CN206362809U - The Test paper component of human growth and differentiation factor 7 15 - Google Patents
The Test paper component of human growth and differentiation factor 7 15 Download PDFInfo
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- CN206362809U CN206362809U CN201621033066.1U CN201621033066U CN206362809U CN 206362809 U CN206362809 U CN 206362809U CN 201621033066 U CN201621033066 U CN 201621033066U CN 206362809 U CN206362809 U CN 206362809U
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Abstract
The utility model discloses the Test paper component of human growth and differentiation factor 7 15.The Test paper component includes:Substrate layer;Sample pad, the sample pad is arranged on the substrate layer;Detecting pad, the detecting pad is arranged on the substrate layer, and it is provided with detection line and control line on the detecting pad, the detection line is located at nearly described sample pad one end, the control line is located remotely from described sample pad one end, the polyclonal antibodies of detection line coating anti alpha GDF 15, the control line is coated with the comlete antigen of growth factor 15;Pad, the pad, which is stacked, to be arranged between the sample pad and the detecting pad, and immunomagnetic beads is contained on the pad, and the immunomagnetic beads is coupled anti-human growth and differentiation factor 15 antibody;Adsorptive pads, the adsorptive pads engagement is arranged on the one end of the detecting pad away from the sample pad.The Test paper component detection speed is fast, and sensitivity and accuracy is high, required sample size is few.
Description
Technical field
The utility model is related to field of biological detection, in particular it relates to the Test paper group of human growth and differentiation factor 7-15
Part.
Background technology
China's sudden death incidence is about the people of 8.8 people/100,000-30/100,000, the people of average out to 15/100,000 every year.Wherein, it is young
The ratio of people increases year by year, and the sudden death less than 45 years old accounts for half, and 20% was accounted for less than 30 years old.Young people is typically considered
" healthy ", its sudden cardiac death causes tremendous influence to emotion, family and society." Pekinese's health white paper " is aobvious within 2012
Show, Beijing young people's health status causes anxiety, data display:Young Patients morbidity of acute coronary syndrome increase in 35 years old to 45 years old
Three one-tenth, acute cardiovascular disease continues to increase to Beijing Young Patients health threat.Acute chest pain is typically young popular feeling source
Property the most important onset symptoms of sudden death, the cause of disease of acute chest pain has a variety of, wherein common Severe acute disease includes:Acute Coronary Syndrome
Levy, dissection of aorta, pulmonary embolism and cardiomyopathy etc..And in the inpatient using acute coronary syndrome as performance
In, it is Stress cardiomyopathy to have 1%-2%, and current people are not enough to the sick understanding, and its illness rate has been underestimated, often by
Mistaken diagnosis, fail to pinpoint a disease in diagnosis.
Growth conversioning factor -15 (growth differentiation factor 15, GDF-15) expression has tissue
Specificity, GDF-15 high expression in prostate and placenta, expresses seldom in its most hetero-organization, including heart.But
In the case of pathology and environmental stress, such as ischemia/reperfusion injury, the rise of cardiac pressure load, myocardial hypertrophy, heart failure and
When atherosclerosis, GDF-15 expresses rapid rise in cardiac muscle cell, makes it have as good angiocarpy
The primary condition of disease biochemical marker, is an important cardiovascular protection factor, has a variety of lifes in angiocardiopathy
Thing function, but the mark only resides within the experimental study stage at present, its research direction is mainly Acute Coronary Syndrome
Levy and heart failure condition assessment in terms of, there is presently no be applied to clinically.Currently marketed GDF-15 kits master
To be ELISA kit, its long detection time is about 2-3 hours, not be suitable for clinically anxious to Stress cardiomyopathy etc.
The quick diagnosis of critical illness.
Thus, the detection kit of growth conversioning factor -15 has much room for improvement.
Utility model content
The utility model is intended at least solve one of technical problem present in prior art.Therefore, of the present utility model
One purpose is the Test paper component for proposing a kind of human growth and differentiation factor 7-15, and the component can quick, Sensitive Detection GDF-
15, prevention, diagnosis and the treatment occurred for cardiocerebrovasculaevents events has particularly important meaning.
It should be noted that the utility model is the following work based on inventor and completed:
Inventor has found, with ST sections of Elevation Myocardial Infarction (STEMI) patient's comparative analyses, (GDF- of growth conversioning factor -15
15) significantly increase in transient when Stress cardiomyopathy patient is being admitted to hospital, and GDF-15 elevated-levels are relevant with prognosis, we
Consider GDF-15 can as diagnosing stress cardiomyopathy ideal marker, currently marketed GDF-15 kits are mainly
ELISA kit, its long detection time is about 2-3 hours, and the time is exactly life, and time is life, and the kit is not
Suitable for clinically to the quick diagnosis of the Severe acute diseases such as Stress cardiomyopathy.
Immunomagnetic beads is the class new material that immunology and magnetic carrier technology are combined and grown up, and is a kind of magnetic bead
Pan coating specific ligand, can with antibody/antigen is specific combines to form magnetic bead-antibody/antigen compound, then pass through
The effect of externally-applied magnetic field, makes magnetic bead and solution quick separating, the characteristics of magnetic bead has following compared with traditional microwell plate:Surface
Product is bigger, can combine more protein moleculars;It can be connected by covalent bond with probe molecule, be the micro- of material than polystyrene
The physisorption of orifice plate is more firm;It is a kind of small-sized, flowing solid phase carrier, reaction is reached that dynamic is flat faster
Weighing apparatus, so as to accelerate reaction speed;The density that surface is combined is high, fluorescence signal is more concentrated;Can be with different probe molecule knots
Close, make it possible different determinands in the same sample of detection;The flexibility of the outward appearance of magnetic bead and coating process is bigger, can be with
Selected according to different requirement of experiment.Inventor utilizes the above-mentioned characteristic of immunomagnetic beads, and immunomagnetic beads is coupled into anti-human life
The long antibody of differentiation factor -15, can be with quick detection GDF-15, so as to quickly be examined Severe acute diseases such as Stress cardiomyopathies
It is disconnected.
Thus, according to one side of the present utility model, the utility model provides a kind of human growth and differentiation factor 7-15
Test paper component.According to embodiment of the present utility model, the Test paper component of human growth and differentiation factor 7-15 includes:Lining
Bottom;Sample pad, the sample pad is arranged on the substrate layer;Detecting pad, the detecting pad is arranged on the substrate layer,
And detection line and control line are provided with the detecting pad, the detection line is located at nearly described sample pad one end, the control line
Described sample pad one end is located remotely from, the detection line is coated with anti alpha-GDF-15 polyclonal antibodies, the control line coating growth
The comlete antigen of the factor -15;Pad, the pad, which is stacked, to be arranged between the sample pad and the detecting pad, and described
Immunomagnetic beads is contained on pad, the immunomagnetic beads is coupled anti-human GDF-15 antibody;Adsorptive pads, the water suction
Pad engagement is arranged on the one end of the detecting pad away from the sample pad.
According to the Test paper component of the human growth and differentiation factor 7 of the utility model embodiment -15, by anti-GDF-15 antibody
It is coupled on immunomagnetic beads, the enrichment density of anti-GDF-15 antibody is high, and GDF is entered by the antibody on detecting pad and pad
Row double antibodies sandwich is detected, it is to avoid the result of false positive, also, the Test paper component detection speed is fast, it is only necessary to which more than ten minutes are
It can complete detection, and sensitivity and accuracy is high, required sample size is few.
Optionally, the sample pad contains sample treatment liquid, and the sample treatment liquid is ox blood clear solution, and the ox blood
Clear solution contains at least one selected from polyethylene glycol, polyvinylpyrrolidone, bovine serum albumin(BSA), surfactant and salt
Kind.
Optionally, the anti-human GDF-15 antibody is monoclonal antibody.
Optionally, a diameter of 20-40nm of the immunomagnetic beads.
Optionally, the substrate layer is made up of polyvinyl chloride.
Optionally, the sample pad is made up of polyester fiber.
Optionally, the detecting pad is made up of nitrocellulose membrane.
Optionally, the pad is made up of glass fibre.
Optionally, the adsorptive pads are made up of blotting paper.
Optionally, pad 1.5-3.0mm overlapping with the sample pad and the detecting pad respectively.
Additional aspect and advantage of the present utility model will be set forth in part in the description, partly by from following description
In become obvious, or by it is of the present utility model practice recognize.
Brief description of the drawings
Of the present utility model above-mentioned and/or additional aspect and advantage will from description of the accompanying drawings below to embodiment is combined
Become substantially and be readily appreciated that, wherein:
Fig. 1 shows the Test paper component of human growth and differentiation factor 7-15 according to the utility model one embodiment
Structural representation;
Fig. 2 shows the Test paper component of human growth and differentiation factor 7-15 according to the utility model one embodiment
Vertical section structure schematic diagram.
Embodiment
Embodiment of the present utility model is described below in detail, the example of the embodiment is shown in the drawings, wherein from beginning
Same or similar element or element with same or like function are represented to same or similar label eventually.Below by ginseng
The embodiment for examining accompanying drawing description is exemplary, is only used for explaining the utility model, and it is not intended that to of the present utility model
Limitation.
According to one side of the present utility model, the utility model provides a kind of detection of human growth and differentiation factor 7-15
Test paper component.With reference to Fig. 1, according to embodiment of the present utility model, the Test paper component of human growth and differentiation factor 7-15 is entered
Row is illustrated.The Test paper component includes:Substrate layer 100, sample pad 200, detecting pad 300, pad 400 and adsorptive pads
500.Each part is introduced below:
Substrate layer 100:According to embodiment of the present utility model, substrate layer 100 is located at the bottommost of Test paper component, uses
In to carrying gas components.
According to embodiment of the present utility model, substrate layer 100 is made up of polyvinyl chloride.Thus, quality is solid, water proofing property
It is good, it is indeformable in a liquid.
Sample pad 200:According to embodiment of the present utility model, sample pad 200 is arranged on substrate layer 100, testing sample
It is added dropwise in sample pad.
According to embodiment of the present utility model, sample pad 200 is made up of polyester fiber.Because there is polyester fiber outward appearance to endure
Include, heat endurance is good, but hygroscopicity it is slightly worse the characteristics of, using polyester fiber formation sample pad, sample pad is unlikely to deform, and property is steady
It is fixed, also, sample is under the suction of blotting paper, it is easy to and flowed to blotting paper direction, reduce sample attached in sample pad
, make the accuracy of testing result higher.
According to embodiment of the present utility model, sample pad 200 contains sample treatment liquid, and sample treatment liquid is that cow's serum is molten
Liquid, and the ox blood clear solution contains selected from polyethylene glycol, polyvinylpyrrolidone, bovine serum albumin(BSA), surfactant and salt
At least one of class, wherein, polyethylene glycol, polyvinylpyrrolidone, bovine serum albumin(BSA), surfactant and salt can bars
The stability of part immunomagnetic beads, makes the sensitivity of detection structure and accuracy higher.
Detecting pad 300:According to embodiment of the present utility model, detecting pad 300 is also disposed on substrate layer 100, and the inspection
Survey on pad 300 and be provided with detection line 310 and control line 320, detection line 310 is located at one end of nearly sample pad 300, and control line position
In away from the one end of sample pad 200, i.e., first pass through detection line 310 during sample flow in sample pad, then by control line 320, its
In, the coating anti alpha-GDF-15 polyclonal antibodies of detection line 310, the anti alpha-GDF-15 polyclonal antibodies can be with the GDF- in sample
15 combine, and the coating comlete antigen of growth factor-1 5 of control line 320, and the comlete antigen of growth factor-1 5 can be with idol on magnetic bead
The anti-human GDF-15 antibody binding of connection.
" polyclonal antibody " described herein, referred to as " many anti-", refers to by multiple subcutaneously (sc), intraperitoneal (ip) or intramuscular
(im) injection corresponding antigens and auxiliary agent, the mixture of the discrete antibody produced in animal." monoclonal resists term used herein
Body ", referred to as " monoclonal antibody ", refers to the antibody obtained from substantially homologous antibody population, that is, the antibody individual for constituting the colony is all identical, and
The mixture of discrete antibody.
According to embodiment of the present utility model, anti alpha-GDF-15 polyclonal antibodies, and with GDF-15 binding site and magnetic
The anti-human GDF-15 antibody being coupled on pearl is different, and the two efficiently avoid the generation of Competitive assays, and control
Line is coated with the comlete antigen specific recognition anti alpha-GDF-15 polyclonal antibodies of growth factor-1 5, and is not present with monoclonal antibody non-specific
Property combine, so formed 4 components sandwich shape conjugate, high specificity.
According to embodiment of the present utility model, detecting pad 300 is made up of nitrocellulose membrane.Because nitrocellulose is to egg
The materials such as white matter have non-specific adsorption ability, and anti-human GDF-15 antibody is specifically tied with the GDF-15 in sample
Closing the compound formed can adsorb on detecting pad, make testing result sensitiveer, accurate.
Pad 400:According to embodiment of the present utility model, pad 400, which is stacked, is arranged on sample pad 200 and detecting pad
Between 300, and immunomagnetic beads is contained on the pad 400, the immunomagnetic beads is coupled anti-human GDF-15 antibody, should
Anti-human GDF-15 antibody and the GDF-15 specific bonds in sample.
According to embodiment of the present utility model, anti-human GDF-15 antibody is monoclonal antibody.With specific single
Clonal antibody recognizes that protein loci is single, it is ensured that the specificity of testing result, at utmost as capture GDF-15 antibody
Avoid false positive results.
According to embodiment of the present utility model, a diameter of 20-40nm of immunomagnetic beads.Thus, the magnetic bead of the particle size range,
Magnetic response is strong, and specific surface area is big, is distributed, is easy to GDF-15 fully to be combined with immunomagnetic beads.
According to embodiment of the present utility model, pad 400 is made up of glass fibre.Because glass fibre is inert material
Matter, adhesion protein, can not effectively prevent GDF-15 or GDF compounds to be trapped in detection error caused on pad.
According to embodiment of the present utility model, pad 400 respectively with sample pad 200 and the overlapping 1.5- of detecting pad 300
3.0mm, is connected sample pad 200 and detecting pad 300 by pad 400, the combined pad of the sample in sample pad is flowed to detecting pad
On, realize sample detection.
Adsorptive pads 500:According to embodiment of the present utility model, the engagement of adsorptive pads 500 is arranged on detecting pad remote 300 from sample
One end of pad 200, for providing the driving force that the sample in sample pad 200 is moved to detecting pad 300.
According to embodiment of the present utility model, adsorptive pads 500 are made up of blotting paper.Thus, material source is extensive, water imbibition
Good, the driving force that driving sample flows to detecting pad direction is strong.
For the ease of understanding the Test paper component of human growth and differentiation factor 7-15 of the present utility model, it is detected at this
Principle is explained, and specking has immunomagnetic beads on pad, wherein, magnetic bead surfaces were coupled with resisting that antigen-specific reacts
Human growth and differentiation factor 7-15 antibody of son, another anti alpha-GDF-15 being fixed with detection line with material idiosyncrasy to be detected is more
Be fixed with clonal antibody, control line can with magnetic bead surfaces antibody specific bond the comlete antigen of growth factor-1 5.In sample pad
A few drop samples in upper drop, sample is chromatographed towards blotting paper direction, and the magnetic bead on pad is soaked and disperseed wherein, magnetic bead by solution
Chromatographed together on nitrocellulose membrane also with solution.If needing to be detected the antibody on material, magnetic bead with to be detected in solution
Antigen-antibody reaction occurs for material, forms the compound of antigen and immunomagnetic beads;When this compound passes through detection line, antigen
Other sites catch immunomagnetic beads in detection line with the antibody response on film, form macroscopic band;It is not detected
The immunomagnetic beads that line is caught continues to chromatograph forward, when reaching control line, antibody of the secondary antibody on control line with immunomagnetic beads surface
With reference to by magnetic bead seizure on the control line.Therefore, if there is band in detection line and on control line, detection sample is sun
Property;If only there is band on control line, sample is feminine gender.
Wherein, Test paper component uses double-antibody method, and capture people's Growth and Differentiation is used as using monoclonal antibody specific
The antibody of the factor -15, identification protein loci is single, it is ensured that the specificity of testing result, at utmost avoids false positive, then
How anti-as detection antibody using specificity, the compatibility of the detection antibody and sub -15 antibody of human growth and differentiation factor 7 is high, fully knows
The cyclin Y not being combined with the monoclonal antibody on solid phase carrier, is not only further eliminated in detection process
False positive results, and the sensitivity of detection is higher, in addition, the site of sub -15 antibody of the how anti-identification human growth and differentiation factor 7 of monoclonal antibody
Difference, the two efficiently avoid the generation of Competitive assays, and control line coating growth factor-1 5 comlete antigen specificity is known
Not resist more, and non-specific binding is not present with monoclonal antibody, so the conjugate of the sandwich shape of four components is formed, specificity
By force thus, can easy, accurate, sensitive and efficient detection human growth and differentiation factor 7-15, especially human blood using the kit
Human growth and differentiation factor 7-15 in cleer and peaceful blood plasma.
Below with reference to specific embodiment, the utility model is illustrated, it is necessary to which explanation is, these embodiments are only
It is illustrative, and it is not intended that to limitation of the present utility model.
Scheme of the present utility model is explained below in conjunction with embodiment.It will be understood to those of skill in the art that under
The embodiment in face is merely to illustrate the utility model, and should not be regarded as limiting scope of the present utility model.It is unreceipted in embodiment
Particular technique or condition, carried out according to the technology or condition described by document in the art or according to product description.
Agents useful for same or the unreceipted production firm person of instrument, be can by the conventional products of acquisition purchased in market, can for example purchase from
Sigma companies.
Embodiment 1
In the present embodiment, it is introduced to preparing the method for Test paper component of human growth and differentiation factor 7-15, specifically such as
Under:
1st, experimental raw
Anti- GDF-15 antibody and related comlete antigen (are purchased from An Di biotech firms, article No.:1860), nitrocellulose
Film, blotting paper, sample pad, 6cm × 30cm PVC boards, surface modification has the 30nm magnetic beads of carboxyl, and (10mg/ml is purchased from the U.S.
QceanNano Tech companies), 2- (N- morpholines) ethyl sulfonic acid monohydrate (MES, purchased from Alafa companies), N- hydroxysuccinimidyl acyls
Imines (NHS, purchased from PIERCE companies), Tween-20 (is purchased from BBI companies), 1- ethyls-(3- dimethylaminopropyls) phosphinylidyne two
Inferior amine salt hydrochlorate (EDC, purchased from the biochemical development in science and technology Co., Ltd of Shanghai Yan length), SuperMagSeparator TM Magneto separate framves
(1.4T, QceanNano Tech companies, the U.S.), the ELIASAs of Wallac Victor 1400 (PE, the U.S.), TGL-16B is at a high speed
Centrifuge (peace booth scientific instrument, Shanghai).
2nd, immunomagnetic beads is prepared
Detection GDF-15 immunomagnetic beads is prepared using EDC/NHS, including activates, be coupled, closing, preserving several steps,
Flow is as follows:
(1) activate:The carboxyl on EDC activated magnetic beads surface, forms O-acylisourea derivatives, in O-acylisourea
NHS is added in derivative, the derivative for suppressing generation hydrolyzes the stable NHS esters of generation in the solution, while activated carboxyl, is obtained
Activated magnetic beads, comprise the following steps that:
(a) 2mg carboxyls magnetic bead is taken in 2ml centrifuge tubes, and adding 500 μ l activation buffers, (pH5.0,0.01M MEST are molten
Liquid (0.05%Tween-20), is mixed, and being placed in Magneto separate frame makes magnetic bead be adsorbed completely, and minipump takes out clear liquid.
(b) 500 μ l activation buffers are added and relaunders magnetic bead twice, 485 μ l activation buffers are added into magnetic bead again
EDC (original content 0.5g/ml) and NHS (original content 0.25g/ml) 2.5mg respectively are added, are well mixed in vortex oscillator,
The carboxyl of room temperature activation 2mg magnetic bead surfaces 30 minutes.
(c) after activating, the unreacted activator of removing is washed with MEST buffer, then changes centrifuge tube, and uses MEST
Twice of buffer washings magnetic bead.
(2) it is coupled:By the amino reaction generation immunomagnetic beads of activated magnetic beads and antibody, comprise the following steps that:
(a) by the magnetic bead after activation, with 500 μ l coupling buffer, (500 μ l, pH9.0,0.005M borate tween delay
Twice of fliud flushing (0.05%Tween-20) washing magnetic bead;
(b) magnetic bead after washing adds the magnetic bead that 475 μ l coupling buffers are resuspended after washing.
(c) magnetic bead being resuspended after washing is added to 25 μ l 6mg/ml anti-GDF-15 antibody so that magnetic bead surfaces are activated
Carboxyl with anti-GDF-15 antibody amino react at room temperature 3 hours, by antibody coupling in magnetic bead surfaces, obtain immunomagnetic beads.It is even
Reaction supernatant is collected after the completion of connection and detects coupling protein amount with BCA kits.
(3) close:Immunomagnetic beads is closed using BSA solution, is comprised the following steps that:
(a) 500 μ l 1%BSA is added, wherein, BSA is dissolved in borate Tween buffer in advance, to immunomagnetic beads
The activated group that surface is not reacted completely is closed, and by BSA physical absorption, closes other spaces site,
To reduce the non-specific adsorption that may occur in experiment afterwards.
(b) capping 30 minutes, the immunomagnetic beads closed at room temperature.
(4) preserve and wash the immunomagnetic beads four times after closing with BST and (pipe need to be changed for the first time, by magnetic bead when washing every time
It is resuspended), magnetic bead is finally resuspended in 500 μ l and preserved in liquid.Preserve liquid formula be:PH9.0,0.005M borate buffer body
System, 0.05%Tween-20, NaN3,0.1%BSA.
3rd, the making of test strips
(1) cut:Nitrocellulose membrane is cut into 0.5cm × 2.5cm square sizes with scissors as detecting pad, glass fibers are cut
0.5cm × 0.5cm square sizes are tieed up as pad, glass fibre 0.5cm × 1.5cm square sizes is cut as pad, cuts
Water suction paper size is 0.5cm × 1.5cm, cuts the supporter that PVC bottom plates make each part for 0.5cm × 8cm square sizes.
(2) making of detecting pad:With liquid-transfering gun with 2 μ l/cm amount by 1mg/m anti alpha-GDF-15 antibody-solutions (antibody
It is dissolved in PBS, cane sugar content prepares detection line 1%) to put on nitrocellulose membrane in solution, then with liquid-transfering gun with 2 μ l/
By the 1mg/m comlete antigen solution of growth factor-1 5, (antigen is dissolved in PBS cm amount, and cane sugar content is 1%) point in solution
In on nitrocellulose membrane, preparing control line, detection line is parallel with control line, and distance therebetween is 1cm.Then, put
Toast 30 minutes, be put in after taking-up stand-by in drying basin in drying in 37 DEG C of closed vacuum drying ovens.
(3) making of pad:The immunomagnetic beads drawn with liquid-transfering gun after 20 μ lBST dilutions is uniformly put on pad,
Pad is put in dry in 37 DEG C of closed vacuum drying ovens and toasted 30 minutes, is put in after taking-up in drying basin, it is standby.
(4) making of sample pad:Different effect material is dissolved in BS solution as sample treatment liquid, wherein, sample
In treatment fluid, PEG, PVP and BSA final concentration are 1%, and final concentration of the 0.5% of the Tween-20 of surfactant, take one
The sample pad of fixed number amount is soaked in a period of time in treatment fluid, and the sample pad after immersion is put in and dries closed 37 DEG C of vacuum baking
Take out and be put in drying basin after being toasted 2 hours in case, be for available sample pad.
(5) detecting pad, pad and the sample pad for obtaining step (2)-(4) are bonded on PVC bottom plates, obtain people's growth
The Test paper component of differentiation factor -15, its structural representation is as shown in Figure 1.
Embodiment 2
What the Test paper component of human growth and differentiation factor 7-15 obtained using embodiment 1 and Andygene companies were produced
GDF-15ELISA kits detect, and result is compared that specific method is as follows simultaneously to sample:
1st, the collection and preservation of sample:
(1) serum:The whole blood sample for being collected in serum separator tube is placed in room temperature and stayed overnight within 2 hours or 4 DEG C, is then centrifuged for
1000g × 20 minute, take supernatant to dispense, and the supernatant dispensed are placed in into -80 DEG C of preservations, but should avoid multigelation.
(2) blood plasma:With EDTA or heparin as anti-coagulants collect specimen, and by sample after acquisition 30 minutes in 4
DEG C centrifugation 1000g × 15 minute, take supernatant to dispense, will dispenses and preserved in -80 DEG C, but should avoid multigelation.
2nd, sample detection
(1) the Test paper component detection of human growth and differentiation factor 7-15:
(a) standard curve:GDF-15 standard items are diluted using PBS, obtain final concentration be respectively 10,25,50,100,
250 and 500ng/l standard solution, takes the μ L of above-mentioned standard product solution 100 to be loaded to the sample pad of Test paper component respectively
Detected, the magnetic signal in detection line and control line is detected with instrument after being dried etc. test strips.It is dense with magnetic signal and GDF-15
Spend for parameter, do quantitative measurement standard curve, detect sample concentration.
(b) it is that sample is detected by the blood preparation of 39 cardiacs, 30 μ L sample serum is taken respectively, will
120 μ L PBS is diluted, and after concussion is mixed, is taken 100 μ L to be loaded to the sample pad of Test paper component and is detected, wait examination
Paper slip detects the magnetic signal in detection line and control line with instrument after drying.
(2) GDF-15ELISA kits are detected,
The GDF-15ELISA kits produced using Andygene companies, are operated according to the specification of the kit,
The serum specimen with a collection of patient is detected using the kit.
3rd, experimental result
The result detected using the Test paper component and GDF-15 kits of human growth and differentiation factor 7-15 to sample
It is as follows:
Examined using the SPSS t that above-mentioned two groups of data are carried out with matched-pair design quantitative data, it is as a result as follows,
P=0.051>0.05 two groups of no significant differences, it is believed that two kinds of measuring method results are identical.And use
The detection of GDF-15ELISA kits needs 3 hours or so, and the Test paper component of human growth and differentiation factor 7-15 only needs more than ten
Minute, in the case of result identical, the detection time significantly saved.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means to combine specific features, structure, material or the spy that the embodiment or example are described
Point is contained at least one embodiment of the present utility model or example.In this manual, to the schematic table of above-mentioned term
State and be not necessarily referring to identical embodiment or example.Moreover, specific features, structure, material or the feature of description can be
Combined in an appropriate manner in any one or more embodiments or example.
While there has been shown and described that embodiment of the present utility model, it will be understood by those skilled in the art that:
These embodiments can be carried out with a variety of changes, modification in the case where not departing from principle of the present utility model and objective, replaced
And modification, scope of the present utility model limits by claim and its equivalent.
Claims (10)
1. a kind of Test paper component of human growth and differentiation factor 7-15, it is characterised in that including:
Substrate layer;
Sample pad, the sample pad is arranged on the substrate layer;
Detecting pad, the detecting pad is arranged on the substrate layer, and is provided with detection line and control line on the detecting pad, institute
State detection line and be located at nearly described sample pad one end, the control line is located remotely from described sample pad one end, the detection line coating
Anti alpha-GDF-15 polyclonal antibodies, the control line is coated with the comlete antigen of growth factor-1 5;
Pad, the pad, which is stacked, to be arranged between the sample pad and the detecting pad, and is contained on the pad
Immunomagnetic beads, the immunomagnetic beads is coupled anti-human GDF-15 antibody;And
Adsorptive pads, the adsorptive pads engagement is arranged on the one end of the detecting pad away from the sample pad.
2. Test paper component according to claim 1, it is characterised in that the sample pad contains sample treatment liquid, institute
Sample treatment liquid is stated for ox blood clear solution.
3. Test paper component according to claim 1, it is characterised in that the anti-human GDF-15 antibody
For monoclonal antibody.
4. Test paper component according to claim 1, it is characterised in that a diameter of 20-40nm of the immunomagnetic beads.
5. Test paper component according to claim 1, it is characterised in that the substrate layer is made up of polyvinyl chloride.
6. Test paper component according to claim 1, it is characterised in that the sample pad is made up of polyester fiber.
7. Test paper component according to claim 1, it is characterised in that the detecting pad is made up of nitrocellulose membrane.
8. Test paper component according to claim 1, it is characterised in that the pad is made up of glass fibre.
9. Test paper component according to claim 1, it is characterised in that the adsorptive pads are made up of blotting paper.
10. Test paper component according to claim 1, it is characterised in that the pad respectively with the sample pad
1.5-3.0mm overlapping with the detecting pad.
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CN111638365A (en) * | 2020-04-30 | 2020-09-08 | 吉林省格瑞斯特生物技术有限公司 | GDF15 quick quantitative detection device and preparation method thereof |
CN113702647A (en) * | 2021-08-31 | 2021-11-26 | 普十生物科技(北京)有限公司 | Human growth differentiation factor 15 instant detection kit, preparation method and application thereof |
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2016
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111638365A (en) * | 2020-04-30 | 2020-09-08 | 吉林省格瑞斯特生物技术有限公司 | GDF15 quick quantitative detection device and preparation method thereof |
CN113702647A (en) * | 2021-08-31 | 2021-11-26 | 普十生物科技(北京)有限公司 | Human growth differentiation factor 15 instant detection kit, preparation method and application thereof |
CN113702647B (en) * | 2021-08-31 | 2024-03-15 | 普十生物科技(北京)有限公司 | Human growth differentiation factor 15 instant detection kit, preparation method and application thereof |
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