CN104714017A - Method for quantitatively detecting procalcitonin - Google Patents

Method for quantitatively detecting procalcitonin Download PDF

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CN104714017A
CN104714017A CN201510065249.5A CN201510065249A CN104714017A CN 104714017 A CN104714017 A CN 104714017A CN 201510065249 A CN201510065249 A CN 201510065249A CN 104714017 A CN104714017 A CN 104714017A
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procalcitonin
antibody
pct
antigen
fluorescence signal
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王键
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BEIJING HOMA BIOLOGICAL ENGINEERING Co Ltd
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BEIJING HOMA BIOLOGICAL ENGINEERING Co Ltd
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    • 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

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Abstract

The invention relates to the biological technical field, and discloses a method for quantitatively detecting procalcitonin. According to the method, a to-be-detected sample is mixed with a fluorescently labeled anti-procalcitonin first antibody, and a reaction liquid containing a procalcitonin antigen antibody complex is obtained; the reaction liquid in a reaction medium is respectively subjected to a reaction with an anti-procalcitonin second antibody and an procalcitonin antigen, and fluorescence signals are respectively detected; the content of procalcitonin in the obtained to-be-detected sample is calculated according to fluorescence signal values. The method applies the technique of combination of a double-antibody sandwich method and a competition method, has stability, linearity and accuracy all significantly improved, and has good application prospects.

Description

A kind of method of quantitative detection Procalcitonin
Technical field
The invention belongs to biological technical field, particularly a kind of method of quantitative detection Procalcitonin.
Background technology
Procalcitonin (procalcitonin, PCT) be peptide material before calcitonin without hormonal activity, calcitonin is only subject to just producing when hormonal stimulates at thyroid C cell, and Procalcitonin can be subject to secreting after proinflammatory stimulation is particularly subject to bacterial stimulation by the dissimilar cell of a lot of organ.
Procalcitonin a kind ofly specificity can distinguish the important symbol thing of the inflammatory reaction that bacteriological infection and other reason cause, and when serious bacterial, fungi, parasitic infection and pyemia and MOFE, its level in blood plasma raises.Clinical data shows, and illustrates to there is clinical relevant bacteriological infection during PCT concentration >0.1ng/mL, needs to adopt microbiotic to treat; As PCT concentration >0.5ng/mL, consider that patient may have the danger developing into severe septicemia or septic shock.PCT reflects the active degree of systemic inflammatory response.The factor affecting PCT level comprises size and type, the kind of bacterium, the degree of inflammation and the immunoreactive situation of infected organ.Detect the change of PCT concentration in case, be conducive to judging body bacteriological infection situation, judge body health situation, to clinical and basic research is all significant.
After the eighties in last century, immunology detection technology develops rapidly along with the maturation of monoclonal antibody, artificial synthetic polypeptide, gene engineering expression antigen and various labelling technique, immunoturbidimetry, rate nephelometry (INA), latex intensified turbidimetry (ITA) and chemiluminescence analytical technique replace traditional immunoprecipitation, immune agglutination thing gradually, make detection more quick, the application detecting (POCT) in recent years fast makes immunology operation more simple, convenient.
At present, panimmunity detection technique is applied to PCT and detects, both can be qualitative, can also be quantitative.Common method has radio immunoassay (RIA), enzyme-labeled immunity analytic approach (ELISA), colloidal gold method, quantitative chemical luminescence immunoassay (CLIA), but above method equal existing defects in application process.Radio-immunity (RIA), enzyme-labeled immunity (ELISA) sensitivity are higher, but complicated operation, keeping life are short; Collaurum is simple and quick, but can not quantitatively detect; The detection of quantitative chemical electrochemiluminescent immunoassay needs the long period and testing cost is high.Therefore a kind of method researching and developing detection Procalcitonin newly is also needed, so that fast, accurately detect the Procalcitonin in sample.
Summary of the invention
Goal of the invention of the present invention is a kind of method providing quantitative detection Procalcitonin, and its stability, linear, accuracy are all significantly increased.
In order to realize goal of the invention of the present invention, the present invention adopts following technical scheme:
Detected sample is mixed with fluorescently-labeled anti-Procalcitonin first antibody, obtains the reactant liquor containing Procalcitonin antigen-antibody complex; By this reactant liquor in reaction medium respectively with anti-Procalcitonin second antibody, Procalcitonin antigen-reactive, detect fluorescence signal respectively; According to typical curve and the content recording Procalcitonin in fluorescence signal value acquisition detected sample.
As preferably, described fluorescence labeling fluorescein used is Alexa Fluor line fluorescent element.
As preferably, described reaction medium is test strips.
More preferably, described anti-Procalcitonin second antibody, Procalcitonin antigen are fixed on the diverse location of same reaction medium.
Quantitative detection Procalcitonin method provided by the invention is a quantitative measurement technology based on immunofluorescence technique, combine double antibody sandwich method and competition law, sample to be tested and the damping fluid containing fluorescently-labeled anti-Procalcitonin first antibody are carried out being mixed to get reactant liquor in reagent bottle, the PCT antigen of the fluorescence labeling PCT antibody in mixed liquor and sample combines and forms antigen antibody complex; Extract reaction solution and react with anti-Procalcitonin second antibody in reaction medium, catch the Procalcitonin and fluorescently-labeled anti-Procalcitonin antibody complex that contain in testing sample, detect, collect the first fluorescence signal; Get gained reactant liquor in reaction medium with Procalcitonin antigen-reactive, the fluorescently-labeled anti-Procalcitonin antibody in competition binding reactant liquor, detect, collect the second fluorescence signal; According to the first fluorescence signal, the second fluorescence signal and typical curve, calculate the content obtaining Procalcitonin in detected sample.
Preferred scheme is described reaction medium is test strips, and described anti-Procalcitonin second antibody, Procalcitonin antigen are fixed on the diverse location of reaction medium, utilize immunochromatographic method to detect.Immunochromatographic method (immunochromatography) its principle is a certain zone special antibody being first fixed on nitrocellulose filter, after sample is immersed in cellulose nitrate one end of this drying, due to capillary action, sample will move forward along this film, when moving to the region being fixed with antibody, in sample corresponding antigen namely with this antibody generation specific binding, realize specific immune detection with immune colloid gold or immunofluorescence label.
Position test strips securing anti-Procalcitonin second antibody is detection line, drip to test strips when mixing sample and be diffused on the detection line of nitrocellulose filter by capillary action, the PCT second antibody on the tested survey line of fluorescently-labeled antigen antibody complex caught.PCT antigen in sample to be tested is more, and the compound on detection line gathers more, and the signal of fluorescence antibody is stronger.
Position test strips securing anti-Procalcitonin is nature controlling line, drips to test strips and be diffused on the nature controlling line of nitrocellulose filter by capillary action when mixing sample, fluorescently-labeled antigen antibody complex catch by the PCT antigen on nature controlling line.PCT antigen in blood sample is more, and the compound on nature controlling line gathers fewer, and the signal of fluorescence antibody is more weak.
By the standard items test strip of concentration known, the fluorescence signal accumulated value of detection line and nature controlling line collected respectively separately by instrument, calculates corresponding ratio with respective accumulated value.Concentration known and corresponding ratio are brought into computing formula and draws typical curve, finally calculate corresponding concentration value according to typical curve.
The technology that the approach application of quantitative detection Procalcitonin provided by the invention double-antibody method combines with competition law.Particularly, quantitatively detect in the process of Procalcitonin in test strips, detection line utilizes double-antibody method, and nature controlling line utilizes competition law.By a large amount of creative works successfully by two kinds of technological incorporation together, Procalcitonin content specificity is good, the range of linearity is good, accuracy is high for quantitatively detecting in testing sample for the results show the method for the present inventor.
Generally, double-antibody method is used for macromolecular detection; Competition law is used for micromolecular detection, and Small molecular belongs to haptens, does not have two or more site binding antibodies, therefore uses competition law.Procalcitonin is macromolecular substances, and in existing detection method, many adopt sandwich method principle to detect, but in actual application, find that the linear dependence of sandwich method is undesirable, and have impact on its accuracy, stability, the term of validity is also relatively short.At present, also not about utilizing two kinds of different Cleaning Principle for detecting the relevant report of Procalcitonin, this greatly, easily causes non-specific binding relevant with Procalcitonin molecular weight.
In some embodiments of the invention, in detection method provided by the invention, fluorescently-labeled anti-Procalcitonin antibody fluorescein used is Alexa Fluor line fluorescent element.
In some embodiments of the invention, the fluorescein that fluorescently-labeled anti-Procalcitonin antibody is used is Alexa 647.
Preferably, in reagent provided by the invention, anti-Procalcitonin antibody, Procalcitonin antigen are fixed on two diverse locations in same reaction medium.More preferably, in reagent provided by the invention, described reaction medium is specially test strips, is fixed with anti-Procalcitonin antibody, Procalcitonin antigen in test strips, and anti-Procalcitonin antibody, Procalcitonin antigen are separately fixed at two different positions, are respectively detection line, nature controlling line.
In other embodiment of the present invention, kit provided by the invention comprises PCT reaction plate, containing have cured the detection line of anti-Procalcitonin antibody and have cured the nature controlling line of Procalcitonin antigen.
In other embodiment of the present invention, the invention provides a kind of kit, it comprises PCT and detects damping fluid, PCT reaction plate, sampler and ID chip; PCT detects damping fluid and is sub-divided in Reagent Tube; PCT reaction plate has independent packaging, inside has drying agent; PCT detects damping fluid and PCT reaction plate is respectively self-assembled into box, separately transport.
The invention provides a kind of method of quantitative detection Procalcitonin, method provided by the invention comprises: get detected sample, mixes, obtain reactant liquor with fluorescently-labeled anti-Procalcitonin antibody; Extract reaction solution and react with anti-Procalcitonin second antibody, catch the Procalcitonin and fluorescently-labeled anti-Procalcitonin antibody complex that contain in testing sample, detect, collect the first fluorescence signal; Get gained reactant liquor and Procalcitonin antigen-reactive, the fluorescently-labeled anti-Procalcitonin antibody in competition binding reactant liquor, detect, collect the second fluorescence signal; According to the first fluorescence signal, the second fluorescence signal, calculate the content obtaining Procalcitonin in detected sample.Experimental result confirms, the stabilization of kit of quantitative detection Procalcitonin provided by the invention is high, good linearity, accuracy are high, easy to use.
Procalcitonin of the present invention (PCT) quantitative determination reagent kit (fluorescence immune chromatography method) is detected the components such as damping fluid, PCT reaction plate, sampler, ID chip and forms by PCT.Its preparation technology is summarized as follows:
(1) PCT detects damping fluid: detect containing the fluorescently-labeled anti-human PCT monoclonal antibody of 5%-25% in damping fluid, be BSA solution containing 5%-15% stabilizing agent, antiseptic is 0.1%-0.3% sodium azide solution.
(2) PCT reaction plate: reaction plate contains the detection line that have cured anti-human PCT monoclonal antibody and the nature controlling line that have cured anti-human PCT antigen.
Draw from principle, the method for the invention has merged sandwich method and competition law two kinds of methods, more effectively can prevent the generation of detection leakage phenomenon; The fusion of two kinds of methods in addition effective algorithm makes the term of validity time longer; The fusion of two kinds of methods, linear relationship R2 value is better.Instant invention overcomes competition law and be applied to macromolecular technological difficulties, its stability, linear, accuracy are all significantly increased, and have a good application prospect.
Accompanying drawing explanation
Fig. 1 shows embodiment 4 experimental result, and wherein, the PCT reaction plate of straight line 1 comprises, and detection line is anti-human PCT monoclonal antibody, and nature controlling line is the RLU value linearity curve of anti-human PCT antigen, R 2=0.995; The PCT reaction plate of straight line 2 comprises, and detection line is anti-human PCT monoclonal antibody, and nature controlling line is the RLU value linearity curve of sheep anti mouse, R 2=0.987.
Fig. 2 and Fig. 3 is the normal distribution of embodiment 5 control experiment result.
Embodiment
The invention discloses a kind of method of quantitative detection Procalcitonin, those skilled in the art can with reference to present disclosure, use the method, realize its application, special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are all deemed to be included in the present invention.The method of the invention and application are described by preferred embodiment, related personnel obviously can not depart from content of the present invention, spirit and scope this paper preparation method and application are changed or suitably change with combination, realize and apply the technology of the present invention.
In order to enable those skilled in the art understand technical scheme of the present invention better, below in conjunction with embodiment, set forth the present invention further:
Embodiment 1: the preparation of Procalcitonin immue quantitative detection reagent box
Procalcitonin (PCT) immue quantitative detection reagent box comprises: PCT detects damping fluid, PCT reaction plate, sampler, ID chip.Detecting damping fluid is sub-divided in Reagent Tube; Reaction plate has independent packaging, inside has drying agent; Detection damping fluid and reaction plate are respectively self-assembled into box, separately transport.
(1) PCT detects damping fluid: be made up of fluorescently-labeled anti-human PCT monoclonal antibody, stabilizing agent BSA solution, antiseptic Sodium azide and damping fluid.Detect containing the fluorescently-labeled anti-human PCT monoclonal antibody of 5%-25% in damping fluid, be BSA solution containing 5%-15% stabilizing agent, antiseptic is 0.1%-0.3% sodium azide solution.Concrete configuration method is as follows:
1. the process of antibody: the anti-human PCT monoclonal antibody of 40 μ L, adds marking fluid 40 μ L, leaves standstill 5min after mixing.
2. antibody is connected with fluorescein: in liquid upon mixing, adds 2 μ L Alexa 647 fluoresceins, after mixing, room temperature mixing 1h on blood mixer.
3. cessation reaction: step 2. in liquid in add the stop buffer of 1 μ L, after mixing, room temperature mixing 1h on blood mixer.
4. step 3. in add the reactant liquor of 1.8mL, and the 10%BSA mixing of 18 μ L, namely synthesizing fluorescently labeled anti-human PCT monoclonal antibody conserving liquid.
5. PCT detects damping fluid: with volume basis, the ratio of fluorescently-labeled anti-human PCT monoclonal antibody conserving liquid and reactant liquor is 1:9, is adding Sodium azide, makes the final concentration of Sodium azide reach 0.3%, finally carries out lucifuge process with masking foil.
6. gained PCT is detected the every 500 μ L packing one bottle of damping fluid, and with aluminium-foil paper sealing, be placed in 2-8 DEG C and preserve.
(2) PCT reaction plate: PCT reaction plate contains the detection line that have cured anti-human PCT monoclonal antibody and the nature controlling line that have cured anti-human PCT antigen, and its preparation method is:
1. NC70 film is cut into every bar 30cm, and is pasted onto on backboard.The backboard pasting NC70 film prepares line.
2. rule:
Detection line: get anti-human PCT monoclonal antibody, with coating buffer 5 times dilution, scribe widths 0.5 μ L/cm;
Nature controlling line: get PCT antigen, with coating buffer 2 times dilution, scribe widths 0.5 μ L/cm.
Scribing position: the top 1.4cm of the top 1.1cm of nature controlling line distance NC70 film, detection line distance NC70 film.
3. backboard pulls rear placement 37 DEG C of constant temperature ovens, the binding time 14h of antibody stabilization.
4. the thieving paper cut and glass, be pasted onto respectively on backboard.
5. the backboard pasted, on cutting cutter, cuts out and is slit into the every bar of 4mm.
6. the test strips after cutting puts into test card, and buckles the lid that gets stuck.
7. in each aluminium foil bag, after putting into a test card and a drying agent, sealing.
8. adhesive label on the aluminium foil bag sealing mouth.
Preferably, marking fluid: 0.1M-0.2M carbonate solution; Fluorescence conserving liquid: 0.01M phosphate solution; Coating buffer: 0.02M phosphoric acid solution; The source of antibody, antigen: source is all Hstest; Antibody concentration: 4.6mg/ml and 7mg/ml; Antigen concentration: 1mg/ml, fluorescein is Alexa 647.
Embodiment 2: Procalcitonin immue quantitative detection reagent box Quality Control correlation parameter of the present invention
(1) outward appearance
Kit outward appearance is clean and tidy, and letter symbol is clear; Reaction plate is neatly complete, impulse-free robustness, without damaged, and material adhesion-tight; Liquid clarification in kit, without precipitation and floccus.
(2) mould bar width
Mould bar width >=2.5mm.
(3) creep speed
Liquid creep speed should be not less than 10mm/min.
(4) blank limit
Should not higher than 0.1ng/ml.
(5) accuracy
Type inspection: use Roche Holding Ag's sterling to carry out recovery test, the recovery is within the scope of 85%-115%.
Factory inspection: by business correctness quality-control product test kit, relative deviation should little 15%.
(6) range of linearity
In 0.1ng/ml ~ 50ng/ml measurement range, linearly dependent coefficient r >=0.990, relative deviation is no more than ± and 15%.
(7) repeatability
Detect with two samples of variable concentrations, each duplicate detection 10 times, its coefficient of variation CV should be not more than 10%.
(8) difference between batch
Detect same increment originally with three lot number kits, each lot number detects 3 times, calculates the mean value often criticizing 3 times respectively, then between three batches of kits, relative extreme difference should be not more than 15%.
(9) specificity
Detecting containing concentration is the human serum albumins of 40g/L, and its testing result is not 0.1ng/ml PCT value higher than detectable concentration.
(10) stability
Kit regulation storage requirement under be saved to the term of validity (18 months) after detect, result meets (3), (4), (5), (6), (7) item requirement.
Embodiment 3: the accuracy of Procalcitonin immue quantitative detection reagent box detects
Experiment material and source thereof:
The Procalcitonin immue quantitative detection reagent box that embodiment 1 prepares;
Control group used kit, corresponding fluorescence immunity analyzer derive from outsourcing, and method is fluorescence immune chromatography method;
Concrete scheme: kit prepared by embodiment 1 and outsourcing kit, does the contrast of low value quality-control product and the detection of high level quality-control product.Each concentration detects 20 times respectively, calculates relative deviation, target value rate.
In testing sample, the concentration of PCT is respectively 0.49ng/mL (corresponding Quality Control scope is 0.39-0.59ng/mL) and 9.46ng/mL (corresponding Quality Control scope is 7.76-11.2ng/mL).
Experimental technique
Detecting instrument is bold and generous biological dry type fluorescence immunity analyzer D-HMF, and testing result is in table 1.
Table 1 experimental group and control group are to the testing result of different detection samples
From table 1, experimental result is known, experimental group is basically identical to PCT concentration contained in the testing result of different testing sample and testing sample, and testing result is all within the scope of target value, and relative deviation is less, be respectively 10.2%, 6.4%, quantitatively can detect the content of PCT in testing sample.And control group is comparatively large to PCT concentration deviation contained in the testing result of different testing sample and testing sample, be respectively 28.6%, 20.1%; And be respectively 20%, 45% in target value rate.Experimental result illustrates, Procalcitonin immue quantitative detection reagent box accuracy provided by the invention is high, can be used in the PCT quantitatively detected in testing sample.
Embodiment 4: the range of linearity of Procalcitonin immue quantitative detection reagent box detects
Experiment material:
Procalcitonin immue quantitative detection reagent box prepared by embodiment 1.
Concrete scheme:
For comparative experiments group reagent box and control group kit are in the strengths and weaknesses of linearity test, the detection of 0.1ng/mL, 1ng/mL, 3.125ng/mL, 6.25ng/mL, 12.5ng/mL, 25ng/mL, 50ng/mL7 concentration is done in the range of linearity, each concentration does 3 repetitions, after drawing average, bring the corresponding R of formulae discovery into by average 2.
Experimental group kit: detection line fixes anti-human PCT monoclonal antibody, nature controlling line fixes anti-human PCT antigen.
Control group kit: PCT reaction plate comprises, detection line fixes anti-human PCT monoclonal antibody, and nature controlling line is fixed mouse source goat-anti muroid common two and resisted.
Experimental technique:
The PCT got in the Procalcitonin immue quantitative detection reagent box that testing sample embodiment 1 prepares detects damping fluid and mixes, and extracts reaction solution and is added drop-wise on PCT reaction plate, fully detects after reaction.
Detecting instrument is bold and generous biological dry type fluorescence immunity analyzer D-HMF, through four parameter fittings and linear fit the data obtained in table 2, table 3.
Table 2 PCT detection line result of linear detection
Experimental group ng/mL RLU value 1 RLU value 2 RLU value 3 RLU average
0.1 503 449 537 496
1 673 804 703 727
3.125 2159 1586 1684 1810
6.25 3453 3829 3317 3533
12.5 6553 6111 6426 6363
25 12196 11856 9830 11294
50 25490 26400 24512 25467
The testing result that table 3 PCT nature controlling line is linear
Control group ng/mL RLU value 1 RLU value 2 RLU value 3 RLU average
0.1 387 388 380 385
1 594 600 585 593
3.125 1486 1492 1471 1483
6.25 2371 2373 2360 2368
12.5 5824 5817 5804 5815
25 8842 8838 8828 8836
50 15640 15643 15628 15637
Be experimental group according to the results are shown in Figure 1, Fig. 1 cathetus 1 in table 2, table 3, detection line fixes anti-human PCT monoclonal antibody, and nature controlling line fixes the RLU value corresponding relation curve of anti-human PCT antigen, R 2=0.995; Straight line 2 is control group, and detection line fixes anti-human PCT monoclonal antibody, and nature controlling line fixes the anti-RLU value corresponding relation curve of mouse source goat-anti muroid two, i.e. R2=0.987.As can be known from the results, the range of linearity of kit provided by the invention is obviously better than control group.
Embodiment 5:
Experimental program: in the range of linearity, the routine serum sample of random detection 130, every increment originally all uses experimental group kit, (nature controlling line is different from experimental group for domestic contrast agents box, namely nature controlling line is fixed mouse source goat-anti muroid common two and is resisted), (nature controlling line is identical with experimental group for external contrast agents box, namely nature controlling line fixes anti-human PCT antigen) contrast detection simultaneously, and for homologous serum, often kind of kit all do two parallel, and calculate corresponding average.Experimental group kit and the thick kit of state's internal reference do with external kit average the contrast distributed just very much respectively, the results are shown in Table 4 and table 5.
Table 4
Table 5
The normal distribution of the external control group made according to above data and domestic control group, experimental group respectively as shown in Figures 2 and 3.Experimental result: normal distribution can be found out, adopt the experimental group data of the inventive method and external contrasting data normal distribution to be better than the normal distribution of domestic contrasting data and external contrasting data, namely the detected value of experimental group kit is more accurate.
Below be only the preferred embodiment of the present invention, it should be pointed out that above-mentioned preferred implementation should not be considered as limitation of the present invention, protection scope of the present invention should be as the criterion with claim limited range.For those skilled in the art, without departing from the spirit and scope of the present invention, can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.

Claims (5)

1. quantitatively detect a method for Procalcitonin, it is characterized in that, comprise the following steps:
Detected sample is mixed with fluorescently-labeled anti-Procalcitonin first antibody, obtains the reactant liquor containing Procalcitonin antigen-antibody complex; By this reactant liquor in reaction medium respectively with anti-Procalcitonin second antibody, Procalcitonin antigen-reactive, detect fluorescence signal respectively; The content obtaining Procalcitonin in detected sample is calculated according to recording fluorescence signal value.
2. method according to claim 1, it is characterized in that, described computing method are specially: the standard items detecting concentration known, collect two kinds of fluorescence signal values of standard items respectively and add up, calculate corresponding ratio with respective accumulated value and draw typical curve, the fluorescence signal value of testing sample is substituted into typical curve, draws the content of Procalcitonin in testing sample.
3. method according to claim 1, is characterized in that, described fluorescence labeling fluorescein used is Alexa Fluor line fluorescent element.
4. method according to claim 1, is characterized in that, described reaction medium is test strips.
5. method according to claim 4, is characterized in that, described anti-Procalcitonin second antibody, Procalcitonin antigen are fixed on the diverse location of reaction medium.
CN201510065249.5A 2015-02-06 2015-02-06 Method for quantitatively detecting procalcitonin Pending CN104714017A (en)

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CN106443007A (en) * 2016-08-31 2017-02-22 青岛汉唐生物科技有限公司 Quantitative detection kit of serum amyloid protein A
CN107782900A (en) * 2016-08-31 2018-03-09 朱海燕 The Test paper component of human growth and differentiation factor 7 15
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CN112924669A (en) * 2021-02-08 2021-06-08 南京农业大学 Multicolor flow measurement immunochromatography test strip based on three primary colors of optics and preparation and detection methods thereof

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