CN109187943A - The preparation method of anti-interference coating in a kind of anti-interference reagent cup and reagent cup - Google Patents
The preparation method of anti-interference coating in a kind of anti-interference reagent cup and reagent cup Download PDFInfo
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- CN109187943A CN109187943A CN201810973454.5A CN201810973454A CN109187943A CN 109187943 A CN109187943 A CN 109187943A CN 201810973454 A CN201810973454 A CN 201810973454A CN 109187943 A CN109187943 A CN 109187943A
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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
- G01N33/5302—Apparatus specially adapted for immunological test procedures
- G01N33/5304—Reaction vessels, e.g. agglutination plates
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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
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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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 problem of the invention discloses a kind of preparation method of anti-interference coating in anti-interference reagent cup and reagent cup, solving existing sealer can participate in the overall process immune response, also reduce the immune response of antigen-antibody while inhibiting interference.The preparation method of anti-interference coating in the present invention, including use CB that sealer is diluted to 0.5~1.5mg/mL, it is added in the cup pipe of reagent cup, 2~8 DEG C are placed 20~24 hours, and cup pipe is then cleaned drying more than twice using purified water；Cup pipe is placed in 40 DEG C or less drying.The present invention can be such that the interfering substance in sample is captured by anti-interference coating, and therefore, the sample for eliminating interference enters in subsequent immune response, and immune response ability can effectively get a promotion.
Description
Technical field
The present invention relates to fluorescence immune chromatography fields, and in particular to anti-interference in a kind of anti-interference reagent cup and reagent cup
The preparation method of coating.
Background technique
Immunodiagnosis is the theory of applied immunology, the various diseases of technology and methods diagnosis and measurement immune state.It is immune
Diagnostic reagent kind in diagnostic kit is most, is widely used in hospital, blood station, medical center, be mainly used for hepatitis detection,
Venereal disease detection, lesion detection, pregnant inspection etc..
It is divided into radioimmunology, enzyme-linked immunization, fluorescence method, chemoluminescence method, colloidal gold method, emulsion method by tracer
Deng.
Fluorescence immune chromatography technology is the film detection technique based on antigen and antibody specific immune response.The technology is with fixation
Having the fibre strip chromatographic material of detection line (coated antibody or envelope antigen) and nature controlling line (antiantibody) is stationary phase, test fluid
For mobile phase, fluorescent labeled antibody or antigen are fixed on connection gasket, move up analysans in chromatography strip by capillarity
It is dynamic.For having the macromolecular antigen (albumen, virus, pathogenic bacteria etc.) of multiple antigenic determinants, " sandwich " type is generallyd use
Double antibodies sandwich immune chromatography method, i.e. determinand are first in conjunction with fluorescent labeled antibody in the case where flowing phase separation, when arrival detection line
Shi Zaiyu coated antibody combines " sandwich " type for forming double antibodies sandwich.Instrument reads the fluorescence signal for being detected region, calculates
Target substance concentration.
It is known that immune detection, especially sandwich formats, are easy the interference by many endogenous antibody, wherein wrapping
Include anti-animal species antibody, rheumatoid factor and other autoantibodies or heterophile antibody.Interference antibody can be any class
Type antibody (IgG, IgA, IgM or IgE), can be in conjunction with other kind antibody or other cellular components and agent formulations.These
Endogenous antibody causes false positive or false negative by changing the binding ability of target object to be checked or detection reagent antibody, provides
The testing result not being consistent with patient's true clinical situation.
And in the prior art, inhibit following two categories mainly to interfere usually using business sealer:
1, rheumatoid factor (RF), RF are typical IgM class antibody, can recognize the Fc section of IgG.RF not only with human IgG knot
It closes, also in conjunction with the immunoglobulin of other kinds.RF is common in rheumatoid patient, autoimmune disease patient or other are strong
Kang Renqun.Controlling RF and interfering relatively effective method is that anti-RF associated antibodies are added in closed system.
2, heterophile antibody, heterophile antibody refer to the human antibody in conjunction with animal's antibody, antigen or autoantigen,
More acurrate description refers to the polyreactive antibody that those antigens unknown or unknown with definition react.It is found in 40% crowd
There are such antibody.The affinity of these antibody is low, and reactivity is weak, but still bridging capture can resist in the case where object to be checked lacks
Body and binding antibody, and binding site corresponding with object effective competition to be checked.By the way that the pure of normal serum or identical kind is added
Interference can be preferably minimized by changing antibody.These antibody can be saturated the binding site of heterophile antibody, so that interference be blocked to generate.
Because traditional sealer application method is that directly sealer is added in reaction system, at this point, sealer meeting
It participates in the overall process immune response, also reduces the immune response of antigen-antibody while inhibiting interference.
Summary of the invention
The immune response the technical problems to be solved by the present invention are: existing sealer can participate in the overall process is inhibiting interference
The problem of also reducing the immune response of antigen-antibody simultaneously, provides anti-interference painting in a kind of anti-interference reagent cup and reagent cup
The preparation method of layer, mode through the invention can be such that the interfering substance in sample is captured by anti-interference coating, therefore, eliminate
The sample of interference enters in subsequent immune response, and immune response ability can effectively get a promotion.
The present invention is achieved through the following technical solutions:
The preparation method of anti-interference coating in a kind of reagent cup, comprising:
Sealer is diluted to 0.5~1.5mg/mL using CB, is added in the cup pipe of reagent cup, 2~8 DEG C are placed 20~24
Hour, cup pipe is then cleaned into drying more than twice using purified water；Cup pipe is placed in 40 DEG C or less drying.
The present invention is by the setting of anti-interference coating, and in the process, the interfering substance coated in sample captures, and by
The interfering substance and sealer of capture are each attached in glass pipe, and subsequent immune response and inspection can't be entered with sample liquid
It in survey, therefore is immunoreacted ability and is greatly enhanced, effect is very significant.
Further, the CB is alkaline buffer, and the time toasted in baking oven is 20~24 hours.
Further, the CB includes NaHCO37.56g/L NaCO31.06g/L, NaCl 7.36g/L；The pH value of CB is
9.0。
Further, the sealer uses the Super ChemiBlock of Merck-millipore company
Heterophile。
Further, the cup pipe is dried in 37 DEG C of baking oven.
A kind of anti-interference reagent cup, including pedestal are sequentially arranged suction nozzle loading hole on pedestal, sample from left to right
Chamber, the first Reagent Tube, the second Reagent Tube, reagent strip；Sample-adding window and detection window are correspondingly arranged on above reagent strip；Described
The anti-interference coating of above method preparation is provided on one Reagent Tube and the second Reagent Tube.
Compared with prior art, the present invention having the following advantages and benefits:
1, after the interfering substance in sample of the present invention is captured by anti-interference coating, therefore the sample for eliminating interference enters back into
In the immune response in face, immune response ability can effectively get a promotion；
2, the relatively traditional chromatography method of coating of the present invention, in the case where background is constant, reaction is improved 20%~
30%.
Detailed description of the invention
Attached drawing described herein is used to provide to further understand the embodiment of the present invention, constitutes one of the application
Point, do not constitute the restriction to the embodiment of the present invention.In the accompanying drawings:
Fig. 1 is the structural schematic diagram of anti-interference reagent cup in the present invention.
1- pedestal, 2- suction nozzle loading hole, 3- sample chamber, the first Reagent Tube of 4-, the second Reagent Tube of 5-, 6- are loaded window, 7- inspection
Survey window.
Specific embodiment
To make the objectives, technical solutions, and advantages of the present invention clearer, below with reference to embodiment and attached drawing, to this
Invention is described in further detail, and exemplary embodiment of the invention and its explanation for explaining only the invention, are not made
For limitation of the invention.
Embodiment 1
The preparation method of anti-interference coating in a kind of reagent cup, comprising: sealer is diluted to 1mg/mL using CB, is added
In Reagent Tube, 2~8 DEG C are placed 20~24 hours, and Reagent Tube is then cleaned drying more than twice using purified water；By reagent
Pipe is placed in 37 DEG C of baking ovens and dries, and baking time is usually 20~24 hours in 37 DEG C of baking ovens.
A kind of anti-interference reagent cup, including pedestal 1 are sequentially arranged suction nozzle loading hole 2 on the pedestal 1, sample from left to right
This chamber 3, the first Reagent Tube 4, the second Reagent Tube 5, reagent strip；Sample-adding window 6 and detection window are correspondingly arranged on above reagent strip
7；The preparation side such as anti-interference coating in a kind of above-mentioned reagent cup is provided on first Reagent Tube 4 and the second Reagent Tube 5
The anti-interference coating that method is prepared.
CB described in the present embodiment refers to a kind of common alkaline buffer, and the pH value of the buffer is 9.0, specifically matches
Side are as follows: NaHCO37.56g/L NaCO31.06g/L, NaCl 7.36g/L.Sealer in the present embodiment uses Merck-
The Super ChemiBlock Heterophile of millipore company.
Specific use process is as follows for the anti-interference reagent cup of one of present invention:
Sample in sample chamber 3 is drawn using the suction nozzle in suction nozzle loading hole 2, is added the samples in the first Reagent Tube 4
After being reacted with the first reagent, then the reaction solution in the first Reagent Tube 4 is drawn with suction nozzle and is added in the second Reagent Tube 5, when first
After the liquid reactions in reaction solution and the second Reagent Tube 5 in Reagent Tube 4, draws be added drop-wise on sample-adding window 6 out again, addition
Onto the reagent strip of 6 lower section of sample-adding window, liquid is moved up in reagent strip and is moved at detection 7 position of window, is by detecting instrument
Signal value can be detected at detection 7 position of window.
Embodiment 2
The present embodiment the difference from embodiment 1 is that, the concentration that sealer adds in the present embodiment is different, in the present embodiment
It is provided that
Sealer is diluted to 0.5mg/mL using CB, is added in Reagent Tube, 2~8 DEG C are placed 20~24 hours, are then made
Reagent Tube is cleaned more than twice with purified water, drying；Reagent Tube is placed in 37 DEG C of baking ovens and is dried.
Embodiment 3
The present embodiment the difference from embodiment 1 is that, the concentration that sealer adds in the present embodiment is different, in the present embodiment
It is provided that
Sealer is diluted to 1.5mg/mL using CB, is added in Reagent Tube, 2~8 DEG C are placed 20~24 hours, are then made
Reagent Tube is cleaned more than twice with purified water, drying；Reagent Tube is placed in 37 DEG C of baking ovens and is dried.
The present invention is using a kind of anti-interference reagent cup made of embodiment 1 and traditional reagent cup respectively in identical reaction
It is detected in system, the operating process of detection is identical with parameter.Without anti-interference coating in traditional reagent cup, but
The sealer of optimal dosage is directly added into when detection in the reaction system.Testing result is as shown in table 1 below.
Table 1
By above-mentioned table 1: the relatively traditional chromatography method of coating improves reaction in the case where background is constant
20%~30%.
Above-described specific embodiment has carried out further the purpose of the present invention, technical scheme and beneficial effects
It is described in detail, it should be understood that being not intended to limit the present invention the foregoing is merely a specific embodiment of the invention
Protection scope, all within the spirits and principles of the present invention, any modification, equivalent substitution, improvement and etc. done should all include
Within protection scope of the present invention.
Claims (6)
1. the preparation method of anti-interference coating in a kind of reagent cup characterized by comprising
Sealer is diluted to 0.5~1.5mg/mL using CB, is added in the cup pipe of reagent cup, 2~8 DEG C of placements 20~24 are small
When, cup pipe is then cleaned into drying more than twice using purified water；Cup pipe is placed in 40 DEG C or less drying.
2. the preparation method of anti-interference coating in a kind of reagent cup according to claim 1, which is characterized in that the CB is
Alkaline buffer, the time toasted in baking oven are 20~24 hours.
3. the preparation method of anti-interference coating in a kind of reagent cup according to claim 2, which is characterized in that the CB packet
Include NaHCO37.56g/L NaCO31.06g/L, NaCl 7.36g/L；The pH value of CB is 9.0.
4. the preparation method of anti-interference coating in a kind of reagent cup according to claim 1, which is characterized in that the closing
Agent uses the Super ChemiBlock Heterophile of Merck-millipore company.
5. the preparation method of anti-interference coating in a kind of reagent cup according to claim 1, which is characterized in that the cup pipe
It is dried in 37 DEG C of baking oven.
6. a kind of anti-interference reagent cup, including pedestal (1), the suction nozzle loading hole being sequentially arranged on pedestal (1) from left to right
(2), sample chamber (3), the first Reagent Tube (4), the second Reagent Tube (5), reagent strip；Sample-adding is correspondingly arranged on above reagent strip
Window (6) and detection window (7)；It is characterized in that, being provided on first Reagent Tube (4) and the second Reagent Tube (5) such as right
It is required that the anti-interference coating of 1~5 described in any item method preparations.
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| CN201810973454.5A CN109187943B (en) | 2018-08-24 | 2018-08-24 | Anti-interference reagent cup and preparation method of anti-interference coating in reagent cup |
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| CN201810973454.5A CN109187943B (en) | 2018-08-24 | 2018-08-24 | Anti-interference reagent cup and preparation method of anti-interference coating in reagent cup |
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Cited By (1)
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|---|---|---|---|---|
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110672839A (en) * | 2019-09-09 | 2020-01-10 | 元升生物科技（上海）有限公司 | Anti-interference method for biochemical reagent |
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