgiosome glycoprotein-73GP73 antibody and sheep anti-mouse antibody respectively;
3) by two cover sample pad, steps 1) prepare two cover gold mark pad, step 2) prepare two cover nitrocellulose filters, two cover adsorptive pads be pasted onto successively on base plate, cutting obtains Test paper card; Finally Test paper is snapped fits into the obtained detection kit that gets stuck.
Preferably, each 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, each 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 the detection kit of described quantitative detection lipoprotein AFP, golgiosome glycoprotein-73GP73 in the purposes of AFP, golgiosome glycoprotein-73GP73 detection field.
Beneficial effect of the present invention is:
AFP, golgiosome glycoprotein-73GP73 are detected by fluorescent micro-ball immune chromatography technology by the detection kit of quantitative detection AFP provided by the present invention, golgiosome glycoprotein-73GP73 first, have high sensitivity and high specific concurrently, the content of AFP, golgiosome glycoprotein-73GP73 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 each self-corresponding pretreated gold mark pad is sprayed respectively, the obtained gold mark pad comprising AFP antibody, golgiosome glycoprotein-73GP73 antibody respectively with AFP antibody, the golgiosome glycoprotein-73GP73 antibody-solutions of fluorescent microsphere mark; In solution, the mass ratio of fluorescent microsphere and antibody is 5:1, and the concentration of solution is 10mg/ml, and quantity for spray is 4ul/cm;
2) on the detection line and nature controlling line of the nitrocellulose filter of AFP Test paper card, AFP antibody and sheep anti-mouse antibody is sprayed respectively; The detection line and nature controlling line of the nitrocellulose filter of golgiosome glycoprotein-73GP73 Test paper card spray golgiosome glycoprotein-73GP73 antibody and sheep anti-mouse antibody respectively; Obtained two kinds of bags by after nitrocellulose filter, the concentration of spray solution is 1mg/ml, and quantity for spray is 1ul/cm;
3) by two cover sample pad, steps 1) prepare two cover gold mark pad, step 2) prepare two cover nitrocellulose filters, two cover adsorptive pads be pasted onto successively on base plate, cutting obtains Test paper card; Finally Test paper is snapped fits into the obtained detection kit that gets stuck.
Typical curve:
Be 0 respectively by concentration, 5,20,100,200,400,500,600,800, the AFP buffer solution of 1000ug/L drips 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, and X-axis is standard items actual value.
Be 0 by concentration respectively, 2, 5, 20, 60, 100, 200, 400, 600, 800, golgiosome glycoprotein-73GP73 the buffer solution of 1000ng/ml drips 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.
AFP, golgiosome glycoprotein-73GP73 anti-interference and specific detection:
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, obtain the detection data of the AFP detected in sample, golgiosome glycoprotein-73GP73 content, AFP detection obtained again, golgiosome glycoprotein-73GP73 content data and true AFP, golgiosome glycoprotein-73GP73 content data contrast, and obtaining accuracy affects deviate.
Sample 1:20ug/L AFP, 1ng/ml golgiosome glycoprotein-73GP73,50mg/L haemoglobin, 50mg/dL triglyceride;
Sample 2:50ug/L AFP, 3ng/ml golgiosome glycoprotein-73GP73,500mg/L haemoglobin, 10mg/dL triglyceride;
Sample 3:100ug/L AFP, 6ng/ml golgiosome glycoprotein-73GP73,100mg/L haemoglobin, 20mg/dL triglyceride;
Sample 4:400ug/L AFP, 150ng/ml golgiosome glycoprotein-73GP73,150mg/L haemoglobin, 30mg/dL triglyceride;
Sample 5:800ug/L AFP, 500ng/ml golgiosome glycoprotein-73GP73,200mg/L haemoglobin, 40mg/dL triglyceride;
Blank: 50mg/L haemoglobin, 50mg/dL triglyceride;
The AFP content data of the detection that sample 1-5 obtains is respectively 20.1ug/L, 49.9ug/L, 99.9ug/L, 400.05ug/L, 800.01ug/L; Golgiosome glycoprotein-73GP73 content data is respectively 1.01ng/ml, 2.99ng/ml, 6.02ng/ml, 149.99ng/ml, 500.03ng/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 each self-corresponding pretreated gold mark pad is sprayed respectively, the obtained gold mark pad comprising AFP antibody, golgiosome glycoprotein-73GP73 antibody respectively with AFP antibody, the golgiosome glycoprotein-73GP73 antibody-solutions of fluorescent microsphere mark; In solution, the mass ratio of fluorescent microsphere and antibody is 5:1, and the concentration of solution is 10mg/ml, and quantity for spray is 4ul/cm;
2) on the detection line and nature controlling line of the nitrocellulose filter of AFP Test paper card, AFP antibody and sheep anti-mouse antibody is sprayed respectively; The detection line and nature controlling line of the nitrocellulose filter of golgiosome glycoprotein-73GP73 Test paper card spray golgiosome glycoprotein-73GP73 antibody and sheep anti-mouse antibody respectively; Obtained three kinds of bags by after nitrocellulose filter, the concentration of spray solution is 1mg/ml, and quantity for spray is 1ul/cm;
3) by two cover sample pad, steps 1) prepare two cover gold mark pad, step 2) prepare two cover nitrocellulose filters, two cover adsorptive pads be pasted onto successively on base plate, cutting obtains Test paper card; Finally Test paper is snapped fits into the obtained detection kit that gets stuck.
The sensitivity of AFP, golgiosome glycoprotein-73GP73 and detectability contrast experiment:
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 AFP known sample getting 1 ~ 20ug/L gradient concentration carries out sensitivity detection, arranges a gradient at interval of 0.5ug/L, and each gradient arranges 40 samples, record testing result.The lowest detection of the kit of result display prepared by embodiment 1 is limited to 1.2ug/L; And the lowest detectable limit of kit prepared by embodiment 2 is higher than 20ug/L.
Adopt 5%BSA normal saline solution as dummy, dummy should not contain measured object.Golgiosome glycoprotein-73GP73 the known sample getting 1 ~ 20pg/ml gradient concentration carries out sensitivity detection, and arrange a gradient at interval of 0.5pg/ml, each gradient arranges 40 samples, record testing result.The lowest detection of the kit of result display prepared by embodiment 1 is limited to 2.5pg/ml; And the lowest detectable limit of kit prepared by embodiment 2 is higher than 20pg/ml.
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 AFP, the detection kit of golgiosome glycoprotein-73GP73, comprise mutual independently AFP Test paper card, golgiosome glycoprotein-73GP73 Test paper card test paper card, described Test paper card comprises base plate all separately, 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, the gold mark pad of described AFP Test paper card comprises AFP antibody, the gold mark pad of described golgiosome glycoprotein-73GP73 Test paper card comprises golgiosome glycoprotein-73GP73 antibody, described each nitrocellulose filter is coated with detection line and nature controlling line, AFP antibody on described gold mark pad, golgiosome glycoprotein-73GP73 antibody adopts fluorescent microsphere mark.
2. detection kit as claimed in claim 1, it is characterized in that, on described each 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. detection kit as claimed in claim 1, is characterized in that, the detection line of described AFP Test paper card is coated with AFP antibody; The detection line of described golgiosome glycoprotein-73GP73 Test paper card is coated with golgiosome glycoprotein-73GP73 antibody.
4. detection kit as claimed in claim 1, is characterized in that, each nature controlling line wraps by sheep anti-mouse antibody.
5. detection kit as claimed in claim 1, it is characterized in that, described each 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. detection kit as claimed in claim 1, it is characterized in that, described each gold mark pad adopts damping fluid process, and 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. detection kit according to claim 6, it is characterized in that, also comprise increased response agent 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. detection kit 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 two parallel test card draw-in grooves, and described test card is embedded in described test card draw-in groove, is covered with two testing windows and two wells on described, matching with the detection line of two test cards and the position of nature controlling line respectively in the position of described two testing windows, matches with the position of the sample pad of two test cards in the position of described two wells.
9. detection kit as claimed in claim 8, is characterized in that, be also provided with the translot of connection two wells between described two wells.
10. the preparation method of the detection kit according to the arbitrary claim of claim 1 ~ 9, specifically comprises the steps:
1) each self-corresponding gold mark pad is sprayed respectively, the obtained gold mark pad comprising AFP antibody, golgiosome glycoprotein-73GP73 antibody respectively with AFP antibody, the golgiosome glycoprotein-73GP73 antibody-solutions of fluorescent microsphere mark;
2) on the detection line and nature controlling line of the nitrocellulose filter of AFP Test paper card, AFP antibody and sheep anti-mouse antibody is sprayed respectively; The detection line and nature controlling line of the nitrocellulose filter of golgiosome glycoprotein-73GP73 Test paper card spray golgiosome glycoprotein-73GP73 antibody and sheep anti-mouse antibody respectively;
3) by two cover sample pad, steps 1) prepare two cover gold mark pad, step 2) prepare two cover nitrocellulose filters, two cover adsorptive pads be pasted onto successively on base plate, cutting obtains Test paper card; Finally Test paper is snapped fits into the obtained detection kit that gets stuck.
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| CN107345969A (en) * | 2016-05-05 | 2017-11-14 | 中国医学科学院基础医学研究所 | Purposes of the serum markers comprising AFP, GP73 and CEACAM1 in diagnosing hepatic diseases |
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