CN109856403A - A kind of S100- β albumen time-resolved fluoroimmunoassay chromatography detection kit - Google Patents
A kind of S100- β albumen time-resolved fluoroimmunoassay chromatography detection kit Download PDFInfo
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- CN109856403A CN109856403A CN201811572355.2A CN201811572355A CN109856403A CN 109856403 A CN109856403 A CN 109856403A CN 201811572355 A CN201811572355 A CN 201811572355A CN 109856403 A CN109856403 A CN 109856403A
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Abstract
The invention discloses a kind of S100- β albumen time-resolved fluoroimmunoassay to chromatograph detection kit, is related to fluorescence immune chromatography technical field in Medical Immunology.After the present invention uses lanthanide chelate mark fluorescent microballoon, and then mark S100- β monoclonal antibody.Because lanthanide chelate stoke displacement is big, exciting light and transmitting light are easily distinguished, the interference of fluorescence that substance generates in exciting light and sample to be tested etc. can be excluded；In addition, the exciting light spectrum width of lanthanide chelate, and emission spectrum is narrow, this can more further decrease local fluorescence, improve sensitivity；Time-resolved fluoroimmunoassay chromatographic technique is further used, the interference of non-specific fluorescence must can be effectively excluded, it is so very big that improve sensitivity；Also, this kit has the advantages that easy to operate quick, testing cost is lower, can make up the deficiency of existing products in markets.
Description
Technical field
The present invention relates to fluorescence immune chromatography technical fields in Medical Immunology, and in particular to a kind of S100- β albumen time
Resolved fluorometric immunochromatographytest test kit.
Background technique
S100- β albumen is called nervous centralis differential protein, and S100- β albumen is described as " the C reaction egg of brain by some scholars
It is white ".S100 albumen is a kind of acid calbindin, and molecular weight 10000-12000 is primarily present in each portion of central nervous system
Starlike Deiter's cells cytosol in, because its in saturated ammonium sulfate can 100% dissolution due to gain the name.S100 albumen is by α
Form three kinds of different forms with two kinds of subunits of β: S100 β is primarily present in Deiter's cells and peripheral neverous system
Schwann is intracellular, in certain neuronal cells and melanocyte, fat cell.Maincenter star spongiocyte, glue of dashing forward less
Cell plastid, the Schwann and satellite cell of periphery contain abundant S100 β albumen.Its gene expression dose and cell Proliferation and
Differentiation state is related, and S100 β is secreted by the spongiocyte that intracerebral activates, and is one of the mark of star spongiocyte activation, is
Most active ingredient in S100 β protein family.S100 β is generated by spongiocyte, acts on neuron and surrounding environment,
It is one of intermediary's substance that Deiter's cells and neuron connect each other, in the growth course of brain and when cerebral injury, S100 β
It is a kind of neurotrophic factor, appropriate expression plays nutrition protective effect to brain tissue.When Deiter's cells expression is excessive,
The deterioration of accelerans system inflammation, and lead to neurological dysfunction.The expression of S100 β has close with cerebral injury severity
Relationship is cut, therefore it has different physiological roles: 1) promoting neurite outgrowth；2) enhance ATP enzyme activity, influence micro-pipe,
The depolymerization of microfilament；3) adjusting of intraor extracellular calcium ion level is participated in as intracellular normal components；4) it can induce in vitro
Neuron and glial cell apoptosis；5) there is neurotrophic effect, Deiter's cells can be with paracrine and autocrine
S100 β albumen, acts on neuron and Deiter's cells, to promote the reparation of growth and the damage of nerve, but high level
S100 β albumen but can produce neurotoxicity.
Measurement S100- β can be used for the disease of auxiliary judgment Yu following related neurological:
1) Alzheimer disease；Expression of the S100 β albumen in brain tissue increases with age；S100 β table
There is close contact with cerebral injury in patients range up to horizontal height；Alzheimer patient's its cerebrospinal fluid after encephalatrophy has occurred
Middle S100 β level is positively correlated with brain atrophy；
2) cranial vascular disease；Numerous studies prove that cranial vascular disease can cause blood-brain barrier permeability to increase, and make brain ridge
S100 β content increases in liquid and serum；
3) blood serum designated object of cerebral ischemic injury；Ischemic brain damage can lead to nerve cell oedema, denaturation, necrosis,
Deiter's cells secretion S100 β injured neuron is repaired, nutrition.Ischemic causes the permeability of blood-brain barrier to increase again
Add, S100 β albumen is made to enter cerebrospinal fluid and serum by blood-brain barrier, makes S100 β is horizontal in blood and cerebrospinal fluid to increase；Blood
The level of S100 β albumen can be used as the periphery mark of Acute ischemia rePerfusion in clear, facilitate Acute ischemia rePerfusion
Diagnosing and treating.
Currently, the kit that market is used to detect S100- β albumen is based primarily upon immunochromatographic method, colloidal gold method, enzyme-linked exempts from
Epidemic disease adsorption measurement (ELISA), latex enhancing immune turbidimetry etc..Wherein, immunochromatographic method and colloidal gold method are easy to operate fast
Victory, but sensitivity is lower, vulnerable to interference of stray light；ELISA method kit has operating process cumbersome, and detection time is long, generates
It is more to have the shortcomings that the waste liquid of sample contamination；Latex enhancing immune turbidimetry higher cost, and need expensive mating
Automatic analytical system is detected.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of S100- β albumen time-resolved fluoroimmunoassay chromatographies to detect
Kit, it is intended to after using lanthanide chelate mark fluorescent microballoon, and then mark S100- β monoclonal antibody.Because of group of the lanthanides member
Plain chelate stoke displacement is big, easily distinguishes exciting light and transmitting light, can exclude what substance in exciting light and sample to be tested generated
The interference of fluorescence etc.；In addition, the exciting light spectrum width of lanthanide chelate, and emission spectrum is narrow, this can more further decrease this
Ground fluorescence improves sensitivity；Time-resolved fluoroimmunoassay chromatographic technique is further used, must can effectively be excluded non-specific glimmering
The interference of light, it is so very big that improve sensitivity；Also, this kit has the advantages that easy to operate quick, testing cost is lower,
The deficiency of existing products in markets can be made up.
In order to solve the above technical problems, the technical solution used in the present invention is:
A kind of S100- β albumen time-resolved fluoroimmunoassay chromatography detection kit, including test strip, the detection
Test strips include PVC board, sample pad, bonding pad, NC film and water absorption pad；It is characterized in that, the test strip is with PVC board
For solid phase support bottom plate；The sample pad, bonding pad, NC film and water absorption pad make adjacent closely to connect between the two in this order
Touching, then sticks in PVC board；The bonding pad is coated with the S100- β monoclonal antibody of time-resolved fluorescence microballoon label
Ⅱ；The time-resolved fluorescence microspherulite diameter is 200nm-300nm, is marked with lanthanide series；
Detection line and nature controlling line are set on the NC film；Detection line is the S100- β monoclonal antibody being sprayed on NC film
Ⅰ；Nature controlling line is the sheep anti-mouse antibody being sprayed on NC film；The detection line and nature controlling line are parallel to each other and are equipped with gap.
A further technical solution lies in the sample pad, bonding pad, NC film and water absorption pad make both adjacent in order
Between overlapping edges.
A further technical solution lies in the test strip is having a size of 80mm × 4mm；Wherein PVC board having a size of
80mm×4mm；Sample pad is having a size of 25mm × 4mm；Bonding pad is having a size of 20mm × 4mm；NC film is having a size of 20mm × 4mm；It inhales
Water cushion is having a size of 21mm × 4mm；The sample pad and bonding pad overlapping edges；The NC film and water absorption pad overlap；The combination
Pad is overlapped with NC film；Overlapping part length is 2mm.
A further technical solution lies in, the sample pad, bonding pad, NC film and water absorption pad in order from top to bottom or
It successively sticks in PVC board from bottom to top.
A further technical solution lies in further include the shell equipped with cavity；The cavity of the enclosure interior places detection
Test strips；Observation window and well are set on the shell.
A further technical solution lies in the shell includes upper housing and lower case；The upper housing and lower case button
It closes, and internal formation cavity；The observation window and well are located on upper housing.
A further technical solution lies in the lanthanide series is europium.
A further technical solution lies in detection lines and nature controlling line spacing 6mm ± 0.1mm on the NC film；The inspection
Survey line is close to bonding pad；The nature controlling line is close to water absorption pad.
A further technical solution lies in further include the ID card containing calibration curve.
A further technical solution lies in I spouting liquid of S100- β monoclonal antibody is 0.8ul/cm, concentration 0.5-
2mg/ml；Sheep anti-mouse antibody spouting liquid is 0.8ul/cm, concentration 0.1-0.5mg/ml.
A further technical solution lies in the ID card is the identification code of mobile device identification；The identification code is two
Tie up code or bar code；It is arranged on shell.
The beneficial effects of adopting the technical scheme are that being intended to micro- using lanthanide chelate mark fluorescent
After ball, and then mark S100- β monoclonal antibody.Because lanthanide chelate stoke displacement is big, exciting light and transmitting are easily distinguished
Light can exclude the interference of fluorescence that substance generates in exciting light and sample to be tested etc.；In addition, the excitation of lanthanide chelate
Spectral width, and emission spectrum is narrow, this can more further decrease local fluorescence, improve sensitivity；Further use time resolution
Fluorescence immune chromatography technology must can effectively exclude the interference of non-specific fluorescence, so very big that improve sensitivity；Also, this examination
Agent box has the advantages that easy to operate quick, testing cost is lower, can make up the deficiency of existing products in markets.
Detailed description of the invention
The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.
Fig. 1 is that detection blocks interior test strips in time-resolved fluoroimmunoassay detection kit embodiment one of the invention
Structural schematic diagram:
Fig. 2 is that detection blocks interior test strips in time-resolved fluoroimmunoassay detection kit embodiment two of the invention
Structural schematic diagram:
Fig. 3 is that detection blocks interior test strips in time-resolved fluoroimmunoassay detection kit embodiment three of the invention
Structural schematic diagram；
Fig. 4 is schematic diagram of housing structure in time-resolved fluoroimmunoassay detection kit of the present invention；
Fig. 5 is kit calibration graph prepared by present example one；
Fig. 6 is that one linear dependence of present example analyzes result figure；
Fig. 7 is kit prepared by present example one and Wuhan Easydiagnosis Biomedicaine Co., Ltd.'s production
The correlativity analysis chart of S100- β albumen (S100- β) detection kit (immunochromatographic method) testing result；
Wherein: 1, PVC board；2, sample pad；3, bonding pad；4, NC film；5, detection line；6, nature controlling line；7, water absorption pad；8, it marks
Know code；9, well；10, observation window；11, upper housing；12, lower case.
Specific embodiment
With reference to the attached drawing in the embodiment of the present invention, technical solution in the embodiment of the present invention carries out clear, complete
Ground description, it is clear that described embodiment is only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
In the following description, numerous specific details are set forth in order to facilitate a full understanding of the present invention, but the present invention can be with
Implemented using other than the one described here other way, those skilled in the art can be without prejudice to intension of the present invention
In the case of do similar popularization, therefore the present invention is not limited by the specific embodiments disclosed below.
The invention discloses a kind of S100- β albumen time-resolved fluoroimmunoassay to chromatograph detection kit, including Test paper
Item, the test strip include PVC board 1, sample pad 2, bonding pad 3, NC film 4 and water absorption pad 7；It is characterized in that, described
Test strip is with PVC board 1 for solid phase support bottom plate；The sample pad 2, bonding pad 3, NC film 4 and water absorption pad 7 are suitable by this
Sequence make it is adjacent be in close contact between the two, then stick in PVC board 1；The bonding pad 3 is coated with time-resolved fluorescence microballoon
The S100- β monoclonal antibody II of label；The time-resolved fluorescence microspherulite diameter is 200nm-300nm, is marked with group of the lanthanides member
Element；
Detection line 5 and nature controlling line 6 are set on the NC film 4；Detection line (T line) 5 is the S100- β that is sprayed on NC film 4 mono-
Clonal antibody I；Nature controlling line (C line) 6 is the sheep anti-mouse antibody being sprayed on NC film 4；The detection line 5 and nature controlling line 6 are mutually flat
It goes and is equipped with gap.
In the preferred embodiment of the present invention, as shown in Figure 1, the sample pad 2, bonding pad 3, NC film 4 and water absorption pad 7 are pressed
Sequence makes adjacent overlapping edges between the two.
In the preferred embodiment of the present invention, as shown in Figure 1, the test strip is having a size of 80mm × 4mm；Wherein PVC board
1 having a size of 80mm × 4mm；Sample pad 2 is having a size of 25mm × 4mm；Bonding pad 3 is having a size of 20mm × 4mm；NC film 4 having a size of
20mm×4mm；Water absorption pad 7 is having a size of 21mm × 4mm；The sample pad 2 and 3 overlapping edges of bonding pad；The NC film 4 and suction
Water cushion 7 overlaps；The bonding pad 3 is overlapped with NC film 4；Overlapping part length is 2mm.
In the preferred embodiment of the present invention, as shown in Fig. 2, the sample pad 2, bonding pad 3, NC film 4 and water absorption pad 7 are pressed
Sequence successively sticks in PVC board from top to bottom.
In the preferred embodiment of the present invention, as shown in figure 3, the sample pad 2, bonding pad 3, NC film 4 and water absorption pad 7 are pressed
Sequence successively sticks in PVC board from bottom to top.
In the preferred embodiment of the present invention, as shown in figure 4, further including the shell equipped with cavity；The cavity of the enclosure interior
Place test strip；Observation window 10 and well 9 are set on the shell.
In the preferred embodiment of the present invention, as shown in figure 4, the shell includes upper housing 11 and lower case 12；The upper casing
Body 11 and lower case 12 fasten, and internal formation cavity；The observation window 10 and well 9 are located on upper housing.
In the preferred embodiment of the present invention, the lanthanide series is europium.
In the preferred embodiment of the present invention, as shown in Figure 1,6 spacing 6mm of detection line 5 and nature controlling line on the NC film 4 ±
0.1mm；The detection line 5 is close to bonding pad；The nature controlling line 6 is close to water absorption pad.
In the preferred embodiment of the present invention, as shown in figure 4, further including the ID card containing calibration curve.
In the preferred embodiment of the present invention, I spouting liquid of S100- β monoclonal antibody is 0.8ul/cm, concentration 0.5-
2mg/ml；Sheep anti-mouse antibody spouting liquid is 0.8ul/cm, concentration 0.1-0.5mg/ml.
In the preferred embodiment of the present invention, as shown in figure 4, the ID card is the identification code of mobile device identification；The mark
Knowing code 8 is two dimensional code or bar code；It is arranged on shell.
Embodiment one
Kit includes detection card, the ID card containing calibration curve in the present embodiment；Detection card includes that shell and setting exist
The intracorporal test strip of shell；Test strip the preparation method is as follows:
1. solution needed for preparing
1) 10%BSA solution
2) 20mg/ml EDC solution
3) 20mg/ml NHS solution
4) 0.02mol/L MES solution, pH5.6
5) 0.02mol/L phosphate solution, pH7.4
6) 0.01mol/L PB solution, pH7.4
7) liquid is redissolved
8) 0.01mol/L, PBS pH7.4 solution (needing matching while using)
9) coating buffer (needing matching while using)
2. the activation of time-resolved fluorescence microballoon
It washes ball: taking the fluorescent microsphere of 100ul to be added in the pH=5.6MES buffer of 900ul, mix, 4 DEG C of centrifugations
(11000g, 15min) abandons supernatant, and pH=5.6MES buffer is added and redissolves to original volume, is vibrated and is mixed with vortex mixer.
Activate microballoon: NHS, EDC and the NHS final concentration for being separately added into the EDC and concentration 20mg/ml of concentration 20mg/ml are equal
For 0.5mg/ml, room temperature is activated 15 minutes, and 4 DEG C of centrifugations (11000g, 15min) abandon supernatant, with 0.02mol/L pH7.4 phosphoric acid
Salt buffer is redissolved to 500ul.
3. time-resolved fluorescence microballoon marks S100- β monoclonal antibody II
Antibody is added: S100- β monoclonal antibody II is diluted with 0.02mol/L pH7.4 phosphate buffer, keeps it dense eventually
Degree is 1.0mg/ml, and microballoon is mixed with antibody-solutions 1:1 after dilution after being redissolved with 0.02mol/L pH7.4 phosphate buffer
It closes, is incubated at room temperature 2h.
Closing: after incubation time arrives, measuring appropriate 10%BSA solution with liquid-transfering gun, makes the final concentration of BSA to 1%, room
Temperature closing 30min.
Centrifugation: after closing, with centrifugal force 11000g, the high speed centrifugation 15min at 4 DEG C.After centrifugation, gently take
Whether test tube out, observation test tube have wall built-up phenomenon, and whether test tube wall has bright spot.It is drawn and supernatant and is discarded with liquid-transfering gun, when absorption
The precipitating of test tube bottom should not be contacted.
It redissolves: redissolution liquid is added and is settled to 1ml, and defines this as protocorm liquid.The S100- β monoclonal redissolved is resisted
II protocorm liquid of body is mixed with redissolution liquid in the ratio of 1:0.6, and is mixed.
4. the preparation of bonding pad
II solution of S100- β monoclonal antibody for being marked the time-resolved fluorescence microballoon with three-dimensional stroke of film gold spraying instrument, with
Discharge rate is 6ul/cm, and nozzle movement speed is that the standard of 80mm/s is sprayed on bonding pad.The bonding pad prepared is by such as
Lower method drying: drying temperature are as follows: 37 DEG C, humidity < 30%, the time: 60 ± 5min, the bonding pad after drying are put in containing dry
In the aluminium foil bag of drying prescription, sealing.
5.NC film draws film
Patch NC film (nitrocellulose filter): NC film is cut into 20mm × 300mm by process requirements amount, is attached to away from PVC board
Along 2.6cm.PVC bottom plate is laid on work top；Open the protective film that PVC bottom plate NC film pastes place, the NC that will be cut out
Film uniformly promotes from left to right, pastes it securely in PVC board.
Antibody is prepared: antibody prepares each component method for determination of amount are as follows: 1) antibody dosage (V)=dose volume × is matched
Concentration processed/antibody indicates concentration；2) coating buffer dosage (V)=dose volume × 0.1；3) 0.01mol/L PBS PH7.4 dosage
(V)=amount of preparation-antibody-coating buffer dosage.C line sheep anti-mouse antibody compound concentration should be 0.2mg/ml, T line S100- β monoclonal
I compound concentration of antibody is 1.0mg/ml.
NC film draws film: drawing film gold spraying instrument using three-dimensional, T line antibody and C line antibody are sprayed on NC film.The T line is anti-
Body, spouting liquid 0.8ul/cm, scribing line speed are 100mm/s, concentration 1mg/ml；The C line antibody, spouting liquid 0.8ul/
Cm, scribing line speed are 100mm/s, concentration 0.2mg/ml.T line is drawn away from NC film top 7.0mm ± 0.1mm at, C line stroke away from
At NC film top 7.0mm ± 0.1mm, T line and C line are at a distance of 6mm ± 0.1mm.
The drying of the NC film prepared: drying temperature are as follows: 37 DEG C, humidity < 30%, the time: 60 ± 5min, after dry
It is stuck in the aluminium foil bag containing desiccant greatly, seals.
6. assembling slitting
Water absorption pad is pasted: opening the protective film that PVC bottom plate water absorption pad pastes place, slight roll promotes water absorption pad, makes it
It is even to attach in PVC board, and water absorption pad covers NC film 2mm；
Bonding pad is pasted: opening the protective film that PVC bottom plate labeling pad pastes place, the bonding pad cut is attached on
In PVC board, the same water absorption pad of method.Bonding pad covers NC film 2mm；
PVC board the lowermost sample pad protective film is opened in sample pad stickup, sample pad is adhered in PVC board, method is same
Water absorption pad, sample pad cover bonding pad 2mm.
7. slitting
Using the automatic cutting machine of micro computer, assembled detection kilocalorie is cut into the examination that width is 4.0mm ± 0.1mm
Paper slip.The test strip is having a size of 80mm × 4mm；Wherein PVC board is having a size of 80mm × 4mm；Sample pad is having a size of 25mm
×4mm；Bonding pad is having a size of 20mm × 4mm；NC film is having a size of 20mm × 4mm；Water absorption pad is having a size of 21mm × 4mm.
8. the performance test pair S100- β albumen time-resolved fluoroimmunoassay chromatography kit:
The drafting of 8.1 calibration curves
The preparation of calibration object: calibration object is various concentration S100- β proteantigen, is diluted to obtain 10ng/mL with calf serum
Calibration object mother liquor, then diluted to obtain the calibration object of various concentration with calf serum.Each concentration calibration product use intra-company school
Quasi- product are traced to the source and assignment process, carry out assignment to calibration object.
Detection card is chromatographed using S100- β albumen time-resolved fluoroimmunoassay manufactured in the present embodiment, with the blue vigorous biology in Guangzhou
The AFS-1000 dry type fluorescence immunity analyzer of Science and Technology Ltd.'s production is test platform, tests the calibration of the various concentration
Product, each concentration retest 3 times.The fluorescence signal and T/C value of C line, T line are read, experimental result such as the following table 1:
1 calibration object test result of table
Using the average value of calibration object concentration and fluorescence signal T/C ratio as reference axis, calibration curve is drawn.Calibration graph
As shown in figure 5, wherein R2Value is 0.9999.It is realized by calibration curve and quantitative survey is carried out to S100- β protein concentration in sample
It is fixed.
8.2 time-resolved fluorescences detect card performance test
Minimum detectability: using zero-dose calibration object or Sample dilution to be detected as sample, replication 20 times, obtains
20 measurement results out calculate its average value (M) and standard deviation (SD), obtain M+2SD, as minimum detectability, result is
0.08ng/mL。
The range of linearity: the calibration object of 6 concentration in 0.08-10.0ng/mL concentration range, each concentration calibration product weight are taken
Repetition measurement tries three times, and measurement mean concentration and theoretical concentration are carried out linear analysis, analyze result such as Fig. 6, linear equation y=
0.9999x+0.1489, R2Value is 0.9999, shows that kit of the present invention is good in 0.08-10.0ng/mL intervals linear correlation
It is good.
Repeatability: taking the detection card prepared in same lot number embodiment 1, respectively detectable concentration in 0.5 ± 0.05ng/ml and
The quality-control product of 5 ± 0.5ng/ml each 10 times, the average value (M) and standard deviation (S) of measured value are calculated separately, calculates the coefficient of variation
(CV), CV value≤15%.The quality-control product of two concentration is tested, test result is respectively as follows: low concentration quality-control product average value and is
0.05ng/mL, CV value are 5.2%；High concentration quality-control product average value is that 4.98, CV value is 3.7%, and test result, which is all satisfied, to be wanted
It asks, shows that the detection card repeatability of 1 scheme of embodiment preparation is good.
Accuracy: low concentration human serum sample is detected, and replication 3 times respectively, calculates its average value C0；Take height
1 part of sample of value replication 3 times respectively, calculates its average value Cs；High level sample is added separately in 9 parts of human serum, is mixed
It after even, is detected as the step of detecting sample to specifications, replication 3 times respectively, calculates its average value C, calculate
The rate of recovery.Wherein, V0For human serum volume；VsBody is added for high level sample
Product.The rate of recovery for recycling sample 8.2ng/mL is 101.5%；The rate of recovery of 5.88ng/mL is 96.2%；The rate of recovery is in 90%-
Between 110%, illustrate that the detection card accuracy of example scheme preparation is good.
Difference between batch: taking the kit prepared according to example scheme of 3 batches of different lot numbers, and every batch of respectively takes 3 detection cards, surveys
Concentration is tried each 3 times in the quality-control product of 0.5 ± 0.05ng/ml and 5 ± 0.5ng/ml, calculates separately the equal of 3 testing results of every batch of
Value.Wherein three batches of kits detect 0.5 ± 0.05ng/ml concentration quality-control product, and relative deviation R is 8.22% between batch；Three batches of reagents
Box detects 5 ± 0.5ng/ml concentration quality-control product, and relative deviation R is 4.97% between batch；Relative deviation R is respectively less than 10% between batch, table
The bright kit prepared according to embodiment 1 and 2 scheme of embodiment, difference between batch meet the requirements.
The detection of 8.3 clinical samples
200 parts of clinical serum sample of acquisition is produced using the present invention and Wuhan Easydiagnosis Biomedicaine Co., Ltd.
S100- β albumen (S100- β) detection kit (immunochromatographic method), the AFS- that Lan Bo Biotechnology Co., Ltd produces in Guangzhou
The detection of sample is carried out on 1000 dry type fluorescence immunity analyzer test platforms.By same sample, the detection knot of different kits
Fruit carries out linear analysis, as shown in figure 5, R2 value is 0.9997, shows that the present invention is with uniformity with commercialized product testing result,
Suitable for clinical detection.
Claims (10)
1. a kind of S100- β albumen time-resolved fluoroimmunoassay chromatographs detection kit, including test strip, the detection examination
Paper slip includes PVC board, sample pad, bonding pad, NC film and water absorption pad；It is characterized in that, the test strip is with PVC board
Solid phase support bottom plate；The sample pad, bonding pad, NC film and water absorption pad make in this order it is adjacent be in close contact between the two,
Then it sticks in PVC board；The bonding pad is coated with the S100- β monoclonal antibody II of time-resolved fluorescence microballoon label；Institute
Stating time-resolved fluorescence microspherulite diameter is 200nm-300nm, is marked with lanthanide series；
Detection line and nature controlling line are set on the NC film；Detection line is the S100- β monoclonal antibody I being sprayed on NC film；Quality Control
Line is the sheep anti-mouse antibody being sprayed on NC film；The detection line and nature controlling line are parallel to each other and are equipped with gap.
2. a kind of S100- β albumen time-resolved fluoroimmunoassay according to claim 1 chromatographs detection kit, feature
It is, the sample pad, bonding pad, NC film and water absorption pad make adjacent overlapping edges between the two in order.
3. a kind of S100- β albumen time-resolved fluoroimmunoassay according to claim 1 or 2 chromatographs detection kit, special
Sign is that the test strip is having a size of 80mm × 4mm；Wherein PVC board is having a size of 80mm × 4mm；Sample pad having a size of
25mm×4mm；Bonding pad is having a size of 20mm × 4mm；NC film is having a size of 20mm × 4mm；Water absorption pad is having a size of 21mm × 4mm；Institute
State sample pad and bonding pad overlapping edges；The NC film and water absorption pad overlap；The bonding pad and NC film overlap；Clinch bit length
Degree is 2mm.
4. a kind of S100- β albumen time-resolved fluoroimmunoassay chromatography inspection described in any one of -3 claims according to claim 1
Test agent box, which is characterized in that further include the shell equipped with cavity；The cavity of the enclosure interior places test strip；Institute
It states and observation window and well is set on shell.
5. a kind of S100- β albumen time-resolved fluoroimmunoassay according to claim 1 chromatographs detection kit, feature
It is, the shell includes upper housing and lower case；The upper housing and lower case fasten, and internal formation cavity；The sight
It examines window and well is located on upper housing.
6. a kind of S100- β albumen time-resolved fluoroimmunoassay according to claim 1 chromatographs detection kit, feature
It is, the lanthanide series is europium.
7. a kind of S100- β albumen time-resolved fluoroimmunoassay according to claim 2 chromatographs detection kit, feature
It is, detection line and nature controlling line spacing 6mm ± 0.1mm on the NC film；The detection line is close to bonding pad；The Quality Control
Line is close to water absorption pad.
8. a kind of S100- β albumen time-resolved fluoroimmunoassay according to claim 1 or 4 chromatographs detection kit, special
Sign is, further includes the ID card containing calibration curve.
9. a kind of S100- β albumen time-resolved fluoroimmunoassay according to claim 1 or 4 chromatographs detection kit, special
Sign is that I spouting liquid of S100- β monoclonal antibody is 0.8ul/cm, concentration 0.5-2mg/ml；Sheep anti-mouse antibody spouting liquid
For 0.8ul/cm, concentration 0.1-0.5mg/ml.
10. a kind of S100- β albumen time-resolved fluoroimmunoassay according to claim 8 chromatographs detection kit, feature
It is, the ID card is the identification code of mobile device identification；The identification code is two dimensional code or bar code；It is arranged in shell
On.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811572355.2A CN109856403A (en) | 2018-12-21 | 2018-12-21 | A kind of S100- β albumen time-resolved fluoroimmunoassay chromatography detection kit |
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Cited By (1)
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Application publication date: 20190607 |