CN113721015A - Automatic microorganism detection device, system and method - Google Patents
Automatic microorganism detection device, system and method Download PDFInfo
- Publication number
- CN113721015A CN113721015A CN202111051010.4A CN202111051010A CN113721015A CN 113721015 A CN113721015 A CN 113721015A CN 202111051010 A CN202111051010 A CN 202111051010A CN 113721015 A CN113721015 A CN 113721015A
- Authority
- CN
- China
- Prior art keywords
- magnetic field
- reaction vessel
- magnetic
- liquid level
- liquid column
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 124
- 244000005700 microbiome Species 0.000 title claims abstract description 54
- 238000000034 method Methods 0.000 title claims abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 163
- 238000006243 chemical reaction Methods 0.000 claims abstract description 111
- 239000003593 chromogenic compound Substances 0.000 claims description 35
- 238000004140 cleaning Methods 0.000 claims description 33
- 238000005086 pumping Methods 0.000 claims description 26
- 239000002699 waste material Substances 0.000 claims description 11
- 241000894006 Bacteria Species 0.000 claims description 10
- 230000009471 action Effects 0.000 claims description 9
- 229920012699 PC light diffusion Polymers 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 5
- 238000007599 discharging Methods 0.000 claims description 5
- 230000014759 maintenance of location Effects 0.000 claims 5
- 230000028993 immune response Effects 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 9
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 32
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 17
- 229910052697 platinum Inorganic materials 0.000 description 16
- 239000000243 solution Substances 0.000 description 13
- 239000000523 sample Substances 0.000 description 9
- 150000001875 compounds Chemical class 0.000 description 8
- 230000005389 magnetism Effects 0.000 description 7
- 230000000813 microbial effect Effects 0.000 description 7
- 239000012488 sample solution Substances 0.000 description 6
- 230000036046 immunoreaction Effects 0.000 description 5
- 239000011259 mixed solution Substances 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 4
- 230000006698 induction Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 3
- 238000013459 approach Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 240000007651 Rubus glaucus Species 0.000 description 1
- 235000011034 Rubus glaucus Nutrition 0.000 description 1
- 235000009122 Rubus idaeus Nutrition 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical group [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000000696 magnetic material Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5302—Apparatus specially adapted for immunological test procedures
- G01N33/5304—Reaction vessels, e.g. agglutination plates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
-
- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
-
- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54346—Nanoparticles
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Food Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Nanotechnology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
本发明提供一种微生物自动化检测装置、系统及方法，包括有工作台、两个固定件、反应容器、磁场旋转部件、两个液位检测部件、磁吸截留部件、背景光源部件和图像采集部件；两个固定件分别固定安装在工作台两侧，磁场旋转部件用于在中心处形成磁感线平行分布的磁场；反应容器一端安装在其中一个固定件上，反应容器另一端穿过磁场旋转部件的中心与另一个固定件连接；两个液位检测部件分别安装在磁场旋转部件的两侧，液位检测部件与反应容器连接；磁吸截留部件位于两个液位检测部件之间；图像采集部件位于背景光源部件上方。本发明实现了对微生物的自动化检测，降低了对仪器设备和专业操作人员的依赖，缩短了微生物检测时间，提高了微生物检测灵敏度。
The invention provides an automatic microorganism detection device, system and method, comprising a workbench, two fixing parts, a reaction container, a magnetic field rotating part, two liquid level detection parts, a magnetic suction interception part, a background light source part and an image acquisition part ; Two fixed parts are fixedly installed on both sides of the worktable, and the magnetic field rotating part is used to form a magnetic field with parallel magnetic field lines at the center; one end of the reaction vessel is installed on one of the fixed parts, and the other end of the reaction vessel rotates through the magnetic field The center of the part is connected with another fixed part; two liquid level detection parts are installed on both sides of the magnetic field rotating part, and the liquid level detection part is connected with the reaction vessel; the magnetic suction interception part is located between the two liquid level detection parts; image The collection part is located above the background light part. The invention realizes the automatic detection of microorganisms, reduces the dependence on instruments and equipment and professional operators, shortens the microorganism detection time, and improves the microorganism detection sensitivity.
Description
Technical Field
The invention relates to the technical field of microorganism detection, in particular to a microorganism automatic detection device, system and method.
Background
Background in food samples is complex, the concentration of food-borne pathogenic bacteria is usually low, and the conventional detection method is difficult to directly detect the food samples. The double-antibody sandwich technology is a biological detection technology based on antigen-antibody immune combination, a capture probe combined with a specific antibody is used for capturing target bacteria, then a signal probe combined with another antibody is used for marking the target bacteria to form an immune capture probe-target bacteria-immune signal probe double-antibody sandwich structure, and a signal probe is used for converting a corresponding bacteria concentration signal into a detectable physical signal such as light, heat, magnetism, force, sound, electricity and the like to indirectly detect the concentration of the target bacteria.
However, most of the existing double-antibody sandwich technologies have low automation degree when applied to microbial detection, and need to rely on various instruments and professional operators, so that the forming efficiency of the double-antibody sandwich structure is low, and the forming efficiency of the double-antibody sandwich structure can influence the detection time and the detection sensitivity, so that the detection time is long and the sensitivity is not high. Therefore, how to improve the efficiency of forming the double-antibody sandwich structure is an important issue to be solved in the industry at present, which is to reduce the detection time and improve the detection sensitivity.
Disclosure of Invention
The invention provides a microbial automatic detection device, a system and a method, which are used for solving the defects that the double-antibody sandwich technology in the prior art has low automation degree, needs to depend on various instruments and professional operators, has low forming efficiency of a double-antibody sandwich structure, long detection time and low sensitivity, realizes the automatic detection of microbes, reduces the dependence on various instruments and equipment and professional operators, improves the forming efficiency of the double-antibody sandwich structure, shortens the detection time of the microbes and improves the detection sensitivity of the microbes.
The invention provides an automatic microorganism detection device, which comprises a workbench, two fixing pieces, a reaction container, a magnetic field rotating part, two liquid level detection parts, a magnetic interception part, a background light source part and an image acquisition part, wherein the workbench is provided with a plurality of fixing pieces;
the two fixing pieces are respectively and fixedly arranged on two sides of the workbench, the magnetic field rotating part is arranged between the two fixing pieces, and the magnetic field rotating part is used for forming a magnetic field with magnetic induction lines distributed in parallel at the center of the magnetic field rotating part;
one end of the reaction vessel is mounted on one of the fixing pieces, and the other end of the reaction vessel penetrates through the center of the magnetic field rotating component and then is connected with the other fixing piece;
the two liquid level detection components are respectively arranged on two sides of the magnetic field rotating component and are connected with the reaction vessel;
the magnetic interception component is arranged on one side of the reaction container and is positioned between the two liquid level detection components;
the background light source component is arranged on the workbench and is positioned below the reaction container;
the image acquisition component is positioned above the background light source component, and the reaction container is positioned between the background light source component and the image acquisition component.
According to the automatic microorganism detection device provided by the invention, the magnetic field rotating part comprises a rotary fixed seat, a rotary driving part and an annular Halbach array magnet, the rotary fixed seat is installed between the two liquid level detection parts, the rotary driving part and the annular Halbach array magnet are both installed on the rotary fixed seat, and the rotary driving part is connected with the annular Halbach array magnet.
According to the automatic microorganism detection device provided by the invention, the annular Halbach array magnet comprises a plurality of magnets distributed in a Halbach array and a magnetic locking ring, and the magnetic locking ring wraps the magnets.
According to the automatic microorganism detection device provided by the invention, the liquid level detection part comprises a liquid level detection fixing frame and a liquid level sensor, the liquid level detection fixing frame is arranged on one side of the reaction container, the liquid level sensor is arranged on the liquid level detection fixing frame, and the liquid level sensor is connected with the reaction container.
According to the automatic microorganism detection device provided by the invention, the magnetic attraction interception part comprises an upper driving part, a lower driving part and a permanent magnet, the upper driving part and the lower driving part are fixedly arranged between the two liquid level detection parts, one end of the permanent magnet is connected with the upper driving part and the lower driving part, and the other end of the permanent magnet is positioned above the reaction container.
According to the automatic microorganism detection device provided by the invention, the background light source component comprises a PC light diffusion plate and a plurality of light-emitting LEDs, the light-emitting LEDs are fixedly arranged on the workbench, the PC light diffusion plate covers the light-emitting LEDs, and the PC light diffusion plate is positioned below the image acquisition component.
According to the automatic microorganism detection device provided by the invention, the image acquisition part comprises an image acquisition driving part and a camera, the image acquisition driving part is fixedly arranged on one side of the reaction container, the camera is arranged on the image acquisition driving part, and the reaction container is positioned between the camera and the background light source part.
The invention also provides a microbial automatic detection system, which comprises a detection box and a microbial automatic detection device, wherein a cavity is arranged in the detection box, the microbial automatic detection device is arranged at the upper half part of the cavity, the workbench is connected with the inner wall surface of the cavity, a pumping module and a control module are arranged at the lower half part of the cavity, the pumping module is connected with the reaction container, and the control module is connected with the pumping module.
According to the automatic microorganism detection system provided by the invention, the automatic microorganism detection system further comprises a waste liquid pool, the waste liquid pool is fixedly arranged on the detection box, one end of the pumping module is connected with the reaction container, and the other end of the pumping module is connected with the waste liquid pool.
The invention also provides a microbial automatic detection method, which comprises the following steps:
respectively injecting the mixed liquid column, the cleaning liquid column and the chromogenic substrate liquid column into a reaction container, and separating the mixed liquid column, the cleaning liquid column and the chromogenic substrate liquid column through an air column;
the rotating component of the rotating magnetic field controls the times of reciprocating motion of the mixed liquid column in the magnetic field interval to the target, so that the target bacteria form a double-antibody sandwich structure under the action of immunoreaction;
enabling the magnetic interception component to be close to the reaction container, discharging the mixed liquid column from the reaction container, controlling the cleaning liquid column to move to a magnetic field interval, and then enabling the magnetic interception component to be far away from the reaction container;
controlling the cleaning liquid column to reciprocate for a target number of times in a magnetic field interval, then enabling the magnetic attraction interception component to be close to the reaction container, and discharging the cleaning liquid column from the reaction container;
pumping the chromogenic substrate liquid column to a magnetic field interval, controlling the magnetic attraction interception component to be far away from the reaction container, controlling the magnetic attraction interception component to be close to the reaction container after the chromogenic substrate changes color, pumping the chromogenic substrate liquid column to the position below the image acquisition component, and then starting a background light source component and taking a picture;
and processing and analyzing the image to obtain a saturation value of a color-changing chromogenic substrate area in the image, and calculating to obtain the bacterial content in the sample to be detected.
According to the automatic microorganism detection device, the automatic microorganism detection system and the automatic microorganism detection method, the driving pump is connected with the reaction container, and the liquid column in the reaction container can be driven to reciprocate by the driving pump. After the mixed liquid column is driven to pass through the magnetic field rotating component repeatedly for a plurality of times, the nano nickel wires are captured in the center of the magnetic field rotating component under the action of the magnetic field and distributed in a chain shape, and rotate along with the rotation of the magnetic field. The target bacteria are rapidly combined with the nano nickel wire and the gold-core platinum nano cluster under the action of immunoreaction to form a double-antibody sandwich structure of the nano nickel wire-target bacteria-gold-core platinum nano cluster, and the double-antibody sandwich structure is fixed at the center of the annular magnetic field rotating component along with the nano nickel wire. The magnetic attraction interception part is close to the reaction container, then the driving pump drives the mixed liquid column to be discharged out of the reaction container, and after the mixed solution is discharged, the magnetic compound is adsorbed in the reaction container. And then driving the cleaning liquid column to move to a position between the two liquid level detection parts to redissolve the double-antibody sandwich structure, then magnetically attracting and intercepting the parts to be far away from the reaction container, and enabling the cleaning liquid column to reciprocate for a plurality of times between the two liquid level detection parts to clean the free gold-core platinum nanoclusters. The magnetic attraction interception component is close to the reaction vessel again, then the cleaning liquid column is discharged out of the reaction vessel, and after the cleaning liquid column is discharged, the magnetic compound is adsorbed in the reaction vessel. Then driving the chromogenic substrate liquid column to move to a rotating part of an annular magnetic field, after a double-antibody sandwich structure is redissolved, enabling a magnetic attraction interception component to be far away from a reaction container, catalyzing the chromogenic substrate by using gold-core platinum nanoclusters positioned on the double-antibody sandwich structure to enable the chromogenic substrate to be changed from colorless to blue, enabling the magnetic attraction interception component to be close to the reaction container, moving the chromogenic substrate liquid column to the magnetic attraction interception component, after the double-antibody sandwich structure is fully adsorbed, driving the chromogenic substrate liquid column after color change to be below an image acquisition component, starting a background light source component, photographing the chromogenic substrate liquid column by the image acquisition component, processing and analyzing the photograph to obtain the saturation value of a blue chromogenic substrate area in the image, and further calculating the bacterial content in a sample to be detected according to a preset saturation-bacterial concentration curve. And further, the automatic detection of the microorganisms is realized, the dependence on various instruments and equipment and professional operators is reduced, the forming efficiency of the double-antibody sandwich structure is improved, the microorganism detection time is shortened, and the microorganism detection sensitivity is improved.
Drawings
In order to more clearly illustrate the technical solutions of the present invention or the prior art, the drawings needed for the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and those skilled in the art can also obtain other drawings according to the drawings without creative efforts.
FIG. 1 is a schematic structural diagram of an automated microorganism detection apparatus provided in the present invention;
FIG. 2 is a second schematic structural view of an automated microorganism detection apparatus provided in the present invention;
FIG. 3 is a sectional view of a ring-shaped Halbach array magnet of the automated microorganism detection apparatus provided in the present invention;
FIG. 4 is a schematic structural diagram of an automated microbial detection system provided by the present invention;
FIG. 5 is a flow chart of the automated microorganism detection method provided by the present invention;
reference numerals:
1: a work table; 2: a fixing member; 3: a reaction vessel;
4: a magnetic field rotating member; 5: a liquid level detection section; 6: the interception component is magnetically attracted;
7: a background light source section; 8: an image acquisition component; 9: a detection box;
41: a rotating fixed seat; 42: a rotary drive member; 43: an annular Halbach array magnet;
51: a liquid level detection fixing frame; 52: a liquid level sensor; 71: a PC light diffuser plate;
61: an upper and lower driving member; 62: a permanent magnet; 91: a cavity;
81: an image acquisition driving member; 82: a camera; 94: a waste liquid tank;
92: a pumping module; 93: a control module; 431: a magnet;
432: a lock magnetic ring.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The automated microorganism detection apparatus, system and method of the present invention will be described with reference to fig. 1 to 5.
As shown in attached figure 1, the automatic microorganism detection device comprises a workbench 1, two fixing pieces 2, a reaction vessel 3, a magnetic field rotating part 4, two liquid level detection parts 5, a magnetic attraction interception part 6, a background light source part 7 and an image acquisition part 8.
Specifically, two mounts 2 are fixedly installed at both sides of the table 1, respectively, and a magnetic field rotating member 4 is installed between the two mounts 2, the magnetic field rotating member 4 being for forming a magnetic field having induction lines distributed in parallel at the center of the magnetic field rotating member 4. One end of the reaction vessel 3 is mounted on one of the fixed members 2, and the other end of the reaction vessel 3 passes through the center of the magnetic field rotating member 4 and then is connected to the other fixed member 2.
Two liquid level detection parts 5 are respectively arranged at two sides of the magnetic field rotating part 4, and the liquid level detection parts 5 are connected with the reaction vessel 3. The magnetic interception component 6 is arranged at one side of the reaction vessel 3, and the magnetic interception component 6 is positioned between the two liquid level detection components 5. The background light source unit 7 is mounted on the table 1, and the background light source unit 7 is located below the reaction vessel 3. The image pickup section 8 is located above the background light source section 7, and the reaction vessel 3 is located between the background light source section 7 and the image pickup section 8.
When in use, the mixed solution of the nano nickel wire modified with the anti-salmonella polyclonal antibody, the sample solution to be detected and the gold-core platinum nano-cluster solution modified with the anti-salmonella monoclonal antibody is injected into the reaction vessel 3 to form a mixed liquid column, 1 percent skim milk is injected into the reaction vessel 3 to form a cleaning liquid column, and TMB-H is added2O2Injecting the mixture into a reaction container to form a chromogenic substrate liquid column, and separating the mixed liquid column, the cleaning liquid column and the chromogenic substrate liquid column by an air column.
Then, the magnetic field rotating member 4 starts to operate, and a magnetic field having lines of magnetic induction distributed in parallel is formed at the center of the magnetic field rotating member 4. Then be connected with reaction vessel 3 through a driving pump, and then can drive the liquid column in reaction vessel 3 through the driving pump and carry out reciprocating motion, and two liquid level detection part 5 can the position of real-time detection liquid column, and then can control the liquid column at two liquid level detection part 5 reciprocating motion, and can learn the number of times of liquid column reciprocating motion through the number of times that liquid level detection part 5 detected the liquid column.
After the mixed liquid column is driven to pass through the magnetic field rotating component repeatedly for a plurality of times, the nano nickel wires are captured in the center of the magnetic field rotating component under the action of the magnetic field and distributed in a chain shape, and rotate along with the rotation of the magnetic field. The target bacteria, the nano nickel wire and the gold-core platinum nano cluster are rapidly combined under the action of immunoreaction to form a double-antibody sandwich structure of the nano nickel wire-target bacteria-gold-core platinum nano cluster, and the double-antibody sandwich structure is fixed at the center of the magnetic field rotating component 4 along with the nano nickel wire.
Make magnetism earlier to inhale interception part 6 and be close to reaction vessel 3, then the drive pump drives mixed liquid column discharge reaction vessel 3, and magnetism is inhaled simultaneously and is held up part 6 and inhale magnetism to the magnetic composition in the mixed liquid, and then prevents that the magnetic composition in the mixed solution from being discharged. After the mixed solution discharges, the drive cleaning solution column removes between two liquid level detection part 5, magnetic composite is adsorbed in reaction vessel 3 this moment, then magnetism is inhaled and is held back part 6 and keep away from reaction vessel 3, then drive cleaning solution column continuous reciprocating motion between two liquid level detection part 5, wash free gold core platinum nanometer cluster, make magnetism inhale and hold back part 6 and be close to reaction vessel 3, then will wash liquid column discharge reaction vessel 3, magnetism is inhaled and is held back part 6 and prevent magnetic composite along with the discharge of cleaning solution column simultaneously.
Then driving the chromogenic substrate liquid column to move to the magnetic field rotating component 4, enabling the magnetic attraction interception component 6 to be far away from the reaction container 3, catalyzing the chromogenic substrate by utilizing a gold-core platinum nanocluster positioned on a double-antibody sandwich structure, enabling the chromogenic substrate to be changed from colorless to blue, enabling the magnetic attraction interception component 6 to be close to the reaction container 3, then driving the chromogenic substrate liquid column after color change to be below the image acquisition component 8, starting the background light source component 7, photographing the chromogenic substrate liquid column by the image acquisition component 8, then processing and analyzing the photograph to obtain the saturation value of the blue chromogenic substrate area in the image, and further calculating the bacterial content in the sample to be detected according to a preset saturation-bacterial concentration curve.
And further, the automatic detection of the microorganisms is realized, the dependence on various instruments and equipment and professional operators is reduced, the forming efficiency of the double-antibody sandwich structure is improved, the microorganism detection time is shortened, and the microorganism detection sensitivity is improved.
In an alternative embodiment of the present invention, the fixing member 2 is, for example, a mounting frame having an upper end provided with a coupling hole to be fitted to the reaction vessel 3. It should be understood that the securing member 2 may be of any other suitable construction.
Wherein, in an alternative embodiment of the present invention, the reaction vessel 3 is, for example, a glass tube. It should be understood that the reaction vessel 3 may be any other suitable vessel.
Further, as shown in fig. 1 and fig. 2, the magnetic field rotating component 4 includes a rotating fixing base 41, a rotating driving component 42 and an annular halbach array magnet 43, the rotating fixing base 41 is installed between the two liquid level detecting components 5, the rotating driving component 42 and the annular halbach array magnet 43 are both installed on the rotating fixing base 41, and the rotating driving component 42 is connected with the annular halbach array magnet 43. When the device is used, the annular Halbach array magnet 43 is driven to rotate by the rotary driving piece 42, a magnetic field with parallel magnetic induction lines is formed at the center of the annular Halbach array magnet 43, magnetic materials such as nano nickel wires in the reaction container 3 are further induced to rotate in a chain-shaped distribution near the center of the magnetic field, the whole cross section is swept during rotation, the mixed liquid column is driven to flow back and forth between the two liquid level detection parts 5, a sample repeatedly passes through the rotary magnetic chain limited at the center of the Halbach ring, and the collision and combination probability of the magnetic substances and target microorganisms in the flowing sample solution is increased. The defects that a chain formed by a traditional magnetic grid is short and cannot be distributed in the whole channel, and partial sample solution cannot be combined with a magnetic substance are overcome.
As shown in fig. 2 and fig. 3, the annular halbach array magnet 43 includes a plurality of magnets 431 distributed in a halbach array and a magnetic-lock ring 432, the magnet 431 is wrapped by the magnetic-lock ring 432, and the direction of the arrow in fig. 3 is the magnetizing direction corresponding to each of the magnets 431. In use, the locking magnet ring 432 may enhance the magnetic field strength within the annular magnetic field.
Wherein in an alternative embodiment of the invention, the rotary drive 42 is, for example, a stepper motor. It should be appreciated that the rotary drive 42 may be any other suitable drive.
Further, as shown in fig. 1 and fig. 2, the liquid level detection unit 5 includes a liquid level detection fixing frame 51 and a liquid level sensor 52, the liquid level detection fixing frame 51 is installed at one side of the reaction vessel 3, the liquid level sensor 52 is installed on the liquid level detection fixing frame 51, and the liquid level sensor 52 is connected to the reaction vessel 3. When using, level sensor 52 detects the liquid column in reaction vessel 3 in real time, when the level sensor 52 of one of them liquid level detection part 5 detected the liquid column, then drive the liquid column toward another liquid level detection part 5 one side motion, when the level sensor 52 of another liquid level detection part 5 detected the liquid column, drive the liquid column toward one of them liquid level detection part 5 one side motion, and then realized the control to liquid column reciprocating motion in reaction vessel 3, and can calculate the number of times of liquid column reciprocating motion, microorganism automated inspection has been realized, the dependence to professional technical personnel has been reduced.
Further, as shown in fig. 1 and fig. 2, the magnetic attraction and interception member 6 includes an upper driving member 61 and a lower driving member 61, and a permanent magnet 62, wherein the upper driving member 61 is fixedly installed between the two liquid level detection members 5, one end of the permanent magnet 62 is connected with the upper driving member 61, and the other end of the permanent magnet 62 is located above the reaction vessel 3. When the device is used, when the mixed liquid column needs to be discharged out of the reaction container 3, the permanent magnet 62 is driven to move downwards through the upper driving piece 61 and the lower driving piece 61, so that the permanent magnet 62 approaches the reaction container 3, magnetic attraction is carried out on the magnetic compound in the mixed liquid, and the magnetic compound in the mixed liquid is prevented from being discharged. Then the cleaning liquid column moves to a Halbach magnetic field rotating part for a redissolution step, and then the permanent magnet 62 is driven by the upper and lower driving pieces 61 to move upwards to be far away from the reaction vessel 3 for a cleaning step. After the cleaning liquid column is cleaned, the permanent magnet 62 is driven to move downwards by the upper and lower driving members 61, so that the permanent magnet 62 approaches the reaction vessel 3 again, and then the cleaning liquid column is discharged out of the reaction vessel 3, thereby preventing the magnetic compound from being discharged out along with the cleaning liquid column.
In an alternative embodiment of the present invention, the upper and lower driving members 61 are, for example, micro-slides based on stepping motors. It should be appreciated that the up-down drive 61 could be any other suitable drive.
Wherein, in an alternative embodiment of the present invention, permanent magnet 62 is, for example, a cylindrical N52 ndfeb permanent magnet 62. It should be appreciated that the permanent magnet 62 may be any other suitable shape or material of the permanent magnet 62.
Further, as shown in fig. 1 and 2, the background light source unit 7 includes a PC light diffusion plate 71 and a plurality of light emitting LEDs, the light emitting LEDs are fixedly mounted on the table 1, the PC light diffusion plate 71 covers the light emitting LEDs, and the PC light diffusion plate 71 is located below the image capturing unit 8. When the LED lamp is used, the light-emitting LEDs serve as light sources to emit light rays, the light rays are uniform after passing through the PC light diffusion plate 71, and the phenomenon that the local light rays of the lamp beads are too strong to influence the image quality is avoided.
In an alternative embodiment of the present invention, the light emitting LED is, for example, a white LED lamp with a rated voltage of 3.3V. It should be understood that any other suitable size LED lamp may be used as the light emitting LED.
Further, as shown in fig. 1 and fig. 2, the image capturing unit 8 includes an image capturing driving unit 81 and a camera 82, the image capturing driving unit 81 is fixedly installed at one side of the reaction container 3, the camera 82 is installed on the image capturing driving unit 81, and the reaction container 3 is located between the camera 82 and the background light source unit 7. When the device is used, the camera 82 is driven to move up and down through the image acquisition driving part 81, so that the distance between the camera 82 and the reaction container 3 is adjusted, and the camera 82 can better shoot in the reaction container 3.
In an alternative embodiment of the present invention, the image capturing driving member 81 is, for example, a micro slide table based on a stepping motor. It should be understood that any other suitable drive may be used as the image acquisition drive 81.
Wherein, in an alternative embodiment of the present invention, the camera 82 is, for example, a CMOS macro camera 82 with 200 ten thousand pixels, and the object distance is greater than 20 mm.
On the other hand, as shown in fig. 1 and fig. 4, the invention further provides a microorganism automatic detection system, which comprises a detection box 9, wherein a cavity 91 is arranged in the detection box 9, the microorganism automatic detection device is installed at the upper half part of the cavity 91, the workbench 1 is connected with the inner wall surface of the cavity 91, the lower half part of the cavity 91 is provided with a pumping module 92 and a control module 93, the pumping module 92 is connected with the reaction container 3, and the control module 93 is connected with the pumping module 92. When the device is used, the upper part and the lower part of the cavity 91 are separated by the workbench 1, so that a darkroom is formed at the upper half part of the cavity 91, a stable light source environment can be provided for image acquisition and image analysis, the control module 93 is used for controlling the pumping module 92, and then the liquid column flow in the reaction container 3 is controlled through the pumping module 92.
Further, as shown in fig. 4, the automatic microorganism detection system further includes a waste liquid tank 94, the waste liquid tank 94 is fixedly mounted on the detection box 9, one end of the pumping module 92 is connected to the reaction container 3, and the other end of the pumping module 92 is connected to the waste liquid tank 94. In use, after the mixed liquid column and the cleaning liquid column in the reaction vessel 3 are used up, the mixed liquid column and the cleaning liquid column are pumped to the waste liquid tank 94 by the pumping module 92 for storage.
Wherein, in an alternative embodiment of the present invention, the pumping module 92 is, for example, a peristaltic pump. It should be appreciated that any other suitable pump may be used as the pumping module 92.
Wherein, in an alternative embodiment of the present invention, the control module 93 is, for example, a raspberry pi based control system. It should be appreciated that any other suitable control system may be used as the control module 93.
In another aspect, as shown in fig. 5, the present invention further provides a method for automatically detecting microorganisms, comprising:
s1, respectively injecting the mixed liquid column, the cleaning liquid column and the chromogenic substrate liquid column into the reaction container, and separating the mixed liquid column, the cleaning liquid column and the chromogenic substrate liquid column through an air column;
specifically, the mixed liquid column comprises a nano nickel wire modified with an anti-salmonella polyclonal antibody, a sample solution to be detected and a gold-core platinum nano-cluster solution modified with an anti-salmonella monoclonal antibody, and the mixed liquid column comprises the nano nickel wire modified with an anti-salmonella polyclonal antibody, a sample solution to be detected and the gold-core platinum nano-cluster solution modified with an anti-salmonella monoclonal antibodyIn the method, the dosage of the nano nickel wire is preferably 40 micrograms, the dosage of the sample solution to be detected is correspondingly 50 microliters, and the dosage of the gold-core platinum nano-cluster solution modified with the anti-salmonella monoclonal antibody is preferably 16 microliters. The column of washing liquid consisted of 50 microliters of 1% skim milk. The color-developing substrate liquid column is composed of 50 microliter TMB-H2O2And (4) forming.
S2, rotating the magnetic field rotating component, controlling the times of reciprocating motion of the mixed liquid column in the magnetic field interval to the target, so that the target bacteria form a double-antibody sandwich structure under the action of immunoreaction;
specifically, the nano nickel wires are captured in the center of the magnetic field rotating component and distributed in a chain shape under the action of the magnetic field, and rotate along with the rotation of the magnetic field. The target bacteria are combined with the nano nickel wire and the gold-core platinum nano cluster under the action of immunoreaction to form a nano nickel wire-target bacteria-gold-core platinum nano cluster double-antibody sandwich structure, and the nano nickel wire is fixed at the center of the magnetic field rotating component.
S3, enabling the magnetic interception component to be close to the reaction container, discharging the mixed liquid column from the reaction container, controlling the cleaning liquid column to move to a magnetic field interval, and then enabling the magnetic interception component to be far away from the reaction container;
specifically, the magnetic attraction interception component is close to the reaction container, so that the magnetic compound in the mixed liquid can be attracted magnetically, and the magnetic compound in the mixed solution is prevented from being discharged.
S4, controlling the reciprocating motion of the cleaning liquid column in the magnetic field interval for a target number of times, then enabling the magnetic interception component to be close to the reaction container, and discharging the cleaning liquid column from the reaction container;
specifically, the cleaning liquid column passes through the magnetic field rotating component repeatedly for a plurality of times to clean the free gold-core platinum nanoclusters, and the magnetic attraction interception component can prevent the magnetic compound from being discharged along with the cleaning liquid column.
S5, pumping the chromogenic substrate liquid column to a magnetic field interval, controlling the magnetic attraction interception component to be far away from the reaction container, controlling the magnetic attraction interception component to be close to the reaction container after the chromogenic substrate changes color, pumping the chromogenic substrate liquid column to the position below the image acquisition component, and then starting a background light source component and taking a picture;
specifically, the gold-core platinum nanocluster on the double-antibody sandwich structure can catalyze the chromogenic substrate, so that the chromogenic substrate is changed from colorless to blue.
S6: and processing and analyzing the image to obtain a saturation value of a color-changing chromogenic substrate area in the image, and calculating to obtain the bacterial content in the sample to be detected.
And further, the automatic detection of the microorganisms is realized, the dependence on various instruments and equipment and professional operators is reduced, the forming efficiency of the double-antibody sandwich structure is improved, the microorganism detection time is shortened, and the microorganism detection sensitivity is improved.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111051010.4A CN113721015B (en) | 2021-09-08 | 2021-09-08 | Automatic microorganism detection device, system and method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111051010.4A CN113721015B (en) | 2021-09-08 | 2021-09-08 | Automatic microorganism detection device, system and method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113721015A true CN113721015A (en) | 2021-11-30 |
| CN113721015B CN113721015B (en) | 2023-03-21 |
Family
ID=78682756
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111051010.4A Active CN113721015B (en) | 2021-09-08 | 2021-09-08 | Automatic microorganism detection device, system and method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113721015B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117420292A (en) * | 2023-11-07 | 2024-01-19 | 青岛华晶生物技术有限公司 | Incubator |
Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1598579A (en) * | 2003-09-18 | 2005-03-23 | 陕西西大北美基因股份有限公司 | Microfluid analytical system using magnetic microsphere as medium and ivestigating method thereof |
| WO2011062557A1 (en) * | 2009-11-23 | 2011-05-26 | Haiqing Gong | Improved microfluidic device and method |
| US20140099236A1 (en) * | 2011-05-09 | 2014-04-10 | Konica Minolta, Inc. | Microchip solution sending system |
| CN103760371A (en) * | 2013-12-23 | 2014-04-30 | 苏州科雷芯电子科技有限公司 | Method and device for rapidly detecting raw milk bacteria |
| WO2017035262A1 (en) * | 2015-08-24 | 2017-03-02 | Gpb Scientific, Llc | Methods and devices for multi-step cell purification and concentration |
| US9788763B1 (en) * | 2014-07-09 | 2017-10-17 | Verily Life Sciences Llc | Methods for magnetic particle capture and separation |
| US20170362563A1 (en) * | 2014-12-15 | 2017-12-21 | The Regents Of The University Of California | Method and device for separation of particles and cells using gradient magnetic ratcheting |
| CN108344615A (en) * | 2018-01-24 | 2018-07-31 | 杭州安弼晟生物科技有限公司 | A kind of circulating tumor cell capture systems and control method |
| US20190339261A1 (en) * | 2016-12-28 | 2019-11-07 | Ge Healthcare Bio-Sciences Ab | Magnetic Immunoglobulin-Binding Particles |
| CN110734853A (en) * | 2019-11-05 | 2020-01-31 | 中国农业大学 | Microorganism detection device and detection method |
| US10712262B1 (en) * | 2019-02-21 | 2020-07-14 | East China Jiaotong University | Rare earth solution image capture device and method |
| CN112752847A (en) * | 2018-09-28 | 2021-05-04 | 奥克泰生物科技股份有限公司 | Magnetic separation |
| US11000780B1 (en) * | 2019-12-06 | 2021-05-11 | AnuCell Biosystems Limited | Torus reactor for a combined cell isolator and bioreactor |
-
2021
- 2021-09-08 CN CN202111051010.4A patent/CN113721015B/en active Active
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1598579A (en) * | 2003-09-18 | 2005-03-23 | 陕西西大北美基因股份有限公司 | Microfluid analytical system using magnetic microsphere as medium and ivestigating method thereof |
| WO2011062557A1 (en) * | 2009-11-23 | 2011-05-26 | Haiqing Gong | Improved microfluidic device and method |
| US20140099236A1 (en) * | 2011-05-09 | 2014-04-10 | Konica Minolta, Inc. | Microchip solution sending system |
| CN103760371A (en) * | 2013-12-23 | 2014-04-30 | 苏州科雷芯电子科技有限公司 | Method and device for rapidly detecting raw milk bacteria |
| US9788763B1 (en) * | 2014-07-09 | 2017-10-17 | Verily Life Sciences Llc | Methods for magnetic particle capture and separation |
| US20170362563A1 (en) * | 2014-12-15 | 2017-12-21 | The Regents Of The University Of California | Method and device for separation of particles and cells using gradient magnetic ratcheting |
| WO2017035262A1 (en) * | 2015-08-24 | 2017-03-02 | Gpb Scientific, Llc | Methods and devices for multi-step cell purification and concentration |
| US20190339261A1 (en) * | 2016-12-28 | 2019-11-07 | Ge Healthcare Bio-Sciences Ab | Magnetic Immunoglobulin-Binding Particles |
| CN108344615A (en) * | 2018-01-24 | 2018-07-31 | 杭州安弼晟生物科技有限公司 | A kind of circulating tumor cell capture systems and control method |
| CN112752847A (en) * | 2018-09-28 | 2021-05-04 | 奥克泰生物科技股份有限公司 | Magnetic separation |
| US10712262B1 (en) * | 2019-02-21 | 2020-07-14 | East China Jiaotong University | Rare earth solution image capture device and method |
| CN110734853A (en) * | 2019-11-05 | 2020-01-31 | 中国农业大学 | Microorganism detection device and detection method |
| US11000780B1 (en) * | 2019-12-06 | 2021-05-11 | AnuCell Biosystems Limited | Torus reactor for a combined cell isolator and bioreactor |
Non-Patent Citations (1)
| Title |
|---|
| LI XUE等: "An impedance biosensor based on magnetic nanobead net and MnO2 nanoflowers for rapid and sensitive detection of foodborne bacteria", 《BIOSENSORS AND BIOELECTRONIC》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117420292A (en) * | 2023-11-07 | 2024-01-19 | 青岛华晶生物技术有限公司 | Incubator |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113721015B (en) | 2023-03-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104483499B (en) | Full-automatic specific protein analyser | |
| JP6348666B2 (en) | Light irradiation multi-sample parallel reaction device | |
| CN108845154B (en) | Immunity quantitative analyzer | |
| CN104614521B (en) | Microfluidic chip-based immunoaggregation detection method, chip and system | |
| CN205333638U (en) | POCT chemiluminescent immunoassay analytic system | |
| CN105203746A (en) | POCT (Point of Care Testing) chemiluminescence immunoassay system and method | |
| CN113721015A (en) | Automatic microorganism detection device, system and method | |
| JP6072891B2 (en) | Bioanalytical device and biomolecule analyzer | |
| CN110883001A (en) | A kind of analyzer magnetic bead cleaning device | |
| CN112424603B (en) | Method and apparatus for performing heterogeneous immunoassays | |
| CN111239128A (en) | Vaginal secretion detector and detection method thereof | |
| CN204462161U (en) | Protein analyzer liquid feeding mixed structure | |
| CN111721948B (en) | Biochemical and immunological integrated analyzer and detection method | |
| CN208140724U (en) | Automatic lmunoassays analyzer | |
| CN115326695B (en) | Indoor marine corrosion test device | |
| JP3213798B2 (en) | Chemical analyzer | |
| JP2013238420A (en) | Sample analysis apparatus and sample analysis method | |
| CN109459287B (en) | Continuous magnetic separation device and method based on micro-fluidic chip | |
| CN117990557B (en) | Rapid detection device and detection method for soil water content | |
| US11579215B2 (en) | Sample analyzer | |
| CN204964275U (en) | Automatic metaplasia thing sample pretreatment device | |
| CN203232040U (en) | Reaction disc | |
| JP3803399B2 (en) | Immunoassay device | |
| CN107807075A (en) | A kind of erythrocyte sedimentation rate automatic detection device | |
| KR20230106600A (en) | Inspection device and inspection method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |