CN114200134B - Automatic rapid detection device, system and method for microorganism detection - Google Patents
Automatic rapid detection device, system and method for microorganism detection Download PDFInfo
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- CN114200134B CN114200134B CN202111422337.8A CN202111422337A CN114200134B CN 114200134 B CN114200134 B CN 114200134B CN 202111422337 A CN202111422337 A CN 202111422337A CN 114200134 B CN114200134 B CN 114200134B
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- 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
- G01N33/56916—Enterobacteria, e.g. shigella, salmonella, klebsiella, serratia
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- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- 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
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- 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
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
- G01N33/587—Nanoparticles
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- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- G01N2333/255—Salmonella (G)
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- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/10—Detection of antigens from microorganism in sample from host
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Abstract
The invention provides an automatic rapid detection device, system and method for microorganism detection, comprising a microfluidic chip and a rotary cavity; the micro-fluidic chip is provided with a through hole matched with the rotary cavity, the micro-fluidic chip is also provided with a plurality of liquid storage cavities, the liquid storage cavities are communicated with the through hole, the rotary cavity is provided with a first communication structure and a second communication structure which are communicated with the rotary cavity, and the rotary cavity is installed in the through hole and can rotate relative to the through hole, so that the first communication structure can be communicated with any one of the liquid storage cavities. The invention realizes the automatic operation of various immune reactions, can carry out rapid automatic detection on microorganisms, does not need professional operation, saves a great deal of manpower and time, shortens the detection time of microorganisms, reduces the influence of artificial operation errors on detection results, and improves the detection sensitivity of microorganisms.
Description
Technical Field
The invention relates to the technical field of microorganism detection, in particular to an automatic rapid detection device, system and method for microorganism detection.
Background
The background in the food sample is complex, the concentration of food-borne pathogenic bacteria is usually low, and the conventional detection method is difficult to directly detect the food sample. The double antibody sandwich technology is a biological detection technology based on antigen-antibody immune combination, utilizes a capture probe combined with a specific antibody to capture target bacteria, then uses a signal probe combined with another antibody to mark the target bacteria, forms an immune capture probe-target bacteria-immune signal probe double-antibody sandwich structure, and uses the signal probe to convert corresponding bacterial concentration signals into light, heat, magnetism, force, sound, electricity and other detectable physical signals to indirectly detect the target bacterial concentration. However, when the existing double-antibody sandwich technology is applied to microorganism detection, most of the double-antibody sandwich technology depends on various instruments and professional operators, the degree of automation is not high, and the double-antibody sandwich technology is not suitable for detection scenes with limited conditions such as a base layer.
In recent years, the biosensor based on the microfluidic chip has been greatly developed due to the advantages of simple operation, low cost, small sample size, high sensitivity, rapid response speed, field detection and the like. However, the existing microfluidic biosensor still needs a complex pumping system when in use, and generally each reagent inlet needs an independent pump for sample addition, and needs professional personnel to perform manual operation, so that manpower cannot be further liberated, and human operation errors are reduced. Therefore, how to increase the automation degree of microorganism detection to realize rapid microorganism detection is an important issue to be solved in the industry.
Disclosure of Invention
The invention provides an automatic rapid detection device, system and method for detecting microorganisms, which are used for solving the defects that the degree of automation is low, a large amount of manpower and time are consumed, detection accuracy is low due to manual operation of professionals, and the like when a microfluidic biosensor in the prior art is used, so that the automation operation of various immune reactions is realized, the microorganisms can be rapidly and automatically detected, the operation of professionals is not needed, a large amount of manpower and time are saved, the microorganism detection time is shortened, the influence of manual operation errors on detection results is reduced, and the microorganism detection sensitivity is improved.
The invention provides an automatic rapid detection device for microorganism detection, which comprises a microfluidic chip and a rotary cavity, wherein the microfluidic chip is arranged on the rotary cavity;
the micro-fluidic chip is provided with a through hole matched with the rotary cavity, the micro-fluidic chip is also provided with a plurality of liquid storage cavities, and the liquid storage cavities are communicated with the through hole
The rotary cavity is provided with a first communication structure and a second communication structure which are communicated with the rotary cavity, and the rotary cavity is installed in the through hole and can rotate relative to the through hole, so that the first communication structure can be communicated with any one of the liquid storage cavities.
According to the automatic rapid detection device for microorganism detection provided by the invention, the microfluidic chip further comprises a light source assembly, and the light source assembly is positioned below the liquid storage cavity.
According to the automatic rapid detection device for detecting microorganisms, the microfluidic chip further comprises a magnetic component, and the magnetic component is located below one of the liquid storage cavities.
According to the automatic rapid detection device for microorganism detection provided by the invention, the rotary cavity comprises an upper cavity and a lower cavity, and the lower end of the upper cavity is provided with a clamping groove matched with the upper end of the lower cavity.
According to the automatic rapid detection device for microorganism detection provided by the invention, the bottom of the lower cavity is provided with the notch.
The invention also provides an automatic rapid detection system for detecting microorganisms, which comprises a detection box, wherein a driving piece, a pumping piece and the automatic rapid detection device for detecting microorganisms are arranged in the detection box;
the automatic rapid detection device for detecting microorganisms is fixedly arranged in the detection box;
the driving piece is connected with the rotating cavity and is used for driving the rotating cavity to rotate;
the pumping member communicates with the second communication structure.
According to the automatic rapid detection system for microorganism detection provided by the invention, the automatic rapid detection system for microorganism detection further comprises an image acquisition component, wherein the image acquisition component is fixedly arranged in the detection box and is positioned above one of the liquid storage cavities.
According to the automatic rapid detection system for microorganism detection provided by the invention, the detection box comprises a box body and a darkroom cover, and the darkroom cover is rotatably connected with the box body.
According to the automatic rapid detection system for microorganism detection provided by the invention, the automatic rapid detection system for microorganism detection further comprises a microcontroller, wherein the microcontroller is arranged in the box body and is electrically connected with the image acquisition component.
The invention also provides an automatic rapid detection method for detecting microorganisms, which comprises the following steps:
respectively injecting a sample to be detected and various solutions required by detection into corresponding liquid storage cavities;
rotating the rotating cavity to enable the rotating cavity to be communicated with any liquid storage cavity through a first communication structure;
extracting the solution in the liquid storage cavity into the rotating cavity or conveying the solution in the rotating cavity into the liquid storage cavity.
According to the automatic rapid detection device, system and method for detecting microorganisms, solutions such as detection samples and detection reagents required by microorganism detection are respectively injected into corresponding liquid storage cavities. The rotary cavity is rotated by applying an external force to the rotary cavity, and the second communication structure is connected with the pumping equipment, so that positive pressure or negative pressure is formed in the rotary cavity through the pumping equipment. Through rotating rotatory cavity for first communication structure can communicate with arbitrary stock solution chamber, and when first communication structure and stock solution chamber communicate, pumping equipment forms malleation or negative pressure in the stock solution intracavity through the second communication structure, and then takes out the solution in the stock solution chamber into rotatory cavity or carries the solution in the rotatory cavity into the stock solution chamber. The staff can carry out corresponding transfer and mixing on different solutions according to detection requirements, so that the operations of sample adding, mixing, incubation, separation and enrichment, cleaning and the like in the conventional microorganism immunodetection experiment are realized, and the application of pumps and valves is reduced. And furthermore, the automatic operation of various immune reactions is realized, the microorganisms can be rapidly and automatically detected, the operation of professional personnel is not needed, a large amount of manpower and time are saved, the microorganism detection time is shortened, the influence of artificial operation errors on the detection result is reduced, and the microorganism detection sensitivity is improved.
Drawings
In order to more clearly illustrate the invention or the technical solutions of the prior art, the following description will briefly explain the drawings used in the embodiments or the description of the prior art, and it is obvious that the drawings in the following description are some embodiments of the invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram of an automated rapid detection device for microbiological detection according to the present invention;
FIG. 2 is a cross-sectional view of a rotating chamber of an automated rapid detection device for microbiological detection provided by the present invention;
fig. 3 is a schematic structural diagram of a microfluidic chip of the automated rapid detection device for detecting microorganisms according to the present invention;
FIG. 4 is a schematic diagram of the structure of an automated rapid detection system for microbiological detection provided by the present invention;
FIG. 5 is a schematic view of a portion of an automated rapid detection system for microbiological detection provided by the present invention;
FIG. 6 is a flow chart of an automated rapid detection method for microbiological detection provided by the present invention;
reference numerals:
1: a microfluidic chip; 2: rotating the cavity; 3: a detection box;
4: an image acquisition component; 5: a microcontroller; 11: a through hole;
12: a liquid storage cavity; 13: a light source assembly; 14: a magnetic attraction component;
15: a magnet; 21: a first communication structure; 22: a second communication structure;
23: an upper cavity; 24: a lower cavity; 25: a clamping groove;
26: a notch; 31: a driving member; 32: a pumping member;
33: a case; 34: a darkroom cover.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings, and it is apparent that the described embodiments are some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The automated rapid detection apparatus, system and method for microbiological detection of the present invention are described below in conjunction with fig. 1-6.
As shown in fig. 1, 2 and 3, an automated rapid detection device for microorganism detection includes a microfluidic chip 1 and a rotary chamber 2.
Specifically, the micro-fluidic chip 1 is provided with a through hole 11 matched with the rotary cavity 2, the micro-fluidic chip 1 is also provided with a plurality of liquid storage cavities 12, and the liquid storage cavities 12 are communicated with the through hole 11
The rotary cavity 2 is provided with a first communication structure 21 and a second communication structure 22 which are communicated with the rotary cavity 2, and the rotary cavity 2 is installed in the through hole 11 and can rotate relative to the through hole 11, so that the first communication structure 21 can be communicated with any one liquid storage cavity 12 of the liquid storage cavities 12.
In use, solutions such as a detection sample and a detection reagent required for microorganism detection are injected into the respective liquid storage chambers 12. By applying an external force to the rotary chamber 2, the rotary chamber 2 is rotated while the second communication structure 22 is connected to the pumping device, by which positive or negative pressure is formed in the rotary chamber 2. By rotating the rotary cavity 2, the first communication structure 21 can be communicated with any liquid storage cavity 12, and when the first communication structure 21 is communicated with the liquid storage cavity 12, the pumping equipment forms positive pressure or negative pressure in the liquid storage cavity 12 through the second communication structure 22, so that the solution in the liquid storage cavity 12 is pumped into the rotary cavity 2 or the solution in the rotary cavity 2 is conveyed into the liquid storage cavity 12. The staff can carry out corresponding transfer and mixing on different solutions according to detection requirements, so that the operations of sample adding, mixing, incubation, separation and enrichment, cleaning and the like in the conventional microorganism immunodetection experiment are realized, and the application of pumps and valves is reduced. And furthermore, the automatic operation of various immune reactions is realized, the microorganisms can be rapidly and automatically detected, the operation of professional personnel is not needed, a large amount of manpower and time are saved, the microorganism detection time is shortened, the influence of artificial operation errors on the detection result is reduced, and the microorganism detection sensitivity is improved.
When the automatic rapid detection device for detecting microorganisms is applied to the double-antibody sandwich technology for detecting microorganisms:
firstly, immune magnetic beads modified with anti-salmonella polyclonal antibody, sample solution to be detected, gold core platinum nanocluster solution (signal probe) modified with anti-salmonella monoclonal antibody, cleaning solution and TMB-H 2 O 2 Chromogenic substrates are injected into the different reservoirs 12 separately.
Then the rotary cavity 2 is rotated, so that the rotary cavity 2 is communicated with the liquid storage cavity 12 through the first communication structure 21, and the sample to be tested, the immunomagnetic beads and the signal probe are sequentially sucked into the rotary cavity 2. One of the liquid storage cavities 12 is selected as a mixing cavity, the rotating cavity 2 is rotated to enable the rotating cavity 2 to be communicated with the mixing cavity, mixed solution in the rotating cavity 2 is pumped back and forth between the rotating cavity 2 and the mixing cavity for a plurality of times, mixing and incubation are achieved, and the double-antibody sandwich structure of the immune magnetic bead-target bacteria-gold core platinum nanocluster is formed under the immune reaction effect.
Then, one of the liquid storage cavities 12 is selected as a capturing cavity, the rotating cavity 2 is rotated to enable the rotating cavity 2 to be communicated with the capturing cavity, the mixed solution is pumped into the capturing cavity, the mixture is kept stand in the capturing cavity for a period of time, and the double-antibody sandwich structure is captured by using a magnet arranged below the capturing cavity. One of the liquid storage cavities 12 is selected as a waste liquid cavity, after the double-antibody sandwich structure is captured and attached to the bottom of the capturing cavity, the solution in the capturing cavity is sucked into the rotating cavity 2, and the rotating cavity 2 is rotated to pump the solution in the capturing cavity to the waste liquid cavity, and the double-antibody sandwich structure is remained in the capturing cavity.
The rotating chamber 2 is then rotated such that the rotating chamber 2 communicates with the liquid storage chamber 12 in which the cleaning liquid is stored, drawing the cleaning liquid into the rotating chamber 2. And then pumping the cleaning liquid back and forth between the rotary cavity 2 and the capturing cavity for a plurality of times, and cleaning the double-antibody sandwich structure adsorbed in the capturing cavity.
One of the liquid storage cavities 12 is selected as a detection cavity, the rotary cavity 2 is rotated to pump a chromogenic substrate to a capturing cavity, and the gold core platinum nanocluster on the double-antibody sandwich structure is used as a catalyst to catalyze TMB-H 2 O 2 The redox reaction takes place to turn the originally colorless transparent solution into blue, and then the blue solution is pumped into the detection chamber and photographed. And finally, processing and analyzing the image to obtain a saturation value of the blue solution image, and calculating the target bacteria content in the sample to be detected according to a pre-calibrated saturation-bacteria concentration curve. And further, the rapid automatic detection of the microorganisms is realized, the operation of professionals is not needed, a large amount of manpower and time are saved, the microorganism detection time is shortened, the influence of artificial operation errors on the detection result is reduced, and the microorganism detection sensitivity is improved.
In an alternative embodiment of the present invention, the number of the liquid storage chambers 12 may be increased or decreased according to actual requirements.
In an alternative embodiment of the present invention, the first communication structure 21 is, for example, a through hole. It should be appreciated that the first communication structure 21 may be any other suitable structure.
Wherein in an alternative embodiment of the present invention, the second communication structure 22 is, for example, a connection tube. It should be appreciated that the second communication structure 22 may be any other suitable structure.
Wherein, as shown in fig. 1 and 2, the first communication structure 21 is located below the second communication structure 22. In use, the rotating chamber 2 can transfer and mix the solution between the first communication structure 21 and the liquid storage chamber 12, avoiding the solution from entering the second communication structure 22.
Wherein in an alternative embodiment of the invention the first communication structure 21 is for example located in a lower part of the rotation chamber 2 and the second communication structure 22 is for example located in an upper part of the rotation chamber 2. It should be appreciated that the first communication structure 21 and the second communication structure 22 may also be located at any other suitable location on the rotating chamber 2.
Further, as shown in fig. 1 and fig. 2, the rotating cavity 2 includes an upper cavity 23 and a lower cavity 24, and a clamping groove 25 matched with the upper end of the lower cavity 24 is provided at the lower end of the upper cavity 23. When in use, the upper end of the lower cavity 24 is clamped into the clamping groove 25 of the upper cavity 23, so that the upper cavity 23 and the lower cavity 24 can be connected through interference fit to form the rotary cavity 2, and the rotary cavity 2 is manufactured by dividing the rotary cavity 2 into the upper cavity 23 and the lower cavity 24, so that the manufacturing difficulty is reduced compared with the manufacturing difficulty of directly manufacturing the rotary cavity 2.
Wherein, as shown in fig. 1 and 2, the bottom of the lower cavity 24 is provided with a notch 26. When in use, the motor can be quickly connected with the lower cavity 24 through the notch 26, so that the motor can drive the lower cavity 24 to rotate, and the automation of microorganism detection is conveniently realized.
Further, as shown in fig. 1, the microfluidic chip 1 further includes a light source assembly 13, and the light source assembly 13 is located below the liquid storage cavity 12. When the device is used, the light source assembly 13 emits light to the liquid storage cavity 12 which has completed the reaction, so that an illumination or fluorescent material excitation light source is provided for obtaining the image in the liquid storage cavity 12, the image data in the liquid storage cavity 12 after the reaction can be directly and accurately obtained, and the image data in the liquid storage cavity 12 can be analyzed to obtain the microorganism concentration. In practical application, the light source assembly 13 can be installed below the liquid storage cavity 12 serving as the detection cavity, so that illumination can be accurately provided for the detection cavity.
Wherein in an alternative embodiment of the invention the light source assembly 13 is for example an LED light source. It should be appreciated that the light source assembly 13 may be any other suitable light source.
Further, as shown in fig. 1, the microfluidic chip 1 further includes a magnetic component 14, where the magnetic component 14 is located below one of the liquid storage cavities 12. When the magnetic detection device is used, the liquid storage cavity 12 above the magnetic component 14 is used as a capturing cavity, and then a magnetic attraction force is formed at the capturing cavity, after the mixed solution obtained by mixing the detection sample and the detection solution is conveyed into the capturing cavity, the magnetic component 14 can adsorb reactants in the mixed solution into the capturing cavity, and further the capturing of the reactants is realized, so that the subsequent detection of the reactants is facilitated.
As shown in fig. 1, the magnetic attraction assembly 14 includes at least one magnet 15, the magnet 15 is installed in the microfluidic chip 1, and the magnet 15 is located below one of the liquid storage cavities 12. In use, the liquid storage cavity 12 above the magnet 15 is used as a capturing cavity, the mixed solution of the detection sample and the detection solution is conveyed into the capturing cavity, and reactants in the mixed solution can be captured through the magnet 15.
On the other hand, as shown in fig. 4 and 5, the present invention further provides an automated rapid detection system for detecting microorganisms, which comprises a detection box 3, wherein a driving member 31 and a pumping member 32 are arranged in the detection box 3, and an automated rapid detection device for detecting microorganisms is fixedly installed in the detection box 3;
the driving piece 31 is connected with the rotating cavity 2, and the driving piece 31 is used for driving the rotating cavity 2 to rotate;
the pumping member 32 communicates with the second communication structure 22.
When in use, the driving piece 31 drives the rotary cavity 2 to rotate, so that the first communication structure 21 on the rotary cavity 2 can be communicated with any liquid storage cavity 12, and the pumping piece 32 is used for pumping liquid in the liquid storage cavity 12 into the rotary cavity 2 or conveying liquid in the rotary cavity 2 into the liquid storage cavity 12 when the rotary cavity 2 is communicated with the liquid storage cavity 12. The method ensures that staff can carry out corresponding transfer and mixing on the solution in each liquid storage cavity 12 according to the detection requirement, realizes the operations of sample adding, mixing, incubation, separation and enrichment, cleaning and the like in the conventional microorganism immunodetection experiment, and reduces the application of pumps and valves. And furthermore, the automatic operation of various immune reactions is realized, the microorganisms can be rapidly and automatically detected, the operation of professional personnel is not needed, a large amount of manpower and time are saved, the microorganism detection time is shortened, the influence of artificial operation errors on the detection result is reduced, and the microorganism detection sensitivity is improved.
In an alternative embodiment of the invention, the driving member 31 is, for example, a rotating electric machine. It should be appreciated that the drive 31 may be any other suitable rotational drive.
In an alternative embodiment of the present invention, pumping member 32 is, for example, a liquid delivery pump. It should be appreciated that pumping member 32 may be any other suitable liquid delivery device.
Further, as shown in fig. 4, the detection box 3 includes a box body 33 and a darkroom cover 34, and the darkroom cover 34 is rotatably connected to the box body 33. When the automatic rapid detection device for detecting microorganisms is used, the automatic rapid detection device for detecting microorganisms is arranged on the upper surface of the box body 33, then the darkroom cover 34 is rotated, so that the darkroom cover 34 is covered on the box body 33, a darkroom is formed between the box body 33 and the darkroom cover 34, the detection of microorganisms and the acquisition of image data in the darkroom are facilitated, the interference of external light to experiments is avoided, and the errors of the experiments are reduced.
As shown in fig. 4 and fig. 5, the microfluidic chip 1 is fixedly mounted on the upper surface of the case 33, and the microfluidic chip 1 is located between the case 33 and the darkroom cover 34. When in use, the darkroom cover 34 is closed, a darkroom is formed between the box body 33 and the darkroom cover 34, the micro-fluidic chip 1 is positioned in the darkroom, and the microorganism detection is performed in the darkroom, so that the sensitivity of the microorganism detection is improved.
Further, as shown in fig. 4 and fig. 5, the automated rapid detection system for detecting microorganisms further includes an image acquisition assembly 4, wherein the image acquisition assembly 4 is fixedly installed in the detection box 3, and the image acquisition assembly 4 is located above one of the liquid storage cavities 12. When the device is used, the liquid storage cavity 12 positioned below the image acquisition component 4 is used as a detection cavity, after the solution in the detection cavity is reacted, the image acquisition component 4 photographs the detection cavity, then the image acquisition component 4 transmits the photographed image data to a corresponding processor, and the concentration of microorganisms can be known through the processing and analysis of the image data by the processor.
Wherein in an alternative embodiment of the invention, the image acquisition assembly 4 is fixedly mounted on the darkroom cover 34, for example, and the image acquisition assembly 4 is located just above the detection chamber when the darkroom cover 34 is closed. It should be appreciated that the image acquisition assembly 4 may be mounted in any other suitable location that enables image acquisition of the detection chamber.
Further, as shown in fig. 5, the automated rapid detection system for detecting microorganisms further includes a microcontroller 5, wherein the microcontroller 5 is mounted in the case 33, and the microcontroller 5 is electrically connected to the image acquisition assembly 4. When the device is used, one liquid storage cavity 12 is selected as the last detection cavity, after the reaction in the detection cavity is completed, the image acquisition component 4 shoots the detection cavity, then the image data in the detection cavity is acquired and transmitted to the microcontroller 5, and the microcontroller 5 analyzes and processes the image data to obtain quantized data, so that the concentration of microorganisms can be obtained.
Wherein in an alternative embodiment of the invention the microcontroller 5 is for example a raspberry pie. It should be appreciated that the microcontroller 5 may also be any other suitable controller.
In another aspect, as shown in fig. 6, the present invention further provides an automated rapid detection method for detecting microorganisms, including:
s1, respectively injecting a sample to be detected and various solutions required by detection into corresponding liquid storage cavities;
specifically, according to the type of microorganism to be detected and the detection method, the corresponding detection solutions are respectively injected into the corresponding liquid storage cavities 12. For example, when the double antibody sandwich method is used for detecting the bacterial concentration of a sample, the sample to be detected, the immunomagnetic beads, the signal probe, the chromogenic substrate and the cleaning liquid are respectively injected into different liquid storage cavities 12 of the microfluidic chip 1.
S2, rotating the rotary cavity to enable the rotary cavity to be communicated with any liquid storage cavity through a first communication structure;
specifically, by driving the rotary cavity 2 to rotate by using a driving device such as a stepping motor, the rotary cavity 2 is controlled to rotate by a corresponding angle, so that the first communication structure 21 on the rotary cavity 2 can be communicated with different liquid storage cavities 12, and the rotary cavity 2 can be communicated with different liquid storage cavities 12.
S3, extracting the solution in the liquid storage cavity into the rotating cavity or conveying the solution in the rotating cavity into the liquid storage cavity;
specifically, the liquid pumping device is communicated with the rotary cavity 2 to form positive pressure or negative pressure in the rotary cavity 2, so that the solution in the liquid storage cavity 12 can be pumped into the rotary cavity 2 or the solution in the rotary cavity 2 can be conveyed into the liquid storage cavity 12, further, different solutions can be correspondingly transferred and mixed according to detection requirements, and the application of pumps and valves is reduced. And furthermore, the automatic operation of various immune responses can be realized, the microorganisms can be rapidly and automatically detected, the operation of professional personnel is not needed, a large amount of manpower and time are saved, the microorganism detection time is shortened, the influence of human operation errors on the detection result is reduced, and the microorganism detection sensitivity is improved.
For example, when the automated rapid detection method for microorganism detection according to the present invention is applied to a double antibody sandwich method for detecting a bacterial concentration of a sample, a sample to be detected, an immunomagnetic bead, and a signaling probe are sequentially inhaled into the rotary chamber 2. One of the liquid storage cavities 12 is selected as a mixing cavity, the rotating cavity 2 is rotated to enable the rotating cavity 2 to be communicated with the mixing cavity, mixed solution in the rotating cavity 2 is pumped back and forth between the rotating cavity 2 and the mixing cavity for a plurality of times, mixing and incubation are achieved, and the double-antibody sandwich structure of the immune magnetic bead-target bacteria-gold core platinum nanocluster is formed under the immune reaction effect.
Then, one of the liquid storage cavities 12 is selected as a capturing cavity, the rotating cavity 2 is rotated to enable the rotating cavity 2 to be communicated with the capturing cavity, the mixed solution is pumped into the capturing cavity, the mixture is kept stand in the capturing cavity for a period of time, and the double-antibody sandwich structure is captured by using a magnet arranged below the capturing cavity. One of the liquid storage cavities 12 is selected as a waste liquid cavity, after the double-antibody sandwich structure is captured and attached to the bottom of the capturing cavity, the solution in the capturing cavity is sucked into the rotating cavity 2, and the rotating cavity 2 is rotated to pump the solution in the capturing cavity to the waste liquid cavity, and the double-antibody sandwich structure is remained in the capturing cavity.
The rotating chamber 2 is then rotated such that the rotating chamber 2 communicates with the liquid storage chamber 12 in which the cleaning liquid is stored, drawing the cleaning liquid into the rotating chamber 2. And then pumping the cleaning liquid back and forth between the rotary cavity 2 and the capturing cavity for a plurality of times, and cleaning the double-antibody sandwich structure adsorbed in the capturing cavity.
One of the liquid storage cavities 12 is selected as a detection cavity, the rotary cavity 2 is rotated to pump a chromogenic substrate to a capturing cavity, and the gold core platinum nanocluster on the double-antibody sandwich structure is used as a catalyst to catalyze TMB-H 2 O 2 The redox reaction takes place to turn the originally colorless transparent solution into blue, and then the blue solution is pumped into the detection chamber and photographed. And finally, processing and analyzing the image to obtain a saturation value of the blue solution image, and calculating the target bacteria content in the sample to be detected according to a pre-calibrated saturation-bacteria concentration curve.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the 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 scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (9)
1. An automatic rapid detection device for detecting microorganisms is characterized by comprising a microfluidic chip and a rotary cavity;
the micro-fluidic chip is provided with a through hole matched with the rotary cavity, and is also provided with a plurality of liquid storage cavities which are communicated with the through hole;
the rotary cavity is provided with a first communication structure and a second communication structure which are communicated with the rotary cavity, and the rotary cavity is arranged in the through hole and can rotate relative to the through hole, so that the first communication structure can be communicated with any one liquid storage cavity of a plurality of liquid storage cavities;
the first communication structure is positioned below the second communication structure, and the second communication structure is connected with pumping equipment;
the rotary cavity comprises an upper cavity and a lower cavity, wherein the lower end of the upper cavity is provided with a clamping groove matched with the upper end of the lower cavity, and the upper cavity is connected with the lower cavity through interference fit.
2. The automated rapid detection device for microbiological detection of claim 1, wherein the microfluidic chip further comprises a light source assembly positioned below the reservoir.
3. The automated rapid detection device for microbiological detection according to claim 1 or 2, wherein the microfluidic chip further comprises a magnetic assembly located below one of the reservoir chambers.
4. An automated rapid detection device for microbiological detection according to claim 1 or 2, wherein the bottom of the lower cavity is provided with a notch.
5. An automated rapid detection system for the detection of microorganisms, comprising a detection tank, wherein a driving member, a pumping member and the automated rapid detection device for the detection of microorganisms according to any one of claims 1 to 4 are disposed inside the detection tank;
the automatic rapid detection device for detecting microorganisms is fixedly arranged in the detection box;
the driving piece is connected with the rotating cavity and is used for driving the rotating cavity to rotate;
the pumping member communicates with the second communication structure.
6. The automated rapid detection system for microbiological detection of claim 5 further comprising an image acquisition assembly fixedly mounted within the detection case, the image acquisition assembly being positioned above one of the liquid storage cavities.
7. The automated rapid detection system for microbiological detection of claim 6 wherein the detection case comprises a case and a darkroom cover rotatably coupled to the case.
8. The automated rapid detection system for microbiological detection of claim 7 further comprising a microcontroller mounted within the housing, the microcontroller being electrically connected to the image acquisition assembly.
9. An automated rapid detection method for microbiological detection, characterized in that it is based on an automated rapid detection system for microbiological detection according to any one of claims 5 to 8, comprising:
respectively injecting a sample to be detected and various solutions required by detection into corresponding liquid storage cavities;
rotating the rotating cavity to enable the rotating cavity to be communicated with any liquid storage cavity through a first communication structure;
extracting the solution in the liquid storage cavity into the rotating cavity or conveying the solution in the rotating cavity into the liquid storage cavity;
sequentially sucking a sample to be detected, an immunomagnetic bead and a signal probe into the rotating cavity, selecting one of the liquid storage cavities as a mixing cavity, rotating the rotating cavity, so that the rotating cavity is communicated with the mixing cavity, and pumping the mixed solution in the rotating cavity back and forth between the rotating cavity and the mixing cavity for a plurality of times to realize mixing and incubation.
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