CN113721015A - Automatic microorganism detection device, system and method - Google Patents
Automatic microorganism detection device, system and method Download PDFInfo
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Abstract
The invention provides a microbial automatic detection device, a system and a method, comprising a workbench, two fixing pieces, a reaction container, a magnetic field rotating part, two liquid level detection parts, a magnetic attraction interception part, a background light source part and an image acquisition part; the two fixing pieces are respectively and fixedly arranged on two sides of the workbench, and the magnetic field rotating part is used for forming a magnetic field with magnetic induction lines distributed in parallel at the center; one end of the reaction vessel is arranged on one of the fixed pieces, and the other end of the reaction vessel penetrates through the center of the magnetic field rotating part and is connected with the other fixed piece; the two liquid level detection components are respectively arranged at two sides of the magnetic field rotating component and are connected with the reaction vessel; the magnetic attraction interception component is positioned between the two liquid level detection components; the image acquisition part is positioned above the background light source part. The invention realizes the automatic detection of the microorganism, reduces the dependence on instruments and equipment and professional operators, shortens the detection time of the microorganism and improves the detection sensitivity of the microorganism.
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)
1. An automatic microorganism detection device is characterized by comprising 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;
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.
2. The automated microorganism detection apparatus according to claim 1, wherein the magnetic field rotation unit comprises a rotary holder, a rotary driver and an annular halbach array magnet, the rotary holder is mounted between the two liquid level detection units, the rotary driver and the annular halbach array magnet are both mounted on the rotary holder, and the rotary driver is connected with the annular halbach array magnet.
3. The automated microorganism detection apparatus according to claim 2, wherein the annular halbach array magnet comprises a plurality of magnets arranged in a halbach array and a magnetic lock ring enclosing the magnets.
4. The automated microorganism detection apparatus according to any one of claims 1 to 3, wherein the liquid level detection unit comprises a liquid level detection holder and a liquid level sensor, the liquid level detection holder is mounted on one side of the reaction vessel, the liquid level sensor is mounted on the liquid level detection holder, and the liquid level sensor is connected to the reaction vessel.
5. The automatic microorganism detection device according to any one of claims 1 to 3, wherein the magnetic attraction trapping 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 installed 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 located above the reaction container.
6. The automated microbial detection apparatus of any one of claims 1-3, wherein the background light source comprises a PC light diffusion plate and a plurality of light emitting LEDs, the light emitting LEDs are fixedly mounted on the worktable, the PC light diffusion plate covers the light emitting LEDs, and the PC light diffusion plate is located below the image acquisition component.
7. The automated microorganism detection apparatus according to any one of claims 1 to 3, wherein the image capturing part comprises an image capturing driving part and a camera, the image capturing driving part is fixedly installed at one side of the reaction container, the camera is installed on the image capturing driving part, and the reaction container is located between the camera and the background light source part.
8. An automatic microorganism detection system, which is characterized by comprising a detection box and an automatic microorganism detection device as claimed in any one of claims 1 to 7, wherein a cavity is arranged in the detection box, the automatic microorganism 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.
9. The automated microbial detection system of claim 8, further comprising a waste liquid pool, wherein the waste liquid pool is fixedly mounted 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.
10. An automated microbial detection method, comprising:
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.
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