CN104777298B - A kind of detection means - Google Patents
A kind of detection means Download PDFInfo
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- CN104777298B CN104777298B CN201510103101.6A CN201510103101A CN104777298B CN 104777298 B CN104777298 B CN 104777298B CN 201510103101 A CN201510103101 A CN 201510103101A CN 104777298 B CN104777298 B CN 104777298B
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- G—PHYSICS
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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Abstract
The present invention relates to it is a kind of to detect in sample whether the detection means containing target analytes, including：Support to include detection zone on the carrier of liquid sample flow, the carrier；Described detection zone includes specific binding molecule, it is characterised in that include allowing the structure of fluid flows decrease in described detection zone downstream.This device can improve the sensitivity of detection.
Description
Technical field
The invention belongs to medical diagnosis class article, be exactly specifically can with quick detection sample whether contain target analysis
A kind of detection means of thing.
Background technology
Examined in any one, detection zone is all very important part.In immunological test, these detections
Reaction on region generally is combined to form to complete by immunity principle, and the result in detection zone can be shown as on test bar
Colored line.In addition to these basic requirements, the sensitivity for improving detection is one of target that product is updated.
The content of the invention
The test device that the present invention is provided, can be very good to improve the sensitivity of detection means.On the one hand, the present invention is provided
A kind of immunoassay device, including the carrier of liquid flowing is supported, detection zone is included on carrier, in the downstream of detection zone
Structure including slowing down liquid flowing.The effect of this structure for slowing down liquid flowing is exactly to allow more liquid to rest on detection
Region allows liquid to rest on for a long time in detection zone, and the material allowed in liquid can fill with the reagent in detection zone
Divide reaction to improve the sensitivity of detection.
In a mode, detection zone is located on film, includes slowing down liquid flowing on the film in detection zone downstream
Structure.When sample solution reaches detection zone, solution temporal persistence is on detection zone so that on sample and detection zone
Reagent fully reacts for a long time.After brief stay, liquid is acted on due to the capillary pulling power of film itself, allows liquid to continue
Downstream is flowed to, whole detection is finally completed.In a word, the effect of this structure for slowing down liquid flowing is to enable solution in detection zone
On reagent more fully reacted.In a preferred mode, the width for making way for the film in detection zone downstream is less than
The width of detection zone, when liquid flows to detection zone, due to such structure, allow liquid flows decrease or to
The flow in detection zone downstream is reduced, and so allows liquid phase by detection zone to resting in detection zone more.With liquid
The flowing of body, liquid is eventually through narrow section downstream diffluence.
In another preferred mode, hydrophobic agents are handled on the subregion in the downstream of detection zone, detection is allowed
The hydrophobic ability of region downstream part is substantially greater than the hydrophobic ability of detection zone in itself.So, when liquid flows to detection zone
When, due to the detection zone downstream hydrophobic ability different from detection zone, the liquid in detection zone is by downstream
Hydrophobic part stop, flows decrease, so allow liquid phase to rest on more in detection zone with the reagent in detection zone more connect
A little times are touched, react more abundant, so as to improve the sensitivity of detection.The hydrophobic agents handled in detection zone downstream can be with
It is conventional hydrophobic agents, particularly, the hydrophobic agents that can be used on nitrocellulose filter.
On the other hand there is provided the detection means for including immunodiagnosis test strips, for detecting the target in sample point
Analyse thing.In some embodiments, analyte is antibody, such as resisting HIV (HIV) antibody or anti-hepatitis C
Viral (HCV) antibody.In other embodiments, analyte is antigen, such as human chorionic gonadotrophin (hCG).
The diagnosis test paper is preferably included containing detection zone, and detection zone includes that target analytes can be specifically bound
Fixed bonding agent.For example, if target analytes are antibody, positive detection area preferably includes fixed antigen.If
Target analytes are antigen, then positive detection area preferably includes fixed antibody.Include slowing down liquid in the downstream of detection zone
The structure of flowing.
It is preferred that, test strips include label derived region, and label includes the first knot for being capable of combining target analyte
Synthesis point and the second display composition.The label source area for being designed to the label of combining target analyte is located at detection zone
Upstream；Come source region upstream positioned at label and be designed to receive the sample application zone of fluid sample.
The sample used in experiment can be any liquid.It is preferred that sample include, for example, blood, serum, blood plasma, saliva
Liquid and urine.Sample is added to sample application zone, then carrys out source region outflow from label.Label flows through film, contacts combined
To all analytes in positive detection area, visible signal is produced.
In a preferred embodiment, test strips include the detection zone with fixed HIV antigens, anti-for detecting
HIV antibody.In this embodiment, the experiment can be used for the HIV of diagnosis patient.In a further preferred embodiment,
Test strips include the detection zone with fixed HCV antigens, for detecting the HCV antigen/antibody combination in sample.In this embodiment,
The experiment can be used for the HCV infection of diagnosis patient.In still another embodiment, test strips include resisting with fixed anti-hCG
The detection zone of body.In this embodiment, the experiment can be used for determining whether individual is pregnant.
Label carrys out source region and preferably includes dry label.For example, it can be labeling pad that label, which carrys out source region, mark
Note thing is dried thereon.Labeling pad is preferably connected with membrane fluid.In some embodiments, after being loaded to sample application zone, mark
Note thing pad is moved to be connected with membrane fluid.In another embodiment, label carrys out the part that source region is film, and label exists
Dry thereon.
In another embodiment, label is suspended in buffer solution.By the way that the buffer solution containing label is added to
Label comes in source region to make label flow to detection zone.
In another embodiment, test strips also include the check plot for being located at detection zone downstream on film.Check plot is preferred
Ground includes the control bonding agent for being capable of binding label.The structure for slowing down liquid flowing is located between detection zone and check plot.
Test strips can include one or more kinds of other assemblies.Buffer liquid pad can carry out source region fluid with label
Connection, so can make label carry out source region outflow from label to application lavation buffer solution on buffer liquid pad, through film, contact
To detection zone.Water sucting belt and adsorptive pads can be located at the downstream of film, when buffer solution is by film for absorbing excessive buffer solution.
Drying agent chip can also be close to film, for absorbing excessive liquid and moisture.
Test strips can be optionally placed in shell such as plastic casing.Shell is preferably included positioned at detection zone and right
According to the window above area, and the sample window positioned at detection zone upstream.
The method of the analyte in detection fluid sample is additionally provided, including sample is added to sample window or sample application zone,
Sample flows successively through a kind of label that label comes in source region, and the label combining target analyte and can produce instruction knot
The visible signal of conjunction.
Beneficial effect
The sensitivity of detection can be improved using the device of the present invention.
Brief description of the drawings
Figure 1A is the test strips dimensional structure diagram in an embodiment of the invention, and Figure 1B is illustrated by Figure 1A
The top plan view that assembles of test strips.
Fig. 2 be another embodiment of the present invention in test strips detect film dimensional structure diagram.
Fig. 3 be another embodiment of the present invention in test strips detect film dimensional structure diagram.
Fig. 4 be another embodiment of the present invention in test strips detect film tangent plane structural representation.
Description of reference numerals
It is loaded pad 18；Label pad 12；Detect film 15；Detection zone 11；Control zone 10；Fluid flows decrease region
13103；Outer object 114.
Definition
Unless otherwise defined, all scientific and technical terminologies here all have and the usual institute of those skilled in the art in the invention
The identical connotation known.It can be used in the implementation of the present invention and described herein similar or suitable all methods, device and material
Material.
Term " label " refers to be designed to combining target analyte and produces the composition of detectable signal.Label
Generally comprise the binding constituents with mark substance markers.The binding constituents enable label combining target analyte, optionally tie
Close control compound.Label produces detectable signal, preferably visible signal.In one embodiment, only label with
Visible signal is just produced when analyte is combined.In a preferred embodiment, target analytes are antibody and label includes
It is capable of the binding constituents of binding antibody.For example, the first binding constituents can be albumin A, although other can also be used to tie
Close the molecule of antibody.In another embodiment, target analytes are that the binding constituents of antigen and label include tying
Close the antibody of antigen.Antibody is preferably monoclonal antibody.
Label is can to adhere to binding constituents or mark and can produce any molecule or the change of detectable signal
Compound.A kind of preferred label is collaurum.Or, label can also be the latex particle of such as dyeing, collargol or
Other colloidal metals, colloid be black or other compositions well known in the art.Preferably, the particle diameter of label is without interference with binding constituents
The ability of combining target analyte.For example, when binding constituents are albumin A, the particle diameter of label is preferably from about 5 nanometers to about
120 nanometers, more preferably from about 20 nanometers to about 80 nanometers.
" mark buffer solution " middle preparation of the label can stablized and preserved to label preferably.In an embodiment party
In case, as described in detail, after preparing label in mark buffer solution, label can the drying in " labeling pad ".
The label dried in substrate is considered as " diffusible to combine ".If can cause diffusion or flow, for example by its with
Buffer fluid contacts, then label is " diffusible to combine ".For example, can spread by drying and be attached to label come in source region
Label can use buffer solution so that its membrane flow along test strips.
" contrasting marking thing " is such a label, and wherein binding constituents are specific for control compound.When
May all labels all combined with the analyte in detection line and be not used as control when, preferably use contrasting marking thing.It is right
According to compound be usually in analyzed sample it is known, and the control bonding agent that control take can be combined.Optionally, it is right
Binding constituents according to label are probably specific for detecting compound present in the check plot of film.
Label is preferably in " label carrys out source region ".Label carrys out source region and generally comprises diffusible combination therewith
Label, such as dry label.Or, label carrys out source region and potentially included to be suspended in buffer solution such as lavation buffer solution or mark
Remember the label in thing buffer solution.In one embodiment, it is labeling pad that label, which carrys out source region, including dry mark
Thing, is connected with detection membrane fluid.In another embodiment, it is to detect a part for film, label that label, which carrys out source region,
Dried thereon.In another embodiment, it is buffer liquid pad that label, which carrys out source region, can upward be applied comprising mark
The buffer solution of thing, it is connected with detection membrane fluid.Label comes the upstream that source region is normally at the detection zone of detection film, so that
Add after the solution comprising label or after the diffusible label dissolving combined, label flows through detection film, with inspection
Survey area's contact.
Label is preferably dissolved with " lavation buffer solution " As described in detail below.Briefly, lavation buffer solution is
Preferably comprise the cushioning liquid of " sealer ".Sealer is well known in the art, including reduction non-specific interaction is for example non-
Any molecule or compound that specific antibody is combined.It is preferred that sealer be protein, such as casein and bovine serum albumin(BSA)
(BSA).Other available sealers can be bought, and those skilled in the art should appreciate that.
" detection film " is a kind of solid support, including detection zone, and optional check plot.Detect that film can be any solid
Body holder, analyte binding agent and control bonding agent can be attached to above.Detection film is preferably nitrocellulose.
Detection film provides side stream passages for liquid.If liquid such as fluid sample or buffer solution can be from a component flow directions
Another component, then the other assemblies for claiming detection film and test strips are " fluid communication ".If the component of test strips connects each other
Touch, then they are to be in fluid communication.But, do not need two specific components it will be appreciated by those skilled in the art that being in fluid communication
Directly contact.
" detection zone ", " detection band " and " detection line ", which can be exchanged, to be used, all referring to analyte binding agent institute on detection film
The region of attachment.
" analyte binding agent ", " specific binding ligand " or " binding constituents " is to be designed to specific binding target point
Analyse any molecule or compound of thing.For example, but be not limited to, analyte binding agent can include antigen, antibody, acceptor, its
His polypeptide, peptide, haptens, agglutinin, nucleic acid include oligonucleotides or small molecule.In one embodiment, analyte is combined
Agent is a kind of antigen, and it is specific for the test antibodies in sample.In another embodiment, analyte is combined
Compound is the specific antibody of target antigen.
" check plot ", " control band " and " control line " can exchange and use, and be compareed all referring on film accompanying by bonding agent
Region.
" control bonding agent " is can to specifically bind any molecule or compound of label or contrasting marking thing.
" test strips " be refer to for detect in sample whether the intact device containing analyte.Test strips are preferably extremely
It is few to include the detection film with detection zone.As described below, test strips can be placed in shell such as plastic casing.Or, examination
Paper slip can also be without shell.If placed in shell, test strips and shell can be referred to as " detection card " together.
" detection card window " and " sample window " refer to detect the hole in card shell, can be loaded by it.Detect card window
Mouth is preferably located in the top of detection zone and check plot.Card window is detected it is also preferred that allowing viewing test result.
" sample application zone " is the position for applying sample in detection means or test strips.Sample application zone is preferably located in label source
The upstream in area.Sample can be applied directly to sample application zone, such as by pipette, or apply indirectly.
The term " antibody " used is its most wide connotation, includes, but not limited to, e.g. complete antibody and single-chain antibody, anti-
Body fragment and chimeric antibody, as long as they retain the binding specificity needed.
It is described in detail
In the following detailed description, the subsidiary reference word of legend is a part here, and it is to illustrate this
Invention can the mode of actable specific concrete scheme illustrate.We, which are not precluded from the present invention, can also carry out other specific
Scheme and the structure for changing the present invention in the case of the use scope without prejudice to the present invention.
Carrier
The carrier of liquid flowing is supported to refer to that liquid from an area can flow to another area on carrier.In a tool
In the mode of body, liquid can flow to detection zone from label derived region.
On the one hand, the flowing of this liquid can be the difference of carrier material itself and allow being moved of liquid active.
In one concrete scheme, there is provided the carrier material for supporting liquid flowing comprising a kind of absorbent material for device." carrier material " is
Refer to a kind of material for supporting liquid flowing and transport.In a concrete scheme, carrier material is a kind of absorbent material.Liquid flow
It is to be acted on realizing by means of capillary motion to cross the present apparatus.It can include detection zone on carrier and label carrys out source region
Domain.Certainly sample area and suction zone can also be included.In different concrete schemes, carrier material can be homogenous material
The bar of composition or the test strips being made up of a variety of absorbent materials interacted in a liquid, such as Fig. 1 are carried
Body may include sample application pad 18, label pad 12, the nitrocellulose membrane group of glass fibre composition of glass fibre or filter paper composition
Into detecting pad 15 and filter paper composition adsorptive pads 17, they couple successively can allow liquid to flow to adsorptive pads from sample pad
On.Processing has labeled material in label pad；15 include detection zone 11 on detecting pad, or preferred include control
Region 10, includes slowing down the structure 13 of liquid flowing in the downstream of detection zone." water suction " material refers to that those can stablize
Ground absorbs moisture and moisture is acted on the material transported by capillary motion wherein.It is fine that the example of absorbent material includes nitrification
Dimension, filter paper, glass fibre, polyester and other suitable materials.Meanwhile, this carrier material also includes only providing one or several
The carrier of single capillary, liquid is moved by these single capillarities.
In a preferred mode, the reagent handled on carrier exists in a dry state, when liquid is applied to
When on carrier, carrier is moistened by liquid.
Slow down the structure of flow rate of liquid
Liquid flows what is always carried out with certain speed on carrier, abbreviation flow velocity.Different carrier materials have difference
Flow rate of liquid, when carrier material is homogenous material, liquid but can lead in the speed also relative constancy of upper flowing
Cross the flow velocity for setting some structures to allow liquid on carrier on carrier and be in non-constant state.
For example, shown in Fig. 4, carrier is oppressed in 13 position with hard foreign object 114 on the carrier material of uniform material, allows
Carrier 13 position produce contraction, when liquid from 152 flow to 111 while when, due to allow carrier quality no longer
Uniformly, liquid flows decrease when flowing through 13, but still there is partially liq to be flowed by 13 positions to 111 parts, this
Sample allows more liquid to rest on 152 parts, but with the extension of time, the liquid being eventually located on 152 positions is by 13
Flow to 111 positions.In addition, hard foreign object can be detect card shell on a part, hard foreign object can be plastics or
Metal.
In addition to above physical method, some organic solvents can also be handled on the regions 13, allow the structure of film to become
Change, such as some organic solvents, such as alcohol can destroy the structure of some carriers, and such as nitrocellulose uses ethanol postincubation
The place depression for the carrier crossed, liquid is when the place of depression is flowed through, and flow rate of liquid is slowed.Alternatively, it is also possible to another
The carrier of some outer materials comes connecting detection region 11 and control zone 13 (optionally having), liquid in the place in region 13
Flow velocity on carrier 13 is less than flow velocity of the liquid on the carrier of detection zone 11, can also so obtain same effect.
The detection zone of traditional immunoassay device is frequently located on the single carrier of material, such as nitrocellulose filter
On, the speed that liquid flows on carrier also relative constancy, and the present invention subtracts on the carrier containing detection zone including some
The structure of slow flow rate of liquid, allows liquid phase to resting on a little times that contacted more with reagent thereon in detection zone, reaction is more
Plus fully, so as to improve the sensitivity of detection.
It is preferred that, these structures for slowing down flow rate of liquid are located at the downstream of detection zone.In a specific mode, such as
Fig. 1, test strips include including the He of detection zone 11 on sample application region 18, label derived region 12 and detection film 15, detection film
Control zone 10, the hydrophobic region 13 for including being treated with hydrophobic agents in the downstream of detection zone, and connect with detection film
The suction zone 17 connect, these regions head and the tail are connected, and allow liquid can be from sample application region 18 by marked region 12 and detection film
Flowed on 15 in suction zone 17.When in use, when liquid flows to detection zone, due to depositing for hydrophobic region 13
Allowing flow rate of liquid to slow down at hydrophobic region, allowing more liquid to contact the reagent in detection zones or contact detection zone
The longer time, reaction can be allowed more abundant, the sensitivity of detection is improved.Then liquid passes through slowly hydrophobic region and flowed to
The control zone in downstream.
The detection film of some commercializations is hydrophilic in itself, can so be handled in detection zone downstream with hydrophobic agents
It is allowed to become water repellent region, and the detection film of some commercializations is hydrophobic in itself, detection zone can be processed into
It is hydrophilic, without the region of processing detection region downstream, the hydrophobic property for allowing it to retain itself.
Those any reagents that can change carrier hydrophobic performance can apply to the present invention.Particularly, one
A little hydrophobic agents processing change the reagent of hydrophobic performance on nitrocellulose filter.The method on film of processing can be
Apply, draw or sprayed with automatic spraying machine, these methods are that those skilled in the art easily realize.Here dredge
Water-based reagent can be sulfydryl polysiloxane emulsion, emulsifying wax wax emulsion, butadiene-styrene latex etc..
In another specific mode, can also make way for detection zone downstream detection film relative narrowness some, this
Sample can also allow the flows decrease of liquid.As shown in Fig. 2 the direction of the liquid flowing shown in arrow, in detection zone 11
Downstream allow detection film to want narrower with respect to detection zone, the narrow connecting detection region of part 103 and control zone 11.Example
Such as, the width in detection zone downstream is the 1/2-1/10 of detection zone width, or 1/4-1/8；Or 1/5.When liquid flows to detection
When region, due to unexpected narrow in downstream, due to the flow velocity of narrow 103 certain (material and inspection when narrow
When the material of the carrier in survey region is the same or when material immobilizes), but the ability of conveying liquid declines, no
Timely the liquid from detection zone can be transported in control zone 11, so allow more liquid to rest on detection zone
On, allow more materials, such as analyte substance of interest and/or mark substance, target analytes and mark substance in fluid sample
The compound substance of formation, the reagent in detection zone 11 is contacted with the more time, and such as fixed antibody or antigen is improved
The sensitivity of detection.Certainly, in a selectable mode, such as Fig. 3, the downstream of detection zone 11 gradually becomes narrow and also can
Play similar effect.In the concrete mode shown in Fig. 2 and 3, it is furthermore preferred that constituting the material of the carrier of narrow 103
Material different from constituting the carrier of detection zone 11, is so selected, and is allowed liquid to be less than liquid by the flow velocity of narrow and is led to
The flow velocity crossed in detection zone, so more can make more liquid rest on detection zone, so that allow reagent more fully to react,
Improve the sensitivity of detection.
It is of course also possible to individually with different liquids flow velocity support liquid flow carrier come constitute detection zone with
The region in detection zone downstream.For example detection zone individually can be constituted with the fast carrier of flow rate of liquid, use liquid velocity ratio
The small carrier of detection zone carrier flow velocity constitutes the region in detection zone downstream, allows the liquid can to flow to detection from detection zone
The downstream in region.When in use, when the liquid from detection zone flows to the region in detection zone downstream, liquid flow
Speed slows down；Allow this when liquid can be with more time or relatively many volumes are rested in detection zone；So allow liquid
In some materials, such as analyte or mark substance can more contact detection zone in quantity or on the time
On secure bond molecule, obtain higher sensitivity.
The above method can be used individually, can also be used in combination.In addition, the control in the above embodiment
Region alternatively, in other modes, it is convenient to omit control zone.
The type of target analytes
Any analyte substance of interest can be analyzed with the present invention.The example of the target analytes of stable detection of the present invention can be used
Attached bag includes (but not only including) human chorionic gonadotrophin (hCG), and lutropin (LH), ovarian stimulation is plain (FSH),
The medicine of hepatitis C virus (HCV), hepatitis B (HBV), hepatitis B surface antigen, AIDS virus and any abuse.Analyte
It can be detected in any liquid or liquefied sample, such as urine, saliva, saliva, blood, blood plasma, or serum.Its
The example of its target analytes also has flesh ammonia acid anhydride acid, and bilirubin, nitrite, protein (nonspecific), blood is white thin
Born of the same parents, blood glucose, heavy metal and toxin, bacterial component (for example, the special protein and sugar of certain types of bacterium, for example greatly
Enterobacteria 0157:H7, staphylococcus aureus, salmonella, C.perfringens, campylobacter, listeria monocytogenes,
V. parahaemolyticus, or wax bacillus).The analyte of any other suitable lateral flow test format can use the present apparatus
Detection.
The type of sample
Any kind of sample can be tested with the device of the present invention, including body fluid is (for example, urine and other bodies
Liquid, and clinical sample).Fluid sample may originate from solid or semisolid sample, including excrement, biological tissue and food
Sample.These solids and semisolid sample can be transformed into fluid sample by any suitable method, such as in one kind
Mixed in appropriate liquid, stamp broken, macerated, be incubated, dissolving or enzymolysis solid sample (for example, water, phosphate buffer or
Other buffer solutions)." biological sample " includes the sample from animal living, plant and food, also including urine, saliva, blood
With blood constituent, cerebrospinal fluid, vaginal swab, seminal fluid, excrement, sweat, secretion, tissue, organ, tumour, tissue and organ
It is the conditioned media of culture, cell culture and there, either people or animal.Foodstuff samples include finished food
Thing composition and last product, meat, cheese, wine, milk and drinking water.Plant sample include from any plant, plant tissue,
The sample of the conditioned media of plant cell cultures and there." environmental sample " is that those are derived from the sample of environment (for example, lake water
The sample of sample or other water bodys, sewage sample, pedotheque, groundwater sample, seawater sample, the sample of runoff water).
Sewage and related waste can also be included in environmental sample.
Experiment
1. experiment material：
Jin Biao：Antihuman hemoglobin mark gold mark；T lines：Antihuman hemoglobin；
Sample pad solution：Tris:6mg/ml；Casein:5mg/ml；PVP:5mg/ml；
Sample：Positive sample PBS extracts the FOB samples by confirming as the positive；' negative ' specimens buffer for PBS
The process that liquid is extracted confirms as the FOB samples of feminine gender
Water-repelling agent：Sulfydryl polysiloxane emulsion BD-307；
NC films, polymer PET and glass：
| Material | Model | Lot number |
| Detect film 15 | Millipore HF13520mm | R9PN67501 |
| Label pad 12：Polymer PET | Polymer PET | EP09020052 |
| It is loaded pad 18：Glass | CMA-435 glasses | EG09120050-T2 |
2. experimental method：
The processing of gold standard pad 12:
Gold solution is processed into OD60, and is sprayed on 2.0ul/cm discharge rate on polymer PET.37 DEG C of oven for drying are stayed overnight.
The processing solution of NC film T lines 11:Antihuman hemoglobin is diluted to 1mg/ml with PBS solution, and with 1ul/cm spray
Amount is sprayed on NC films, is stayed overnight in 37 DEG C of oven for drying.
The processing of sample pad 18:It is sprayed on sample pad solution on the wide glasses of 18cm, 37 DEG C of oven for drying are stayed overnight.
Processing 1:
The NC films being baked, NC is being sprayed onto above T lines with sulfydryl polysiloxane emulsion at about 0.2cm with 1.0ul/cm discharge rate
On film, hydrophobic region 13 is formed.And be placed again into 37 DEG C of oven for drying and stay overnight.(Figure 1A)
Processing 2：
It is being sprayed onto above T lines at about 0.5cm with 0.2cm with PBS solution with 1.0ul/cm discharge rate on NC films.And again
37 DEG C of oven for drying are put into stay overnight.
Processing 3：
At about 0.2cm, the wide osculums of each about 0.2cm are being cut off in film both sides with scissors, centre is stayed above the T lines of film
About 0.5cm bridge, enables solution to pass through from middle bridge 103.(Fig. 2)
Control：
Other modes are identical with processing 1-3, but without any processing to the downstream of detection line 11.
The assembling of reagent strip
According to above method, sample-adding pad 18, label pad 12, and detecting pad 15 and adsorptive pads 17 are first according to Fig. 1 method
Position connection, obtains correspondence and processing 1-3 and the reagent strip compareed.Then on the sample-adding pad 18 of 12 reagent strips of each processing
It is added dropwise 2 and drips positive FOB solution, ' negative ' specimens is then added dropwise again to each remaining 10 reagent strip of processing, then at 3 minutes
Inner reference standard colorimetric card reads testing result, and each processing is repeated 3 times.It is shown in Table 3.
3. experimental result：
| Positive sample | ' negative ' specimens 1 | ' negative ' specimens 2 | ' negative ' specimens 3 | |
| Control | +4 | - | - | - |
| Processing 1 | +8 | - | - | - |
| Processing 2 | +4 | - | - | - |
| Processing 3 | +8 | - | - | - |
4. summarize：
Handled on film after water-repelling agent, a strong hydrophobic surface has been formed, when the solution in sample and Jinsui River reaches this
When local, solution temporal persistence is in this place so that sample fully reacts with T lines.After brief stay, liquid
Due to pulling force effect, this layer of hydrophobic surface can be crossed, or is passed through from the bottom and both sides of film, race plate is finally completed.In a word, this
The effect of layer hydrophobic surface is biased sample solution is fully reacted in the capture antigen of T lines, and reaches that final T lines are fully anti-
The effect answered.
Similarly, in order that sample is reached after T lines so that sample can have the sufficient reaction time with T lines, use narrow bridge
Method, that is, allow the width of the part in T lines downstream to be less than width at T lines, this method has also reached same effect.
Claims (7)
1. it is a kind of to detect in sample whether the detection means containing target analytes, including:Support liquid sample flow
Include detection zone on carrier, the carrier；Described detection zone includes specific binding molecule, it is characterised in that in institute
The detection zone downstream stated includes allowing the structure of fluid flows decrease；The described structure for slowing down flow rate of liquid is in detection zone
Downstream includes hydrophobic parts, wherein, the hydrophobic ability of the downstream hydrophobic parts is more than the hydrophobic ability of detection zone.
2. device as claimed in claim 1, wherein, described hydrophobic parts are treated by hydrophobic agents.
3. device as claimed in claim 2, wherein, described hydrophobic agents are sulfydryl polysiloxane emulsion.
4. the device as described in one of claim 1-3, wherein, described detection zone is located on nitrocellulose filter.
5. it is a kind of to detect in sample whether the detection means containing analyte, including:Support the load of liquid sample flow
Include detection zone on body, the carrier；Described detection zone includes specific binding molecule, it is characterised in that described
The width in detection zone downstream is smaller than the width of detection zone；Detection zone downstream material is different from detection zone material, liquid
Flow velocity of the body in the small part of detection zone downstream width is less than flow velocity of the liquid in detection zone.
6. device as claimed in claim 5, wherein, the width in detection zone downstream is the 1/2-1/10 of detection zone width.
7. device as claimed in claim 6, wherein, the width in detection zone downstream is the 1/5 of detection zone width.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103163304A (en) * | 2011-12-19 | 2013-06-19 | 天津中新科炬生物制药有限公司 | Method for rapidly detecting follicle stimulating hormone (FSH) in urine sample in a quantitative mode |
| CN102768275A (en) * | 2012-07-10 | 2012-11-07 | 北京陆桥技术有限责任公司 | Staphylococcal enterotoxin assay kit, method for preparing same and application |
| CN106796226B (en) * | 2014-10-02 | 2019-10-15 | 索尼公司 | Target substance assay kit, target substance measurement system, immune chromatograph assay kit and immune chromatograph measure system |
| CN104777194B (en) * | 2015-04-21 | 2017-10-27 | 苏州市玮琪生物科技有限公司 | Anti-interference sensing electrode and micro-fluidic test paper runner flow rate control method |
| CN106525838A (en) * | 2016-12-07 | 2017-03-22 | 百奥森（江苏）食品安全科技有限公司 | Prednisolone determination method and prednisolone determination card |
| CN108535472B (en) * | 2018-02-27 | 2021-09-24 | 上海艾瑞德生物科技有限公司 | Detection test strip capable of remarkably improving lateral flow immunochromatography |
| CN109444400B (en) * | 2018-10-29 | 2022-01-28 | 陕西科技大学 | Lateral flow chromatography test paper and preparation method and application thereof |
| CN113466461B (en) * | 2021-08-20 | 2022-03-18 | 广东省第二人民医院（广东省卫生应急医院） | Phospholipase A2 lateral chromatography detection test strip, detection card and application thereof |
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| CN104777298A (en) | 2015-07-15 |
| CN102236022A (en) | 2011-11-09 |
| CN102236022B (en) | 2015-04-22 |
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