CN107748160B - POCT ready-to-use micro-fluidic chip and application method - Google Patents
POCT ready-to-use micro-fluidic chip and application method Download PDFInfo
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- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 21
- 238000003860 storage Methods 0.000 claims abstract description 12
- 239000012472 biological sample Substances 0.000 claims abstract description 5
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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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
- G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
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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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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Abstract
The invention discloses a POCT ready-to-use micro-fluidic chip and an application method thereof. The chip comprises a plurality of parallel coaxial sleeve structures, a core tube positioned on an inner layer comprises a plurality of mutually isolated cabin sections and corresponding slits for reagent loading and functional operation, a tube sleeve positioned on an outer layer is provided with a plurality of slits or through holes with openings or semi-openings, and biological analysis is carried out through the relative movement of the core tube and the sleeve, including axial movement, rotation and combined operation thereof. The chip of the invention has the following advantages: the requirements of reagent preloading and storage and transportation can be met, and the reagent can be used only by filling a sample before analysis; the analysis operation is simple, and external complex pump valve control is not needed; the parallel multi-step operation of certain biological analysis can be realized in a fully integrated mode, so that the automation of an analysis instrument is facilitated; no complex micro-channel, simple processing and convenient batch production. The chip can be used for extracting and analyzing biological samples such as protein, nucleic acid and the like.
Description
Technical Field
The invention belongs to the technical field of detection, particularly relates to a microfluidic technology and a POCT product technology, and particularly relates to a POCT ready-to-use microfluidic chip and an application method.
Background
POCT (Point-of-care testing, Point-of-care testing or Point-of-care testing) refers to a testing method performed at a sampling site and used for rapidly obtaining a testing result by using a portable analyzer and a matching reagent.
Traditionally, biological detection processes are mostly performed in separate reaction tubes or plates, and the loading of reagents and samples is generally performed in a "pipetting" manner by a professional or an automated workstation. For biological detection involving a multi-step operation process, manual operation is time-consuming and labor-consuming, and the risk of unstable results caused by human factors exists; although the automatic pipetting workstation can meet the requirement of high-flux rapid and accurate analysis, the system is complex, the equipment is heavy and expensive and the maintenance requirement is high due to the pump valve pipeline control and the mechanical arm shifting operation. Therefore, such biological tests traditionally relying on complex pipetting operations are mostly limited to be performed in professional laboratories or large test centers, and are difficult to adapt to the requirements of POCT applications.
Microfluidics (microfluidics), a system science technique for manipulating or processing micro-fluids using microchannels, can provide a very advantageous fluid manipulation tool for biological analysis for POCT applications. The method can realize flexible combination and scale integration of functional units (such as sample preparation, reaction, separation and detection) required by biological analysis on a chip with a few square centimeters, greatly reduces the consumption of reagents/samples, accelerates the reaction analysis speed, and is easy for high-throughput operation, automation and portability. These advantages are not in line with the characteristics of miniaturization of instruments, simplicity and convenience in operation, timeliness of results, sample quantification and the like required by POCT products, and therefore, more and more POCT products take microfluidic chips as core technology platforms.
However, the existing microfluidic chip POCT products still face many challenges, including: (1) the fluid operation mode is still relatively complex and mostly depends on precise pump valve operation; (2) the special requirements of the chip on the material or structure (such as biocompatibility and micromachining) lead to difficult mass production and high cost; (3) integration level, test flux and detection performance are balanced, most of the existing microfluidic POCT products have multiple detection capabilities and cannot realize quantitative detection or full integration, and the detection indexes of the full integration and the quantitative detection function are limited by single application.
Disclosure of Invention
The invention aims to provide a POCT ready-to-use micro-fluidic chip which is low in cost, simple in production process and free from the limitation of single detection index and application and an application method thereof.
In order to overcome the defects in the prior art, the invention adopts the following technical scheme:
a POCT ready-to-use micro-fluidic chip comprises a pipe sleeve chip, wherein a plurality of parallel core pipes with coaxial sleeve structures are formed on a pipe sleeve positioned in the pipe sleeve chip; the pipe sleeve chip comprises an operation section positioning hole, an initial limiting section, a core pipe action section, a core pipe termination limiting section and a limiting screw, wherein the operation section positioning hole, the initial limiting section, the core pipe action section and the core pipe termination limiting section are positioned on the pipe sleeve; the core tube consists of an operation section and a main body section, the main body section and the operation section are integrated or fixedly connected, the main body section and the operation section are structurally and integrally arranged in a coaxial series cylinder, and the diameter of the cylinder of the main body section is slightly larger than that of the cylinder of the operation section; the bottom of the main body section is provided with a plurality of slits, the peripheral surface of the operation section is provided with a rotating position mark, wherein one surface on the same side with the slits indicates loading, one surface on the completely different side from the slits indicates analysis, and two surfaces between the loading and the analysis are both stored.
Furthermore, a collecting pool, an operation hole and an optical detection hole are arranged at the bottom of the pipe sleeve chip; the top surface of the pipe sleeve chip is provided with a rotating position indicating window, a sample adding hole, a sealing sheet clamping groove and a sample adding hole sealing sheet. The material of the overlapping part of the sample adding hole sealing sheet and the sample adding hole can be a plastic hydrophobic film easy to puncture or an aluminum foil containing a hydrophobic layer.
The inner diameter of the positioning hole of the operating rod is equivalent to the outer diameter of the operating section of the core tube, and the positioning hole is used for positioning and butting the external operating rod and the operating section of the core tube of the chip; the inner diameter of the core tube action section is equivalent to the outer diameter of the core tube main body section, and a sealing structure is formed by matching with the core tube for pre-storing a reagent, recycling waste liquid and providing a space required by the operation and the movement of the core tube; an inner diameter mutation area between the core tube action section and the operating rod positioning hole forms an initial limiting section which is used for positioning the operation initial position of the core tube and preventing the core tube from sliding out of the operating rod positioning hole; the sample adding holes are of through hole structures and are positioned on the top surface of the core tube action section, the number of the sample adding holes is twice of that of the cabin sections of the core tube, namely each cabin section corresponds to two sample adding holes and is respectively used for a reagent/sample filling port and exhaust operation during filling, and the hole pitch proportion of the sample adding holes is corresponding to the length of the corresponding cabin sections; the collecting tank is positioned at the bottom of the action section of the core tube, has a semi-open structure, and the open side of the collecting tank can be communicated with the slit of the core tube and is used for collecting and transferring a specific reagent; the operation hole and the detection hole are positioned at the bottom of the core tube action section and are of a half-opening structure, and the opening direction faces the outside of the chip and is used for positioning the operation unit and the detection unit; the stop limiting section and the limiting screw are used for limiting the operation stop position of the core tube, and the core tube can be inserted into the tube sleeve chip through one side of the stop limiting section when the limiting screw is not installed; the pipe sleeve chip is provided with a sealing sheet clamping groove which is positioned above the sampling hole and is embedded with the sampling hole sealing sheet for permanently or temporarily sealing the sampling hole.
Furthermore, the main body section is also provided with a plurality of cabin sections with different volumes and a light-emitting detection cabin, and the cabin sections are divided into a sample cabin, a primary washing cabin and a secondary washing cabin.
Furthermore, the thickness (partition structure) between the cabin sections is not less than the length of the pipe sleeve chip collecting pool parallel to the axial direction, so that the better separation and series flow blocking effects are achieved.
The width of the slit is not more than 1 mm, the inner side surface of the slit is smooth, and an included angle between the inner side surface of the slit and the radial direction of the core tube is an acute angle, so that a better separation effect is achieved.
The cross section of the collecting tank parallel to the axis direction is in an equiangular trapezoid shape with smooth transition corners, one side close to the axis is a wide side, and the effective thickness of the bottom of the collecting tank is not more than 1 mm, so that better separation and cabin sealing effects are achieved.
Furthermore, the core tube and the pipe sleeve chip are made of hydrophobic polymer materials or materials subjected to surface hydrophobic treatment on solution contact surfaces.
Further, the core tube is made of black or dark opaque materials, and the tube sleeve is made of optical transparent materials or partially contains the optical transparent materials.
Further, the POCT ready-to-use microfluidic chip is applied to biological sample analysis, and the biomolecules are preferably protein and nucleic acid.
Further, the biological sample analysis includes biomolecule extraction, and qualitative and/or quantitative analysis.
A use method of a POCT ready-to-use microfluidic chip is characterized by comprising the following steps:
(1) when the slit openings of the core tube are rotated to coincide with the sampling holes on the top surface of the sleeve, aqueous phase solution containing samples or reagents can be respectively added into each cabin section in the core tube;
(2) when the slit openings of the core tube are rotated to be staggered by 90 degrees with the sampling holes on the top surface of the sleeve, the sealing storage and transportation of the aqueous phase solution of the sample or the reagent in each cabin section in the core tube can be realized;
(3) when the slit openings of the core tube are rotated to be staggered 180 degrees with the sampling hole on the top surface of the sleeve, the core tube and the sleeve move relatively, so that aqueous phase solution of samples or reagents in all cabin sections in the core tube is sequentially fused, exchanged or separated with substances in a collecting pool on the bottom surface of the sleeve, and then multi-step biological analysis is completed.
Furthermore, a certain amount of oil phase is injected into each cabin section before the reagent is added, so that better biocompatibility, sealing and lubricating effects are achieved.
The POCT ready-to-use micro-fluidic chip and the application method provided by the invention have scientific and reasonable design and the following advantages:
(1) the POCT ready-to-use microfluidic chip provided by the invention can be preloaded with a reagent, only a sample to be detected needs to be added when the POCT ready-to-use microfluidic chip is used, the use is convenient, and the analysis time is saved.
(2) The POCT ready-to-use microfluidic chip provided by the invention can realize complex multistep biological analysis by simple relative displacement operation, and reduces the requirements of the current microfluidic chip on operation control support equipment.
(3) The POCT ready-to-use microfluidic chip provided by the invention has no complex microfluidic pipeline, is simple to process and low in cost, and is convenient for batch production.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without inventive exercise:
FIG. 1 is a schematic view of an embodiment of a POCT ready-to-use microfluidic chip according to the present invention;
FIG. 2 is a schematic view of a chip-in-tube structure plate of a POCT ready-to-use microfluidic chip;
FIG. 3 is a schematic diagram of a core tube structure of a unit of the POCT ready-to-use microfluidic chip;
fig. 4 is a schematic cross-sectional view of a core tube of the POCT ready-to-use microfluidic chip of fig. 3;
fig. 5 is a schematic diagram of the rotational operation of the POCT ready-to-use microfluidic chip;
FIG. 6 is an axial operation schematic diagram of the POCT ready-to-use microfluidic chip for magnetoenzymeimmune chemiluminescence analysis;
FIG. 7 is a schematic axial diagram of the POCT ready-to-use microfluidic chip for "sample in and out" nucleic acid analysis;
the device comprises a pipe sleeve chip 1, a pipe sleeve chip 2, a core pipe 1-1, an operation section positioning hole 1-2, an initial limiting section 1-3, a collecting pool 1-4, an operation hole 1-5, a detection hole 1-6, a core pipe action section 1-7, a core pipe termination limiting section 1-8, a limiting screw 1-9, a rotation position indicating window 1-10, a sample adding hole 1-11, a sealing sheet clamping groove 1-12, a sample adding hole sealing sheet 2-1, a core pipe main body 2-2, a slit 2-3, an operation section 2-4, a partition structure 2-5, an operation section slot 2-6, an operation section slot 2-7, a core pipe slit opening edge w, a core pipe slit opening width α and a radial included angle of the slit edge of the core pipe.
Detailed Description
The present invention will be described in detail with reference to the following specific examples, which are provided for illustration and not for limitation of the invention.
Example 1
A POCT ready-to-use micro-fluidic chip and a method for magnetic enzyme immunoassay chemiluminescence analysis thereof are disclosed, as shown in figures 1 and 2, a transparent Teflon pipe sleeve chip 1 comprises an operation section positioning hole 1-1, an initial limiting section 1-2, a core pipe action section 1-6, a core pipe termination limiting section 1-7 and a limiting screw 1-8, which are used for positioning, rotating and pushing operation of a core pipe, the top surface of the pipe sleeve chip 1 is provided with a rotating position indication window 1-9, a sample adding hole 1-10, a sealing sheet clamping groove 1-11 and a sample adding hole sealing sheet 1-12, the bottom of the pipe sleeve chip 1 is provided with a collection pool 1-3, a magnetic operation hole 1-4 and an optical detection hole 1-5, as shown in figures 3 and 4, a black Teflon pipe 2 is composed of an operation section 2-3 and a main body section 2-1 provided with a series of slits 2-2 at the top, wherein the main body section 2-1 is provided with a volume cabin section 2-4, a primary washing cabin, a secondary washing cabin and a luminescence detection cabin, a magnetic bead, a horse radish enzyme immunoassay chip is provided with an operation section 2-2 incubation operation section, a horse radish antibody storage section with a radial direction, a horse radish antibody storage section with a width equal to a horse radish antibody storage section with a horse radish antibody storage depth equal to horse radish antibody storage section equal to a horse radish antibody storage section equal to horse, a horse radish antibody storage section with a horse.
The using method comprises the following steps:
firstly, as shown in FIG. 5I, placing sample adding holes 1-10 on a pipe sleeve chip 1 with openings facing upwards, confirming that a rotating position indicating window 1-9 displays sample adding, and injecting a reagent and an oil phase with a certain proportion into each cabin section 2-4 of a core tube 3 in sequence according to analysis requirements; then, as shown in fig. 5 II, the core tube 2 is rotated to the rotating position indication window 1-9 to display "storage", and the sampling hole sealing piece 1-12 is inserted into the sealing piece clamping groove 1-11 to be sealed and stored for standby; before use, the core tube 2 is rotated to the rotating position indicating window 1-9 to display 'sample adding', the openings of sample adding holes 1-10 on the tube sleeve chip 1 are placed upwards, sample adding hole sealing pieces 1-12 are torn off, and a sample to be detected is injected into a core tube sample cabin section 2-4; for analysis, in III shown in FIG. 5, the core tube 2 is first rotated to the "analysis" position.
The operation method for magnetoenzyme immunoassay chemiluminescence analysis is as follows:
(1) after the antigen and the antibody are fully reacted, applying a magnetic field to the magnetic operation hole 1-4 to enable the compound of the magnetic bead-the antigen-the horseradish peroxidase to pass through the slit 2-2 and gather at the collecting pool 1-3, as shown in I in figure 6;
(2) driving the core tube 2 to move relative to the tube sleeve chip 1, maintaining the magnetic field until the partition between the sample chamber and the primary washing chamber passes through the collection pool 1-3, and separating the compound of the magnetic beads, the antigen and the horseradish peroxidase from the waste liquid in the sample chamber, as shown in II in figure 6;
(3) driving the core tube 2 to move relative to the tube sleeve chip 1 until the collecting pool 1-3 is positioned below the primary washing chamber, removing the magnetic field, and fusing the compound of the magnetic bead-antigen-horseradish peroxidase with the primary washing solution under the action of surface tension, as shown in III in fig. 6;
(4) driving the core tube 2 to move relative to the tube sleeve chip 1, maintaining the magnetic field until the partition between the primary washing chamber and the secondary washing chamber passes right above the collecting pool 1-3, and separating the compound of the magnetic beads, the antigens and the horseradish peroxidase from the waste liquid of the primary washing chamber;
(5) driving the core tube 2 to move relative to the tube sleeve chip until the collecting pool 1-3 is positioned below the secondary washing cabin, removing the magnetic field, and fusing the compound of the magnetic beads, the antigens and the horseradish peroxidase with secondary washing liquid under the action of surface tension;
(5) driving the core tube 2 to move relative to the tube sleeve chip 1, maintaining the magnetic field until the partition between the secondary washing chamber and the luminescence detection chamber passes through the collection pool 1-3, and separating the compound of the magnetic beads, the antigens and the horseradish peroxidase from the waste liquid of the secondary washing chamber;
(6) the core tube 2 is driven to move relative to the tube sleeve chip 1 to the collection pool 1-3 and is positioned below the luminescence detection cabin, the magnetic field is removed, the compound of the magnetic bead-antigen-horseradish peroxidase is fused with the substrate reaction liquid under the action of surface tension to trigger chemiluminescence, and at the moment, a photoelectric conversion device for detecting chemiluminescence is positioned in an optical detection hole 1-5, as shown in IV in figure 6.
Example 2
A POCT ready-to-use micro-fluidic chip and a method for isothermal nucleic acid amplification analysis of 'sample in result out' thereof are disclosed, as shown in FIG. 1 and FIG. 2, a tube-in-tube chip 1 made of transparent polypropylene comprises an operation section positioning hole 1-1, an initial limiting section 1-2, a core tube action section 1-6, a core tube termination limiting section 1-7, a limiting screw 1-8, which are used for positioning, rotating and pushing operation of a core tube, a top surface of the tube-in-tube chip is provided with a rotating position indication window 1-9, a sample adding hole 1-10, a sealing sheet clamping groove 1-11 and a sample adding hole sealing sheet 1-12, a bottom of the tube-in-tube chip 1 is provided with a collection pool 1-3, a magnetic operation hole 1-4 and an optical detection hole 1-5, as shown in FIG. 3, a dark polypropylene tube 2 is composed of an operation section 2-3 and a main body section 2-1 provided with a series of slits 2-2, wherein the main body section 2-1 is provided with a volume 2-4-sample compartment, a primary washing compartment, a secondary washing compartment and an amplification detection compartment, a magnetic bead for amplification of cells, a magnetic bead.
The using method comprises the following steps:
firstly, as shown in I in figure 5, placing the sample adding holes 1-10 on the pipe sleeve chip 1 with the openings facing upwards, confirming that the rotating position indicating windows 1-9 display sample adding, and injecting an oil phase and a reagent in a certain proportion into each cabin section of a core pipe in sequence according to analysis requirements; then, as shown in II in fig. 5, the core tube 2 is rotated to the rotating position indication window 1-9 to display "storage", and the sampling hole sealing piece 1-12 is inserted into the sealing piece clamping groove 1-11 to be sealed and stored for standby; before use, the core tube 2 is rotated to the rotating position indicating window 1-9 to display 'sample adding', the openings of sample adding holes 1-10 on the tube sleeve chip 1 are placed upwards and pierce sample adding hole sealing sheets 1-12 above a sample cabin, and a sample to be detected is injected into a sample cabin section 2-4 of the core tube; for analysis, the core tube is first rotated to the "analysis" position, as shown in III in fig. 5.
The operation method for isothermal nucleic acid amplification analysis of 'sample in and out' is as follows:
(1) after the nucleic acid released by cell lysis is fully combined with the functionally modified magnetic beads, applying a magnetic field to the magnetic operation hole 1-4 to enable the magnetic particles carrying the nucleic acid to pass through the slit 2-2 and gather at the collection pool 1-3, as shown in I in FIG. 7;
(2) driving the core tube 2 to move relative to the tube sleeve chip, maintaining the magnetic field until the partition between the sample chamber and the primary washing chamber passes through the collection pool 1-3, and separating the magnetic particles carrying the nucleic acid from the waste liquid in the sample chamber, as shown in II in FIG. 7;
(3) driving the core tube 2 to move relative to the tube sleeve chip 1 until the collecting pool 1-3 is positioned below the primary washing cabin, removing the magnetic field, and fusing the magnetic particles carrying the nucleic acid with the primary washing solution under the action of surface tension, as shown in III in fig. 7;
(4) driving the core tube 2 to move relative to the tube sleeve chip 1, maintaining the magnetic field until the partition between the primary washing chamber and the secondary washing chamber passes through the collection pool 1-3, and separating the magnetic particles carrying the nucleic acid from the waste liquid of the primary washing chamber;
(5) driving the core tube 2 to move relative to the tube sleeve chip until the collecting pool 1-3 is positioned below the secondary washing cabin, removing the magnetic field, and fusing the magnetic particles carrying the nucleic acid with the secondary washing liquid under the action of surface tension;
(5) driving the core tube 2 to move relative to the tube sleeve chip 1, maintaining the magnetic field until the partition between the secondary washing chamber and the isothermal amplification detection chamber passes through the collection pool 1-3, and separating magnetic particles carrying nucleic acid magnetically from the waste liquid of the secondary washing chamber;
(6) driving the core tube 2 to move relative to the tube sleeve chip until the collecting pool 1-3 is positioned below the isothermal amplification detection cabin, removing the magnetic field, fusing magnetic particles carrying nucleic acid with amplification reaction premix liquid under the action of surface tension, and dissociating and releasing the nucleic acid from the surface of the magnetic particles into the amplification premix liquid;
(7) applying a magnetic field to the magnetic operation holes 1-4 to separate the magnetic particles from the amplification reaction solution and collect the magnetic particles in the collection pool 1-3, as shown in FIG. 7-IV;
(8) and driving the core tube 2 to move relative to the tube sleeve chip 1 until the optical detection holes 1-5 are positioned under the isothermal amplification detection cabin, heating the isothermal amplification detection cabin to the temperature required by isothermal amplification, and continuously acquiring fluorescence signals in the amplification process by a fluorescence excitation and acquisition module positioned at the optical detection holes 1-5 to generate a real-time quantitative amplification curve.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (8)
1. The utility model provides a POCT ready-to-use micro-fluidic chip, includes the pipe box chip, is located the pipe box of pipe box chip inside and forms the nuclear core tube of a plurality of parallel coaxial bushing structures, its characterized in that:
the pipe sleeve chip comprises an operation section positioning hole, an initial limiting section, a core pipe action section, a core pipe termination limiting section and a limiting screw, wherein the operation section positioning hole, the initial limiting section, the core pipe action section and the core pipe termination limiting section are positioned on the pipe sleeve; the core tube consists of an operation section and a main body section, and the bottom of the main body section is provided with a plurality of slits; a collecting pool, a magnetic operation hole and an optical detection hole are arranged at the bottom of the pipe sleeve chip; the top surface of the pipe sleeve chip is provided with a rotating position indicating window, a sampling hole, a sealing sheet clamping groove and a sampling hole sealing sheet; the sample adding holes are of a through hole structure and are positioned on the top surface of the core tube action section, the number of the sample adding holes is twice that of the core tube cabin sections, namely each cabin section corresponds to two sample adding holes and is respectively used for a reagent/sample filling port and exhaust operation during filling, and the hole pitch proportion of the sample adding holes is corresponding to the length of the corresponding cabin section; the collecting tank is positioned at the bottom of the action section of the core tube, has a semi-open structure, and the open side of the collecting tank can be communicated with the slit of the core tube and is used for collecting and transferring a specific reagent; the magnetic operation hole and the detection hole are positioned at the bottom of the core tube action section and are of a half-opening structure, and the opening direction faces the outside of the chip and is used for positioning the operation unit and the detection unit; the sealing piece clamping groove is positioned above the sampling hole and is embedded with the sampling hole sealing piece to be used for permanently or temporarily sealing the sampling hole; applying a magnetic field to the magnetic operation hole to enable the complex of the magnetic bead-antigen-horseradish peroxidase to pass through the slit and be gathered at the collection pool.
2. The POCT-ready microfluidic chip of claim 1, wherein:
the main body section is also provided with a plurality of cabin sections with different volumes and a light-emitting detection cabin, and the cabin sections are divided into a sample cabin, a primary washing cabin and a secondary washing cabin.
3. The POCT-ready microfluidic chip of claim 2, wherein:
the thickness between the cabin sections is not less than the length of the pipe sleeve chip collecting pool parallel to the axis direction;
the width of the slit is not more than 1 mm, the inner side surface of the slit is smooth, and an included angle between the inner side surface of the slit and the radial direction of the core tube is an acute angle;
the cross section of the collecting tank parallel to the axis direction is in an equiangular trapezoid shape with smooth transition corners, one side close to the axis is a wide side, and the effective thickness of the bottom of the collecting tank is not more than 1 mm.
4. The POCT-ready microfluidic chip of claim 1, wherein:
the core tube and the tube sleeve chip are made of hydrophobic polymer materials or materials with the surfaces subjected to hydrophobic treatment on solution contact surfaces.
5. The POCT-ready microfluidic chip of claim 1, wherein:
the core tube is made of dark opaque material, and the tube sleeve is made of optical transparent material or partially comprises the optical transparent material.
6. The POCT ready-to-use microfluidic chip according to any one of claims 1 to 5, in which: the POCT ready-to-use microfluidic chip is applied to biological sample analysis.
7. Use of the POCT-ready microfluidic chip according to claim 6, wherein:
the biological sample analysis comprises extraction of biological molecules, and qualitative analysis and/or quantitative analysis.
8. The use method of the POCT ready-to-use microfluidic chip as claimed in any one of claims 1 to 5, comprising the following steps:
(1) when the slit openings of the core tube are rotated to coincide with the sampling holes on the top surface of the sleeve, aqueous phase solution containing samples or reagents can be respectively added into each cabin section in the core tube;
(2) when the slit openings of the core tube are rotated to be staggered by 90 degrees with the sampling holes on the top surface of the sleeve, the sealing storage and transportation of the aqueous phase solution of the sample or the reagent in each cabin section in the core tube can be realized;
(3) when the slit openings of the core tube are rotated to be dislocated by 180 degrees with the sampling hole on the top surface of the sleeve, the magnetic field is applied or removed through the magnetic control hole, the fusion, exchange or separation of the aqueous phase solution of the sample or the reagent in each cabin section and the substances in the collection pool is realized by combining the relative motion of the core tube and the sleeve and utilizing the accurate magnetic field application position, and then the multi-step biological analysis is completed.
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| CN201710861093.0A CN107748160B (en) | 2017-09-21 | 2017-09-21 | POCT ready-to-use micro-fluidic chip and application method |
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| CN107748160B true CN107748160B (en) | 2020-06-02 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111111798A (en) * | 2019-06-04 | 2020-05-08 | 厦门大学 | Micro-fluidic detection chip |
| CN117607223B (en) * | 2024-01-22 | 2024-04-09 | 南昌航空大学 | Self-driven micro-fluidic system based on monolithic column enrichment and separation |
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| CA2606916A1 (en) * | 2005-05-06 | 2006-11-16 | Platypus Technologies, Llc | Liquid crystal based analyte detection |
| US20140193807A1 (en) * | 2006-04-18 | 2014-07-10 | Advanced Liquid Logic, Inc. | Bead manipulation techniques |
| US8637317B2 (en) * | 2006-04-18 | 2014-01-28 | Advanced Liquid Logic, Inc. | Method of washing beads |
| KR101135084B1 (en) * | 2008-12-23 | 2012-04-16 | 한국전자통신연구원 | Microfluidic device and microfluidic analysis equipment |
| US20130288254A1 (en) * | 2009-08-13 | 2013-10-31 | Advanced Liquid Logic, Inc. | Droplet Actuator and Droplet-Based Techniques |
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