CN203745473U - Detection device - Google Patents
Detection device Download PDFInfo
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- CN203745473U CN203745473U CN201320612882.8U CN201320612882U CN203745473U CN 203745473 U CN203745473 U CN 203745473U CN 201320612882 U CN201320612882 U CN 201320612882U CN 203745473 U CN203745473 U CN 203745473U
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Abstract
The utility model provides a testing device, and particularly provides a rapid detection device. The detection device comprises a first container, an antigen solution in the first container, an optional second container, a labeled antibody X' which is arranged in the second container and labeled by a detectable marker, and a detecting piece which comprises a sample receiving part and a reaction part. After the rapid detection device is used, the step of mixing a sample and an antigen only stably existing in a liquid environment in advance can be omitted, and the step of diluting the antibody in the sample can be avoided, so that high sensitivity and high specificity of detection can be realized; the detection device is simple, convenient and rapid and is convenient for a user to use.
Description
Technical field
The utility model belongs to biological immunology, relates to particularly a kind of device for quick testing, for detection of the antibody of the antigen of stable existence in buffer solution only.
background technology
Be applied to along with immunoassay is increasingly extensive taking the clinical diagnosis industry (diagnostic industry) as main and non-clinical field, immunoassay develops to both direction: a class is full-automatic immunoassay; Another kind of is if nitrocellulose membrane is as the tachysynthesis analysis of carrier taking porosint.The former need expensive full-automatic instrument and with the strict supporting various kits of instrument, at present can only be in the application of medical inspection center, though also can faster provide result, but still need certain hour, be not suitable for the area away from medical treatment and inspection center, more can not be used for the needs of " patient bed other inspection " (real time clinical decision) and generaI investigation.On the basis of EIA enzyme immunoassay, the main method for quick taking nitrocellulose filter as carrier grows up rapidly and widely.
Fast detecting should be detection method a kind of cheapness, that be easy to use, based on film, and the vision that it can provide analyte to be present in fluid sample proves.This detection can, with test strips independently, also can be carried out with the form being contained in plastic casing.Usually, detect the fluid sample that only needs 200 μ l, generally in 2-5 minute, complete.In clinical detection, sample can be urine, blood, serum, saliva or other body fluid.In non-clinical detection, sample can be by the solution of the ready a small amount of such as soil, dust, plant or food, then directly tests by film test strips similarly.This detection is without any need for instrument, can be used to clinical, laboratory, outdoor and family, and often can be detected by inexperienced personnel.
Also disunity very of these class methods name on document and market at present.In fact it belong to quick spot immune binding analysis (dot immunobinding assay, DIBA), and this analysis starts from the eighties, mainly contains following two kinds of patterns:
1, spot immune diafiltration analysis (dot immunofiltrtion assay, DIFA) immune response is to be fixed with (the flow through type) that the nitrocellulose filter of part carries out by vertically penetrating.
2, spot immune chromatographic analysis (dot immunochromatography assay, DICA) analysis principle is identical with DIFA, just reaction liquid flow be not directly to penetrate mobile, but the lateral flow of chromatography effect (lateral flow type) starts application in nineteen ninety.
In above-mentioned two kinds of patterns, the kind of nitrocellulose filter and the difference of form building form are:
It is disk film that immunity percolation is analyzed the nitrocellulose filter adopting in (penetrating filterable form) method, and aperture 0.2-0.4 μ m is simpler, forms by being loaded with the cellulose nitrate diaphragm of part (antibody or antigen) and the water suction bedding and padding of bottom.
The nitrocellulose filter adopting in immunochromatographiassays assays (lateral flow ejector half) is narrow long type, according to the requirement of sensitivity and flow velocity, can select the different suitable film in aperture, comprising from near-end to far-end (from left to right): (1) sample pad (sample pad), (2) be loaded with glass fibre element film bar or the polyester film bar (conjugate release membrane) of solid phase labelling part (antibody or antigen), (3) suitable aperture, be coated with the part line of catching analyte that shows positive findings, nitrocellulose filter bar (nitrocellulose memtrane) with negative control group line, (4) adsorptive pads (absorbent pad), this four parts linking is fixed on (back sheet) on the book plastic back plate that does not disturb immune response and flow velocity.
The label adopting in above-mentioned two kinds of patterns can be selected from: the detectable such as the microballoon (latex particles) of colloidal metal (collaurum, electroselenium), nano particle (as metal nanoparticle or carbon nanomaterial), decentralized dyestuff (disperse dyes) or dye marker.Aforementioned tagging material respectively has relative merits, can be according to following Standard Selection, as: sensitivity, needed color, pattern and the operation steps analyzed, preparation, ensure repeatability, anxious qualitative and scale and complexity and repeatability, and the required amount of ligand of label etc.Although wherein enzyme has high sensitivity; the advantages such as the reproducible and easy scale of label; but it needs washing; add the steps such as substrate; need the time of reaction also long; technology is also had to ask for something, therefore fail as the main mark thing of quick diagnosis (initial immunity percolation analysis in 1985 is with the enzyme thing that serves as a mark).Current most widely used label is collaurum, comprise: colloidal gold immunity percolation analysis (gold immumofiltraition assay, GIFA) or drip golden immunoassay and highly-pathogenic avian influenza (gold immunochromatography assay, GICA).Because GICA is easier fast, become one of current topmost tachysynthesis analytical approach.
It is basic identical with ELISA with operation steps that colloidal gold immunity percolation is analyzed (GIFA) principle: add sample in being fixed with part (antibody or antigen, this sentences antibody is example, on nitrocellulose filter down together), in film, form Antibody-antigen complex by diafiltration, wash after diafiltration, then add the colloidal gold labeled monoclonal antibody of liquid.In the time that result is positive, on film, is fixed with " antibody-antigen-colloidal gold labeled monoclonal antibody compound " and presents punctation.
Highly-pathogenic avian influenza CICA principle is identical with GIFA, just operation steps is different, as changing lateral chromatography into by vertical diafiltration, reaction liquid flow direction flows: sample is added in sample pad (1), sample flows to adsorptive pads (4) direction, after being loaded with solid phase colloidal gold labeled monoclonal antibody film (2), golden labelled antibody is redissolved completely, antigen and golden labelled antibody form " golden labelled antibody-antigenic compound ", continue to flow to forward the antibody line place that is fixed with capture antigen on nitrocellulose filter bar (3).In the time that result is positive, on golden labelled antibody-antigenic compound, will form " golden labelled antibody-antigen-insolubilized antibody " compound, present red positive band; As in sample without antigen, be not hunted down, just do not develop the color, result is negative.Unnecessary free gold labelled antibody continues to move forward to and is fixed with the anti-place of anti-golden marked body two, is fixed and presents red contrast.
GICA and GIFA difference are that the latter is single immunochromatography bar, does not need other reagent, single stepping.It is greatly to shorten in the reaction time that the two and ELISA do not exist together, only need within several minutes, just complete ELISA need the result that a few hours could show.Because in GIFA and GICA, the antibody of high concentration is concentrated and is fixed in the micropore of porosint (as cellulose), and determined antigen is in the time of diafiltration or chromatography, and flow through micropore and fixing high concentration antibody close contact, complete immune association reaction very soon.The immune inspissation (immunoconcentraiton) in basic collaurum tachysynthesis binding analysis that Here it is taking film.
In ELISA, determined antigen will, through diffusion in liquid phase, be combined with the antibody that is adsorbed on solid phase surface gradually.Meanwhile, labelled antibody also needs same diffusion bond process to form antibody-antigen-labelled antibody compound, therefore it is longer to take time.
In existing technology, while detecting for the antibody of antigen, antigen potentially unstable under drying regime that some contains three grades or quaternary structure, these antigens can significantly reduce specificity and sensitivity under drying regime, typical example is, the virus-like particle (VLP) of anti-human papilloma virus (anti-HPV) (as 16,18 and 58 types).
While at present detecting anti-human papillomavirus antibody, employing be the virus surface proteins (or virus-like particle) that is arranged in liquid.In the method, sample must add in damping fluid and then mixing material is added in lateral flow proving installation.This existing methodical shortcoming is, before the mixed liquor that contains sample is added to lateral flow proving installation, must first through one, sample be joined to damping fluid in and the process that mixes, still need to meet certain reaction time simultaneously.In addition, if only contain a small amount of antibody in sample, because those detection methods on market conventionally need first sample is diluted with damping fluid, if the insufficient sensitivity of detection method so just easily causes and detects unsuccessful (false negative).
Publication number be WO2012097788 PCT Patent Application Publication a kind of fast detecting and device, can be used for qualitative and/or quantitative test and be present in anti-human papilloma virus (HPV) antibody in body fluid.Described in this utility model, test provides a kind of humoral sample that is mixed with reagent fast, described reagent itself contains the fluid of physiologically active and at least one HPV specific antigen of scheduled volume of scheduled volume, subsequently potpourri is analyzed, utilized measurable variation and/or the variation that can be observed by user detects.The equipment that carries out test fast provides a container (2), be used for accepting described reagent (14), the humoral specimen wherein gathering from patient can be added to described container (2) and with reagent (14) and mix.In addition, this equipment also provides an analytic system (4), and it utilizes measurable variation and/or the change that can be observed by user is analyzed, and described analytic system is designed to can accept to exist the substrate of scheduled volume in container (2).In this technical scheme, before adding testing sample to pick-up unit, still in advance sample and liquid antigenic solution be mixed.Therefore, increased on the one hand the step being pre-mixed, on the other hand still in the time of reaction by Sample Dilution, same so easily causing detect unsuccessful (false negative).
Therefore, in this area, detect in the urgent need to exploitation new quick, highly sensitive and be particularly useful for the pick-up unit of the antibody of anti-labile antigen.
Utility model content
The purpose of this utility model is for a kind of device for fast detecting is provided.
Provide a kind of pick-up unit at the utility model, described pick-up unit comprises:
The first container and be positioned at the antigenic solution of described the first container, the antigen Y in described antigenic solution can with antibody X specific binding to be measured, wherein said antigen Y is unlabelled antigen Y or the labelled antigen Y' with detectable;
Optional second container and be positioned at described second container by the labelled antibody X' of detectable institute mark; With
One detection piece, described detection piece comprises a sample receiving portion and a reacting part, wherein
Described sample receiving portion is used for accepting testing sample solution, and described sample receiving portion is loaded with the trapping agent for catching described antibody X to be measured;
Described reacting part is provided with control line region, and described control line region is provided with the control line for representing to have detected; Described reacting part is also provided with the detection line for detection of described antibody X to be measured, described detection line region is loaded with insolubilized antibody X', and described insolubilized antibody X' can react with described antigen Y and labelled antibody X' formation " insolubilized antibody X'-antigen Y-labelled antibody X' " ternary complex; Or described insolubilized antibody X' reacts formation " insolubilized antibody X' – labelled antigen Y' " binary complex with described labelled antigen Y'.
In another preference, described detection piece also comprises a bond release portion, and be loaded with releasable bond in described bond release portion, described bond comprises by the antibody X' of detectable institute mark and optional by the Quality Control thing of detectable institute mark; And described bond release portion is located between described sample receiving portion and reacting part.
In another preference, described detection piece also comprises an antigenic solution receiving portion, and described sample receiving portion and described antigenic solution receiving portion are one.
In another preference, described detection piece also comprises an antigenic solution receiving portion, and described antigenic solution receiving portion is located at the upstream of sample receiving portion; And/or
Described detection piece also comprises an imbibition portion, and described imbibition portion is located at the downstream of described reacting part, promotes flow direction reacting part.
In another preference, described sample receiving portion is also provided with covalently bound anti erythrocyte antibody; And/or the control line region of described reacting part is loaded with Quality Control antibody.
In another preference, described reacting part is provided with detection line, and described detection line region is loaded with the insolubilized antibody X' that is selected from lower group: anti-HPV16L1 monoclonal antibody, anti-HPV18L1 monoclonal antibody, anti-HPV58L1 monoclonal antibody.
In another preference, described reacting part is located at below sample receiving portion; And described pick-up unit also comprises:
Second container and the labelled antibody X' solution that is positioned at described second container.
In another preference, the releasable bond that described bond release portion is loaded with comprises the antibody X' of golden mark and the Quality Control thing antibody of golden mark, and the control line region of described reacting part is also loaded with Quality Control antibody; And/or
The Quality Control thing that simultaneously comprises golden mark in described golden labelled antibody X' solution, the control line region of described reacting part is loaded with Quality Control antibody.
In another preference, described antibody X to be measured is one or more, is designated as respectively Xn, and n is positive integer; Correspondingly, the antigen Y containing in described antigenic solution is one or more, is designated as respectively Yn, and n is positive integer; Labelled antibody X' is one or more, is designated as respectively X'n, and n is positive integer;
And the detection line region of described reacting part is provided with the n bar detection line corresponding to each antibody X to be measured, wherein n article of detection line is corresponding is respectively loaded with the insolubilized antibody X'n corresponding to antibody Xn to be measured,
Wherein, described each insolubilized antibody X'n reacts formation " insolubilized antibody X'n-antigen Yn-labelled antibody X'n " ternary complex with corresponding antigen Yn and labelled antibody X'n;
" insolubilized antibody X'-antigen Y-labelled antibody X' " ternary complex; Or described insolubilized antibody X' reacts formation " insolubilized antibody X' – labelled antigen Y' " binary complex with described labelled antigen Y'.
In another preference, described pick-up unit is kit form, and wherein said kit comprises described pick-up unit and operation instruction.
In should be understood that within the scope of the utility model, above-mentioned each technical characterictic of the present utility model and can combining mutually between specifically described each technical characterictic in below (eg embodiment), thus form new or preferred technical scheme.As space is limited, tire out and state no longer one by one at this.
Brief description of the drawings
Fig. 1 is the vertical view of lateral flow device for immunochromatography in embodiment 1;
Fig. 2 is the schematic diagram of the interior reagent bar of lateral flow device for immunochromatography in embodiment 1;
Fig. 3 is the combination situation of the sample receiving portion of lateral flow device for immunochromatography inside in embodiment 1 and the schematic diagram of negative findings;
Fig. 4 is the combination situation of the reacting part of lateral flow device for immunochromatography inside in embodiment 1 and the schematic diagram of negative findings;
Fig. 5 is the combination situation of the sample receiving portion of lateral flow device for immunochromatography inside in embodiment 2 and the schematic diagram of positive findings;
Fig. 6 is the combination situation of the reacting part of lateral flow device for immunochromatography inside in embodiment 2 and the schematic diagram of positive findings;
Fig. 7 is the vertical view of lateral flow device for immunochromatography in embodiment 3;
Fig. 8 is the schematic diagram of the interior reagent bar of lateral flow device for immunochromatography in embodiment 3;
Fig. 9 is the combination situation of the reacting part of lateral flow device for immunochromatography inside in embodiment 3 and the schematic diagram of positive findings;
Figure 10 is the vertical view of immunity percolation test unit in embodiment 4;
Figure 11 is the schematic diagram of the sample receiving portion of immunity percolation test unit in embodiment 4;
Figure 12 is the schematic diagram of the reacting part of immunity percolation test unit in embodiment 4;
Figure 13 combines the reacting part reagent strip of specificity component and the schematic diagram of positive findings in immunity percolation test unit in embodiment 4;
Figure 14 is the schematic diagram of positive findings in embodiment 4;
Figure 15 is the schematic diagram of negative findings in embodiment 5.
Embodiment
The inventor is through extensive and deep research, provide first a kind of fast, for the pick-up unit of the antibody of anti-labile antigen.While using this pick-up unit to detect, the dilution to testing sample while having avoided detecting, thereby can sensitivelyer correctly draw testing result.Complete on this basis the utility model.
Particularly, in first aspect of the present utility model, provide a kind of pick-up unit to comprise:
The first container and be positioned at the antigenic solution of described the first container, the antigen Y in described antigenic solution can with antibody X specific binding to be measured, wherein said antigen Y is unlabelled antigen Y or the labelled antigen Y' with detectable;
Optional second container and be positioned at described second container by the labelled antibody X' of detectable institute mark; With
One detection piece, described detection piece comprises a sample receiving portion and a reacting part, wherein
Described sample receiving portion is used for accepting testing sample solution, and described sample receiving portion is loaded with the trapping agent for catching described antibody X to be measured;
Described reacting part is provided with control line region, and described control line region is provided with the control line for representing to have detected; Described reacting part is also provided with the detection line for detection of described antibody X to be measured, described detection line region is loaded with insolubilized antibody X', and described insolubilized antibody X' can react with described antigen Y and labelled antibody X' formation " insolubilized antibody X'-antigen Y-labelled antibody X' " ternary complex; Or described insolubilized antibody X' reacts formation " insolubilized antibody X' – labelled antigen Y' " binary complex with described labelled antigen Y'.
In another preference, described trapping agent comprises part, antibody or its combination of being combined with antibody X to be measured.
In another preference, described detection piece comprises detection lug, detector bar, check-out console.
In another preference, described testing sample solution can flow to described reacting part from described sample receiving portion by lateral flow chromatography effect or transudation.
In another preference, described detectable comprises: the microballoon (latex particles) of colloidal metal (collaurum, electroselenium), nano particle (carbon granule), fluorescence molecule, chromophore, decentralized dyestuff (disperse dyes) or dye marker.
In another preference, described pick-up unit is kit form, and described kit comprises described pick-up unit, and operation instruction.
In another preference, described detection piece also comprises a bond release portion, and be loaded with releasable bond in described bond release portion, described bond comprises by the antibody X' of detectable institute mark and optional by the Quality Control thing of detectable institute mark (sheep polyclonal antibody as anti-in rabbit); Described bond release portion is located between described sample receiving portion and reacting part.
In another preference, the control line region of described reacting part is loaded with Quality Control antibody.
In another preference, described Quality Control antibody is for the Quality Control thing in conjunction with discharging, and described Quality Control antibody comprises (but being not limited to): solid phase goat-anti rabbit polyclonal antibody etc.
In another preference, the bond release portion of described detection piece is also loaded with by the Quality Control thing of detectable institute mark.Described Quality Control thing comprises (but being not limited to): the anti-sheep polyclonal antibody of rabbit.
In another preference, in the time that liquid (as testing sample solution) flows to bond release portion from sample receiving portion, described releasable bond is released, and then flows to reacting part together.
In another preference, described detection piece also comprises an antigenic solution receiving portion, and described antigenic solution receiving portion is located at the upstream of sample receiving portion, the antigenic solution that the makes an addition to described antigenic solution receiving portion described sample receiving portion that can flow to and flow through; And/or
Described detection piece also comprises an imbibition portion, and described imbibition portion is located at the downstream of described reacting part, promotes flow direction reacting part.
In another preference, described sample receiving portion and described antigenic solution receiving portion are one, and described sample receiving portion had both been accepted testing sample, also accepted antigenic solution.
In another preference, described antibody X to be measured comprises the antibody for pathogen (as virus, conveyor screw, bacterium), preferably described antibody comprises the antibody for HPV, more preferably comprises the antibody that is selected from lower group: HPV16 antibody, HPV18 antibody or HPV58 antibody;
Described antigenic solution corresponds to: comprise at least 5, the damping fluid of the virus-like particle of preferably >=100 HPV16L1 capsid proteins; And/or comprise by least 5, the damping fluid of the virus-like particle of preferably >=100 HPV18L1 capsid proteins, and/or comprise by least 5 the damping fluid of the virus-like particle of preferably >=100 HPV58L1 capsid proteins.
In another preference, described damping fluid is Tris BSA damping fluid.
In another preference, the material of described sample receiving portion comprises: glass fibre element film, nitrocellulose filter or dacron film; And the trapping agent of the antibody X to be measured that described sample receiving portion is loaded with is to resist with two of the covalently bound albumin A of described sample receiving portion, Protein G or anti-antibody X to be measured.
In another preference, described two resist for goat anti-human igg antibody or rabbit anti-human igg's antibody.
In another preference, described sample receiving portion is also provided with covalently bound anti erythrocyte antibody.
In another preference, the material of described reacting part is nitrocellulose filter or nylon membrane; And/or
Described reacting part is provided with detection line, and the insolubilized antibody X' that described detection line region is loaded with comprises: anti-HPV16L1 monoclonal antibody, anti-HPV18L1 monoclonal antibody, anti-HPV58L1 monoclonal antibody; And described labeling antibody X' corresponds to respectively: the anti-HPV58L1 monoclonal antibody of the anti-HPV16L1 monoclonal antibody of colloid gold label, the anti-HPV18L1 monoclonal antibody of colloid gold label, colloid gold label.Preferably, described monoclonal antibody is antibody or the mouse-anti body that is derived from mouse.
In another preference, described reacting part is located at below sample receiving portion, and described testing sample solution flows to reacting part by transudation from sample receiving portion; And described pick-up unit also comprises:
Second container and the labelled antibody X' solution that is positioned at described second container.
In another preference, described detection piece comprises: a sample receiving portion, one bond release portion and a reacting part, described bond release portion is located between sample receiving portion and reacting part, testing sample solution can flow to bond release portion from sample receiving portion by lateral flow chromatography effect, and then flows to reacting part;
Described bond release portion is provided with labelled antibody X'.
In another preference, the releasable bond that described bond release portion is loaded with, comprise antibody X' and the Quality Control thing (sheep polyclonal antibody as anti-in the rabbit of golden mark) of golden mark, and the control line region of described reacting part is also loaded with Quality Control antibody (as solid phase goat-anti rabbit polyclonal antibody); And/or
The Quality Control thing (sheep polyclonal antibody as anti-in rabbit) that simultaneously comprises golden mark in described golden labelled antibody X' solution, the control line region of described reacting part is loaded with Quality Control antibody (as solid phase goat-anti rabbit polyclonal antibody).
In another preference, described pick-up unit also comprises an antigenic solution receiving portion, and described antigenic solution receiving portion is located at the upstream of sample receiving portion, and antigenic solution can flow to sample receiving portion by for example lateral flow chromatography effect.
In another preference, the material of described antigenic solution receiving portion comprises glass fibre element film.
In second aspect, a kind of pick-up unit adopting described in above-mentioned first aspect is provided, detect the method for antibody X to be measured in testing sample, the method comprises the following steps:
(1) testing sample solution is added to described sample receiving portion, and make it flow to described reacting part;
(2) antigenic solution is added to sample receiving portion, and make it flow to described reacting part;
(3) labelled antibody X' solution is added to sample receiving portion, and make it flow to described reacting part;
(4) read the reaction result of described reacting part, thereby draw the testing result of antibody X to be measured, wherein,
If the control line region of described reacting part manifests a line, but detection line region does not manifest the detection line corresponding to antibody X to be measured, represents to have described antibody X to be measured in testing sample solution, positive;
If the control line region of described reacting part manifests a line, and detection line region manifests the detection line corresponding to antibody X to be measured, represents not have described antibody X to be measured in testing sample solution, negative.
In the third aspect, a kind of pick-up unit adopting described in above-mentioned first aspect is provided, detect the method for antibody X to be measured in testing sample, the method comprises the following steps:
(1) testing sample solution is added to described sample receiving portion, and make it flow to described reacting part;
(2) antigenic solution is added to sample receiving portion or antigen receiving portion, and make it flow to described reacting part;
(3) read the reaction result of described reacting part, thereby draw the testing result of antibody X to be measured, wherein,
If the control line region of described reacting part manifests a line, but detection line region does not manifest the detection line corresponding to antibody X to be measured, represents to have described antibody X to be measured in testing sample solution, positive;
If the control line region of described reacting part manifests a line, and detection line region manifests the detection line corresponding to antibody X to be measured, represents not have described antibody X to be measured in testing sample solution, negative.
In another preference, in step (2), add after antigenic solution, wait for 5-30 minute after (preferably 8～12 minutes or 10-20 minute), then reading result.
In fourth aspect, a kind of pick-up unit that is preferably applicable to detect Multiple Antibodies based on above-mentioned first aspect is provided, wherein, described antibody X to be measured is one or more, is designated as respectively Xn, n is positive integer; Correspondingly, the antigen Y containing in described antigenic solution is one or more, is designated as respectively Yn, and n is positive integer; Labelled antibody X' is one or more, is designated as respectively X'n, and n is positive integer;
And the detection line region of described reacting part is provided with the n bar detection line corresponding to each antibody X to be measured, wherein n article of detection line is corresponding is respectively loaded with the insolubilized antibody X'n corresponding to antibody Xn to be measured,
Wherein, described each insolubilized antibody X'n reacts formation " insolubilized antibody X'n-antigen Yn-labelled antibody X'n " ternary complex with corresponding antigen Yn and labelled antibody X'n;
" insolubilized antibody X'-antigen Y-labelled antibody X' " ternary complex; Or described insolubilized antibody X' reacts formation " insolubilized antibody X' – labelled antigen Y' " binary complex with described labelled antigen Y'.
In another preference, described label is collaurum (gold mark), and in described antigenic solution, contains two or more antigen: antigen Y1, antigen Y2, ..., antigen Yn, the antigen Y1 in described antigen Y solution, antigen Y2, ..., antigen Yn can be respectively and antibody X1 to be measured, X2, ..., Xn specific binding; Kind and the antigen Y kind quasi-equal of gold labelled antibody, correspond to respectively golden labelled antibody X1', X2' ..., Xn'; Described sample receiving portion is loaded with can catch described antibody X1 to be measured, X2 ..., the trapping agent of Xn; The number of the detection line that described reacting part is provided with and antigen Y kind quasi-equal, and described detection line region respectively correspondence be loaded with insolubilized antibody X1', X2', ..., Xn', described insolubilized antibody X1', X2' ..., Xn' can be accordingly and antigen Y1, antigen Y2 ..., antigen Yn, gold labelled antibody X1', X2' ..., Xn' forms " insolubilized antibody X'n-antigen Yn-gold labelled antibody X'n " ternary complex by sandwich reaction; Wherein n is more than or equal to 2.
Aspect the 5th, a kind of pick-up unit described in above-mentioned first aspect that adopts is provided, detect the method for antibody X to be measured in testing sample, the method comprises the following steps:
(1) testing sample solution is added to described sample receiving portion, and make it flow to described reacting part;
(2) antigenic solution is added to sample receiving portion, and make it flow to described reacting part;
(3) by specificity from high to low, successively by labelled antibody X1', X2' ..., Xn' solution adds to sample receiving portion, and makes it flow to described reacting part;
(4) read the reaction result of described reacting part, thereby draw the testing result of each antibody Xn to be measured, wherein,
If the control line region of described reacting part manifests a line, but detection line region does not manifest the detection line corresponding to antibody Xn to be measured, represents to have described antibody Xn to be measured in testing sample solution, positive;
If the control line region of described reacting part manifests a line, and detection line region manifests the detection line corresponding to antibody Xn to be measured, represents not have described antibody Xn to be measured in testing sample solution, negative.
In another preference, if only a line appears in control line region, positive; If a line all appears in control line region and at least one detection line region, be that the result of target to be measured corresponding to described at least one detection line region is negative.
In another preference, be 8-15min detection time.
The detection piece of sidestream immune chromatography and detection method
Referring to Fig. 1 and Fig. 2.The utility model provides a kind of detection piece of lateral flow immunochromatography, be provided with from left to right: a sample receiving portion 2, one bond release portion 3, one reacting part 4, one imbibition portion 5, wherein, described lateral flow device for immunochromatography also can comprise an antigen receiving portion 1, in order to accept antigenic solution, described antigen receiving portion is located at the left side (upstream) of described sample receiving portion, and be connected with described sample receiving portion, liquid can flow to sample receiving portion from antigen receiving portion by chromatography effect; Above-mentioned antigen receiving portion is in order to accept antigenic solution (the preferably solution of unsettled antigen under drying regime); Above-mentioned sample receiving portion is in order to accept sample; The antibody (or providing by the labelled antibody X' in second container) of anti-determined antigen and the Quality Control agent (sheep polyclonal antibody as anti-in rabbit etc.) for Quality Control should be provided in above-mentioned bond release portion; Above-mentioned reacting part is provided with detection line and/or control line/control line.
In preferred technical scheme, the utility model can also comprise a backing, and above-mentioned antigen receiving portion, sample receiving portion, bond release portion, reacting part, the five parts linkings of imbibition portion are fixed on the backing that does not disturb immune response and flow velocity.The material of described backing is conventional, can include but not limited to plastics.
In preferred technical scheme, the utility model can also comprise a shell, and described shell is provided with corresponding antigen well, sample pipetting volume hole and the area of observation coverage, corresponding antigen receiving portion respectively, the position of sample receiving portion and reacting part.
In the utility model, the material of described antigen receiving portion comprises porosint various routines, that be applicable to sidestream immune chromatography or immunity percolation analysis, and representational example comprises (but being not limited to): glass fibre element film (functionalization or not functionalization) nitrocellulose filter or dacron film.
In the utility model, the material of described sample receiving portion comprises porosint various routines, that be applicable to sidestream immune chromatography or immunity percolation analysis, and representational example comprises (but being not limited to): glass fibre element film, nitrocellulose filter or the dacron film of functionalization.Described sample receiving portion is provided with covalently bound for catching the trapping agent of detection antibody X, and representational trapping agent comprises (but being not limited to): in albumin A, Protein G or anti-sample, two of antibody is anti-.When sample is this time of human blood sample, described two anti-goat anti-human igg antibody or the rabbit anti-human igg's antibody of being preferably.
In the time that sample is whole blood sample, described sample receiving portion is also provided with covalently bound anti erythrocyte antibody.In the time adopting nitrocellulose filter or dacron film, irreversible with being combined into of described anti erythrocyte antibody, for example, goat-anti people polyclonal antibody and rabbit anti erythrocyte antibody.
In the utility model, the material of described bond release portion comprises porosint various routines, that be applicable to sidestream immune chromatography or immunity percolation analysis, and representational example includes but not limited to: nitrocellulose filter or dacron film.Described bond release portion is provided with the antibody of anti-above-mentioned labile antigen epi-position.Preferably, described antibody is for for example, by the antibody of detectable institute mark mark, the antibody of colloid gold label.For example, for the detection of HPV16, in described bond release portion, be preferably loaded with the mouse-anti HPV16L1 monoclonal antibody of colloid gold label and the anti-sheep polyclonal antibody of the rabbit of colloid gold label.
In the utility model, the material of described reacting part comprises porosint various routines, that be applicable to sidestream immune chromatography or immunity percolation analysis, and representational example comprises (but being not limited to): nitrocellulose filter or dacron film.Described reacting part is provided with detection line.Preferably, on described reacting part, be also provided with the control line for representing to have detected.Preferably, on described reacting part, be also provided with negative control group line.
In technique scheme, described imbibition portion is located at downstream, and fluid-absorbing also ensures the proper flow of sample.
The method that adopts lateral flow device for immunochromatography described in technique scheme to detect testing sample comprises the following steps: (1) is added to sample drop in sample receiving portion, and described sample can be selected from but be not limited to people's whole blood or serum or blood plasma; (2) antigenic solution is added drop-wise in antigen receiving portion; (3) wait for 10～15 minutes, reading result, those skilled in the art can understand the rule of judged result in conjunction with the embodiments.
Referring to Fig. 7 and Fig. 8.Another technical scheme that the utility model adopts is: described antigen receiving portion and sample receiving portion unite two into one.This detection piece is provided with from left to right: a sample receiving portion 2a, and bond release portion 3, one reacting part 4, one imbibition portions 5, described sample receiving portion is first accepted to accept antigen after sample, and the material of described sample receiving portion is the glucosan that is combined with Protein G.In the time that the materials'use of described receiving portion is combined with the glucosan of Protein G, owing to not having red blood cell to be detained, found that in sample, the absorption affinity of antibody has improved greatly, therefore can obtain higher sensitivity, particularly for serum or plasma sample.
Equally, in preferred technical scheme, can also comprise a backing, above-mentioned receiving portion, bond release portion, reacting part, the four parts linkings of imbibition portion are fixed on the backing that does not disturb immune response and flow velocity.
In preferred technical scheme, the utility model can also comprise a shell, and described shell is provided with corresponding well and the area of observation coverage, the position of corresponding receiving portion and reacting part respectively.
The method that adopts lateral flow device for immunochromatography described in technique scheme to detect testing sample comprises the following steps: (1) is added to sample drop in sample receiving portion, and described sample can be selected from but be not limited to people's whole blood or serum or blood plasma; (2) antigenic solution is added drop-wise in sample receiving portion (doubling as antigen receiving portion); (3) wait for 10～15 minutes, reading result, those skilled in the art can understand the rule of judged result in conjunction with the embodiments.
The detection piece of immunity percolation and detection method
In the utility model, adoptable another technical scheme is: a kind of immunity percolation fast detecting part.Referring to Figure 10 and Figure 11, this detection piece is provided with from top to bottom: a sample receiving portion (simultaneously having pre-filtered effect) and a reacting part; Described reacting part comprises a nitrocellulose filter, and described nitrocellulose filter is provided with detection line and the control line for representing to have detected.
Described sample receiving portion comprises a filter paper, described filter paper comprises porosint various routines, that be applicable to immunity percolation analysis, and representational example comprises that (but being not limited to) material can be selected from but be not limited to: glass fibre element film, nitrocellulose filter or the dacron film of functionalization.
Described sample receiving portion is provided with covalently bound for catching the trapping agent of detection antibody X, and representational trapping agent comprises (but being not limited to): in albumin A, Protein G or anti-sample, two of antibody is anti-.When sample is this time of human blood sample, described two resist for preferred goat anti-human igg antibody or rabbit anti-human igg's antibody.
In the time that sample is whole blood sample, described sample receiving portion is also provided with covalently bound anti erythrocyte antibody; In the time adopting nitrocellulose filter or dacron film, irreversible with being combined into of described anti erythrocyte antibody, for example, goat-anti people polyclonal antibody and rabbit anti erythrocyte antibody.
In technique scheme, on described detection line, be loaded with mouse-anti HPV16L1 monoclonal antibody; On described control line, be loaded with goat-anti rabbit polyclonal antibody.
The first container and antigenic solution
The utility model device for fast detecting also comprises the first container and is positioned at the antigenic solution of described the first container (the first solution).Antigen Y in described antigenic solution can with antibody X specific binding to be measured, wherein said antigen Y is unlabelled antigen Y or the labelled antigen Y' with detectable.
In the utility model, for antigen, Y is not particularly limited, and antigen is the antigen that derives from various different pathogens (as virus, conveyor screw, bacterium).But the utility model is specially adapted to contain the antigen of three grades or quaternary structure and at the poor antigen of drying regime stability inferior.
Correspondingly, in the utility model, detectable antibody X to be measured does not also limit, this antibody X can be the antigen for various different pathogens (as virus, conveyor screw, bacterium), preferably, described antibody is the antibody for unsettled antigen under drying regime, for example, for the antibody of HPV, more preferably comprises (but being not limited to): HPV16 antibody, HPV18 antibody or HPV58 antibody.
In the utility model, to detect HPV as example (antigen is as L1 capsid protein or VLP), described antigenic solution can comprise: comprise at least 5, preferably >=100 HPV16L1 capsid proteins or the damping fluid by L1 virus-like particle that capsid protein forms; And/or comprise by least 5, preferably >=100 HPV18L1 capsid proteins or by the damping fluid of L1 virus-like particle that capsid protein forms, and/or comprise by least 5 preferably >=100 HPV58L1 capsid proteins or the damping fluid by L1 virus-like particle that capsid protein forms.
In another preference, described damping fluid or buffer system are not particularly limited, as long as it is conducive to the stable preservation of antigen.A kind of representational damping fluid is Tris BSA damping fluid.
Second container and the labelled antibody X' that is positioned at second container
In another kind of scheme of the present utility model, if bond release portion is not provided with releasable labelled antibody X', in the utility model device, should comprise second container and the labelled antibody X' that is positioned at second container.Conventionally, labelled antibody X' exists with solution form, but also can exist with solid-state form, and adds liquid (damping fluid) before using, thereby forms the solution (the second solution) containing described labelled antibody X'.
Correspondingly, in the time detecting, can be before testing sample, antigenic solution be made an addition to detection piece, afterwards or between, the solution containing described labelled antibody X' is made an addition to detection piece.Optimal way is after having added testing sample and antigenic solution, then adds the solution containing described labelled antibody X'.
On detection piece, while adding the described solution containing described labelled antibody X', can make an addition to the independent point of addition position of bond release portion (for example, corresponding to), or be added to the position of share, for example antigenic solution receiving portion or sample receiving portion.
In the utility model, if bond release portion is provided with releasable labelled antibody X', in the utility model device, can not comprise second container and the labelled antibody X' that is positioned at second container.Certainly, in the utility model device, comprise extraly that second container is also feasible.
Label and labelled antibody
In the utility model, antibody can be used conventional method, carries out mark by various detectable.Representational label comprises (but being not limited to): the microballoon (latex particles) of colloidal metal (collaurum, electroselenium), nano particle (as metal nanoparticle or carbon nanomaterial), decentralized dyestuff (disperse dyes) or dye marker.
A kind of particularly preferred label is collaurum.
Major advantage of the present utility model comprises:
(1) use device for fast detecting described in the utility model, sample directly can be added in pick-up unit, omit in advance by sample and the antigen blend step of stable existence in liquid environment only, avoid the step of antibody in dilute sample, thereby realize the high sensitivity detecting, high specific, and more simple and convenient, user-friendly.
(2) pick-up unit described in the utility model does not need dilute sample, and therefore processing sample is easier.
(3) pick-up unit described in the utility model can use a kind of antigenic solution to carry out multiple test, and more convenient user uses.
(4) pick-up unit described in the utility model can enriched antibody, therefore can realize highly sensitive lateral flow immunity test fast.
(5) immunity percolation device for quick testing described in the utility model can be suitable for enriched sample and separation of whole blood sample, therefore can realize highly sensitive detection.
(6) can realize the enrichment of different HPV antibody in same sample due to the utility model, therefore, can realize the multiparameter detection that simultaneously detects different antibodies by adding successively different golden labeling antibody solution, and improve the specificity and the sensitivity that detect simultaneously; Also can optionally first make HPV16 and HPV58 inactivation, thereby realize only enrichment and detect the antibody of anti-HPV18, get rid of thereby realize the interference that cross reaction brings, and successfully realize and detecting in high sensitivity by the not high antibody of specificity.
Below in conjunction with accompanying drawing, the utility model is described in detail, following table is classified the source of all material in embodiment as, the commercial product that the material that the utility model uses is known to the skilled person:
Embodiment 1
The device for fast detecting of the present embodiment comprises the first container and is positioned at the antigenic solution of described the first container, and described antigenic solution is the Tris BSA damping fluid containing at least 100 HPV16L1 capsid protein VLP; And detection piece.
As shown in Figure 1-2, described lateral flow immunochromatography detection piece, is provided with detection piece from left to right: an antigen receiving portion, a sample receiving portion, a bond release portion, a reacting part, an imbibition portion.Wherein, in bond release portion, be loaded with the antibody of anti-determined antigen; Above-mentioned reacting part is provided with detection line and control line.Described lateral flow device for immunochromatography comprises a backing, and the material of described backing is plastics.
Described detection piece also comprises a shell, and described shell is provided with corresponding antigen well, sample pipetting volume hole and the area of observation coverage, corresponding antigen receiving portion respectively, the position of sample receiving portion and reacting part.
The material of described antigen receiving portion is glass fibre element film.
Described sample receiving portion is the porous nitrocellulose filter bar that is loaded with goat anti-human igg antibody and rabbit anti erythrocyte antibody.
Described bond release portion is the dacron film that is loaded with the mouse-anti HPV16L1 monoclonal antibody of colloid gold label and the anti-sheep polyclonal antibody of rabbit of colloid gold label.
Described reacting part is nitrocellulose filter.Described reacting part is provided with detection line, is loaded with mouse-anti HPV16L1 monoclonal antibody on described detection line; On described reacting part, be also provided with control line, on described control line, be loaded with goat-anti rabbit polyclonal antibody.
The method that whether contains HPV16L1 antibody in detection testing sample comprises the following steps:
(1) 20 μ l testing samples are added drop-wise in sample receiving portion, described sample can be selected from but be not limited to people's whole blood or serum or blood plasma;
(2) 100 μ l antigenic solutions are added drop-wise in antigen receiving portion;
(3) wait for 15 minutes, reading result, the rule of judged result is:
Positive findings: if in the situation of tiring higher than threshold value of antibody, HPV16L1VLP antigen and indirect immobilized people's antibody response be (comprising those anti-HPV16L1VLP parts) completely.Therefore do not have HPV16L1VLP can be penetrated into bond release portion.Result is not have detection line to occur.This shows, the antibody titer in sample is higher than certain certain numerical value (higher than threshold value).Threshold value can change and can calculate according to the different antigen Y content in antigenic solution.
Negative findings: if in the situation of tiring lower than threshold value of antibody, HPV16L1VLP antigen and indirect immobilized people's antibody (being trapped in the antibody to be measured of sample receiving portion) do not have complete reaction (in the situation that titre/tire is low) or do not react (in negative sample in the situation that) at all.Therefore, unreacted or all HPV16L1VLP can be penetrated into bond release portion.Result is that a detection line will occur.At detection line place, because HPV16L1VLP comprises multiple identical epitopes, therefore there is sandwich reaction at detection line, form " insolubilized antibody X'-antigen Y-labelled antibody X' " ternary complex.This shows, the antibody titer in sample is lower than certain certain numerical value (lower than threshold value).
According to technique scheme, one group of sample is tested, obtain Fig. 4.
According to above-mentioned judgment rule, known in conjunction with Fig. 3 and Fig. 4, the testing result of the sample detecting is negative.
Embodiment 2:
Adopt the same device for fast detecting of embodiment 1, by embodiment 1 same procedure, another group sample is tested, obtain Fig. 6, according to above-mentioned judgment rule, known in conjunction with Fig. 5 and Fig. 6, the testing result of the sample detecting is positive.
In addition, for the anti-HPV16 antibody sample of concentration known, be the sample of 5000mIU/ml, 1000mIU/ml, 500mIU/ml, 200mIU/ml, 100mIU/ml, 50mIU/ml, 20mIU/ml and 10mIU/ml for series of standards concentration by serial dilution legal system.
Above-mentioned sample is detected by pick-up unit and the detection method of embodiment 1, result shows, the utility model pick-up unit can detect the sample that is low to moderate 50mIU/ml, and the detection sensitivity of existing method (WO2012097788) is minimum for 500mIU/ml (seeing the following form).
The comparison of table 1 detection sensitivity
| Antibody concentration in sample | Embodiment 2 | Existing method (WO2012097788) |
| 5000mIU/ml | Positive | Positive |
| 1000mIU/ml | Positive | Positive |
| 500mIU/ml | Positive | Positive |
| 200mIU/ml | Positive | Negative |
| 100mIU/ml | Positive | Negative |
| 50mIU/ml | Positive | Negative |
| 20mIU/ml | Extremely weak positive | Negative |
| 10mIU/m | Negative | Negative |
Embodiment 3:
The device for fast detecting of the present embodiment comprises the first container and is positioned at the antigenic solution of described the first container, and described antigenic solution is the Tris BSA damping fluid containing the VLP of at least 200 HPV16L1 capsid proteins; And detection piece.
Described detection piece is as shown in referring to Fig. 7 and Fig. 8.Be provided with from left to right: a sample receiving portion, one bond release portion, one reacting part, one imbibition portion, wherein, described sample receiving portion is first accepted to accept antigenic solution after sample solution, the glass fibre of the amido functional group of described receiving portion has been covalent bond goat anti-human igg antibody and rabbit anti erythrocyte antibody.It is faster that this receiving portion that different with embodiment 1 is can accept that the sample of greater number and antigenic solution flow.
In the present embodiment, described receiving portion also can adopt the sample that is used to absorb greater number on the glucosan in conjunction with Protein G.
Described lateral flow immunochromatography part also comprises a shell, and described shell is provided with corresponding well and the area of observation coverage, the position of corresponding receiving portion and reacting part respectively.
Other structures of described detection piece are identical with embodiment 1.
The method that detects testing sample comprises the following steps:
(1) 50 μ l samples are added in sample receiving portion, described sample can be selected from but be not limited to people's whole blood or serum or blood plasma;
(2) then 100 μ l antigenic solutions are added in sample receiving portion;
(3) wait for 10 minutes, reading result, the rule of described judged result is with embodiment 1.
According to technique scheme, one group of sample is tested, obtain Fig. 9.
Known according to above-mentioned judgment rule, the testing result of the sample detecting is positive.
Embodiment 4:
The device for fast detecting of the present embodiment comprises the first container and is positioned at the antigenic solution of described the first container, and described antigenic solution is the Tris BSA damping fluid containing at least 100 virus-like particles that are made up of HPV16L1 capsid protein (VLP); Second container and be positioned at described second container gold mark mouse-anti HPV16-L1 monoclonal antibody; And detection piece.
Described detection piece is as shown in Figure 10～12.Described immunity percolation detection piece is provided with from top to bottom:
One sample receiving portion; Described sample receiving portion comprises a filter paper, the glass fibre element film that the material of described filter paper is functionalization, and on filter paper, immobilization is in conjunction with goat anti-human igg antibody and rabbit anti erythrocyte antibody;
One reacting part; Described reacting part comprises a nitrocellulose filter, and described porous nitrocellulose filter is provided with detection line, and control line for representing to have detected; On described detection line, be loaded with mouse-anti HPV16L1 monoclonal antibody; On described control line, be loaded with goat-anti rabbit polyclonal antibody.
Described in employing technique scheme, the method for immunity percolation device for quick testing detection testing sample comprises the following steps:
(1) add 100 μ l samples waits for until all liq passes through filter paper to pre-service filter paper;
(2) then 100 μ l antigenic solutions are added on pre-service filter paper until all liq passes through filter paper;
(3) add 100 μ l gold mark mouse-anti HPV16-L1 monoclonal antibodies and the anti-sheep Anti-TNF-α of gold mark rabbit liquid solution, wait for until all liq passes through filter paper; Remove filter paper reading result:
If the concentration of anti-HPV16L1 antibody is more than threshold value in testing sample, only there is a line (point) in Quality Control region, positive; If the concentration of not anti-HPV16L1 antibody or anti-HPV16L1 antibody is lower than threshold value, except nature controlling line there will be line (point), detection line also will there will be line (point), negative.
According to technique scheme, one group of sample is tested, obtain Figure 14.
According to above-mentioned judgment rule, known in conjunction with Figure 13 and 14, the testing result of the sample detecting is positive.
The advantage of the present embodiment: this device can be for detection of the antibody in the testing sample after enrichment/concentrated, and the testing sample of high application of sample amount (or even whole blood sample), therefore can reach the sensitivity more much higher than effluent system.Detection for some rare HPV infection type is meaningful especially.
Shortcoming: need to additionally add again a step gold mark solution after sample box antigenic solution adds.
Embodiment 5:
Adopt the same device for fast detecting of embodiment 4, by embodiment 1 same procedure, another group sample is tested, obtain Figure 15.
According to above-mentioned judgment rule, known in conjunction with Figure 15, the testing result of the sample detecting is negative.
Embodiment 6:
The device for fast detecting of the present embodiment is substantially similar to embodiment 1, and difference is:
Comprise 2 the first containers, in 2 the first containers, corresponding antigenic solution is respectively the Tris BSA damping fluid containing the VLP of at least 100 HPV16L1 capsid proteins; Tris BSA damping fluid with the VLP containing at least 100 HPV18L1 capsid proteins.
Described test block is as the test block of embodiment 1, different: to be provided with two detection lines at reacting part, be loaded with anti-HPV116L1VLP monoclonal antibody (T1) as detection line 1 at nitrocellulose filter, be loaded with monoclonal/polyclonal antibody (T2) of anti-HPV58L1VLP as detection line 2 simultaneously.In addition, same in addition: bond release portion is the dacron film that is only loaded with the anti-sheep polyclonal antibody of rabbit of colloid gold label.
HPV antibody in whole blood/blood serum/blood plasma of people, usually can with dissimilar HPV reaction, for example HPV16, HPV18, HPV58 and other types, for example HPV5 in theory.The multiparameter that can simultaneously detect the antibody of HPV58L1 and HPV16L1 in order to realize one-time detection detects, only there are a kind of anti-HPV58L1VLP monoclonal/polyclonal antibody of anti-HPV16L1VLP monoclonal antibody low specificity relative to one and this antibody of high specific cross reaction to occur with HPV16L1VLP, in this case, if carrying out simply the multiparameter of Parallel testing HPV16 and HPV58 antibody detects, so because HPV16V1VLP can be simultaneously in conjunction with HPV16 monoclonal antibody and HPV58 antibody, then golden labeling antibody also can be simultaneously in conjunction with HPV16V1VLP and HPV58V1VLP, therefore cannot correctly detect and whether contain HPV58.
For above problem, feasible solution is: adopt the antibody of the anti-HPV16L1VLP of specificity to make a HPV16L1VLP inactivation, then relatively low specific anti-HPV58L1VLP (monoclonal) antibody can not react and can only react with HPV58L1VLP with HPV16L1VLP.
Introduce the antibody of the specific anti-HPV16L1VLP of above-mentioned employing and (monoclonal/polyclone) antibody of low specificity/non-specific anti-HPV58L1VLP below in conjunction with specific embodiment, detect anti-HPV16L1VLP antibody and anti-HPV58L1VLP antibody.
Detection method comprises the following steps:
(1) people's whole blood sample is added to sample receiving portion;
(2) again the hybrid antigen solution that contains HPV16L1VLP and HPV58L1VLP is added to antigen receiving portion, all HPV16L1VLP can be arrived and combination by anti-HPV16 antibody capture by the people in sample receiving portion, and all HPV58L1VLP will be combined with the antibody of the anti-HPV58L1VLP in detection line 2 regions;
(3) then first add the anti-HPV16L1VLP monoclonal antibody solution of gold mark to sample receiving portion, the anti-HPV16L1VLP monoclonal anti of gold mark reacts with the anti-HPV16 antibody conjugates of HPV16L1VLP-people, make HPV16L1VLP inactivation, there is not visible line in simultaneous reactions portion detection line 1 region, shows the tests positive of the anti-HPV16 antibody of people, add again the anti-HPV58L1VLP antibody-solutions of gold mark, because the anti-HPV58L1VLP antibody of described gold mark cannot be combined with the HPV16L1VLP of sample receiving portion (because the anti-HPV16L1VLP monoclonal anti of gold mark reacts with the anti-HPV16 antibody conjugates of HPV16L1VLP-people, make HPV16L1VLP inactivation), therefore the anti-HPV58L1VLP antibody of described gold mark moves to the detection line 2 of reacting part, and react with the antibody conjugates generation sandwich of the anti-HPV58L1VLP of HPV58L1VLP-herein, thereby form a visible detection line, the testing result that shows the anti-HPV58 antibody of people in this sample is negative.
In another detects, in sample, only contain anti-HPV58 antibody, result is so: the gold first adding is marked anti-HPV16L1VLP antibody-solutions and move to detection line 1 region of reacting part, the HPV16L1VLP antigen in region-anti-HPV16L1VLP antibody conjugates combination therewith, thereby make HPV16L1VLP inactivation, and detection line 1 is shown as visible line, show that the testing result of the anti-HPV16 antibody of people in sample is negative; After the gold that adds mark anti-HPV58L1VLP antibody-solutions and the HPV58L1VLP of sample receiving portion antigen-anti-HPV58L1VLP antibody conjugates is combined, and reacting part detection line 2 regions do not show visible detection line.Therefore, even if only having under the condition of the anti-HPV16 antibody of specificity and the anti-HPV58 antibody of non-specific/low specificity, still can detect by adding successively the gold anti-HPV16L1VLP antibody of mark and gold mark HPV58L1VLP antibody to realize specific multiparameter, detect and contain anti-HPV16 or/and HPV58 simultaneously.
Above-mentioned detection device is lateral flow device for immunochromatography, those skilled in the art can be accordingly, under the prerequisite without any need for creative work, the method is applied to immunity percolation and tests fast, add successively the gold anti-HPV16L1VLP antibody of mark and gold mark HPV58L1VLP antibody.
All documents of mentioning at the utility model are all quoted as a reference in this application, are just quoted separately as a reference as each section of document.In addition should be understood that those skilled in the art can make various changes or modifications the utility model after having read above-mentioned instruction content of the present utility model, these equivalent form of values fall within the application's appended claims limited range equally.
Claims (10)
1. a pick-up unit, is characterized in that, described pick-up unit comprises:
The first container and be positioned at the antigenic solution of described the first container, the antigen Y in described antigenic solution can with antibody X specific binding to be measured, wherein said antigen Y is unlabelled antigen Y or the labelled antigen Y' with detectable;
Optional second container and be positioned at described second container by the labelled antibody X' of detectable institute mark; With
One detection piece, described detection piece comprises a sample receiving portion and a reacting part, wherein
Described sample receiving portion is used for accepting testing sample solution, and described sample receiving portion is loaded with the trapping agent for catching described antibody X to be measured;
Described reacting part is provided with control line region, and described control line region is provided with the control line for representing to have detected; Described reacting part is also provided with the detection line for detection of described antibody X to be measured, described detection line region is loaded with insolubilized antibody X', and described insolubilized antibody X' can react with described antigen Y and labelled antibody X' formation " insolubilized antibody X'-antigen Y-labelled antibody X' " ternary complex; Or described insolubilized antibody X' reacts formation " insolubilized antibody X' – labelled antigen Y' " binary complex with described labelled antigen Y'.
2. pick-up unit according to claim 1, it is characterized in that, described detection piece also comprises a bond release portion, and be loaded with releasable bond in described bond release portion, described bond comprises by the antibody X' of detectable institute mark and optional by the Quality Control thing of detectable institute mark; And described bond release portion is located between described sample receiving portion and reacting part.
3. according to pick-up unit described in claim 1 or 2, it is characterized in that, described detection piece also comprises an antigenic solution receiving portion, and described sample receiving portion and described antigenic solution receiving portion are one.
4. pick-up unit according to claim 1, is characterized in that, described detection piece also comprises an antigenic solution receiving portion, and described antigenic solution receiving portion is located at the upstream of sample receiving portion; And/or
Described detection piece also comprises an imbibition portion, and described imbibition portion is located at the downstream of described reacting part, promotes flow direction reacting part.
5. pick-up unit according to claim 1, is characterized in that, described sample receiving portion is also provided with covalently bound anti erythrocyte antibody; And/or the control line region of described reacting part is loaded with Quality Control antibody.
6. pick-up unit according to claim 1, is characterized in that:
Described reacting part is provided with detection line, and described detection line region is loaded with the insolubilized antibody X' that is selected from lower group: anti-HPV16L1 monoclonal antibody, anti-HPV18L1 monoclonal antibody, anti-HPV58L1 monoclonal antibody.
7. pick-up unit according to claim 1, is characterized in that, described reacting part is located at below sample receiving portion; And described pick-up unit also comprises:
Second container and the labelled antibody X' solution that is positioned at described second container.
8. according to pick-up unit described in claim 1,2 or 4, it is characterized in that: the releasable bond that described bond release portion is loaded with comprises the antibody X' of golden mark and the Quality Control thing antibody of golden mark, and the control line region of described reacting part is also loaded with Quality Control antibody; And/or
The Quality Control thing that simultaneously comprises golden mark in described golden labelled antibody X' solution, the control line region of described reacting part is loaded with Quality Control antibody.
9. pick-up unit according to claim 1, is characterized in that, described antibody X to be measured is one or more, is designated as respectively Xn, and n is positive integer; Correspondingly, the antigen Y containing in described antigenic solution is one or more, is designated as respectively Yn, and n is positive integer; Labelled antibody X' is one or more, is designated as respectively X'n, and n is positive integer;
And the detection line region of described reacting part is provided with the n bar detection line corresponding to each antibody X to be measured, wherein n article of detection line is corresponding is respectively loaded with the insolubilized antibody X'n corresponding to antibody Xn to be measured,
Wherein, described each insolubilized antibody X'n reacts formation " insolubilized antibody X'n-antigen Yn-labelled antibody X'n " ternary complex with corresponding antigen Yn and labelled antibody X'n;
" insolubilized antibody X'-antigen Y-labelled antibody X' " ternary complex; Or described insolubilized antibody X' reacts formation " insolubilized antibody X' – labelled antigen Y' " binary complex with described labelled antigen Y'.
10. according to arbitrary described pick-up unit in claim 1,2,4-7 and 9, it is characterized in that: described pick-up unit is kit form, wherein said kit comprises described pick-up unit and operation instruction.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201320612882.8U CN203745473U (en) | 2013-09-30 | 2013-09-30 | Detection device |
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| CN201320612882.8U CN203745473U (en) | 2013-09-30 | 2013-09-30 | Detection device |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104515850A (en) * | 2013-09-30 | 2015-04-15 | 万冰 | Quick test device and quick test method |
| WO2022114941A1 (en) | 2020-11-24 | 2022-06-02 | Autonomous Organization Of Education "Nazarbayev University" | Method for detecting hpv16 and in vitro diagnostic device for detection |
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2013
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104515850A (en) * | 2013-09-30 | 2015-04-15 | 万冰 | Quick test device and quick test method |
| CN104515850B (en) * | 2013-09-30 | 2019-05-03 | 万冰 | Device for quick testing and method |
| WO2022114941A1 (en) | 2020-11-24 | 2022-06-02 | Autonomous Organization Of Education "Nazarbayev University" | Method for detecting hpv16 and in vitro diagnostic device for detection |
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