CN112304911A - Biosensor system and biological sample detection method - Google Patents

Biosensor system and biological sample detection method Download PDF

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Publication number
CN112304911A
CN112304911A CN202011089473.5A CN202011089473A CN112304911A CN 112304911 A CN112304911 A CN 112304911A CN 202011089473 A CN202011089473 A CN 202011089473A CN 112304911 A CN112304911 A CN 112304911A
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biosensor
electromagnetic particles
detection
optical
electromagnetic
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黄国莲
李相才
程巧霞
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Shaoxing Dalengken Biotechnology Co ltd
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Shaoxing Dalengken Biotechnology Co ltd
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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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/533—Production of labelled immunochemicals with fluorescent label
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
    • 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
    • G01N33/564—Immunoassay; Biospecific binding assay; Materials therefor for pre-existing immune complex or autoimmune disease, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
    • 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
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks

Abstract

The invention relates to a biosensor system comprising: the biosensor comprises a substrate and a magnet assembly, wherein the substrate is provided with a connecting part and a micro-flow channel arranged in the substrate, the magnet assembly comprises a magnetic driving layer coated on the inner wall of the micro-flow channel and a plurality of electromagnetic particles dissociated in the micro-flow channel, the magnetic field intensity of the electromagnetic particles can be respectively changed through electric control, the magnetic driving layer has an acting force for driving away the electromagnetic particles, and the surfaces of the electromagnetic particles are coupled with molecular groups which can react with substances to be detected in a sample and generate optical signals; and the biosensor box comprises a box body, and a magnetic field forming mechanism and an optical detection mechanism which are fixed on the box body, wherein the box body is provided with a connecting part connected with the biosensor, the magnetic field forming mechanism forms a variable magnetic field, and the optical detection mechanism is used for detecting optical signals in the biosensor. The biosensor system can provide more reliable test results.

Description

Biosensor system and biological sample detection method
Technical Field
The invention relates to the technical field of biology, in particular to a biosensor system and a biological sample detection method.
Background
Pathogenic microorganisms are invisible hazards and are always the focus of attention of people, and the pollution is a great safety problem and seriously affects the health of human beings. Estimated by the world health organization, billions of people worldwide are infected with diseases each year by pathogenic microorganisms. The main microorganisms causing the disease include Vibrio parahaemolyticus, Staphylococcus, Salmonella and Bacillus cereus. Chemical pollutants such as lean meat essence, melamine and the like are terrible, and food-borne diseases caused by pathogenic microorganisms are the first problem of food safety. The detection technology of pathogenic microorganisms is a key technical link for preventing and controlling food-borne diseases caused by pathogenic microorganisms. Conventional detection techniques mostly rely on a method for culturing a target microorganism to determine whether a sample is contaminated by the microorganism, and include steps of enrichment, isolated culture, morphological observation, biochemical identification and the like. The method has high accuracy and sensitivity, but has more involved experiments, complicated operation, long required time and low automation degree, and generally, one test program only aims at one bacterium and is not suitable for batch and large-range rapid detection. In order to overcome the defects, people develop various novel technologies for rapidly detecting pathogenic microorganisms based on different principles, which mainly comprise a polymerase chain reaction method, an enzyme-linked immunosorbent assay, a colloidal gold immunochromatography method and the like. The polymerase chain reaction is a technique for in vitro selective amplification of DNA or RNA, namely, an enzymatic aggregation reaction of a template DNA primer and 4 kinds of deoxyribonuceloxib triphosphates under the catalysis of DNA polymerase, and consists of 3 steps of denaturation, annealing and extension. The technological equipment for gene amplification consists of three parts, including template DNA preparing apparatus, high speed micro centrifuge or high speed freezing centrifuge; PCR gene amplification instrument; and thirdly, DNA amplification result interpretation and determination equipment mainly comprises a horizontal low-pressure electrophoresis apparatus, a PCR nucleic acid electrophoresis tank, an ultraviolet transmission apparatus, a DNA micro-fluorometer and the like, is large in equipment and high in price, and is not suitable for field operation. Most of PCR technologies applied in pathogenic microorganism detection are based on DNA level detection technologies, although the PCR technologies are rapid and specific and have high sensitivity, the biggest problems are that false positive and quantification difficulties are caused by nucleic acid pollution and dead bacteria, the detection result is represented as existence or nonexistence, only qualitative judgment can be carried out, and quantitative analysis cannot be carried out. The enzyme-linked immunosorbent assay uses an enzyme-linked immunosorbent assay as a matched instrument, has strong sensitivity and specificity, but needs repeated plate washing, and has complex operation, time and labor waste. The colloidal gold immunochromatography is simple, convenient and quick, the result can be judged only by adding the processed sample into the sample hole of the test paper strip in the gold-labeled quantitative analyzer, but the colloidal gold immunochromatography has the defect of low sensitivity, is generally qualitative measurement, can not read data of a corresponding detection plate, and the excellent gold-labeled diagnostic reagent can only achieve semi-quantification. Various analytical processes are known for detecting analytes in test samples. For example, immunoassays use the mechanisms of the immune system, wherein antibodies and the respective antigens are able to bind to each other. This particular reaction mechanism is used to determine the presence or concentration of the antigen in the test sample. In particular, the antibody or antigen (analyte of interest) is labeled to quantify the interaction between the antibody and the antigen. Common labels are for example fluorescent and chemiluminescent molecules, colored particles (beads) or radioisotopes.
More recently, magnetic labels have been used in immunoassays to detect the presence or quantity of an analyte. The use of magnetic labels, such as magnetic particles (beads), has several advantages. The magnetic particles can be driven by applying a magnetic field, which can facilitate the analysis process. Furthermore, there is no magnetic background signal in the biological test sample that affects the detection of the magnetic particles. However, these immunoassay tests using magnetic labels require means for driving antigen-bound magnetic particles immobilized near the sensor surface of the sensor cartridge, and means for washing away remaining unbound magnetic particles so as not to affect the measurement of the number of bound particles, are complicated in structure, and cannot perform closed detection of magnetic particles, and are limited in application.
Disclosure of Invention
In view of the above, there is a need to provide a biosensor system, which solves the problem of limited application of the existing magnetic particles due to the inability of closed detection.
The present invention provides a biosensor system comprising:
the biosensor comprises a substrate and a magnet assembly arranged in the substrate, wherein the substrate is provided with a connecting part and a micro-flow channel arranged in the substrate, the magnet assembly comprises magnetic driving layers coated at two tail ends of the micro-flow channel and a plurality of electromagnetic particles dissociated in the micro-flow channel, the magnetic field intensity of the electromagnetic particles can be respectively changed through electric control, the magnetic driving layers have acting force for driving away the electromagnetic particles, and the surfaces of the electromagnetic particles are coupled with molecular groups which can react with substances to be detected in a sample and generate optical signals; and
the biosensor box comprises a box body, and a magnetic field forming mechanism and an optical detection mechanism which are fixed on the box body, wherein the magnetic field forming mechanism forms a variable magnetic field to act on the magnetic driving layer and the electromagnetic particles, and the optical detection mechanism is used for detecting optical signals in the biosensor.
Furthermore, the biosensor cartridge further comprises a liquid driving mechanism, the liquid driving mechanism is provided with a connected part, and after the connected part is connected to the connected part, the liquid driving mechanism drives the liquid in the microfluidic channel to flow.
Specifically, the substrate is made of a flexible transparent material and is provided with a sample loading interface communicated with the micro-flow channel, and the sample loading interface is used for injecting sample liquid.
Specifically, the microflow channel includes a plurality of detection sections and is located every adjacent two free section between the detection section, free a plurality of electromagnetic particles in the detection section, free section is close to the tip inner wall coating of detection section has drive the magnetic layer.
Preferably, the cross section of the detection section is flat, the cross section of the free section is circular, and the maximum width of the cross section of the detection section is not greater than that of the free section.
Specifically, the magnetic driving layer is a coating made of an excitation material.
Furthermore, the biosensor system also comprises a processing controller which is electrically connected with the liquid driving mechanism, the light ray detection mechanism and the magnetic field forming mechanism; the liquid driving mechanism comprises a liquid storage tank, a recovery tank, a driving pump and a multi-way control valve, the liquid storage tank, the recovery tank, the multi-way control valve, the driving pump, a micro-flow channel of the biological sensor and the recovery tank can form a fluid channel, and the fluid channel is used for sample liquid, buffer liquid, reaction liquid or cleaning liquid to flow through or for recovery of electromagnetic particles; the optical detection mechanism comprises a light source, an emergent light focusing lens, an optical fiber and an optical detector, wherein the light source and the emergent light focusing lens are respectively arranged corresponding to the biosensor, the emergent light focusing lens is connected with the optical detector, the optical detector is electrically connected with the processing controller, the light source is used for emitting incident light to the detection section, the light focusing lens is used for collecting, focusing and coupling optical signals generated by the detection section, and the optical detector is used for processing the optical signals and outputting the optical signals to the processing controller.
The invention also provides a method for detecting a biological sample by using the biosensor system, which comprises the following steps:
s1, preparing the electromagnetic particles with surface coupling molecular groups and the base material;
s2, pouring the electromagnetic particles into the microfluidic channel, and enabling the magnetic driving layer to generate magnetic driving force through the magnetic field forming mechanism so as to limit the electromagnetic particles in the microfluidic channel;
s3, enabling the solution to be detected to flow through the microfluidic channel, reacting with the molecular groups on the surfaces of the electromagnetic particles, and generating optical signals;
s4, the optical detection mechanism acquires the optical signal, focuses, couples and processes the optical signal and outputs the optical signal to the processing controller;
and S5, the processing controller processes and calculates, and outputs the biological data of the sample.
Optionally, the step S2 may also be:
s21, pouring the electromagnetic particles into a microfluidic channel, and generating driving force for the electromagnetic particles through the magnetic field forming mechanism to enable the electromagnetic particles to be uniformly distributed in the microfluidic channel;
and S22, enabling the lock driving magnetic layer to generate driving force through the magnetic field forming mechanism, and fixing the electromagnetic particles in each detection section.
Optionally, the step S4 may also be:
s41, recovering the reacted electromagnetic particles;
and S42, acquiring the surface optical signal change of the electromagnetic particles before and after reaction through the optical detection mechanism, focusing, coupling and processing the surface optical signal change, and outputting the surface optical signal change to the processing controller.
Has the advantages that:
1. according to the invention, the electromagnetic particles are used for detecting the biological sample, so that the extremely high specific surface area can be provided, and the fixed quantity of the antibody is increased, thus finally increasing the sensitivity and linear range of the detection of the mesoporous biochip biosensor, increasing the flux of the detection substance in the chip, and shortening the detection time and operation steps.
2. The biosensor system provided by the invention detects a biological sample through coupling molecular groups by electromagnetic particles, and fixes the biological sample in a microfluidic channel through an electromagnetic field. The magnetic field intensity of each electromagnetic particle can be changed in an electric control mode, the electromagnetic particles can be driven to be uniformly distributed in the micro-flow channel, and a more reliable test result can be provided through the simple biosensor system.
3. The invention can recycle the electromagnetic particles for multiple use by detecting and analyzing the electromagnetic particles, thereby saving the cost.
Drawings
Fig. 1 is a schematic structural diagram of the overall structure of a biosensor system according to an embodiment of the present invention.
Fig. 2 is a schematic perspective view of a biosensor according to an embodiment of the present invention.
FIG. 3 is a schematic cross-sectional view of an alternative biosensor provided in an embodiment of the present invention.
FIG. 4 is a schematic cross-sectional view of an alternative biosensor provided in an embodiment of the present invention.
FIG. 5 is a schematic cross-sectional view of an alternative biosensor provided in an embodiment of the present invention.
FIG. 6 is a schematic cross-sectional view of an alternative biosensor provided in an embodiment of the present invention.
FIG. 7 is a schematic cross-sectional view of an alternative biosensor provided in an embodiment of the present invention.
FIG. 8 is a schematic cross-sectional view of an alternative biosensor provided in accordance with an embodiment of the present invention.
Fig. 9 is an enlarged view of fig. 7 at a.
Fig. 10 is an enlarged view of fig. 8 at B.
Fig. 11 is an overall architecture diagram of a biosensor system according to an embodiment of the present invention.
FIG. 12 is a logic control diagram of a biosensor system according to an embodiment of the present invention.
Fig. 13 is a flowchart of a method for detecting a biological sample by using the biosensor system according to an embodiment of the present invention.
Fig. 14 is an alternative detailed flowchart of step S2 in fig. 13.
Fig. 15 is an alternative detailed flowchart of step S4 in fig. 13.
1 biosensor, 10 substrates, 100 connecting parts, 101 microfluidic channels, 1010 detection sections, 1011 free sections,
102 a sample loading interface,
11 magnet assembly, 110 magnetic drive layer, 111 electromagnetic particles,
2 biosensor cartridge, 20 cartridge body, 201 end, 2000 fluid channel, 21 magnetic field forming mechanism, 22 optical detection mechanism, 220 light source, 2200 pressing part,
221 outgoing light focusing lens, 2210 pressed part, 222 optical fiber, 223 optical detector,
23 liquid taking mechanism, 230 connected part, 231 liquid storage tank, 232 recovery tank, 233 drive pump, 234 multi-way control valve,
And 3, processing the controller.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, 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 present invention provides a biosensor system, as shown in fig. 1 and 11, comprising:
the biosensor 1 comprises a substrate 10 and a magnet assembly 11 arranged in the substrate 10, wherein the substrate 10 is provided with a connecting part 100 and a micro-flow channel 101 arranged in the connecting part, the magnet assembly 11 comprises a magnetic driving layer 110 coated at two tail ends of the micro-flow channel 101 and a plurality of electromagnetic particles 111 dissociated in the micro-flow channel 101, the magnetic field intensity of each electromagnetic particle 111 can be respectively changed by electric control, the magnetic driving layer 110 has an acting force for driving away the electromagnetic particles 111, and the surfaces of the electromagnetic particles 111 are coupled with molecular groups capable of reacting with substances to be detected in a sample and generating optical signals; and
the biosensor cartridge 2 comprises a cartridge body 20, and a magnetic field forming mechanism 21 and an optical detection mechanism 22 fixed to the cartridge body, wherein the magnetic field forming mechanism 21 forms a variable magnetic field to act on the magnetic driving layer 110 and the electromagnetic particles 111, and the optical detection mechanism 21 is used for detecting an optical signal of the biosensor 1.
The biosensor system provided by the invention detects a biological sample by coupling molecular groups through electromagnetic particles, and fixes the biological sample in a microfluidic channel through an electromagnetic field. The magnetic field intensity of each electromagnetic particle can be changed in an electric control mode, the electromagnetic particles can be driven to be uniformly distributed in the microfluidic channel, and a more reliable test result can be provided through the simple biosensor system.
Specifically, the biosensor cartridge 2 further includes a liquid driving mechanism 23, the liquid driving mechanism 23 has a connected portion 230, and after the connected portion 230 is connected to the connecting portion 100, the liquid driving mechanism 23 drives the liquid in the microfluidic channel 101 to flow.
Specifically, the substrate 10 provided in the present invention is made of a transparent material, which may be a rigid or flexible transparent material, so as to facilitate the observation and detection of the optical detection mechanism 22.
In some embodiments, the substrate 10 is made of an optically transparent flexible material, so as to facilitate sampling, transportation and storage in practical applications; particularly for some virus samples that are genotoxic, or that pose a biological safety hazard, they need to be sealed, sterilized, and disposed of. For practical use, these applications make the substrate 10 flexible, which can be more convenient for practical use and handling.
The substrate 10 is selected with consideration of its optical properties, such as light transmission and background fluorescence. The mesoporous material for making the electromagnetic particles is preferably one of silicon-based mesoporous materials, doped silicon-based mesoporous materials, titanium dioxide-based mesoporous materials, carbon-aluminum-phosphate-based mesoporous materials, aluminum-based mesoporous materials or other mesoporous materials, and other materials such as glass, quartz, polymers and the like can also be used as base materials as long as the materials are suitable for optical detection. The aperture of the micropores in the mesoporous material is generally not more than 500 μm, and is usually 20-300 μm. Preferably, one or more of polydimethylsiloxane, polymethyl methacrylate, epoxy resin, silicone resin, polycarbonate, polystyrene, polyethylene, fluororesin, unsaturated polycarbonate resin, melamine-formaldehyde resin, polyphenylene oxide, polyaluminum, polyoxymethylene, nylon, polyamide-polypropylene, polyvinyl chloride, ABs resin and other polymers and silicon wafer, glass, quartz, ceramics and other materials are compounded.
Further, in these embodiments, since the substrate 10 is made of a flexible material, when the substrate 10 needs to be optically detected, wrinkles are inevitably generated, and thus the optical detection is affected, so when the substrate 10 is connected to the box body 20, the substrate needs to be pulled and fixed, in this embodiment, the connecting portion 200 and the connected portion 230 are in a snap connection, the box body 20 includes two end portions 201 for connection, each end portion is formed with a connecting portion 200, and at least one of the two end portions 201 can move, so that the distance between the two end portions 201 can be changed, and when the substrate 10 is connected between the two end portions 201, the substrate 10 can be pulled and fixed by changing the distance between the two end portions 201, so that the surface thereof becomes flat, and the optical observation and detection are facilitated.
In addition, as shown in FIG. 2, the substrate 10 further has a loading interface 102 connected to the microfluidic channel 101 for facilitating external sample liquid injection, such as injection of a syringe, a sampler, or any other device capable of sampling the sample liquid from the loading interface 102. Specifically, the sample loading interface 102 may be a separate interface or an interface of the connection portion 100, so that the connection and the sample loading of the biosensor 1 are integrated.
Furthermore, as shown in fig. 3 to 8, the microfluidic channel 101 includes a plurality of detection segments 1010 and a free segment 1011 located between every two adjacent detection segments 1010, wherein a plurality of electromagnetic particles 111 are free in the detection segments 1010, and the inner wall of the end of the free segment 1011 near the detection segments 1010 is coated with a magnetic driving layer 110. Like this, form variable magnetic field through magnetic field forming mechanism for produce between driving magnetic layer 110 and the electromagnetism granule 111 and drive away the effect, can equally divide micro-flow channel 101 into a plurality of detection section 1010, be fixed with a certain amount of electromagnetism granule 110 in every detection section 1010, thereby the electromagnetism granule 110 that makes distributes more evenly in micro-flow channel 101, reduces the reunion of electromagnetism granule 110, detects more fully and accurately.
To facilitate movement of the electromagnetic particle 110 within the microfluidic channel 101, in a preferred embodiment, the cross-section of the detection section 1010 is flat, the cross-section of the free section 1011 is circular, and the maximum width of the cross-section of the detection section 1010 is no greater than the maximum width of the cross-section of the free section 1011. In this manner, the flat shape of the sensing segment 1010 further provides for a uniform distribution of the electromagnetic particles 110 within the interior thereof, further reducing agglomeration thereof. More preferably, at the position where the free section 1011 meets the detection section 1010, the cross section of the free section 1011 is slowly changed to be matched with the cross section of the detection section 1010, so that in some embodiments, the electromagnetic particles move in the free section 1011 to facilitate the passage of the sample and other solutions for detection reaction, and the dead angle is reduced.
Specifically, as shown in fig. 3 to 10, the magnetic driving layer 110 is a coating made of an excitation material. The electromagnetic particles 110 are uniformly distributed due to the need to separate the electromagnetic particles 110 within each detection segment 1010. In a further embodiment, the electromagnetic particles 110 are injected into the microfluidic channel 101 from the outside, and are not directly fixed in the microfluidic channel 101, so that the magnetic driving layer 110 is induced by the magnetic field forming mechanism 21, so that the magnetic driving layer 110 generates its own magnetic field to drive the electromagnetic particles 110, whereas in the injection of the electromagnetic particles 110 or in other embodiments, the electromagnetic particles 110 need to be injected into the microfluidic channel 101 or recovered without the magnetic driving layer 110 generating its own magnetic field, so the magnetic driving layer 110 is made of an excitation material.
Further, as shown in fig. 11-12, the biosensor system provided by the present invention further comprises a processing controller 3, wherein the processing controller 3 is electrically connected to the liquid driving mechanism 23, the light detecting mechanism 22 and the magnetic field forming mechanism 21;
specifically, the liquid driving mechanism 23 includes a liquid storage tank 231, a recovery tank 232, a driving pump 233 and a multi-way control valve 234, the liquid storage tank 231, the recovery tank 232, the multi-way control valve 234, the driving pump 233, the microfluidic channel 101 of the biosensor 1 and the recovery tank 232 may form a fluid channel 2000, and the fluid channel 2000 is used for a sample liquid, a buffer liquid, a reaction liquid or a cleaning liquid to flow through, or for recovery of electromagnetic particles; specifically, the reservoir 231 has a plurality of reservoirs for storing the sample liquid, the buffer liquid, the reaction liquid and the cleaning liquid, respectively, each reservoir 231 has an outlet communicated with the multi-way inlet of the multi-way control valve 234, the outlet of the multi-way control valve 234 is communicated with the inlet of the driving pump 233, the outlet of the driving pump 233 extends to be communicated with the connected part 230, and the connected part 230 is communicated with the connecting part 100, so that the multi-way control valve 234 is switched to allow the sample liquid, the buffer liquid, the reaction liquid or the cleaning liquid to flow into the microfluidic channel 101;
the optical detection mechanism 22 includes a light source 220, an emergent light focusing lens 221, an optical fiber 222 and an optical detector 223, the light source 220 and the emergent light focusing lens 221 are respectively disposed corresponding to the biosensor 1, the emergent light focusing lens 221 is connected with the optical detector 223, the optical detector 223 is electrically connected with the processing controller 3, the light source 220 is used for emitting incident light to the detection section 1010, the light focusing lens 221 is used for collecting, focusing and coupling optical signals generated from the detection section 1010, and the optical detector 223 is used for processing the optical signals and outputting the optical signals to the processing controller 3.
The biosensor system provided by the invention has a compact structure of the liquid storage tank 231 and the recovery tank 232. The liquid storage tanks can be arranged in a plurality of ways and are respectively used for storing a solution to be detected, a buffer solution, a solution containing a labeled antibody and a catalytic reaction solution, the outlets of the liquid storage tanks are respectively connected with the inlets of the driving pumps through connecting pipes and control valves, the solution to be detected, the buffer solution, the solution containing the labeled antibody and the catalytic reaction solution are sequentially and respectively injected, and the solution to be detected, the buffer solution, the solution containing the labeled antibody and the catalytic reaction solution are respectively pumped into the micro-flow channels by the driving. When the labeled antibody is a fluorescence labeled antibody and the catalytic reaction solution is not required to be injected, a detection solution storage tank can be left unused. The time intervals between the pumping of different solutions into the microchannel depend on the situation of the previous process, the time intervals are short when the previous process is performed, and the time intervals are long when the previous process is performed slowly.
In the biosensor system provided by the invention, the types of the surface molecular groups of the electromagnetic particles can be suitable for detection of different objects to be detected, the flow speed, flow and time of the detection liquid are controlled by the control processing controller through the control valve and the sample injection pump, the control valve respectively controls the sample injection type according to the test requirement, namely controls the sample injection of the sample solution to be detected, the buffer solution and the labeled antibody solution, and the flow speed, flow and time of the sample injection are controlled through the sample injection pump.
The invention is carried out by using the electromagnetic particles, thereby greatly improving the reaction surface area and the sensitivity. According to the invention, specific antibodies are fixed on the surfaces of electromagnetic particles in advance, when a solution sample to be detected flows into the electromagnetic particles through a pump, pathogenic microorganisms contained in the solution sample react with the antibodies fixed on the surfaces of the electromagnetic particles so as to be captured, and the separation and enrichment of the microorganisms to be detected and the solution are realized; then pumping and flowing through an antibody solution containing enzyme or fluorescent reagent marks, enabling the marked antibody to generate specific immune reaction with the microorganism to be detected, forming an immune complex of a sandwich structure (surface-fixed antibody-microorganism-enzyme or fluorescent reagent marked antibody) on the surface of the electromagnetic particle, the compound on the surface of the electromagnetic particle is irradiated by measuring light to excite the compound to generate corresponding light effect, namely, the fluorescence reagent marked in the sandwich structure compound is excited by the measuring light to generate fluorescence, or the enzyme-catalyzed chromogenic reaction marked in the sandwich structure compound generates measuring light absorption, or the enzyme catalyzes the reaction of the luminescent agent to generate chemiluminescence, then a light detector is utilized to detect transmitted light, chemiluminescence or fluorescence signals coming out of the electromagnetic particles, and the transmitted light, chemiluminescence or fluorescence signals are processed by the optical signal processing controller to output biological data of the object to be detected. The detection of pathogenic microorganisms or other microbial substances is realized by detecting the absorption of an enzyme catalysis reaction product to light, the chemiluminescence intensity or the fluorescence intensity of a fluorescence labeling antibody.
In some embodiments, as shown in fig. 1 and 11, since the substrate 10 needs to be pulled and fixed, a light source 220 is correspondingly disposed outside each detection segment 1010, the light source 220 has a pressing portion 2200, the exiting light focusing lens 221 has a pressed portion 2210, and the pressing portion 2200 and the pressed portion 2210 can be fitted to and clamped outside the detection segment 1010, thereby reducing the influence caused by the variation of the incident angle of the light source path.
Since the electromagnetic particles 110 in each detection segment 1010 may be uniformly distributed, and still have agglomeration phenomenon, or may be impacted to agglomerate when the sample liquid or the reaction liquid flows through, and still be not beneficial for detection, in some embodiments, the magnetic field forming mechanism 21 may further form a variable magnetic field outside each detection segment 1010 to drive the electromagnetic particles 110 to move therein until being uniform and uniform, so as to facilitate detection.
Biological sample detection method
Referring to fig. 13, a method for performing biological sample detection using the biosensor system provided in the above embodiment,
s1, preparing electromagnetic particles with surface coupled molecular groups and a base material;
s2, pouring the electromagnetic particles into the microfluidic channel, and enabling the magnetic driving layer to generate magnetic driving force through the magnetic field forming mechanism to limit the electromagnetic particles in the microfluidic channel;
s3, enabling the solution to be detected to flow through the micro-flow channel, and generating reaction with the molecular groups on the surface of the electromagnetic particles and generating optical signals;
s4, the optical detection mechanism acquires the optical signal, and outputs the optical signal to the processing controller after focusing, coupling and processing;
and S5, processing and calculating by the processing controller, and outputting the biological data of the sample.
In the embodiment of dividing the microfluidic channel into a plurality of detection segments, as shown in fig. 14, the step S2 may further be:
s21, pouring electromagnetic particles into the microfluidic channel, and generating driving force on the electromagnetic particles through the magnetic field forming mechanism to enable the electromagnetic particles to be uniformly distributed in the microfluidic channel;
and S22, enabling the lock driving magnetic layer to generate driving force through the magnetic field forming mechanism, and fixing the electromagnetic particles in each detection section.
In some embodiments, as in fig. 15, the step of S4 may also be:
s41, recovering the reacted electromagnetic particles;
and S42, acquiring the surface optical signal change of the electromagnetic particles before and after reaction by an optical detection mechanism, focusing, coupling and processing the surface optical signal change, and outputting the surface optical signal change to a processing controller.
In some more specific embodiments, the electromagnetic particles provided herein are immune electromagnetic particles. More specific detection methods of the biological sample comprise:
(1) fixing an antibody on the surface of the immune electromagnetic particle, filling the electromagnetic particle into the microfluidic channel, and enabling the magnetic driving layer to generate a magnetic driving force through the magnetic field forming mechanism so as to limit the electromagnetic particle in the microfluidic channel;
(2) pumping a solution to be detected into a micro-flow channel, so that an object to be detected contained in the solution to be detected reacts with the antibody fixed on the surface of the electromagnetic particle to form an antigen-antibody compound which is enriched on the surface of the electromagnetic particle;
(3) pumping a buffer solution into the microchannel, washing the antigen-antibody complex enriched on the surface of the electromagnetic particle, and washing to remove impurities;
(4) pumping a solution containing a fluorescence-labeled antibody or an enzyme-labeled antibody into the microchannel, so that the labeled antibody in the solution and the antigen in the antigen-antibody complex carry out immunoreaction to form an immune complex with a sandwich structure loaded on the antibody-antigen-labeled antibody complex; when the labeled antibody is an enzyme labeled antibody, pumping a catalytic reaction solution after an immunoreaction to perform an enzyme catalytic chromogenic reaction to form a colored substance to absorb detection light, or pumping an enzyme catalytic reaction luminescent agent to perform an enzyme catalytic luminescent reaction to generate chemiluminescence;
(5) pumping a buffer solution into the electromagnetic particle microfluidic channel to wash the free labeled antibody;
(6) the method comprises the steps of irradiating an antibody-antigen-labeled antibody compound in an electromagnetic particle micro-flow channel by using measuring light, exciting a fluorescence labeled antibody to emit fluorescence, or forming a colored substance through an enzyme catalysis color development reaction to absorb the measuring light to form transmitted light, or forming chemiluminescence through an enzyme catalysis luminescence reaction, wherein the detecting light emitted from the electromagnetic particles is focused and coupled to an optical fiber through a lens, and is transmitted to a light detector for detection processing, and a signal after the detection processing is transmitted to a light signal processing controller to output data of a biological sample to be detected, so that the quantitative determination of the object to be detected is realized.
In the above method for rapidly detecting a microorganism sample of the present invention, the reaction conditions in step (2): the pH is 7.0-7.6, and the temperature is 20-42 ℃; the operating condition in the step (3) is that the temperature is 4-42 ℃; the immunoreaction conditions in the step (4) are as follows: the pH is 7.0-7.6, the temperature is 2-42 ℃, and the enzyme catalysis and color development reaction conditions are as follows: the pH is 7.0-7.6, and the temperature is 2-42 ℃; reaction conditions in step (5): the pH is 7.0-7.6 and the temperature is 2-42 ℃. Reaction conditions are as follows: the pH is 7.0-7.6 and the temperature is 2-42 ℃.
In the method for rapidly detecting a microorganism sample according to the present invention, the antibody may be immobilized on the surface of the electromagnetic particle by one or a combination of physical adsorption, chemical bonding and biological affinity.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are also within the scope of the present invention.

Claims (10)

1. A biosensor system, comprising:
the biosensor comprises a substrate and a magnet assembly arranged in the substrate, wherein the substrate is provided with a connecting part and a micro-flow channel arranged in the substrate, the magnet assembly comprises magnetic driving layers coated at two tail ends of the micro-flow channel and a plurality of electromagnetic particles dissociated in the micro-flow channel, the magnetic field intensity of the electromagnetic particles can be respectively changed through electric control, the magnetic driving layers have acting force for driving away the electromagnetic particles, and the surfaces of the electromagnetic particles are coupled with molecular groups which can react with substances to be detected in a sample and generate optical signals; and
the biosensor box comprises a box body, and a magnetic field forming mechanism and an optical detection mechanism which are fixed on the box body, wherein the magnetic field forming mechanism forms a variable magnetic field to act on the magnetic driving layer and the electromagnetic particles, and the optical detection mechanism is used for detecting optical signals in the biosensor.
2. The biosensor system according to claim 1, wherein the biosensor cartridge further comprises a liquid driving mechanism having a connected portion, and the liquid driving mechanism drives the liquid in the microfluidic channel to flow after the connected portion is connected to the connecting portion.
3. The biosensor system according to claim 1 or 2, wherein the substrate is made of a flexible transparent material, and the substrate has a loading interface communicated with the microfluidic channel, and the loading interface is used for injecting a sample liquid.
4. The biosensor system according to claim 1, wherein the microfluidic channel comprises a plurality of detection segments and a free segment located between every two adjacent detection segments, the detection segments free a plurality of electromagnetic particles, and an inner wall of an end of the free segment close to the detection segments is coated with the magnetic driving layer.
5. The biosensor system of claim 4, wherein the cross-section of the detection section is flat, the cross-section of the free section is circular, and the cross-sectional maximum width of the detection section is not greater than the cross-sectional maximum width of the free section.
6. The biosensor system of claim 5, wherein said magnetic drive layer is a coating of an excitation material.
7. The biosensor system according to claim 6, further comprising a process controller, wherein the process controller is electrically connected to the liquid driving mechanism, the light detecting mechanism and the magnetic field forming mechanism;
the liquid driving mechanism comprises a liquid storage tank, a recovery tank, a driving pump and a multi-way control valve, the liquid storage tank, the recovery tank, the multi-way control valve, the driving pump, a micro-flow channel of the biosensor and the recovery tank can form a fluid channel, and the fluid channel is used for sample liquid, buffer liquid, reaction liquid or cleaning liquid to flow through or for recovery of electromagnetic particles;
the optical detection mechanism comprises a light source, an emergent light focusing lens, an optical fiber and an optical detector, wherein the light source and the emergent light focusing lens are respectively arranged corresponding to the biosensor, the emergent light focusing lens is connected with the optical detector, the optical detector is electrically connected with the processing controller, the light source is used for emitting incident light to the detection section, the light focusing lens is used for collecting, focusing and coupling optical signals generated by the detection section, and the optical detector is used for processing the optical signals and outputting the optical signals to the processing controller.
8. A method for performing biological sample testing using the biosensor system of claim 1, comprising:
s1, preparing the electromagnetic particles with surface coupling molecular groups and the base material;
s2, pouring the electromagnetic particles into the microfluidic channel, and enabling the magnetic driving layer to generate a magnetic driving force through the magnetic field forming mechanism so as to limit the electromagnetic particles in the microfluidic channel;
s3, enabling the solution to be detected to flow through the microfluidic channel, reacting with the molecular groups on the surface of the electromagnetic particles, and generating an optical signal;
s4, the optical detection mechanism acquires the optical signal, focuses, couples and processes the optical signal and outputs the optical signal to the processing controller;
and S5, the processing controller processes and calculates, and outputs the biological data of the sample.
9. The method of claim 8, wherein the step of S2 is further:
s21, pouring the electromagnetic particles into a microfluidic channel, and generating driving force for the electromagnetic particles through the magnetic field forming mechanism to enable the electromagnetic particles to be uniformly distributed in the microfluidic channel;
and S22, enabling the lock driving magnetic layer to generate driving force through the magnetic field forming mechanism, and fixing the electromagnetic particles in each detection section.
10. The method of claim 8, wherein the step of S4 is further:
s41, recovering the reacted electromagnetic particles;
and S42, acquiring the surface optical signal change of the electromagnetic particles before and after reaction through the optical detection mechanism, focusing, coupling and processing the surface optical signal change, and outputting the surface optical signal change to the processing controller.
CN202011089473.5A 2020-10-13 2020-10-13 Biosensor system and biological sample detection method Withdrawn CN112304911A (en)

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