CN210720416U - Magnetic particle luminous micro-fluidic chip - Google Patents
Magnetic particle luminous micro-fluidic chip Download PDFInfo
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- CN210720416U CN210720416U CN201921000188.4U CN201921000188U CN210720416U CN 210720416 U CN210720416 U CN 210720416U CN 201921000188 U CN201921000188 U CN 201921000188U CN 210720416 U CN210720416 U CN 210720416U
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Abstract
The utility model is suitable for a micro-fluidic chip luminescence immunoassay technical field provides a magnetic particle luminescence micro-fluidic chip, the chip includes: the device comprises a substrate, a sample adding part, a storage part, a liquid homogeneous phase magnetic label ligand, a liquid label ligand, an air pump, a reaction area, a cleaning solution storage part, a cleaning solution and a connecting part. The embodiment of the utility model provides a through using liquid homogeneous phase magnetic label ligand and liquid mark ligand to set up the storage portion of storing liquid homogeneous phase magnetic label ligand and liquid mark ligand, make liquid homogeneous phase magnetic label ligand and liquid mark ligand save in the storage portion when the chip does not use, because the mobility of liquid is good, during the detection, make liquid homogeneous phase magnetic label ligand, liquid mark ligand and the intensive mixing that carries out of joining sample three, can effectively improve the speed of reaction, thereby improve the sensitivity that detects, repeatability and precision.
Description
Technical Field
The utility model belongs to the technical field of the luminous immunodetection of micro-fluidic chip, especially, relate to a luminous micro-fluidic chip of magnetic particle and reaction method.
Background
Currently, there are two major trends In Vitro Diagnostics (IVD): one is automatic and integrated, namely, the high-precision disease analysis and diagnosis is realized by utilizing full-automatic and high-sensitivity large-scale instruments and equipment of a central laboratory matched with a large-scale hospital, and the adopted reagent is a large-package reagent and can be used for analyzing samples for many times; the other miniaturized and bedside analyzer adopts single-person packaged reagent to realize on-site rapid analysis and diagnosis.
The small hospital or community hospital has insufficient funds and small sample amount, is not suitable for purchasing expensive large-scale equipment, has few samples to be analyzed, and has limited service time after the large packaged reagent is unpacked, so that the reagent is overdue and wasted. The miniaturized analyzer uses a single-person packaged reagent, and can solve the problems of high cost and reagent waste of large-scale equipment in small hospitals or community hospitals.
The microfluidic chip is also called a Lab-on-a-chip (Lab-on-a-chip), and is characterized in that basic operation units related to the fields of biology, chemistry, medicine and the like, such as sample preparation, reaction, separation, detection and the like, are integrated on a chip with a micro-channel with a micron scale, and the whole process of reaction and analysis is automatically completed. The analysis and detection device based on the microfluidic chip has the advantages that: the sample dosage is less, the analysis speed is fast, the portable instrument is convenient to manufacture, and the method is very suitable for real-time and on-site analysis.
However, the single-portion packaged microfluidic chip has the problems of low sensitivity, poor repeatability and obvious interference during detection due to the insufficient reaction.
SUMMERY OF THE UTILITY MODEL
The embodiment of the utility model provides a luminous micro-fluidic chip of magnetic particle aims at solving current micro-fluidic chip reaction inadequately, leads to detection sensitivity low, the repeatability is poor, receives the obvious problem of interference.
The embodiment of the utility model provides a realize like this, provide a luminous micro-fluidic chip of magnetic particle, the chip includes:
a substrate;
a sample adding part arranged on the substrate and used for adding a sample;
the liquid homogeneous phase magnetic label ligand and the liquid label ligand are stored in the storage part;
an air pump disposed on the substrate;
the reaction area is arranged on the substrate and communicated with the sample adding part, and the reaction area can be used for mixing and reacting the sample with the liquid homogeneous phase magnetic label ligand and the liquid label ligand;
a cleaning liquid storage part arranged on the substrate and a cleaning liquid stored in the cleaning liquid storage part;
and connecting parts are arranged between the reaction area and the sample adding part, between the reaction area and the storage part and between the reaction area and the cleaning solution storage part.
Still further, the storage section includes:
a first storage chamber for storing the liquid homogeneous phase magnetic label ligand;
a second storage chamber for storing the liquid marker ligand.
Still further, the storage part includes a mixing storage chamber storing the liquid homogeneous magnetic label ligand and the liquid label ligand.
Still further, the liquid tagged ligand comprises an enzyme or a luminescent agent tagged ligand.
Still further, the enzyme comprises: one or more of horseradish peroxide and alkaline phosphatase; the luminescent agent includes: one or more of acridinium ester, ABEI, fluorescent dye, fluorescent protein and fluorescent microsphere.
Furthermore, the reaction area is provided with a cleaning area, and the chip further comprises:
a detection zone in communication with the wash zone;
a light emitting liquid storage unit and a light emitting liquid stored in the light emitting liquid storage unit;
the cleaning area with be provided with between the detection zone, luminous liquid storage portion with all be provided with connecting portion between the detection zone.
Furthermore, at least one of the sample adding part, the storage part, the reaction region, the cleaning region, the detection region, the cleaning solution storage part, the luminescent solution storage part and each connecting part adopts a micro-channel structure, and at least one dimension of the micro-channel is a micrometer scale.
Still further, the substrate includes:
a middle plate;
the top adhesive tape is arranged at the top of the middle plate;
a bottom adhesive tape arranged at the bottom of the middle plate;
the sample adding part, the storage part, the reaction area and each connecting part are arranged on the middle plate.
Still further, the base plate includes a top plate and a bottom plate.
Furthermore, the reaction area comprises a first reaction partition and a second reaction partition which are communicated with each other, the first reaction partition is arranged on the top plate and is communicated with the sample adding part, and the second reaction partition is arranged on the bottom plate.
The utility model discloses the beneficial effect who reaches: the embodiment of the utility model provides a through using liquid homogeneous phase magnetic label ligand and liquid mark ligand to set up the storage portion of storing liquid homogeneous phase magnetic label ligand and liquid mark ligand, make liquid homogeneous phase magnetic label ligand, liquid mark ligand is stored in the storage portion when the chip does not use, because the mobility of liquid is good, during the detection, make liquid homogeneous phase magnetic label ligand, liquid mark ligand and the intensive mixing that carries out of joining sample three, can effectively improve the speed of reaction, thereby improve the sensitivity that detects, repeatability and precision.
Drawings
Fig. 1 is an exploded view of an embodiment of a magnetic particle luminescent microfluidic chip according to the present invention;
fig. 2 is an exploded view of another embodiment of the magnetic particle luminescent microfluidic chip according to the present invention;
fig. 3 is a flowchart of a reaction method of the magnetic particle luminescent microfluidic chip according to an embodiment of the present invention;
fig. 4 is an exploded view of another embodiment of the magnetic particle luminescent microfluidic chip according to the embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The embodiment of the utility model provides a through using liquid homogeneous phase magnetic label ligand and liquid mark ligand to set up the storage portion of storing liquid homogeneous phase magnetic label ligand and liquid mark ligand, make liquid homogeneous phase magnetic label ligand and liquid mark ligand save in the storage portion when the chip does not use, because the mobility of liquid is good, during the detection, make liquid homogeneous phase magnetic label ligand and add the sample and make the three carry out the intensive mixing, can effectively improve the speed of reaction, thereby improve the sensitivity, repeatability and the precision of detection.
Example one
As shown in fig. 1, the embodiment of the present invention is achieved by providing a magnetic particle luminescent microfluidic chip, which includes:
a substrate;
a sample addition part 21 provided on the substrate and to which a sample is added;
a storage part 23 provided on a substrate, and a liquid homogeneous magnetic label ligand and a liquid label ligand stored in the storage part 23;
an air pump 22 disposed on the substrate;
a reaction area 24 disposed on the substrate and connected to the sample adding part 21, wherein the reaction area 24 can be used for mixing and reacting the sample with the liquid homogeneous magnetic label ligand and the liquid label ligand;
the liquid homogeneous phase magnetic standard ligand comprises magnetic beads, a temperature sensitive material and a solution;
a cleaning liquid storage part 25 provided on the substrate and a cleaning liquid stored in the cleaning liquid storage part 25;
connecting parts 27 are provided between the reaction region 24 and the sample addition part 21, between the reaction region 24 and the storage part 23, and between the reaction region 24 and the cleaning solution storage part 25. Reaction zone 24 is also replaced by a partial connection 27.
The magnetic beads comprise one or more of streptavidin magnetic beads, antibody modified magnetic beads and antigen modified magnetic beads; the temperature sensitive material is a thermoreversible gel comprising one or more of gelatin, agar, alginate, carrageenan, hydroxymethyl cellulose, acacia, guar gum, locust bean gum, pectin, starch, and xanthan gum; the solution is a buffer system containing surfactant and protein.
The embodiment of the utility model provides a through using liquid homogeneous phase magnetic label ligand and liquid mark ligand to set up the storage portion of storing liquid homogeneous phase magnetic label ligand and liquid mark ligand, make liquid homogeneous phase magnetic label ligand and liquid mark ligand save in the storage portion when the chip does not use, because the mobility of liquid is good, during the detection, make liquid homogeneous phase magnetic label ligand, liquid mark ligand and the intensive mixing that carries out of joining sample three, can effectively improve the speed of reaction, thereby improve the sensitivity that detects, repeatability and precision.
Specifically, the cleaning solution is used for cleaning magnetic beads, and removing non-specifically adsorbed analytes, luminescent agent markers and other substances influencing the detection result. The cleaning solution mainly comprises a buffer system, protein and a surfactant, wherein the buffer system comprises but is not limited to borate, phosphate, Tris-HCl, acetate and the like, and the pH range of the cleaning solution is 6.0-10.0. The protein includes but is not limited to bovine serum albumin, casein, etc. Wherein the surfactant includes but is not limited to Tween 20, Tween 80, Triton X-100, polyethylene glycol, polyvinylpyrrolidone, etc. Preferably, in this embodiment, the washing solution is Tris-HCl buffer (pH7.0) containing bovine serum albumin, Tween 20 and Proclin 300.
Specifically, the storage portion 23 is a sealed cavity, and the sealing material is an elastic material or a high-barrier film, specifically, glass, plastic, rubber, aluminum foil or a high-barrier film, wherein the sealing material may be composed of the same material or a combination of multiple materials. Under physical compression, the reservoir 23 may be partially ruptured, thereby releasing the stored material.
Specifically, the storage part 23 and the cleaning liquid storage part 25 may be made of the same or different materials and methods. Preferably, the storage part 23 and the cleaning liquid storage part 25 are sealed with plastic or elastic rubber. It is also preferable that the storage part 23 is sealed with plastic or elastic rubber, and the cleaning liquid storage part 25 is sealed with a high barrier film.
The air pump 22 is a balloon built in the substrate, and the balloon communicates with the connection portion 27, and the air in the balloon is repeatedly pushed or released to repeatedly move the air in the balloon into and out of the connection portion 27. The air pump 22 is used for absorbing or extruding the air of the connecting portion 27, so that the liquid homogeneous magnetic label ligand, the liquid labeled ligand and the sample flow to the reaction area 24, and the liquid homogeneous magnetic label ligand and the liquid labeled ligand are fully mixed with the sample in the reaction area 24 by repeatedly absorbing or extruding the air of the connecting portion 27. However, the operation of the air pump 22 needs to be performed in a sealed environment, and in order to seal the inside of the chip, a sealing cap 133 is further provided at the sample addition port 134 of the sample addition part 21, and after the sample is added to the sample addition part 21, the sealing cap 133 is covered.
Example two
As shown in fig. 2, in addition to the first embodiment, the present invention also provides an embodiment, in which the storage unit 23 includes:
a first storage chamber 231 for storing the liquid homogeneous magnetic label body;
a second storage chamber 232 for storing the liquid marker ligand.
In this embodiment, the first storage chamber and the second storage chamber may be made of the same or different materials and methods. Preferably, the first storage cavity and the second storage cavity are formed by sealing plastic and elastic rubber. In another preferred embodiment, the first storage chamber is sealed with plastic and elastic rubber, and the second storage chamber is sealed with a high-barrier film. The liquid homogeneous phase magnetic label ligand and the liquid label ligand are stored separately, so that the independent performance of each material can be effectively guaranteed without mutual influence.
EXAMPLE III
The storage part 23 includes a mixing storage chamber for storing the liquid homogeneous magnetic label ligand and the liquid label ligand.
In this embodiment, the mixed storage chamber adopts plastics and elastic rubber to seal and forms, mixes both and stores, and the follow-up reaction of being convenient for goes on fast, and is simple convenient.
Example four
In an alternative embodiment of the present invention, the liquid tagged ligand comprises a luminophore-tagged ligand.
The luminescent agent may further include: one or more of acridinium ester, ABEI, fluorescent dye, fluorescent protein and fluorescent microsphere.
The cleaning liquid storage part 25 and the cleaning liquid stored in the cleaning liquid storage part 25;
a detection zone 28 in communication with said wash zone 29;
a light emitting liquid storage part 26 and a light emitting liquid stored in the light emitting liquid storage part 26 for further cleaning magnetic beads or enhancing light emitting signals;
connecting portions 27 are provided between the cleaning region 29 and the detection region 28, and between the luminescent liquid storage portion 26 and the detection region 28.
Specifically, the ligands include: one or more of an antigen, an antibody, a hapten and a nucleic acid.
The luminescent agent binds or competes with the analyte to form a luminescent agent-labeled ligand; the magnetic particle label binds or competes with the analyte to form a magnetic bead labeled ligand, which may be the same or different; the magnetic labeling ligand and the luminescent agent labeling ligand are used by ligands comprising nucleic acid, antigen, monoclonal antibody, polyclonal antibody and hormone receptor, and the analyte comprises DNA, small molecules (drugs or drugs), antigen, antibody, hormone, antibiotic, bacteria or virus and other biochemical markers.
In this embodiment, the liquid labeled ligand may be bound to a liquid homogeneous magnetic labeled ligand (e.g., double antibody sandwich) or may compete with the labeled ligand (e.g., competition). The ligand marked by the luminous agent can be the same as or different from the liquid homogeneous magnetic marker ligand. Preferably, in one embodiment of the present invention, two different antibodies are selected as the liquid labeled ligand and the liquid homogeneous magnetic labeled ligand to detect the analyte in a double antibody sandwich method.
EXAMPLE five
In an alternative embodiment of the invention, the liquid labelling ligand comprises an enzyme-labelled ligand.
The enzyme includes: one or more of horseradish peroxide and alkaline phosphatase.
Specifically, the ligands include: one or more of an antigen, an antibody, a hapten and a nucleic acid.
The enzyme binds or competes with the analyte to form an enzyme-labeled ligand; the magnetic particle label binds or competes with the analyte to form a magnetic bead labeled ligand, which may be the same or different; the ligands used by the magnetic enzyme labeling ligand and the enzyme labeling ligand comprise nucleic acid, antigen, monoclonal antibody, polyclonal antibody and hormone receptor, and the analytes comprise DNA, small molecules (drugs or drugs), antigen, antibody, hormone, antibiotics, bacteria or viruses and other biochemical markers.
In this embodiment, the liquid labeled ligand may be bound to a liquid homogeneous magnetic labeled ligand (e.g., double antibody sandwich) or may compete with the labeled ligand (e.g., competition). Wherein the enzyme-labeled ligand can be the same as or different from the liquid homogeneous magnetic labeling ligand. Preferably, in one embodiment of the present invention, two different antibodies are selected as the liquid labeled ligand and the liquid homogeneous magnetic labeled ligand to detect the analyte in a double antibody sandwich method. In another embodiment of the present invention, an antigen and an antibody are selected as the liquid labeled ligand and the liquid homogeneous magnetic labeled ligand, respectively, to detect the sample by a competitive method.
EXAMPLE six
In an optional embodiment of the present invention, the reaction region 24 is provided with a cleaning region 29, and the chip further comprises:
a detection zone 28 in communication with said wash zone 29;
a light-emitting liquid storage section 26 and a light-emitting liquid stored in the light-emitting liquid storage section 26;
connecting portions 27 are provided between the cleaning region 29 and the detection region 28, and between the luminescent liquid storage portion 26 and the detection region 28. The cleaning liquid storage portion 25 is provided in communication with the cleaning region 29 or in communication with the reaction region 24.
In this embodiment, the chemical luminescence requires a luminescent liquid, and the fluorescence also requires a luminescent source.
Set up detection zone 28, be convenient for after the washing liquid washs, move final complex to detection zone 28 and observe and detect, convenient to use, it is simple convenient.
Specifically, the substrate is further provided with a waste liquid storage portion 30 communicating with the cleaning region 29. The waste liquid after washing and reaction is convenient for collect, the interference of the waste liquid to the detection can be reduced, and the detection precision is effectively improved.
EXAMPLE seven
At least one of the sample addition part 21, the storage part 23, the reaction region 24, the washing region 29, the detection region 28, the washing liquid storage part 25, the luminescent liquid storage part 26 and each connection part 27 adopts a micro-channel structure, and at least one dimension of the micro-channel is a micrometer scale.
The utility model discloses a micro-fluidic chip all reagent components that the testing process is required (liquid mark ligand, liquid homogeneous phase magnetic label ligand, the washing liquid, luminous liquid etc.) all integrate, it is built-in to the micro-fluidic chip, and through ingenious microchannel design, under the operation of supporting instrument, realize the one-key formula operation of micro-fluidic chip (only need press the start key just can realize detecting, need not complicated operation), realize whole blood separation, immunoreaction, the washing separation, chemiluminescence detection, thereby structural design is simple among the current micro-fluidic chip, operate not enough and defect such as complicated when detecting. And the defect that the traditional chemiluminescence apparatus can only detect serum or plasma but not a whole blood sample is overcome.
Example eight
In an alternative embodiment, the substrate comprises:
a middle plate 11;
a top tape 13 disposed on the top of the middle plate 11;
a bottom tape 14 disposed at the bottom of the middle plate 11;
the sample addition part 21, the storage part 23, the reaction region 24, and each connection part 27 are provided on the middle plate 11. The top tape 13 is provided with a first window 131 used in cooperation with the storage part 23, a second window 132 used in cooperation with the cleaning solution storage part 25 and the luminescent solution storage part 26, and a sample addition port 134 used in cooperation with the sample addition part 21, and a sealing cover 133 is arranged at the sample addition port 134. The first window 131 and the second window 132 are provided to facilitate the extrusion of each reservoir portion by the extrusion stem extending from the inside of the first window 131 and the second window 132 into the inside of the chip, so that the liquid substance in each reservoir portion flows out; and set up sealed lid 133, be convenient for seal with application of sample portion 21 to make chip inside keep sealed, be convenient for the air pump to carry out work.
In this embodiment, the molding material of the substrate is a polymer, including but not limited to polystyrene, polyvinyl chloride, polypropylene, epoxy resin, and the like.
The embodiment of the utility model provides an adopt micro-fluidic chip technique, mix, react, separation and detect the sample and integrate on the chip to all reagent components that the reaction is required are integrated to the chip. The operation is simple, and during detection, only the sample needs to be added, the cover is covered, and the chip is placed into a small portable matching instrument.
As shown in fig. 3, the reaction method according to the embodiment of the present invention includes:
adding a sample from the sample addition part 21 and allowing the sample to enter the reaction region 24;
placing the liquid homogeneous magnetic label ligand and the liquid label ligand in the storage part 23 into the reaction area 24;
the sample is fully mixed and reacted with the liquid homogeneous phase magnetic labeling ligand and the liquid labeling ligand in the reaction area 24 through the air pump 22;
and (4) cleaning the reacted compound.
The embodiment of the utility model provides a through using liquid homogeneous phase magnetic label ligand and liquid mark ligand, and set up the storage part 23 of storing liquid homogeneous phase magnetic label ligand and liquid mark ligand, make liquid homogeneous phase magnetic label ligand and liquid mark ligand save in storage part 23 when the chip does not use, detect time measuring, liquid homogeneous phase magnetic label ligand and liquid mark ligand and add the sample three and accomplish the mixture in the reaction zone in one step, the reaction, make and mix, the reaction is rapid, can effectively improve the speed of reaction, thereby improve the efficiency that detects, sensitivity, repeatability and precision.
The reaction zone is provided with a cleaning zone 29, and the compound after the cleaning reaction specifically comprises:
placing a cleaning fluid into the reaction zone 24 to clean the reacted composite; or
The reacted complex is introduced into the cleaning region 29 by the air pump 22, and the cleaning solution is introduced into the cleaning region 29 to clean the reacted complex.
In the embodiment, the magnetic beads are cleaned by the cleaning solution to remove the non-specifically adsorbed analytes, the luminescent agent markers and other substances influencing the detection result; the detection accuracy can be further improved.
The embodiment of the utility model provides a through using liquid homogeneous phase magnetic label ligand and liquid mark ligand to set up the storage part 23 of storing liquid homogeneous phase magnetic label ligand and liquid mark ligand, make liquid homogeneous phase magnetic label ligand and liquid mark ligand save in storage part 23 when the chip does not use, examine time measuring and carry out the intensive mixing with adding the sample three, can effectively improve the speed of reaction, thereby improve sensitivity, repeatability and the precision of detection.
In addition, the liquid labeled ligand may be bound to a liquid homogeneous magnetic label ligand (e.g., double antibody sandwich) or may compete with the labeled ligand (e.g., competition). The ligand marked by the luminescent agent and the ligand marked by the enzyme can be the same as or different from the liquid homogeneous phase magnetic marking ligand. Preferably, in one embodiment of the present invention, two different antibodies are selected as the liquid labeled ligand and the liquid homogeneous magnetic labeled ligand to detect the analyte in a double antibody sandwich method. In another embodiment of the present invention, an antigen and an antibody are selected as the liquid labeled ligand and the liquid homogeneous magnetic labeled ligand, respectively, to detect the sample by a competitive method. Set up detection zone 28, be convenient for after the washing liquid washs, move final complex to detection zone 28 and observe and detect, convenient to use, it is simple convenient. The utility model discloses a micro-fluidic chip all reagent components that the testing process is required (liquid mark ligand, liquid homogeneous phase magnetic label ligand, washing liquid, luminous liquid etc.) all integrate, built-in to micro-fluidic chip to through ingenious microchannel design, under the operation of supporting instrument, realize one-key formula operation of micro-fluidic chip (only need press the start key just can realize detecting, need not complicated operation), realize whole blood separation, immunoreaction, washing separation, detection.
Example nine
As shown in fig. 4, on the basis of the first embodiment, the base plate includes a top plate 12 and a bottom plate 15;
the liquid homogeneous magnetic label ligand storage part 231 and the liquid label ligand storage part 232 are respectively arranged on the top plate 12 and the bottom plate 15, and the positions can be interchanged.
The sample addition part 21 is provided on the top plate 12.
The reaction region 24 includes a first reaction partition 241 and a second reaction partition 242 which are communicated with each other, the first reaction partition 241 is disposed on the top plate 12 and communicated with the sample addition part 21, and the second reaction partition 242 is disposed on the bottom plate 15.
In this embodiment, the molding material of the substrate is a polymer, including but not limited to polystyrene, polyvinyl chloride, polypropylene, epoxy resin, and the like.
The embodiment of the utility model provides an adopt micro-fluidic chip technique, mix, react, separation and detect the sample and integrate on the chip to all reagent components that the reaction is required are integrated to the chip. The operation is simple, and during detection, only a sample needs to be added, the sealing cover is covered, and the chip is placed into a small portable matching instrument.
And set up sealed lid, be convenient for seal with application of sample portion 21 to make chip inside keep sealed, the air pump of being convenient for carries out work.
The reaction method provided by the embodiment of the utility model comprises the following steps:
adding a sample from the sample addition part 21 and allowing the sample to enter the first reaction partition 241;
putting the liquid labeled ligand into the first reaction subarea 241, and enabling the sample and the liquid labeled ligand to be fully mixed and reacted in the first reaction subarea 241 through an air pump 22 to obtain a labeled ligand compound;
placing the labeled ligand complex into second reaction partition 242;
putting the liquid homogeneous phase magnetic standard ligand into a second reaction subarea 242, and mixing and reacting the labeled ligand compound and the liquid homogeneous phase magnetic standard ligand in the second reaction subarea 242 to obtain a final compound;
the final complex after the reaction was washed.
In the embodiment, the liquid-state labeled ligand and the liquid-state homogeneous-phase magnetic labeled ligand are respectively reacted with the sample and the labeled ligand compound through a two-step method, so that the reaction is more sufficient, the accuracy of subsequent detection is effectively improved, and the method is simple and convenient.
The utility model discloses an use liquid homogeneous phase magnetic label ligand and liquid mark ligand to set up liquid homogeneous phase magnetic label ligand storage unit 231 of storage and liquid mark ligand storage unit 232, make liquid homogeneous phase magnetic label ligand and liquid mark ligand save in storage unit separately when the chip does not use, it makes liquid homogeneous phase magnetic label ligand or liquid mark ligand and adds the sample earlier and carries out the intensive mixing to examine time measuring, can effectively improve the speed of reaction, thereby improve the sensitivity that detects, repeatability and precision.
The compound after the washing reaction specifically comprises:
placing a cleaning solution into the second reaction partition 242, and cleaning the final compound after reaction; or
The reacted final composite enters the washing section 29, and a washing liquid is put into the washing section 29 to wash the reacted final composite.
In the embodiment, the magnetic beads are cleaned by the cleaning solution to remove the non-specifically adsorbed analytes, the luminescent agent markers and other substances influencing the detection result; the detection accuracy can be further improved.
The utility model discloses another reaction method of embodiment, include:
adding a sample from the sample addition part and allowing the sample to enter the first reaction partition 241;
putting the liquid homogeneous phase magnetic standard ligand into the first reaction subarea 241, and enabling the sample and the liquid homogeneous phase magnetic standard ligand to be fully mixed and reacted in the first reaction subarea 241 through an air pump 22 to obtain a magnetic standard ligand compound;
placing the magnetic standard ligand complex into the second reaction partition 242;
putting the liquid labeled ligand into the second reaction subarea 242, and mixing and reacting the magnetic labeled ligand compound and the liquid labeled ligand in the second reaction subarea 242 to obtain a final compound;
the final complex after the reaction was washed.
In the embodiment, the liquid-state labeled ligand and the liquid-state homogeneous-phase magnetic labeled ligand are respectively reacted with the sample and the labeled ligand compound through a two-step method, so that the reaction is more sufficient, the accuracy of subsequent detection is effectively improved, and the method is simple and convenient.
The utility model discloses an use liquid homogeneous phase magnetic label ligand and liquid mark ligand to set up liquid homogeneous phase magnetic label ligand storage unit 231 of storage and liquid mark ligand storage unit 232, make liquid homogeneous phase magnetic label ligand and liquid mark ligand save in storage unit separately when the chip does not use, it makes liquid homogeneous phase magnetic label ligand or liquid mark ligand and adds the sample earlier and carries out the intensive mixing to examine time measuring, can effectively improve the speed of reaction, thereby improve the sensitivity that detects, repeatability and precision.
The embodiment of the utility model provides a reaction method still, include:
adding a sample from the sample addition part and allowing the sample to enter the first reaction partition 241;
putting the liquid homogeneous phase magnetic standard ligand into the first reaction subarea 241, and enabling the sample and the liquid homogeneous phase magnetic standard ligand to be fully mixed and reacted in the first reaction subarea 241 through an air pump 22 to obtain a magnetic standard ligand compound;
cleaning the reacted magnetic standard ligand compound;
placing the cleaned magnetic standard ligand compound into a second reaction subarea 242;
and putting the liquid labeled ligand into the second reaction subarea 242, and mixing and reacting the magnetic labeled ligand compound and the liquid labeled ligand in the second reaction subarea 242 to obtain a final compound.
In the embodiment, the liquid-state labeled ligand and the liquid-state homogeneous-phase magnetic labeled ligand are respectively reacted with the sample and the labeled ligand compound by a two-step cleaning method, so that the reaction is more sufficient, the follow-up detection accuracy is effectively improved, and the method is simple and convenient. Washing the magnetic beads by a cleaning solution to remove non-specifically adsorbed analytes, luminescent agent markers and other substances influencing the detection result; the detection accuracy can be further improved.
The utility model discloses an use liquid homogeneous phase magnetic label ligand and liquid mark ligand to set up liquid homogeneous phase magnetic label ligand storage unit 231 of storage and liquid mark ligand storage unit 232, make liquid homogeneous phase magnetic label ligand and liquid mark ligand save in storage unit separately when the chip does not use, it makes liquid homogeneous phase magnetic label ligand or liquid mark ligand and adds the sample earlier and carries out the intensive mixing to examine time measuring, can effectively improve the speed of reaction, thereby improve the sensitivity that detects, repeatability and precision.
In addition, the liquid labeled ligand may be bound to a liquid homogeneous magnetic label ligand (e.g., double antibody sandwich) or may compete with the labeled ligand (e.g., competition). The ligand marked by the luminescent agent and the ligand marked by the enzyme can be the same as or different from the liquid homogeneous phase magnetic marking ligand. Preferably, in one embodiment of the present invention, two different antibodies are selected as the liquid labeled ligand and the liquid homogeneous magnetic labeled ligand to detect the analyte in a double antibody sandwich method. In another embodiment of the present invention, an antigen and an antibody are selected as the liquid labeled ligand and the liquid homogeneous magnetic labeled ligand, respectively, to detect the sample by a competitive method. Set up detection zone 28, be convenient for after the washing liquid washs, move final complex to detection zone 28 and observe and detect, convenient to use, it is simple convenient. The utility model discloses a micro-fluidic chip all reagent components that the testing process is required (liquid mark ligand, liquid homogeneous phase magnetic label ligand, the washing liquid, luminous liquid etc.) all integrate, it is built-in to the micro-fluidic chip, and through ingenious microchannel design, under the operation of supporting instrument, realize the one-key formula operation of micro-fluidic chip (only need press the start key just can realize detecting, need not complicated operation), realize whole blood separation, immunoreaction, the washing separation, chemiluminescence detection, thereby structural design is simple among the current micro-fluidic chip has been avoided, operate not enough and defect such as complicated when detecting. And the defect that the traditional chemiluminescence apparatus can only detect serum or plasma but not a whole blood sample is overcome.
The above description is only exemplary of the present invention and should not be taken as limiting the scope of the present invention, as any modifications, equivalents, improvements and the like made within the spirit and principles of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. A magnetic particle luminescent microfluidic chip, the chip comprising:
a substrate;
a sample adding part arranged on the substrate and used for adding a sample;
the liquid homogeneous phase magnetic label ligand and the liquid label ligand are stored in the storage part;
an air pump disposed on the substrate;
the reaction area is arranged on the substrate and communicated with the sample adding part, and the reaction area can be used for mixing and reacting the sample with the liquid homogeneous phase magnetic label ligand and the liquid label ligand;
a cleaning liquid storage part arranged on the substrate and a cleaning liquid stored in the cleaning liquid storage part;
and connecting parts are arranged between the reaction area and the sample adding part, between the reaction area and the storage part and between the reaction area and the cleaning solution storage part.
2. The magnetic particle luminescent microfluidic chip of claim 1, wherein the storage portion comprises:
a first storage chamber for storing the liquid homogeneous phase magnetic label ligand;
a second storage chamber for storing the liquid marker ligand.
3. The magnetic particle luminescent microfluidic chip of claim 1, wherein the storage portion comprises a mixing storage chamber for storing the liquid homogeneous magnetic label ligand and the liquid label ligand.
4. The magnetic particle luminescent microfluidic chip of any one of claims 1-3, wherein the liquid labeled ligand comprises an enzyme or a luminescent agent labeled ligand.
5. The magnetic particle luminescent microfluidic chip according to claim 4, wherein said enzyme comprises: one or more of horseradish peroxide and alkaline phosphatase; the luminescent agent includes: one or more of acridinium ester, ABEI, fluorescent dye, fluorescent protein and fluorescent microsphere.
6. The magnetic particle luminescent microfluidic chip of claim 5, wherein said reaction region is provided with a cleaning region, said chip further comprising:
a detection zone in communication with the wash zone;
a light emitting liquid storage unit and a light emitting liquid stored in the light emitting liquid storage unit;
the cleaning area with be provided with between the detection zone, luminous liquid storage portion with all be provided with connecting portion between the detection zone.
7. The magnetic particle luminescent microfluidic chip according to claim 6, wherein at least one of the sample addition part, the storage part, the reaction region, the washing region, the detection region, the washing liquid storage part, the luminescent liquid storage part, and each connection part has a microchannel structure, and at least one dimension of the microchannel is a micrometer scale.
8. The magnetic particle luminescent microfluidic chip of claim 1, wherein said substrate comprises:
a middle plate;
the top adhesive tape is arranged at the top of the middle plate;
a bottom adhesive tape arranged at the bottom of the middle plate;
the sample adding part, the storage part, the reaction area and each connecting part are arranged on the middle plate.
9. The magnetic particle luminescent microfluidic chip of claim 1, wherein said substrate comprises a top plate and a bottom plate.
10. The magnetic particle luminescent microfluidic chip of claim 9, wherein said reaction region comprises a first reaction partition and a second reaction partition in communication with each other, and said first reaction partition is disposed on said top plate in communication with said sample addition portion, and said second reaction partition is disposed on said bottom plate.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201921000188.4U CN210720416U (en) | 2019-06-27 | 2019-06-27 | Magnetic particle luminous micro-fluidic chip |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201921000188.4U CN210720416U (en) | 2019-06-27 | 2019-06-27 | Magnetic particle luminous micro-fluidic chip |
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