CN110237876A - A kind of paper chip general channels structure and preparation method thereof - Google Patents
A kind of paper chip general channels structure and preparation method thereof Download PDFInfo
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- CN110237876A CN110237876A CN201910509971.1A CN201910509971A CN110237876A CN 110237876 A CN110237876 A CN 110237876A CN 201910509971 A CN201910509971 A CN 201910509971A CN 110237876 A CN110237876 A CN 110237876A
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
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- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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Abstract
The present invention relates to a kind of paper chip general channels structures and preparation method thereof, belong to disease detection technical field.The structure is prepared, the significant Protein Detection suitable for colloidal gold immunity chromatography using water imbibition filter paper as substrate based on fluid mechanics principle combination wax spray printing technique.The detection line and nature controlling line for being fixed with antibody are respectively set in detection reaction zone and Quality Control reaction zone, by the special shape for designing channel design, fluid to be measured is enabled to flow into reaction zone at a relatively high speed by inlet region and channel region, and make the vortex flow field for the low-speed stable that antibody protein not easily runs off and antigen-antibody reaction sufficiently carries out in reaction zone formation, solve the problems, such as that filter paper does not have ankyrin function, testing result is showed with macroscopic red stripes, and finally from outlet area flow pass structure.Replace the nitrocellulose membrane structure in traditional colloidal gold strip to carry out assembling as reacting environment structure of the invention, improves paper chip assembling and detection efficiency, effectively reduce cost.
Description
Technical field
The invention belongs to disease detection technical field, it is related to a kind of paper chip general channels structure and preparation method thereof, is
Micro-fluidic core chip technology is prepared based on fluid mechanics principle and wax spray printing, designed for detecting the paper chip of significant albumen
General channels structure.
Background technique
Using significant albumen such as test strips detection tumor markers, frequently with nitrocellulose filter as substrate, thereon
Sessile antibody albumen is immunoreacted by antigen-antibodies such as colloidal gold immunity chromatographies and is developed the color, to reach testing goal.Nitric acid is fine
Plain film higher cost is tieed up, test strips structure and assembling are complicated, and are not easy to form the microchannel structure of multi objective joint-detection.Such as
Fruit uses filter paper for substrate, although having good hydrophily and water absorbing capacity, the effect of ankyrin is poor, causes to react
And the antibody protein of detection zone is washed away by fluid, to be unable to normal reaction colour developing.
Summary of the invention
To solve the above problems, the present invention proposes that one kind prepares hydrophobic channel based on fluid mechanics principle combination wax spray printing
Micro-fluidic paper chip universal architecture and preparation method, by filter paper substrate wax spray print specific shape hydrophilic channel,
To control the flow velocity and flow direction of fluid, make it during being immunoreacted, the antibody protein as detection marker is not
It is easy to run off, and can uniformly mix and sufficiently be reacted to reach color developing effect with antigen, to replace in traditional colloidal gold strip
Nitrocellulose membrane structure, and facilitate and realize multi objective joint-detection, paper chip assembling detection and efficiency are improved, is effectively reduced into
This.The present invention is using absorbent filter as substrate, the channel design based on fluid mechanics principle and wax spray printing preparation specific shape,
Significant Protein Detection suitable for colloidal gold immunity chromatography.
Technical solution of the present invention is as follows:
A kind of paper chip general channels structure, be bilateral symmetry, including inlet region 1, channel region 2, reaction zone 3 and go out
Mouth region 4；The inlet region 1 and outlet area 4 is located at the both ends of paper chip general channels structure, channel region 2 and reaction zone 3
Between inlet region 1 and outlet area 4.
Fluid feed channel of the inlet region 1 as paper chip general channels structure, outlet area 4 is as completion reaction
The channel of residual fluid outflow paper chip general channels structure afterwards；
It totally two groups of the channel region 2, is arranged symmetrically, every group of channel region 2 includes that arrival end is wider than the oblique logical of outlet end
Road a2-1 and slanted channel b2-2, for guiding fluid to enter reaction zone 3 with higher flow velocity；The reaction zone 3 includes inspection
Survey reaction zone 3-1 and Quality Control reaction zone 3-2；The arrival end of two slanted channel a2-1 is connected with inlet region 1, two slanted channels
The outlet end of a2-1 is connected with the bottom end of detection reaction zone 3-1, and the two sides for detecting the bottom end reaction zone 3-1 are equipped with connection and lead to
Road is connected by communicating passage with the arrival end of two slanted channel b2-2, and fluid flows into slanted channel a2-1's from inlet region 1
Arrival end, then flowed out from the outlet end of slanted channel a2-1, from the fluid that the outlet end of slanted channel a2-1 is flowed out, a branch
It flows into detection reaction zone 3-1 and is reacted, flowed out in slanted channel b2-2 after reaction from communicating passage, another part is direct
It is flow in slanted channel b2-2 from communicating passage；The arrival end of two slanted channel b2-2 passes through communicating passage and detection reaction zone 3-
1 is connected with two slanted channel a2-1, and the outlet end of two slanted channel b2-2 is connected with the bottom end of Quality Control reaction zone 3-2, matter
The two sides for controlling the bottom end reaction zone 3-2 are equipped with communicating passage, are connected by communicating passage with outlet area 4, from two slanted channel b2-
In the fluid of 2 outlet end outflow, a part flows into Quality Control reaction zone 3-2 and is reacted, and flows out to after reaction from communicating passage
In outlet area 4, another part is directly flow in outlet area 4 from communicating passage；Detection line is equipped in the detection reaction zone 3-1
Nature controlling line 6 is equipped in 5, the Quality Control reaction zone 3-2；The detection line 5 and nature controlling line 6 with paper chip general channels knot
Structure two sides are perpendicular；The labelled antibody of the significant albumen composition of colloidal gold labeled monoclonal antibody-, institute are coated in the detection line 5
The secondary antibody of colloidal gold labeled monoclonal antibody is coated on the nature controlling line 6 stated.
The special construction of reaction zone 3 can make fluid form the vortex flow field of low flow velocity herein, keep it immune in antigen-antibody
In the generating process of reaction, the antibody for detecting marker can be kept fixed position and not be lost and uniformly mix with corresponding antigens
Sufficiently reaction is closed to reach color developing effect.
The two sides of the paper chip general channels structure are using hydrophobic barrier as edge；Between two slanted channel a2-1
For hydrophobic barrier；It is the hydrophobic barrier of triangular shape between at the top of two slanted channel b2-2 and detection reaction zone 3-1；Detection is anti-
Answer the two sides of area 3-1 and Quality Control reaction zone 3-2 using hydrophobic barrier as edge；The top of Quality Control reaction zone 3-2 is triangular shape
Hydrophobic barrier.
The paper chip general channels structure, substrate are water imbibition filter paper, and the material of hydrophobic barrier appearance is paraffin.
The paper chip general channels structure is rectangular configuration, length-width ratio 16:3；Paper chip general channels structure two
The hydrophobic barrier of side, length-width ratio 160:3.
The inlet region 1 and outlet area 4 is rectangular configuration, length-width ratio 25:24.
The slanted channel a2-1 and slanted channel b2-2, vertical height and mean breadth ratio are 28:13, inclination angle
Degree is 40 °.
The detection reaction zone 3-1 and Quality Control reaction zone 3-2 is rectangular configuration, length-width ratio 5:6.
According to demand, increases detection reaction zone 3-1 structure, realize the joint-detection to multiple markers albumen.
The preparation method of the paper chip general channels structure utilizes wax spray printer using water imbibition filter paper as substrate
Carry out paraffin micro-injection, structure needed for printing is formed, then by the filter paper of print structure with 115~120 DEG C of 15~30s of heating,
So that paraffin is impregnated with filter paper completely, forms the hydrophobic barrier that there is hydrophobic property and fluid flow direction can be effectively controlled, it is intermediate
The channel and conversion zone that gap section, which is reaction liquid, to be flowed through by filter paper siphon；It is cooling to paper chip general channels structure
To room temperature, two kinds of antibody are respectively coated and are protected from light natural drying at room temperature in being placed in detection line 5 and nature controlling line 6, it is dry after i.e.
Realize detection function.
Beneficial effects of the present invention: the present invention uses filter paper as substrate, is designed on filter paper based on fluid mechanics principle
Specific channel structure simultaneously carries out wax spray printing, realizes in the immunoreaction process in paper chip, fluid feed channel structure
Enter reaction solution region at a relatively high speed afterwards, and form stable low speed vortex flow field in reaction zone, can both enable and be used as inspection
The antibody for surveying marker is kept fixed position and is not lost, and antigen-antibody can be made to be sufficiently mixed and reacted to reach color developing effect
Function.The present invention, which is changed, is but difficult to by force the shortcomings that ankyrin is immunoreacted in filter paper water absorbing capacity, to replace
Nitrocellulose filter used in traditional paper chip can realize mostly finger by increasing detection reaction zone number as reacting environment
Joint-detection is marked, detection efficiency is improved, effectively reduces cost.
Detailed description of the invention
Fig. 1 is channel design schematic diagram of the invention:
Fig. 2 is the fluid flow line figure that the present invention is obtained using COMSOL software emulation.
Fig. 3 is testing result reference view of the invention, wherein (A) is the positive findings for detecting target antigen,
It (B) is negative findings, (C), (D) are the null result without reference value.
In figure: 1 inlet region；2 channel regions；3 reaction zones；4 outlet areas；5 detection lines；6 nature controlling lines；2-1 slanted channel a；2-2
Slanted channel b；3-1 detects reaction zone；3-2 Quality Control reaction zone.
Specific embodiment
Below in conjunction with attached drawing and technical solution, a specific embodiment of the invention is further illustrated.
As shown in Figure 1, a kind of paper chip general channels structure, filled black part is the hydrophobic barrier portion of wax spray printing
Point, rest part is the channel flowed through for fluid, and channel includes inlet region 1, channel region 2, reaction zone 3 and outlet area 4, and
Detection line 5 and nature controlling line 6.It totally two groups of channel region 2, is arranged symmetrically, every group includes slanted channel a2-1 and slanted channel b2-2；Instead
Answering area 3 includes detection reaction zone 3-1 and Quality Control reaction zone 3-2.
The paper chip general channels structure, long 32mm, wide 6mm；The hydrophobic barrier, long 32mm, wide 0.6mm.
The inlet region 1 and outlet area 4, long 5mm, wide 4.8mm.
The slanted channel a2-1 and slanted channel b2-2, vertical height 2.8mm, mean breadth 1.3mm, tilt angle
It is 40 °.
The detection reaction zone 3-1 and Quality Control reaction zone 3-2, long 2mm, wide 2.4mm, the distance of the two are 8.2mm.
Fig. 2 is the fluid flow line figure that the present invention is obtained using COMSOL software emulation, is utilizing progress fluid flow rate emulation
During, fluid is set with the speed of 8mm/s and reaches detection reaction zone 3-1, Jin Erjing through slanted channel a2-1 by inlet region 1
Slanted channel b2-2 reaches Quality Control reaction zone 3-2.Depth color represents fluid velocity, and streamline represents fluid trend.It can from figure
After finding out that flow field is stablized, inlet region 1, channel region 2 and outlet area 4 all show fair speed and the uniform streamline of shape, and
There is the whirlpool for representing low flow velocity He representing fluid stable, reaching dynamic equilibrium in detection reaction zone 3-1 and Quality Control reaction zone 3-2
Eddy flow line.Analyze simulation result it can be concluded that, the High Speed Flow Field of inlet region 1, channel region 2 and outlet area 4 can guarantee that detection is high
Effect carries out, and the low flow velocity steady flow condition for detecting reaction zone 3-1 and Quality Control reaction zone 3-2 can guarantee to be used as detection marker
Antibody is fixed in position, and antigen-antibody can be made to be sufficiently mixed to react and reach color developing effect.
After auto-CAD Software on Drawing paper chip general channels structure of the invention, using wax spray printer qualitative
Print channel design on filter paper and cut, be placed on 120 DEG C of heating 30s in heating plate, until wax melt after formed dredge
Water barrier, later will be for by colloidal gold labeled monoclonal antibody-target protein complex antibody in sample product (detection antibody) spraying
The secondary antibody (control Quality Control antibody) of colloidal gold labeled monoclonal antibody is done at room temperature in detection line 5, while in 6 fixed pin of nature controlling line
It is dry.Detection line 5 and nature controlling line 6 are the top regions along lower 1mm high of each reaction zone, are a part of reaction zone, material is also
Filter paper.
Specifically used mode are as follows: when being flowed through detection reaction zone 3-1 by inspection sample fluid, fixed detection at detection line 5
Antibody can form double antibodies sandwich immune complex in conjunction with colloidal gold labeled monoclonal antibody-target protein complex；Continue flow through Quality Control
When reaction zone 3-2, fixed secondary antibody can form immune complex in conjunction with colloidal gold labeled monoclonal antibody at nature controlling line 6.Two reaction lifes
At immune complex reach and can show the red of colloidal gold aggregation after a certain concentration.
Testing result is as shown in figure 3, (A) is positive findings (detecting target antigen): detection line 5 has with nature controlling line 6
Colour developing；(B) negative findings (target antigen is not detected): detection line 5 does not develop the color and nature controlling line 6 develops the color；(C), (D) null result:
If nature controlling line 6 does not develop the color, no matter whether detection line 5 develops the color, which does not have reference value, and it is invalid to be considered as.
Claims (6)
1. a kind of paper chip general channels structure, which is characterized in that the paper chip general channels structure is that bilateral symmetry is tied
Structure, including inlet region (1), channel region (2), reaction zone (3) and outlet area (4)；The inlet region (1) and outlet area (4) point
Not Wei Yu paper chip general channels structure both ends, channel region (2) and reaction zone (3) be located at inlet region (1) and outlet area (4) it
Between；
Fluid feed channel of the inlet region (1) as paper chip general channels structure, outlet area (4) is as completion reaction
The channel of residual fluid outflow paper chip general channels structure afterwards；
Totally two groups of the channel region (2), is arranged symmetrically, and every group of channel region (2) includes that arrival end is wider than the oblique logical of outlet end
Road a (2-1) and slanted channel b (2-2), for guiding fluid to enter reaction zone (3)；The reaction zone (3) includes that detection is anti-
Answer area (3-1) and Quality Control reaction zone (3-2)；The arrival end of two slanted channel a (2-1) is connected with inlet region (1), and two is oblique logical
The outlet end of road a (2-1) is connected with the bottom end of detection reaction zone (3-1), and the two sides for detecting the bottom end reaction zone (3-1) are equipped with
Communicating passage is connected by communicating passage with the arrival end of two slanted channel b (2-2), and fluid flows into oblique from inlet region (1)
The arrival end of channel a (2-1), then flowed out from the outlet end of slanted channel a (2-1), it is flowed from the outlet end of slanted channel a (2-1)
In fluid out, a branch, which flows into detection reaction zone (3-1), is reacted, and flows out to slanted channel from communicating passage after reaction
In b (2-2), another part is directly flow in slanted channel b (2-2) from communicating passage；The arrival end of two slanted channel b (2-2)
It is connected by communicating passage with detection reaction zone (3-1) and two slanted channel a (2-1), the outlet of two slanted channel b (2-2)
End is connected with the bottom end of Quality Control reaction zone (3-2), and the two sides of the bottom end Quality Control reaction zone (3-2) are equipped with communicating passage, passes through company
Circulation passage is connected with outlet area (4), and from the fluid that the outlet end of two slanted channel b (2-2) is flowed out, a part flows into Quality Control
Reaction zone (3-2) is reacted, and is flowed out in outlet area (4) after reaction from communicating passage, another part is directly from communicating passage
It flow in outlet area (4)；Detection line (5) are equipped in the detection reaction zone (3-1), in the Quality Control reaction zone (3-2)
Equipped with nature controlling line (6)；The detection line (5) and nature controlling line (6) is perpendicular with paper chip general channels structure two sides；It is described
Detection line (5) on be coated with the labelled antibody of the significant albumen composition of colloidal gold labeled monoclonal antibody-, on the nature controlling line (6)
It is coated with the secondary antibody of colloidal gold labeled monoclonal antibody；
The two sides of the paper chip general channels structure are using hydrophobic barrier as edge；It is between two slanted channel a (2-1)
Hydrophobic barrier；It is the hydrophobic barrier of triangular shape between at the top of two slanted channel b (2-2) and detection reaction zone (3-1)；Detection
The two sides of reaction zone (3-1) and Quality Control reaction zone (3-2) are using hydrophobic barrier as edge；The top of Quality Control reaction zone (3-2) is
The hydrophobic barrier of triangular shape.
2. a kind of paper chip general channels structure according to claim 1, which is characterized in that the paper chip is general logical
Road structure, substrate are water imbibition filter paper, and the material of hydrophobic barrier appearance is paraffin.
3. a kind of paper chip general channels structure according to claim 1 or 2, which is characterized in that
The paper chip general channels structure is rectangular configuration, length-width ratio 16:3；Paper chip general channels structure two sides
Hydrophobic barrier, length-width ratio 160:3；
The inlet region (1) and outlet area (4) is rectangular configuration, length-width ratio 25:24；
The slanted channel a (2-1) and slanted channel b (2-2), vertical height and mean breadth ratio are 28:13, inclination angle
Degree is 40 °；
The detection reaction zone (3-1) and Quality Control reaction zone (3-2) is rectangular configuration, length-width ratio 5:6.
4. a kind of paper chip general channels structure according to claim 1 or 2, which is characterized in that according to demand, increase inspection
Reaction zone (3-1) structure is surveyed, realizes the joint-detection to multiple markers albumen.
5. a kind of paper chip general channels structure according to claim 3, which is characterized in that according to demand, increase detection
Reaction zone (3-1) structure realizes the joint-detection to multiple markers albumen.
6. a kind of preparation method of paper chip general channels structure described in claim 1-5, which is characterized in that filtered with water imbibition
Paper is substrate, using wax spray printer carry out paraffin micro-injection, printing formed needed for structure, then by the filter paper of print structure with
115~120 DEG C of 15~30s of heating make paraffin be impregnated with filter paper completely, and being formed has hydrophobic property and fluid flowing can be effectively controlled
The hydrophobic barrier in direction, the channel and conversion zone that intermediate gap section, which is reaction liquid, to be flowed through by filter paper siphon；To
Paper chip general channels structure is cooled to room temperature, and two kinds of antibody are respectively coated in detection line (5) and nature controlling line (6), are placed in
It is protected from light natural drying at room temperature, realizes detection function after dry.
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| CN111856008A (en) * | 2020-08-03 | 2020-10-30 | 齐鲁工业大学 | Test paper for rapidly detecting various respiratory pathogens and preparation method thereof |
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| CN113671176A (en) * | 2021-08-02 | 2021-11-19 | 卢青松 | Target object immunochromatography test paper |
| CN113984753A (en) * | 2021-11-04 | 2022-01-28 | 长春美泰仪器有限公司 | Formaldehyde detection test paper, preparation method thereof and formaldehyde detection system |
| CN114196519A (en) * | 2021-12-10 | 2022-03-18 | 中国检验认证集团辽宁有限公司 | Disease marker detection chip general structure utilizing body temperature reaction and use method thereof |
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