CN113341127B

CN113341127B - High-sensitivity multi-mycotoxin synchronous rapid detection robot and detection method

Info

Publication number: CN113341127B
Application number: CN202110605318.2A
Authority: CN
Inventors: 杨雪娇; 徐正华; 蔡翔宇; 陈庚超
Original assignee: Huangpu Customs Technical Center
Current assignee: Huangpu Customs Technical Center
Priority date: 2021-05-31
Filing date: 2021-05-31
Publication date: 2022-04-08
Anticipated expiration: 2041-05-31
Also published as: CN113341127A

Abstract

The invention discloses a high-sensitivity multi-mycotoxin synchronous rapid detection robot and a detection method; the device comprises a reaction filtering device, a driving device and a sample processing device, wherein a plurality of reaction tanks and filtering units are uniformly distributed on the edge of the reaction filtering device so as to realize the simultaneous detection of a plurality of mycotoxins; the antibody liquid box of the sample processing device is filled with the radiolabeled antibody, the mycotoxin in a reaction tank sample passing through the reaction filtering device is subjected to specific immunoreaction and sedimentation reaction, precipitates generated by the sedimentation reaction are filtered and separated through the filtering unit, the precipitates are eluted through the eluting device, after the precipitates are dried through the hot air drying device, the radioactive ray intensity is detected by utilizing the radioactive ray excitation fluorescent lens light-emitting principle, and the content of the mycotoxin is quantitatively calculated through the characteristic that the luminous intensity of the fluorescent lens is in direct proportion to the radioactivity size, so that the high-sensitivity rapid quantitative detection of various mycotoxins is realized.

Description

High-sensitivity multi-mycotoxin synchronous rapid detection robot and detection method

Technical Field

The invention relates to the field of mycotoxin detection, in particular to a high-sensitivity multi-mycotoxin synchronous rapid detection robot and a detection method.

Background

Mycotoxins are toxic secondary metabolites produced by certain fungi in the growth process, and more than 300 types are known at present, wherein more than 30 types of mycotoxins have strong toxicity to human beings and animals, and comprise aflatoxin, zearalenone, vomitoxin, ochratoxin, fumonisin and the like; these toxins can contaminate food, crops and their products extensively and cause poisoning of consumers, some can induce gene mutation and produce carcinogenicity, and some show toxicity to specific organs.

At present, two methods, namely a chromatography method and a test paper enzyme-linked immunosorbent assay method, are mainly adopted for detecting the mycotoxins, and the chromatography method has the limitations of complicated sample treatment, harsh experimental conditions, various experimental equipment, low detection efficiency and the like, and cannot be applied to the application scene of simultaneously carrying out rapid quantitative detection on various mycotoxins; the enzyme-linked immunosorbent assay with test paper has limited antibody amount, and can not combine with target mycotoxin in the sample sufficiently, resulting in poor detection result accuracy and failure in accurate quantitative detection of mycotoxin content.

Disclosure of Invention

The invention aims to overcome the defects and provide a high-sensitivity multi-mycotoxin synchronous rapid detection robot capable of rapidly and synchronously detecting multiple mycotoxins.

In order to achieve the purpose, the invention adopts the following specific scheme:

a high-sensitivity multi-mycotoxin synchronous rapid inspection robot comprises a reaction filtering device; the driving device is used for driving the reaction filtering device to rotate;

the reaction filtering device is rotatably arranged right above the driving device; comprises a reaction disc and a filter disc; a plurality of reaction grooves for containing each reaction component are uniformly distributed on the periphery of the reaction disc; the reaction components are all substances added into the reaction tank in the detection process; a plurality of filtering units which correspond to the reaction grooves in position and are used for filtering the reaction liquid after reaction are uniformly distributed on the periphery of the filtering disc;

the sample processing device is fixedly arranged at the top end of the driving device; the sample processing device comprises a plurality of antibody dripping devices for dripping antibody liquid into the reaction groove; antibody liquid with weak radioactivity corresponding to the type of the target mycotoxin to be detected is respectively contained in the plurality of antibody dripping devices; the antibodies in the antibody fluid can react with the corresponding fungal viruses in the test sample in an immune manner; the sample processing device also comprises a sample dripping device for dripping a sample into the reaction tank; the sample processing device also comprises a precipitation titration device which is used for dripping a precipitation reaction reagent into the reaction tank and carrying out precipitation reaction on the antibody-antigen combination obtained after immunoreaction to form a precipitate; the sample processing device also comprises an elution device for eluting the solution remained on the precipitate; the sample processing apparatus further comprises a radioactivity detecting device for detecting the radioactivity of the reaction precipitate.

Furthermore, the driving device also comprises a second fixing frame; a plurality of supporting columns for supporting are arranged on the periphery of the second fixing frame, and the tops of the supporting columns are fixedly connected with an intermediate connecting plate; the top of the intermediate connecting plate is provided with a waste liquid tank, and the bottom of the waste liquid tank is symmetrically provided with second through holes; a third through hole is formed in the center of the top surface of the middle connecting plate; waste liquid boxes are symmetrically arranged between the second fixing frame and the middle connecting plate; the liquid inlet of the waste liquid box is communicated with the second through hole; the driving device further comprises a bracket; the bracket is of a concave structure, and a fourth through hole is formed in the center of a concave plane at the bottom of the bracket; the left side and the right side of the fourth through hole are respectively provided with a first plane which is fixedly connected with the middle connecting plate;

the driving device further comprises a second belt wheel; one end of the second belt wheel is provided with a first cylindrical boss which is rotationally connected with the fourth through hole; the central part of the second belt wheel is provided with a profile hole; a first motor is fixedly installed on the left side of the bottom surface of the second fixing frame, and a first belt wheel is fixedly installed on the output end of the first motor; a second motor is fixedly installed on the right side of the bottom surface of the second fixing frame, and a third belt wheel is fixedly installed on the output end of the second motor; the third belt wheel drives the second belt wheel to synchronously rotate through a second synchronous belt; and a second electromagnet which is superposed with the axis of the molded surface hole is fixedly arranged in the middle of the second fixed frame.

The bottom of a reaction disc of the reaction filtering device is provided with a second cylindrical boss with a hollow structure; a fourth belt wheel is fixedly arranged on the end surface of the bottom of the reaction disc; the reaction disc is rotatably connected into the third through hole through the second cylindrical boss; the first belt pulley drives the fourth belt pulley to synchronously rotate through a first synchronous belt; the central part of the bottom surface of the filter disc is provided with a profile rod connected with the profile hole surface; the bottom of the profile rod is fixedly provided with a permanent magnet with the polarity opposite to that of the second electromagnet; the bottom of the filter disc is also fixedly connected with a piezoelectric ultrasonic vibrating reed used for driving the filter unit to vibrate and stir the liquid in the reaction tank.

Furthermore, the antibody dripping device also comprises an antibody liquid box for containing antibody liquid; a constant flow pump for quantitatively pumping the antibody liquid; a burette for adding an antibody liquid to the reaction tank; a magnetic lifting seat for fixing the burette; and the electromagnetic coil drives the magnetic lifting seat to move up and down.

The invention further comprises a guide cylinder fixedly connected to the top surface of the first fixing frame and a test solution groove for containing test solution, wherein the guide cylinder is arranged on the top surface of the first fixing frame; a limiting surface is arranged in the middle of the inner hole of the guide cylinder; a dropping head capable of sliding up and down is also arranged in the inner hole of the guide cylinder; a second spring is arranged between the guide cylinder and the dripping head; a second one-way valve is also arranged in the dripping head; the top of the dripping head is also provided with a three-way joint; a first one-way valve is arranged on the left side of the three-way joint; a liquid pipe is arranged on the left side of the first one-way valve; the second one-way valve is opposite to the first one-way valve in installation direction; the top of the three-way joint is also fixedly connected with a liquid suction barrel; a piston rod which forms a closed space with the liquid suction barrel is also connected in the liquid suction barrel in a sliding manner; a first spring is arranged between the liquid suction cylinder and the piston rod; the first spring rate is greater than the second spring.

Furthermore, the radioactivity detecting device also comprises an analyzing device for analyzing the area and the brightness of the light spot; the lens body is fixedly connected to the front end of the analysis device; the fluorescent lens is fixed on the front side of the lens body and used for ray induction luminescence; the surface of the fluorescent lens is coated with a fluorescent luminous coating; the convex lens is fixed in the middle of the lens body and used for amplifying the fluorescent light spots; the photosensitive film is used for sensing light imaging of the fluorescent plate.

The invention has the beneficial effects that: the antibody liquid is subjected to radioactive labeling, a plurality of antibody dripping devices are arranged at the same time, and a plurality of rotary reaction tanks are matched, so that the simultaneous addition of a plurality of antibody liquids is realized; dropwise adding the sample into the antibody liquid in each reaction tank by using a sample dropwise adding device to perform specific immunoreaction so as to combine mycotoxin; the antigen-antibody combination product is precipitated through a precipitation titration device, the precipitate is separated by matching with a filtering unit, the precipitate is eluted through an elution device, a fluorescent lens is excited by radioactive rays to emit light, and the light-emitting area and the light intensity are detected, so that high-sensitivity accurate quantitative detection on various mycotoxins is realized simultaneously.

The invention also provides a method for detecting the multi-mycotoxin by using the high-sensitivity multi-mycotoxin synchronous rapid detection robot, which comprises the following steps:

s1: mounting an antibody dripping device on a sample processing device;

s2: the driving device drives the filter disc to move downwards, and the filter units on the periphery of the filter disc are sleeved in the reaction tanks;

s3: the driving device drives the reaction disc and the filter disc to synchronously rotate, and drives each reaction tank and each filter unit to move to the position below the antibody dripping device corresponding to the mycotoxin species to be detected; the antibody dripping device drips excessive volume of antibody liquid into the reaction tank;

s4: the driving device drives the reaction disc and the filter disc to synchronously rotate, and drives each reaction tank and each filter unit to move to the lower part of the sample dripping device; the sample dripping device drips a fixed-volume sample into the reaction tank;

s5: standing for a certain time to mix the sample with the antibody solution and perform an immune combination reaction;

s6: the driving device drives the reaction disc and the filter disc to synchronously rotate, and drives each reaction tank and each filter unit to move to the lower part of the precipitation and titration device; the precipitation titration device is used for dropwise adding a precipitation reaction reagent into the reaction tank, so that the antibody which is immunologically combined with the mycotoxin is precipitated and separated out from the solution;

s7: the driving device drives the filter disc to drive each filter unit to move upwards, and precipitates in the reaction tank are separated from the residual liquid in the reaction tank;

s8: the driving device drives the filter disc to drive the filter unit to move to the lower part of the elution device; the elution device elutes the reaction liquid remained on the surface of the precipitate;

s9: the driving device drives the filter disc to drive the filter unit to move to the lower part of the radioactivity detection device, the radioactivity detection device emits light to generate light spots under the excitation of radioactive rays through the fluorescent lens, the light spots are projected on the photosensitive sheet after being amplified through the convex lens, the light intensity and the area of the light spots are automatically analyzed through the analysis device, and the radioactivity intensity of sediments is detected; and judging the content of the mycotoxin corresponding to the antibody in the sample according to the radioactivity intensity of the precipitate.

Compared with the existing mycotoxin detection method, the method has the beneficial effects that: the method has the advantages that the mycotoxin antibody liquid subjected to weak radioactive labeling treatment is set to perform specific immunoreaction with various mycotoxins in the sample respectively, and corresponding mycotoxins in the sample are combined, so that the rapid detection of various mycotoxins can be realized; the protein in the antibody-antigen binding state is precipitated by an isoelectric titration method, the protein is subjected to radioactivity detection after being filtered and separated, and the concentration of the mycotoxin of the corresponding species in the sample is quantitatively judged by the intensity of the radioactivity, so that the detection sensitivity of the mycotoxin can be remarkably improved.

Drawings

FIG. 1 is an exploded view of the present invention;

FIG. 2 is a perspective view of a sample processing device according to the present invention;

FIG. 3 is a sectional view of an antibody dropping device of the present invention;

FIG. 4 is a cross-sectional view of a sample dropping device according to the present invention;

FIG. 5 is a perspective view of the hot air drying device of the present invention;

FIG. 6 is a cross-sectional view of a radiation detecting device of the present invention;

FIG. 7 is a cross-sectional view of the drive of the present invention;

FIG. 8 is a perspective view in half section of a reaction filtration apparatus of the present invention;

FIG. 9 is a perspective view of the intermediate connecting plate of the present invention;

FIG. 10 is a perspective view in half section of a first pulley of the present invention;

FIG. 11 is a perspective view of the stand of the present invention;

FIG. 12 is a flow chart of the detection method of the present invention;

description of reference numerals: 1. a reaction filtration device; 2. a drive device; 3. a sample processing device; 31. an antibody dropping device; 32. a sample dropping device; 33. a precipitation titration device; 34. an elution device; 35. a hot air drying device; 36. a first fixing frame; 37. a radioactivity detecting device; 311. antibody liquid box; 312. a constant delivery pump; 313. a burette; 314. a magnetic lifting seat; 315. an electromagnetic coil; 321. a test solution tank; 322. a liquid pipe; 323. a first check valve; 324. a liquid suction cylinder; 324a, a liquid inlet hole; 324b, a liquid suction cavity; 325. a piston rod; 326. a first spring; 327. a dropping head; 328. a guide cylinder; 328a and a limit surface; 329. a second spring; 3210. a second one-way valve; 3211. a three-way joint; 3212. a first spring seat; 351. a first mounting plate; 352. a first electromagnet; 353. a third spring; 354. drying the blocks by hot air; 354a, a front protrusion; 354b, a first groove; 354c, a plurality of hot air holes; 354d, a first guide groove; 355. a guide rail; 371. a fluorescent lens; 372. a lens body; 373. a convex lens; 374. a photosensitive sheet; 375. an analysis device; 21. a second fixing frame; 21a, a support column; 22. an intermediate connection plate; 22a, a plurality of first through holes; 22b, a second through hole; 22c, a waste liquid tank; 22d, a third through hole; 23. a first synchronization belt; 24. a waste liquid box; 25. a first pulley; 26. a first motor; 27. a support; 27a, a fourth via; 27b, a first plane; 28. a second pulley; 28a, a profile hole; 28b, a first cylindrical boss; 28c, a belt groove; 29. a second electromagnet; 210. a second synchronous belt; 211. a third belt pulley; 212. a second motor; 11. a permanent magnet; 12. a reaction disc; 12a, a drain hole; 12b, a reaction tank; 12c, a second cylindrical boss; 13. an electromagnetic valve; 14. a piezoelectric ultrasonic vibrating piece; 15. a filter tray; 15a, a filtering tank body; 15b, a filtering unit; 15c, profile bar; 16. a filtration membrane; 17. a fourth pulley.

Detailed Description

The invention will be described in further detail with reference to the following figures and specific examples, without limiting the scope of the invention.

3. As shown in fig. 1 to fig. 11, the high-sensitivity multi-mycotoxin synchronous rapid inspection robot according to the present embodiment includes a reaction filtering device 1; a driving device 2 and a sample processing device 3 for driving the reaction filtering device 1 to rotate;

the reaction filtering device 1 is rotatably arranged right above the driving device 2; comprises a reaction disc 12 and a filter disc 15; a plurality of reaction grooves 12b for accommodating each reaction component are uniformly distributed on the periphery of the reaction disc 12; the reaction components are all substances added into the reaction tank 12b in the detection process; a plurality of filtering units 15b which correspond to the reaction tank 12b in position and are used for filtering the reacted reaction liquid are uniformly distributed on the periphery of the filtering disc 15;

the sample processing device 3 is fixedly arranged at the top end of the driving device 2; the sample processing apparatus 3 includes a plurality of antibody dropping devices 31 for dropping an antibody liquid into the reaction well 12 b; antibody liquid with weak radioactivity corresponding to the type of the target mycotoxin to be detected is respectively contained in the plurality of antibody dripping devices 31; the antibodies in the antibody fluid can react with the corresponding fungal viruses in the test sample in an immune manner; the sample treatment apparatus 3 further comprises a sample dropping device 32 for dropping a sample into the reaction tank 12 b; the sample processing apparatus 3 further includes a precipitation titration apparatus 33 for dropping a precipitation reaction reagent into the reaction tank 12b and causing an antibody-antigen conjugate obtained after the immunoreaction to undergo a precipitation reaction and form a precipitate; the sample processing device 3 further comprises an elution device 34 for eluting the solution remaining on the precipitate; the sample treatment apparatus 3 further includes a radioactivity detecting device 37 for detecting the radioactivity of the reaction precipitate.

The specific working mode of the embodiment is as follows: when the device works, the filter disc 15 is driven by the driving device 2 to stretch downwards until the bottom surface is abutted against the upper surface of the reaction disc 12, and each filter unit 15b at the periphery of the filter disc 15 is sleeved in each reaction tank 12b at the periphery of the reaction disc 12; at the moment, the driving device 2 drives the reaction filtering device 1 in the superposition state to drive the reaction tank 12b to rotate to the position below the antibody dripping device 31 corresponding to the fungal virus to be detected, and corresponding antibody liquid with excessive volume is dripped into the reaction tank 12b through the antibody dripping device 31; after the completion of the dropwise addition of the antibody, the reaction tank 12b in the superposed state is indexed to a position below the sample dropwise addition device 32, and a sample of a fixed volume is dropwise added into the reaction tank 12b by the sample dropwise addition device 32; after the dripping is finished, standing for 1-2min to ensure that the mycotoxin and the antibody in the sample are fully subjected to immunoreaction and combined; after the standing reaction is finished, the reaction tank 12b in the superposed state is transposed to the lower part of the precipitation titration device 33, and a precipitation reaction reagent is dripped into the reaction tank 12b through the precipitation titration device 33, so that the antibody-antigen combination generated by the immunoreaction combination is separated out from the solution to form a precipitate; after the precipitation reaction is finished, the driving device 2 drives the filter disc 15 to drive the filter unit 15b to move upwards, and precipitates are separated from the solution in the reaction tank 12 b; after the separation is completed, the filter disc 15 drives the filter unit 15b to be indexed to the position below the elution device 34, and the solution remained on the precipitate is eluted by the elution device 34; after the elution is completed, the filter disc 15 drives the filter unit 15b to shift to the radioactivity detection device 37, the radioactivity of the precipitate is detected, and the concentration of the mycotoxin in the sample is judged according to the radioactivity intensity of the precipitate.

In this embodiment, preferably, the precipitation reaction can be performed by using an isoelectric precipitation method for antigen-antibody conjugate to precipitate and separate the antigen-antibody conjugate from the solution, wherein the precipitation reaction reagent is an acid solution, the acid solution is preferably a 12% hydrochloric acid solution, and the alkali solution is preferably a 10% sodium hydroxide solution;

in this embodiment, the preferred antibody is preferably a conjugate of a monoclonal antibody against the mycotoxin of interest and bovine serum albumin labeled with carbon 14;

in this embodiment, by simultaneously providing a plurality of sets of reaction tanks 12b and a plurality of filter units 15b, a specific immunoreaction is simultaneously performed on a sample in cooperation with a plurality of types of antibody solutions subjected to weak radioactive treatment, and after an antigen-antibody conjugate in the reaction solution is subjected to precipitation separation by a precipitation titration apparatus 33, the radioactivity intensity of a precipitate is detected by a radioactivity detection apparatus 37, so that a plurality of mycotoxins in the sample are simultaneously quantitatively detected; in the detection process, the sample does not need to be subjected to complicated treatment, the detection efficiency is high, and the detection result is accurate; meanwhile, the antibody liquid is excessively dripped, so that the mycotoxin in the reaction tank 12b is completely reacted, and the defect that the content of the mycotoxin in a sample cannot be accurately and quantitatively detected due to the fact that the mycotoxin cannot be completely combined with the antibody because of excessive mycotoxin when test paper is used for detection is overcome;

based on the above embodiment, further, the driving device 2 further includes a second fixing frame 21; a plurality of supporting columns 21a for supporting are arranged on the periphery of the second fixing frame, and the tops of the supporting columns 21a are fixedly connected with an intermediate connecting plate 22; the top of the intermediate connecting plate 22 is provided with a waste liquid tank 22c, and the bottom of the waste liquid tank 22c is symmetrically provided with second through holes 22 b; a third through hole 22d is formed in the center of the top surface of the middle connecting plate 22; waste liquid boxes 24 are symmetrically arranged between the second fixing frame 21 and the middle connecting plate 22; the liquid inlet of the waste liquid box is communicated with the second through hole 22 b; the drive device 2 further comprises a bracket 27; the bracket 27 is of a concave structure, and a fourth through hole 27a is formed in the center of a concave plane at the bottom of the bracket; the left side and the right side of the fourth through hole 27a are respectively provided with a first plane 27b which is fixedly connected with the middle connecting plate (22);

the drive means 2 further comprise a second pulley 28; a first cylindrical boss 28b rotatably connected with the fourth through hole 27a is arranged at one end of the second belt pulley 28; a profile hole 28a is formed in the central part of the second belt pulley 28; a first motor 26 is fixedly installed on the left side of the bottom surface of the second fixing frame 21, and a first belt pulley 25 is fixedly installed on the output end of the first motor 26; a second motor 212 is fixedly installed on the right side of the bottom surface of the second fixing frame 21, and a third belt wheel 211 is fixedly installed on the output end of the second motor 212; the third belt wheel 211 drives the second belt wheel 28 to synchronously rotate through a second synchronous belt 210; and a second electromagnet 29 which is superposed with the axis of the profile hole 28a is fixedly arranged in the middle of the second fixing frame 21.

In this embodiment, during operation, the first motor 26 serves as a power source for the rotation of the reaction tray 12, the second motor 212 serves as a power source for the rotation of the filter tray 15, and the second electromagnet 29 serves as a power source for the upward movement of the filter tray 15; the second motor 212 drives the second belt pulley 28 to rotate through the third belt pulley 211 and the second synchronous belt 210, and the second belt pulley 28 outputs a rotation torque to the outside through a profile hole 28a formed in the center; after the detection is finished, the reaction tank 12b discharges the reaction waste liquid to the waste liquid tank 22c, and the waste liquid in the waste liquid tank 22c enters the waste liquid box 24 from the liquid inlet of the waste liquid box 24311 through the second through hole 22b for storage, so that the radioactive waste liquid is prevented from polluting the environment.

Based on the above embodiment, further, the bottom of the reaction disk 12 of the reaction filtering device 1 is provided with a second cylindrical boss 12c with a hollow structure, and the end face of the bottom is fixedly provided with a fourth pulley 17; the reaction disc 12 is rotatably connected in the third through hole 22d through the second cylindrical boss 12c, and a fourth belt wheel 17 at the bottom of the reaction disc is synchronously connected with the first synchronous belt wheel 23 through a first synchronous belt 23; the central part of the bottom surface of the filter disc 15 is provided with a profile rod 15c connected with the profile of the profile hole 28a, and the bottom of the profile rod is provided with a permanent magnet 11 with the polarity opposite to that of the second electromagnet 29; the bottom of the reaction tank 12b is also provided with an electromagnetic valve 13 which is communicated with the reaction tank and is used for discharging reaction waste liquid; the bottom of the filtering unit 15b is provided with a filtering membrane 16 for filtering reaction sediments; the bottom is also fixedly connected with a piezoelectric ultrasonic vibration sheet 14 for driving the filter unit 15b to vibrate and stir the tank liquid in the reaction tank 12 b.

In this embodiment, the reaction disk 12 is rotatably connected to the third through hole 22d of the driving device 2 through the second cylindrical boss 12c, and the fourth pulley 17 fixedly mounted at the bottom thereof is located in a space formed between the concave plane of the bracket 27 and the bottom surface of the intermediate connecting plate 22; when the device works, the first motor 26 drives the reaction disc 12 to drive the reaction tank 12b to rotate, and the second motor 212 drives the filter disc 15 to drive the filter unit 15b to rotate; the second electromagnet 29 drives the filter disc 15 to move upwards by generating a magnetic field with the same polarity as the permanent magnet 11; the piezoelectric ultrasonic vibration piece 14 transmits vibration to the reaction tank 12b through the filter disc 15 at a reaction stage after the charging is completed, so that liquid in the reaction tank 12b is stirred, the liquid mixing is accelerated, and the detection efficiency is improved.

Based on the above embodiment, further, the antibody dripping device 31 further includes an antibody liquid box 311 for containing antibody liquid; a dosing pump 312 for quantitatively pumping the antibody liquid; a burette 313 for adding an antibody solution to the reaction tank 12 b; a magnetic lifting seat 314 for fixing the burette 313; and an electromagnetic coil 315 for driving the magnetic elevating base 314 to move up and down.

In this embodiment, the antibody liquid box 311 is communicated with the quantitative pump 312, the front end of the burette 313 is fixedly installed in the magnetic lifting seat 314, and the magnetic lifting seat 314 can drive the burette 313 to move up and down under the driving of the electromagnetic coil 315; in the working process, before titration is carried out, the electromagnetic coil 315 drives the lifting seat to drive the burette 313 to move downwards, so that the distance between the pipe orifice and the reaction tank 12b is closer, and reagent waste caused by splashing in the titration feeding process is avoided.

Based on the above embodiment, further, the sample dropping device 32 further includes a guiding cylinder 328 fixedly connected to the top surface of the first fixing frame 36 and a sample solution groove 321 for holding a sample solution; a limiting surface 328a is arranged in the middle of the inner hole of the guide cylinder 328; a dropping head 327 capable of sliding up and down is further arranged in the inner hole of the guide cylinder 328; a second spring 329 is arranged between the guide cylinder 328 and the dripping head 327; a second one-way valve 3210 is further arranged in the dropping head 327; the top of the dripping head 327 is also provided with a three-way joint 3211; a first one-way valve 323 is arranged at the left side of the three-way joint 3211; a liquid pipe 322 is arranged on the left side of the first one-way valve 323; the second check valve 3210 is installed in the opposite direction to the first check valve 323; the top of the three-way joint 3211 is also fixedly connected with a liquid suction barrel 324; a piston rod 325 forming a closed space with the liquid suction cylinder 324 is also connected in the liquid suction cylinder 324 in a sliding manner; a first spring 326 is further arranged between the liquid suction cylinder 324 and the piston rod 325; the first spring 326 is stiffer than the second spring 329.

In this embodiment, the preferred liquid suction cylinder is made of glass material, and the surface of the liquid suction cylinder is provided with scales;

in this embodiment, when the sample is placed in the sample cup during operation, the top of the piston rod 325 is pressed by hand, and the first spring 326 is compressed because the stiffness of the first spring 326 is greater than that of the second spring 329, and the dropping head 327 slides downwards along the inner hole of the guide cylinder 328 until the lower surface of the middle boss abuts against the limiting surface 328a of the guide cylinder 328; the top of the piston rod 325 is continuously pressed, the first spring 326 is compressed, the piston moves downwards along the liquid suction cylinder 324, and air in the cylinder is discharged from the dripping head 327 through the second one-way valve 3210; after the air is completely discharged, the pressing is cancelled, after the second spring 329 resets the dropping head 327, the volume of the liquid suction cylinder 324 is increased to generate negative pressure in the process that the piston rod 325 moves upwards, and the sample is sucked into the liquid suction cylinder 324 from the sample groove 321 through the liquid pipe 322 via the first one-way valve 323; after the completion of the pipetting, the top of the piston rod 325 is pressed, and after the downward movement of the pipette head 327, the scale on the pipette barrel 324 is observed, and a fixed volume of the sample solution is dripped into the reaction tank 12 b.

The sample in the sample groove 321 is continuously sucked out through the matching of the first one-way valve 323 and the second one-way valve 3210 which are installed reversely; meanwhile, the automatic downward movement of the drip stand head in the dripping process is realized by utilizing the rigidity difference between the first spring 326 and the second spring 329, the splashing is reduced, the drip stand head 327 automatically ascends after the dripping is finished, and the interference between the drip stand head 327 and the filter disc 15 when the drip stand head moves upwards on the filter disc 15 is avoided; the return action of the second spring 329 causes the piston rod 325 to move up and return, thereby drawing the sample into the pipette 324 and maintaining the pipette 324 in a full state.

Based on the above embodiment, further in practical use, the sample processing apparatus 3 further includes a hot air drying device 35 for dehydrating and drying the precipitate; the hot air drying device 35 includes a guide rail 355 fixedly installed on the bottom surface of the first fixing frame 36 for guiding; a hot air drying block 354 of an L-shaped structure slidably fitted over the guide rail 355; a first mounting plate 351 fixedly mounted to the rear of the guide rail 355; a second electromagnet 29 fixedly attached to the front end of the first attachment plate 351; a third spring 353 having a rear end connected to the first mounting plate 351 and a front end connected to the hot air drying block 354; a first guide groove 354d used for being in sliding fit with the guide rail 355 is formed in an upper protruding part of the hot air drying block 354 with the L-shaped structure, a U-shaped first groove 354b is formed in a front protruding part 354a, and a plurality of hot air holes 354c are uniformly distributed in the bottom and the side surface of the first groove 354 b; the hot air drying block 354 is made of a ferromagnetic material.

In this embodiment, the hot air drying device 35 is disposed along the diameter direction of the first fixing bracket 27, when the hot air drying device works, the filtering unit 15b is shifted to the front side of the hot air drying block 354, at this time, the current of the second electromagnet 29 is reduced, so that the magnetic field attraction of the second electromagnet to the hot air drying block 354 is reduced, the hot air drying block 354 moves forward along the guide rail 355 under the elastic force of the third spring 353, so that the filtering unit 15b is located inside the first groove 354 b; the filter unit 15b is dried by hot air through the hot air holes in the first groove 354b, so that the moisture content of precipitates in the filter unit 15b is reduced, and the influence of water vapor volatilized in the subsequent radioactive ray detection process on the detection result is prevented.

In this embodiment, preferably, the gas sprayed from the hot air drying holes is an inert gas to prevent the precipitate from being oxidized during the hot air drying process.

Based on the above embodiment, further, the radioactivity detecting device 37 further includes an analyzing device 375 for analyzing the spot area and brightness; a lens body 372 fixedly connected to the front end of the analysis device 375; a fluorescent lens 371 fixed on the front side of the lens body 372 and used for ray induction luminescence; the surface of the fluorescent lens 371 is coated with a fluorescent luminous coating; a convex lens 373 fixed to the middle of the lens body 372 for amplifying the fluorescent light spot; a photosensitive sheet 374 for sensitive phosphor plate light imaging.

In this embodiment, the filtering unit 15b carrying the dried precipitate during operation is indexed to the lower side of the radioactivity detecting device, and moved upward to reduce the distance between the lens body 372 of the radioactivity detecting device and the filtering unit 15 b; as the bovine serum albumin in the precipitate is marked by carbon 14 and has weak radioactivity, the fluorescent luminescent coating coated on the surface of the fluorescent lens 371 is excited, and light spots are generated on the surface of the fluorescent lens 371; the brightness and the radioactive intensity of the light spot are in positive correlation, and the light rays are amplified by the convex lens 373 and then act on the photosensitive film 374, so that the information such as the area, the luminous intensity and the like of the light spot can be obtained; and analyzing the area of the light spot and the luminous intensity to calculate the radioactive intensity so as to obtain the content of the combined mycovirin in the precipitate.

As shown in fig. 1 to 12, the present embodiment further provides a method for detecting multiple mycotoxins by using the high-sensitivity multiple mycotoxin synchronous rapid inspection robot, including the following steps:

s1: the antibody dropping device 31 is mounted on the sample treatment device 3;

in this step, a structure capable of being quickly disassembled is provided between the antibody liquid box 311 and the quantitative pump 312, so that the antibody liquid box 311 can be conveniently adjusted and replaced;

s2: the driving device drives the filter disc 15 to move downwards, and the filter units 15b on the periphery of the filter disc 15 are sleeved in the reaction tanks 12 b;

in this step, after the filtering unit 15b is inserted into the reaction tank 12b, the filtering membranes 16 on the outer wall and the bottom of the filtering unit are attached to the reaction tank 12b to ensure that the precipitate generated after the reaction is completely collected;

s3: the driving device 2 drives the reaction disc 12 and the filter disc 15 to synchronously rotate, and drives each reaction tank 12b and each filter unit 15b to move to the position below the antibody dripping device 31 corresponding to the mycotoxin species to be detected; the antibody dropping device 31 drops an excessive volume of the antibody liquid into the reaction tank 12 b;

in the step, the antibody liquid is excessively dripped, so that the target mycotoxin is completely combined in the reaction process, and the condition that the accurate quantitative result cannot be obtained due to the fact that part of mycotoxin is not combined by reaction due to the fact that the antibody liquid is insufficient is avoided;

s4: the driving device 2 drives the reaction disc 12 and the filter disc 15 to synchronously rotate, and drives each reaction tank 12b and each filter unit 15b to move to the lower part of the sample dripping device 32; the sample dropping device 32 drops a fixed volume sample into the reaction tank 12 b;

in the step, the adding volume of the sample can be adaptively adjusted according to factors such as the concentration of the sample, the content of impurities and the like under the principle that the adding volume of the antibody liquid with the same concentration is not exceeded;

the standing time in the step is 1-2min at 25 ℃, and the standing time can be adaptively adjusted according to the environmental temperature;

s6: the driving device 2 drives the reaction disc 12 and the filter disc 15 to synchronously rotate, and drives each reaction tank 12b and each filter unit 15b to move to the lower part of the precipitation and titration device; the precipitation titration device 33 is used for dropwise adding a precipitation reaction reagent into the reaction tank 12b, so that the antibody which is immunologically combined with the mycotoxin is precipitated and separated out from the solution;

in the step, the precipitation reaction reagent is acid liquor or alkali liquor; wherein, the acid solution is preferably 10% hydrochloric acid solution, and the alkali solution is preferably 8% sodium hydroxide solution; through adjusting the pH value of the bath solution to the isoelectric point of the antigen-antibody conjugate, the aggregation precipitate between adjacent molecules is separated out from the solution;

s7: the driving device 2 drives the filter disc 15 to drive each filter unit 15b to move upwards, so that the sediment in the reaction tank 12b is separated from the residual liquid in the reaction tank 12 b;

in the step, the upward moving speed of the filter disc 15 is controlled within 2mm/s, so that the phenomenon that the detection result is influenced by the fact that the tank liquor in the filter unit 15b flows out too fast and the precipitate is scattered and carried out is avoided;

s8: the driving device 2 drives the filter disc 15 to drive the filter unit 15b to move below the elution device 34; the elution device 34 elutes the reaction liquid remaining on the surface of the precipitate;

in the step, the pH value of the eluent is equal to the isoelectric point pH value of the antigen-antibody conjugate, so that the influence on the detection result caused by partial dissolution of a precipitate due to deviation of the isoelectric point is avoided;

s9: the driving device 2 drives the filter disc 15 to drive the filter unit 15b to move to the position below the radioactivity detecting device 37, the radioactivity detecting device 37 emits light to generate light spots under the excitation of radioactive rays through the fluorescent lens 371, the light spots are projected on the photosensitive sheet 374 after being amplified through the convex lens 373, the light emitting intensity and the area of the light spots are automatically analyzed through the analyzing device 375, and the radioactivity intensity of sediments is detected; judging the content of mycotoxin corresponding to the antibody in the sample according to the radioactivity intensity of the precipitate;

in this step, the height of the filtering unit 15b should be adjusted according to the amount of the precipitate, so as to ensure that the detection distances between the fluorescence lens 371 and the precipitate are strictly equal, and prevent the difference between the luminous intensity and the area of the fluorescence lens 371 caused by the difference between the detection distances from affecting the experimental result.

The above description is only a preferred embodiment of the present invention, and all equivalent changes or modifications of the structure, characteristics and principles described in the present patent application are included in the protection scope of the present patent application.

Claims

1. A high sensitivity many fungaltoxin examine robot fast in step which characterized in that: comprises a reaction filtering device (1); a driving device (2) for driving the reaction filtering device (1) to rotate and a sample processing device (3); the reaction filtering device (1) is rotatably arranged right above the driving device (2); the reaction filtering device (1) comprises a reaction disc (12) and a filtering disc (15); a plurality of reaction grooves (12b) for containing each reaction component are uniformly distributed on the periphery of the reaction disk (12); the reaction components are all substances added into the reaction tank (12b) in the detection process; a plurality of filtering units (15b) which correspond to the reaction tank (12b) in position and are used for filtering the reaction liquid after the reaction are uniformly distributed on the periphery of the filtering disc (15); the sample processing device (3) is fixedly arranged at the top end of the driving device (2); the sample processing device (3) comprises a plurality of antibody dripping devices (31) for dripping antibody liquid into the reaction tank (12 b); antibody liquid with weak radioactivity corresponding to the type of the target mycotoxin to be detected is respectively contained in the plurality of antibody dripping devices (31); the antibodies in the antibody fluid can react with the corresponding fungal viruses in the test sample in an immune manner; the sample processing device (3) further comprises a sample dropping device (32) for dropping a sample into the reaction tank (12 b); the sample processing device (3) further comprises a precipitation titration device (33) for dropping a precipitation reaction reagent into the reaction tank (12b) and causing the antibody-antigen combination obtained after the immunoreaction to perform a precipitation reaction and form a precipitate; the sample processing device (3) further comprises an elution device (34) for eluting the solution remained on the precipitate; the sample processing device (3) further comprises a radioactivity detection device (37) for detecting the radioactivity of the reaction precipitate; the driving device (2) comprises a second fixing frame (21); a plurality of supporting columns (21a) for supporting are arranged on the periphery of the second fixing frame, and the tops of the supporting columns (21a) are fixedly connected with an intermediate connecting plate (22); the top of the intermediate connecting plate (22) is provided with a waste liquid tank (22c), and the bottom of the waste liquid tank (22c) is symmetrically provided with second through holes (22 b); a third through hole (22d) is formed in the center of the top surface of the middle connecting plate (22); waste liquid boxes (24) are symmetrically arranged between the second fixing frame (21) and the middle connecting plate (22); the liquid inlet of the waste liquid box (24) is communicated with the second through hole (22 b); the drive device (2) further comprises a bracket (27); the bracket (27) is of a concave structure, and a fourth through hole (27a) is formed in the center of a concave plane at the bottom of the bracket; the left side and the right side of the fourth through hole (27a) are respectively provided with a first plane (27b) which is fixedly connected with the middle connecting plate (22); the drive device (2) further comprises a second pulley (28); one end of the second belt wheel (28) is provided with a first cylindrical boss (28b) which is rotatably connected with the fourth through hole (27 a); a profile hole (28a) is formed in the central part of the second belt wheel (28); a first motor (26) is fixedly installed on the left side of the bottom surface of the second fixing frame (21), and a first belt wheel (25) is fixedly installed on the output end of the first motor (26); a second motor (212) is fixedly installed on the right side of the bottom surface of the second fixing frame (21), and a third belt wheel (211) is fixedly installed on the output end of the second motor (212); the third belt wheel (211) drives the second belt wheel (28) to synchronously rotate through a second synchronous belt (210); a second electromagnet (29) which is coincident with the axis of the profile hole (28a) is fixedly arranged in the middle of the second fixing frame (21); a second cylindrical boss (12c) with a hollow structure is arranged at the bottom of the reaction disc (12); a fourth belt pulley (17) is fixedly arranged on the bottom end face of the reaction disc (12); the reaction disc (12) is rotatably connected in the third through hole (22d) through the second cylindrical boss (12 c); the first belt wheel (25) drives the fourth belt wheel (17) to synchronously rotate through a first synchronous belt (23); a profile rod (15c) connected with the profile of the profile hole (28a) is arranged at the central part of the bottom surface of the filter disc (15); the bottom of the profile rod (15c) is fixedly provided with a permanent magnet (11) with the polarity opposite to that of the second electromagnet (29); the bottom of the filter disc (15) is also fixedly connected with a piezoelectric ultrasonic vibrating reed (14) which is used for driving the filter unit (15b) to vibrate and stir the tank liquid in the reaction tank (12 b); the radioactivity detection device (37) also comprises an analysis device (375) for analyzing the area and the brightness of the light spot; a lens body (372) fixedly connected to the front end of the analysis device (375); a fluorescent lens (371) fixed on the front side of the lens body (372) and used for ray induction luminescence; the surface of the fluorescent lens (371) is coated with a fluorescent luminous coating; a convex lens (373) which is fixed in the middle of the lens body (372) and amplifies fluorescent light spots; a photosensitive sheet (374) for sensing a phosphor plate for photoimaging.

2. The high-sensitivity multi-mycotoxin synchronous rapid inspection robot according to claim 1, characterized in that: the antibody dripping device (31) further comprises an antibody liquid box (311) for containing the antibody liquid, a quantitative pump (312) for quantitatively pumping out the antibody liquid, a burette (313) for adding the antibody liquid to the reaction tank (12b), a magnetic lifting seat (314) for fixing the burette (313), and an electromagnetic coil (315) for driving the magnetic lifting seat (314) to move up and down.

3. The high-sensitivity multi-mycotoxin synchronous rapid inspection robot according to claim 1, characterized in that: the sample dripping device (32) also comprises a guide cylinder (328) fixedly connected to the top surface of the first fixing frame (36) and a test solution groove (321) for containing test solution; a limiting surface (328a) is arranged in the middle of the inner hole of the guide cylinder (328); a dropping head (327) capable of sliding up and down is further arranged in the inner hole of the guide cylinder (328); a second spring (329) is arranged between the guide cylinder (328) and the dripping head (327); a second one-way valve (3210) is further arranged in the dripping head (327); the top of the dripping head (327) is also provided with a three-way joint (3211); a first one-way valve (323) is arranged on the left side of the three-way joint (3211); a liquid pipe (322) is arranged on the left side of the first one-way valve (323); the second one-way valve (3210) is installed in the opposite direction to the first one-way valve (323); the top of the three-way joint (3211) is fixedly connected with a liquid suction barrel (324); a piston rod (325) which forms a closed space with the liquid suction cylinder (324) is also connected in the liquid suction cylinder (324) in a sliding manner; a first spring (326) is also arranged between the liquid suction cylinder (324) and the piston rod (325); the first spring (326) is stiffer than the second spring (329).

4. The high-sensitivity multi-mycotoxin synchronous rapid inspection robot according to claim 1, characterized in that: the sample processing device also comprises a hot air drying device for dehydrating and drying the precipitate; the hot air drying device comprises a guide rail which is fixedly arranged on the bottom surface of the first fixing frame and used for guiding; the hot air drying block is in an L-shaped structure and is sleeved on the guide rail in a sliding manner; the first mounting plate is fixedly mounted at the tail part of the guide rail; the second electromagnet is fixedly arranged at the front end of the first mounting plate; the rear end of the third spring is connected with the first mounting plate, and the front end of the third spring is connected with the hot air drying block; a first guide groove used for being in sliding fit with the guide rail is formed in an upper convex part of the hot air drying block of the L-shaped structure, a U-shaped first groove is formed in a front convex part, and a plurality of hot air holes are uniformly distributed in the bottom and the side face of the first groove; the hot air drying block is made of ferromagnetic materials.

5. The method for detecting the multi-mycotoxin by using the high-sensitivity multi-mycotoxin synchronous rapid detection robot as set forth in any one of claims 1 to 4, comprising the following steps: s1: an antibody dropping device (31) is installed on the sample processing device (3); s2: the driving device drives the filter disc (15) to move downwards, and the filter units (15b) at the periphery of the filter disc (15) are sleeved in the reaction tanks (12 b); s3: the driving device (2) drives the reaction disc (12) and the filter disc (15) to synchronously rotate, and drives each reaction tank (12b) and each filter unit (15b) to move to the position below the antibody dripping device (31) corresponding to the mycotoxin species to be detected; the antibody dripping device (31) drips an excessive volume of antibody liquid into the reaction tank (12 b); s4: the driving device (2) drives the reaction disc (12) and the filter disc (15) to synchronously rotate, and drives each reaction tank (12b) and each filter unit (15b) to move to the lower part of the sample dripping device (32); the sample dripping device (32) drips a fixed volume sample into the reaction tank (12 b); s5: standing for a certain time to mix the sample with the antibody solution and perform an immune combination reaction; s6: the driving device (2) drives the reaction disc (12) and the filter disc (15) to synchronously rotate, and drives each reaction tank (12b) and each filter unit (15b) to move to the lower part of the precipitation and titration device; the precipitation titration device (33) is used for dropwise adding a precipitation reaction reagent into the reaction tank (12b) so that the antibody which is immunologically combined with the mycotoxin is precipitated and separated out from the solution; s7: the driving device (2) drives the filter disc (15) to drive each filter unit (15b) to move upwards, and precipitates in the reaction tank (12b) are separated from the residual liquid in the reaction tank (12 b); s8: the driving device (2) drives the filter disc (15) to drive the filter unit (15b) to move to the lower part of the elution device (34); the elution device (34) elutes the reaction liquid remaining on the surface of the precipitate; s9: the driving device (2) drives the filter disc (15) to drive the filter unit (15b) to move to the position below the radioactivity detection device (37), the radioactivity detection device (37) emits light under the excitation of radioactive rays through the fluorescent lens (371) to generate light spots, the light spots are amplified through the convex lens (373) and then projected onto the light-sensitive sheet (374), the light-emitting intensity and the area of the light spots are automatically analyzed through the analysis device (375), and the radioactivity intensity of sediments is detected; and judging the content of the mycotoxin corresponding to the antibody in the sample according to the radioactivity intensity of the precipitate.