CN104215764A - Detection reagent plate used for diagnosis of hypertriglyceridemia - Google Patents
Detection reagent plate used for diagnosis of hypertriglyceridemia Download PDFInfo
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Abstract
The invention relates to a detection reagent plate used for diagnosis of hypertriglyceridemia (HTG). In the detection reagent plate, apolipoprotein A5 (ApoA5) is used as a biochemical index for diagnosis of the HTG to prepare an anti-ApoA5 monoclonal antibody pair. The anti-ApoA5 monoclonal antibody pair, which is marked with a colloidal gold marker, is fixed on a one-step detection reagent plate and is used as a capture antibody. The anti-ApoA5 monoclonal antibody pair, which is not marked, is accurately distributed on an immunochromatographic membrane in a transversal-bar shape and is used as a detection antibody. The content of the ApoA5 in a sample is detected and determined through display of the bars on the immunochromatographic membrane, so that the detection reagent plate is used for etiologic diagnosis of the HTG and can be used in effective prevention and treatment of cardiovascular diseases, such as coronary heart disease, stroke and the like. The detection reagent plate is simple and quick in use. Each part of reagent is packaged individually so that the detection reagent plate can be stored at room temperature and is convenient to store. The detection reagent plate can be used with cooperation of a colloidal gold color development quantitative detector and a detection result is compared with a preset Cutoff value and is reported so that quick quantitative detection and analysis of the ApoA5 through colloidal gold are achieved. Miss-determination and fault-determination which are caused by determination by eyes in color development with the colloidal gold are avoided and sensitivity of detection of the ApoA5 is improved.
Description
Technical field
The present invention relates to the biochemical indicator that ApoA5 (ApoA5) is diagnosed as hypertriglyceridemia (HTG), the technology of preparing that ApoA5 monoclonal antibody is right, the immunochromatography reaction technology of colloid gold label colour developing and utilize the antibody of ApoA5 to carry out the technical field of diagnosing high triglyceride (HTG), particularly one is used for the detection agent plate of diagnosing high triglyceride (HTG).
Technical background
The project detecting blood fat clinically has a lot, and the elementary item of each hospital general inspection has T-CHOL (TC), triglyceride (TG), apolipoprotein (Apo) B, Apo AI, lipoprotein (a) [Lp (a)] etc.Survey data research shows, high triglyceride, high cholesterol, low high-density lipoprotein (HDL)-C (HDL-C) and high low-density lipoprotein-C (LDL-C) together constitute the hazards of coronary heart disease.Apolipoprotein (Apo) abnormal with coronary heart disease and apoplexy closely related.Correctly diagnose as early as possible, be conducive to these serious diseases of early prevention and treatment.Apolipoprotein detects the exploitation of reagent, contributes to effective control of coronary heart disease and apoplexy.
Serum ApoAI can represent HDL level, is proportionate with HDL-C, its clinical meaning also broadly similar.But HDL is a series of grain size and the inhomogenous lipoprotein of composition, and under pathological state, HDL subclass often changes with composition, therefore the fixed and HDL-C of going up and down with different paces of ApoAI changes completely proportional.The ratio of ApoB100 nearly 90% is distributed in LDL, so serum ApoB main representative LDL level, it and serum LDL-C level are obvious positive correlation, and the clinical meaning of ApoB level height is also similar to LDL-C.But, in limited instances, high ApoB100 mass formed by blood stasis can be there is and the normal situation of LDL-C concentration, so these two kinds of apolipoprotein reliabilities and accuracy are very not satisfactory.
ApoA5 belongs to ApoAI/CIII/AIV gene group, at liver expression, relevant with HDL/VLDL.The single nucleotide variations (SNP) of ApoA5 gene promoter directly affects triglyceride levels.Experimental study shows, in the blood plasma of expressing the genetically modified mouse of mankind ApoA5, the concentration of triglyceride drops to 1/3rd of contrast mouse, and adds 4 times in the mouse lacking this gene.ApoA5 has very high Sensitivity and Specificity as the etiological diagnosis index of hypertriglyceridemia.
High levels of triglycerides can make little dense low-density lipoprotein (sLDL) concentration raise, and has stronger atherogenicity effect.Triglyceride levels is higher, and the cholesteryl ester in low-density lipoprotein is more.People are verified, and the cholesterol in atherosclerotic plaque mainly comes from low-density lipoprotein, and low-density lipoprotein is maximum containing cholesterol in blood plasma, is also the lipoprotein that atherogenic is the strongest.In addition, epidemiological study shows, high levels of triglycerides makes high-density lipoprotein concentration decline, and high-density lipoprotein concentration is by the impact of triglyceride levels, and both are negative correlation.Recent research shows, blood triglyceride levels is abnormal abnormal closely related with the body intravascular coagulation factor, plasma triglyceride level raises and body intravascular coagulation factor level can be caused to raise, thus promotion blood clotting, and tissue plasminogen activator's inhibitor (PAI) can be made to synthesize increase, suppress fibrinolysis, form hypercoagulative state, promote the generation of coronary heart disease.Have experimental result to show, triglyceride concentration rising 1mmol/L, the cardiovascular unexpected incidence male sex rises 32%, and women is 76%.After the impact correcting high-density lipoprotein (HDL), the level of significance male sex rises 14%, and women is 37%.These results show that the rising of serum triglyceride concentration is the important risk factor of angiocardiopathy.
Primary hypertriglyceridemiapatients patients often has hypertension and insulin resistance simultaneously.Insulin resistance is often referred to patient and tends to non-insulin dependence abnormal carbohydrate metabolism, and such patient often forms coronary artery disease earlier.Usually lipid triad, hypercoagulative state, hypertension, insulin resistance are called metabolic syndrome.Confirm that primary hypertriglyceridemiapatients patients is often associated with other exception in metabolic syndrome.The clinical meaning of hypertriglyceridemia is this.And seeming particularly important as the index ApoA5 directly affecting triglyceride levels, the single nucleotide polymorphism of ApoA5 is expected to as risk indicateing arm and the therapy target reducing serum lipid concentrations.
ApoA5 is applied to clinical practice to reach the quick diagnosis of the cause of disease of hypertriglyceridemia by the right technology of preparing of monoclonal antibody and colloidal gold immunochromatographimethod technology by the present invention, and the object that the angiocardiopathy such as coronary heart disease and apoplexy is is effectively prevented and treated.
Sandwich double-antibody technique is based upon the newer rear antibody technique of biological technical field on monoclonal antibody technique.Double antibody sandwich method application antibody is to identifying that capture antigen is carried out in two sites of antigen, avoid the competition binding in the same site of antibody and antigen, decrease the steric hindrance of antigenic determinant, so more general antigen capture method is more responsive, specificity is stronger, repeatability is better, is particularly useful for the detection of the little sample of antigenic content.The present invention, in conjunction with the antibody pair for ApoA5 of this technological development, more highlights its significance to micro biochemical index (antigen) in detection HTG patient blood.
Summary of the invention
The object of this invention is to provide a kind of detection agent plate for diagnosing high triglyceride, utilize the biochemical indicator that ApoA5 (ApoA5) is diagnosed as hypertriglyceridemia (HTG), the monoclonal antibody of preparation ApoA5 to and corresponding colloidal gold immunochromatographimethod quick detection reagent plate, detection time is 8-12 minute only, by measuring the concentration of the ApoA5 in human blood, and etiological diagnosis is carried out to hypertriglyceridemia (HTG), thus reach effective control of the angiocardiopathy such as coronary heart disease and apoplexy.
The object of the present invention is achieved like this:
For a detection agent plate for diagnosing high triglyceride, comprise ApoA5 monoclonal antibody and unlabelled ApoA5 monoclonal antibody, sheep anti-mouse igg antibody, polyethylene board, Polyvinylchloride lining form, filter membrane, all-glass paper, polyester film, immunochromatography film and the thieving paper composition of colloid gold label.Detect agent plate surface level to be followed successively by from bottom to top: absorption of sample district, Jin Biao monoclonal antibody district, immobilized antibody district and suction zones, absorption of sample district lay for lay filter membrane and all-glass paper on polyethylene board and Polyvinylchloride lining form, suction zones is thieving paper, it is characterized in that: Jin Biao monoclonal antibody district is: the ApoA5 monoclonal antibody of colloid gold label be evenly sprayed on the polyester film of unit area; Immobilized antibody district is: unlabelled ApoA5 monoclonal antibody and sheep anti-mouse igg antibody are fixed on the immunochromatography film of unit area.
The preparation method of described colloid gold label ApoA5 monoclonal antibody: get collaurum and corresponding ApoA5 monoclonal antibody that appropriate radius is 20-60nm respectively, combination is made by magnetic agitation vibration under the condition of pH7.2-9.2, adding mass concentration is that 2-4% bovine serum albumin(BSA) BSA is as stabilizing agent, adopt centrifuge method to remove unconjugated monoclonal antibody and unstabilized colloid gold particle and agglutinator, the peony bottom centrifuge tube is precipitated as collaurum-ApoA5 monoclonal antibody complex.
The antibody of described anti-ApoA5 is monoclonal antibody pair.
The ApoA5 immunity BALB/c mouse of described anti-ApoA5 monoclonal antibody purifying, obtains the spleen cell of antigenic stimulus, merges the myeloma cell line SP2/0 of this spleen cell and mouse; Utilize HAT Selective agar medium to select hybridoma, and utilize ELISA method Screening and Identification to produce the cell of anti-ApoA5 antibody; Obtain the cell line of the monoclonal antibody of secreting high specific after carrying out three time clonings, the cell line of the monoclonal antibody that the generation that preservation screens is combined with ApoA5 high specific, produces the monoclonal antibody of anti-ApoA5 by mouse ascites.Recycling Protein A-SePharose affinity column is further purified the monoclonal antibody of anti-ApoA5 in ascites, ELISA and Western-Blot identifies the monoclonal antibody of separation and purification.
The screening that the grand antibody of anti-ApoA5 monoclonal antibody is right, the subclone of main employing suppression method screening high-affinity, and then the basis being added experimental result in repeatedly cloning and ELISA is carried out the mensuration of Competitive assays epitope analysis and affinity, obtain the monoclonal antibody cell line of different epi-position, further production ascites purifying obtains volume antibody, carry out individual plant mark, ELISA direct method is adopted to carry out the working concentration titration of labelled antibody, then adopt ELISA A competitive inhibition method to carry out single labeling antibody epi-position to measure, thus filter out the sandwich antibody pair of efficient affinity.
Provided by the present inventionly detecting agent plate for diagnosing high triglyceride (HTG), is using ApoA5 as biochemical indicator to diagnose HTG.This comprises the right method of the monoclonal antibody of preparing above-mentioned ApoA5, and utilizes colloidal gold labeled monoclonal antibody and immune chromatography method to detect the concentration of the ApoA5 in blood or serum.Specifically, be exactly utilize the anti-ApoA5 monoclonal antibody of gold mark as first antibody, be used for capture antigen; Accurately be used in the horizontal stripe shape unmarked ApoA5 monoclonal antibody be distributed on immunochromatography film identifying antigen, by the striped on display immunochromatography film, association colloid gold instrument, judges the content of the ApoA5 detected in sample, thus screening and diagnosis HTG.
Compare with technology with existing index, the present invention has outstanding advantage and practicality:
1, detection specificity is practical by force.ApoA5 belongs to ApoAI/CIII/AIV gene group, at liver expression, relevant with HDL/VLDL.The single nucleotide variations (SNP) of ApoA5 gene promoter directly affects triglyceride levels.ApoA5 has very high Sensitivity and Specificity as the etiological diagnosis index of hypertriglyceridemia.Present invention, avoiding like product in the market can only performance results merely, but can not clearly state the shortcoming of the cause of disease.Hypertriglyceridemia is that ApoA5 gene mutation reason causes once finding, and can carry out corresponding treatment for the cause of disease, thus fundamentally solves the prevention and therapy of the relevant disease of hypertriglyceridemia and initiation thereof.
2, detect quantitatively and fast.The present invention according to colloid gold particle under specific wavelength, to the absorption of light this characteristic relevant to the amount of colloid gold particle with scattering, colloid gold particle is detected to the absorption of light and scattering by the photoelectric sensor in instrument, thus the antigenic content obtained in sample to be checked, thus realization detection reagent is promoted to full basis weight products by traditional qualitative, sxemiquantitative.Native system more domestic other whole detection time quantitatively detect and shorten dramatically, and export from the blood sampling that starts to of instrument to going out result, can complete in 15 minutes, doctor can make Treatment and diagnosis scheme to patient subsequently.
Two high-tech technology (detecting reagent+quantitatively instrument) effectively combine by the present invention, achieve the target of POCT (detection of Point-Of-Care Testing bedside) quick " quantitatively ", achieve effective, accurate statistics analysis and the clinical practice of test data, fill the domestic gaps, and import can be substituted completely.
3, the present invention changes HTG screening and diagnoses the limitation that just can must be detected by professional or instrumentation.Agent plate is simple to operate, and sample can be whole blood, serum or blood plasma, can directly with referring to that blood examination is surveyed; Collaurum instrument is easy to carry, easy and simple to handle.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the invention will be further described.
Fig. 1 is planar structure areal map of the present invention.
Fig. 2 is profile structural drawing of the present invention.
In figure: 1, absorption of sample district; 2, Jin Biao monoclonal antibody district; 3, immobilized antibody district; 4, suction zones; 5, base plate and one deck PVC sheet; 6, immobilization ApoA5 monoclonal antibody immunochromatography film; 7, the polyester film of gold mark ApoA5 monoclonal antibody compound; 8, absorption of sample district, 8a is layer of glass paper, 8b is one deck filter membrane; 9, the suction zones of one deck thieving paper composition.
Fig. 3 is that ApoA5 concentration of the present invention and collaurum develop the color the curve map of fringe intensity.
Embodiment
The invention will be further described by reference to the accompanying drawings.
As shown in Figure 1 and Figure 2, for a detection agent plate for diagnosing high triglyceride, comprise ApoA5 (ApoA5) monoclonal antibody and unlabelled ApoA5 monoclonal antibody, sheep anti-mouse igg antibody, polyethylene board, Polyvinylchloride lining form, filter membrane, all-glass paper, polyester film, immunochromatography film and the thieving paper composition of colloid gold label.
As shown in Figure 1, detect agent plate surface level to be followed successively by from bottom to top: absorption of sample district 1, Jin Biao monoclonal antibody district 2, immobilized antibody district 3, and suction zones 4; As shown in Figure 2, the base plate be made up of polyethylene board and Polyvinylchloride lining form and one deck PVC sheet 5, the absorption of sample district 8 be made up of layer of glass paper 8a and one deck filter membrane 8b, the suction zones 9 of one deck thieving paper composition, is characterized in that: Jin Biao monoclonal antibody district is: the polyester film 7 ApoA5 monoclonal antibody of colloid gold label being evenly sprayed on the gold mark-ApoA5 monoclonal antibody compound that the polyester film of unit area forms; Immobilized antibody district is: unlabelled ApoA5 monoclonal antibody and sheep anti-mouse igg antibody are fixed on the immobilization ApoA5 monoclonal antibody immunochromatography film that the immunochromatography film of unit area forms.
Embodiment one, the production method of hypertriglyceridemia (HTG) diagnostic detect reagent plate:
1, the preparation that the monoclonal antibody of ApoA5 is right
Immune BALB/c mouse is distinguished as antigen with the ApoA5 of purifying, the 21 days immunity second time in interval, once at interval of immunity in 10 days later, immune 3-4 time altogether, then the spleen cell of immune mouse is taken out, with PEG, the myeloma cell line SP2/0 of this spleen cell and mouse is merged, HAT selective medium is utilized to screen hybridoma on 96 porocyte culture plates, the cell line of anti-ApoA5 is identified by ELISA method, the cell line of stably excreting highly specific monoclonal antibody is obtained after carrying out three time clonings, plate is adopted ELISA suppress method at same antigen bag, namely cells and supernatant is added in reference group, cells and supernatant and antigen mixture is added in suppression group, relatively two groups of OD value differences are different thus filter out the subclone of high-affinity.Then experimentally carry out additive process epitope analysis at indirect ELISA, namely add two individual plant cells and supernatant in reference group respectively, in mensuration group, add two strain cells and supernatant simultaneously, compare two groups of OD value (A
1, A
2and A
1+2), by formulae discovery below:
The basis of relatively result of calculation is carried out the mensuration of epitope analysis and affinity, AI value is less than 50% for being added feminine gender, namely the epi-position of the antibody recognition antigen of two cell line secretions is identical, AI value is greater than 50% for being added the positive, represent uncontested between two cell lines, namely the epi-position that two of antibody recognition antigen of two cell lines secretions are different, thus tentatively obtain the monoclonal antibody cell line for the different epi-position of antigen.Further the monoclonal antibody cell line obtained is injected into mouse peritoneal respectively and carries out ascites production, Protein A-Sepharose affinity column purifying ascites is utilized to obtain volume monoclonal antibody, with HRP enzyme, individual plant mark is carried out to the antibody of purifying, the working concentration titration of labelled antibody is carried out by ELISA direct method, then in reference group, enzyme labelled antibody is added with ELISA A competitive inhibition method, enzyme labelled antibody and unmarked test antibodies potpourri is added in mensuration group, relatively two groups of OD values, there is Competitive assays in the expression that reference group OD value is greater than mensuration group OD value, the epi-position being considered as two antibody recognition antigens is identical.Otherwise the epi-position being considered as two antibody recognition antigens when two antibody do not exist Competitive assays is different, thus final screening obtains the sandwich antibody pair of efficient affinity.
2, the method for the anti-ApoA5 monoclonal antibody of colloid gold label
Get collaurum and corresponding ApoA5 monoclonal antibody that appropriate radius is 40nm respectively, combination is made by magnetic agitation vibration under the condition of pH8.0, adding mass concentration is that the bovine serum albumin(BSA) BSA of 3% is as stabilizing agent, adopt centrifuge method to remove unconjugated monoclonal antibody and unstabilized colloid gold particle and agglutinator, the peony bottom centrifuge tube is precipitated as collaurum-ApoA5 monoclonal antibody complex.
3, golden labeling antibody compound is sprayed on polyester film
With polyglycol washing colloids gold-monoclonal antibody complex, centrifugal removing supernatant, obtain peony precipitation, the precipitation buffer solution after purifying, is applied on polyester film with spraying equipment, dries.
4, the bag quilt of immunochromatography film
Rule on nitric acid cellulose fiber film with another monoclonal antibody solution of the anti-ApoA5 monoclonal antibody centering of purifying and sheep anti-mouse igg antibody solution spraying equipment, the length of line is 3mm, and the dripping quantity of every bar line on the nitric acid cellulose fiber film that 3mm is wide is 5-7ng.Two kinds of antibody are respectively ApoA5 and sheep anti-mouse igg antibody from the bottom to top, every bar line interval 3mm.Bag is that 3%BSA solution is closed by good immunochromatography film mass concentration.
5, agent plate equipment
Plastic polyethylene plate, as prop carrier, is stained with one deck polychlorovinyl sheet material above as lining form, the absorption of sample district be made up of one deck filter membrane and layer of glass from the bottom to top, is secondly three layers of polyester film having adsorbed collaurum-monoclonal antibody compound.Then be nitrocellulose membrane, most last layer is water accepting layer, and be made up of one deck filter paper, the special adhesive tape in outside is encapsulated.
The using method of embodiment two, HTG diagnostic detect reagent plate:
1, the use of agent plate
Detection to whole blood: instill in the hole of check-out console with dropper fetching blood 100ul, 8-12 minute observations.Be the positive when there is two bands.Only having during a band is feminine gender.One band all without time represent that check-out console lost efficacy.
Detection to serum, blood plasma: with dropper get centrifugal good serum, blood plasma 100ul vertically instills in the hole of check-out console, 8-12 minute observations.Be the positive when there is two bands.Only having during a band is feminine gender.One band all without time represent that check-out console lost efficacy.
2, upper machine quantitatively detects, two kinds of modes between point instant and full-time:
Immediate mode: after blood sample instillation agent plate, namely agent plate is inserted in instrument, manually boots application program and test.
Full-time mode: after blood sample is put into agent plate, operator oneself timing is after 15 minutes.Blood sample agent plate is inserted in instrument, automatically starts application program and test.
3, testing process:
1) blood sample is instilled agent plate, darkroom put into by reacted test sample;
2) open test procedure and input relevant information and then click instant test or full time test;
3) systematic analysis, marks and calculates test sample test figure;
4) test result exports.
4, typical curve is formulated
Containing 0,31,63,125,250, the ApoA5 standard antigen of 500ng/ml instills agent plate respectively, the collaurum instrument (model JH-2) that application our company produces, agent plate is put into darkroom, open test procedure and input relevant information and then click instant test, the colored intensity data of record colloidal gold reagent plate detection line, with ApoA5 concentration (ng/ml) for horizontal ordinate, striped colored intensity is ordinate, drawing standard curve.
During product batch production, the product of every batch all must prepare corresponding typical curve, is supplied to end user.
5, the ApoA5 concentration in serum is measured
The serum instillation agent plate to be detected of suitable multiple dilution, the collaurum instrument (model JH-2) that application our company produces, agent plate is put into darkroom, open test procedure and input relevant information and then click instant test, the colored intensity data of record colloidal gold reagent plate detection line, according to above-mentioned typical curve, calculate the concentration value of trying to achieve ApoA5 in serum.If be set as automatic mode, detector automatically will record, calculate, and printed report testing result.
6, result judges
According to cutoff value (the cut off value) 215ng/ml of ApoA5 as hypertriglyceridemia (FHTG) etiological diagnosis index, concentration lacks lower than the be judged to be ApoA5 of this value, has the danger of hypertriglyceridemia; Maybe can judge the Etiological of ApoA5 gene mutation as hypertriglyceridemia.Under collaurum instrument automatic mode, detectable concentration value and the Cutoff value preset of ApoA5 compare and automatically report the result of determination of hypertriglyceridemia.
Claims (3)
1., for a detection agent plate for diagnosing high triglyceride, comprise ApoA5 (ApoA5) monoclonal antibody and unlabelled ApoA5 monoclonal antibody, sheep anti-mouse igg antibody, polyethylene board, Polyvinylchloride lining form, filter membrane, all-glass paper, polyester film, immunochromatography film and the thieving paper composition of colloid gold label.Detect agent plate surface level to be followed successively by from bottom to top: absorption of sample district, Jin Biao monoclonal antibody district, immobilized antibody district and suction zones, absorption of sample district lay for lay filter membrane and all-glass paper on polyethylene board and Polyvinylchloride lining form, suction zones is thieving paper, it is characterized in that: Jin Biao monoclonal antibody district is: the ApoA5 monoclonal antibody of colloid gold label be evenly sprayed on the polyester film of unit area; Immobilized antibody district is: unlabelled ApoA5 monoclonal antibody and sheep anti-mouse igg antibody are fixed on the immunochromatography film of unit area.
2. detect agent plate for diagnosing high triglyceride as claimed in claim 1, it is characterized in that the preparation method of described colloid gold label ApoA5 monoclonal antibody: get collaurum and corresponding ApoA5 monoclonal antibody that radius is 20-60nm respectively, combination is made by magnetic agitation vibration under the condition of pH7.2-9.2, adding mass concentration is that 2-4% bovine serum albumin(BSA) BSA is as stabilizing agent, centrifuge method is adopted to remove unconjugated monoclonal antibody and unstabilized colloid gold particle and agglutinator, peony bottom centrifuge tube is precipitated as collaurum-ApoA5 monoclonal antibody complex.
3. detect agent plate for diagnosing high triglyceride as claimed in claim 1, it is characterized in that described detection agent plate and collaurum develop the color instrument with the use of, and compare with the Cutoff value preset and automatically report testing result, achieve the analysis of ApoA5 collaurum Quantitative detection, avoid the erroneous judgement of failing to judge that collaurum colour developing with the naked eye judges to cause, improve the sensitivity that ApoA5 detects.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104597248A (en) * | 2015-01-05 | 2015-05-06 | 中南大学湘雅二医院 | Method and kit for detecting biological activity of apolipoprotein ApoA5 non-diagnostically |
CN107132356A (en) * | 2017-04-27 | 2017-09-05 | 北京航空航天大学 | A kind of colloidal gold test paper card for detecting carcinoma of urinary bladder and preparation method thereof |
-
2014
- 2014-07-28 CN CN201410362304.2A patent/CN104215764A/en active Pending
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104597248A (en) * | 2015-01-05 | 2015-05-06 | 中南大学湘雅二医院 | Method and kit for detecting biological activity of apolipoprotein ApoA5 non-diagnostically |
CN104597248B (en) * | 2015-01-05 | 2016-11-23 | 中南大学湘雅二医院 | The method of a kind of non-diagnostic purpose detection ApoA5 biologic activity and test kit |
CN107132356A (en) * | 2017-04-27 | 2017-09-05 | 北京航空航天大学 | A kind of colloidal gold test paper card for detecting carcinoma of urinary bladder and preparation method thereof |
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Application publication date: 20141217 |