CN115141739A - Air microorganism sampling and detection integrated device and sampling and detection method - Google Patents
Air microorganism sampling and detection integrated device and sampling and detection method Download PDFInfo
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Abstract
The invention provides an air microorganism sampling and detecting integrated device, which relates to the technical field of microorganism detection and comprises the following components: the device comprises an air microorganism sampling device, an elution cavity and a chip; the sampling cavity, the elution cavity and the chip of the air microorganism sampling device are sequentially arranged from top to bottom, and the sampling cavity is communicated with the elution cavity through a first channel between the sampling cavity and the elution cavity; the elution cavity is communicated with the sample adding port of the chip through a second channel between the elution cavity and the sample adding port; the scheme provided by the invention can be used for respectively completing the collection, nucleic acid extraction and detection of air microorganism samples, is simple to operate, avoids the pollution of mixed bacteria caused by complicated operation or aerosol caused by detection reaction, and is convenient and rapid.
Description
Technical Field
The invention relates to the technical field of microorganism detection, in particular to an air microorganism sampling and detecting integrated device and a sampling and detecting method.
Background
Microorganisms in human, animal and plant bodies and soil can be dispersed in the air by droplets, dust, and the like, so that the air contains a certain variety and number of microorganisms. There are generally no pathogenic microorganisms in the air theoretically, but aerosols of pathogenic microorganisms are often suspended in the air near hospitals, veterinary hospitals and livestock houses, and healthy people or animals are often infected by inhalation. Air contaminated by pathogenic microorganisms can often be a source or vehicle of contamination, causing epidemic disease. Therefore, the detection of airborne microorganisms is of great significance for the prevention and control of infectious diseases and the hygiene supervision and protection of the environment.
The air sampling bottle is an important tool in the air microorganism detection process, and is of various types, wherein the impact type air sampling bottle collects microorganism particles in the air into sampling liquid in an air jet mode. In the sampling process, after the sampling liquid is added into the sampling bottle, the air inlet device is started, air enters from the air inlet of the sampling bottle, microbial particles in the air impact the sampling liquid of the sampling bottle, and the microbial particles are captured due to the adhesion of the liquid.
Nucleic acid extraction technology is the basic technology of molecular biology research, is the first step of nucleic acid test, and is also the most critical step. The magnetic bead method nucleic acid extraction technology adopts nanometer magnetic bead microbeads, and the surfaces of the magnetic bead microbeads are marked with a functional group which can perform adsorption reaction with nucleic acid. The magnetic bead method for extracting nucleic acid can be widely applied to many fields such as genome research, HPV detection, paternity test, archaeology and the like. The magnetic bead method is superior to the traditional method for extracting nucleic acid, such as: the Chelex100 method, the organic method, the silicon dioxide method, the salting-out method and the like are simpler and more convenient, and the magnetic bead method is irreplaceable in the aspects of nucleic acid purification, micro-detection, PCR amplification and the like.
The paper-based micro-fluidic chip is a new micro-fluidic analysis technology platform, has the advantages of low cost, simple and easy processing, convenient use and carrying and the like, and has great application prospect in the application fields of clinical diagnosis, food quality control, environmental monitoring and the like. The paper-based microfluidic chip adopts paper as a substrate to replace materials such as silicon, glass, high polymers and the like, and the analysis device is called a micro lab on paper and is also called a microfluidic paper analysis device, and has extremely wide application in the fields of clinical diagnosis, food quality, environmental monitoring and the like.
The air microorganism detection technology in the prior art usually needs to sequentially carry out operations such as air microorganism sample collection, microorganism culture, nucleic acid extraction, microorganism detection reaction and the like, so that detection and identification of air microorganisms are completed, the operations are all completed by utilizing different experimental devices, the operation is complex, more instruments and equipment are needed, the consumed time is long, non-sampling bacterial pollution or aerosol pollution is easily introduced or caused in the operation, and the rapid detection of the air microorganisms is not facilitated.
Disclosure of Invention
The invention aims to provide an air microorganism sampling and detecting integrated device and a sampling and detecting method, which aim to solve the problems in the prior art.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides an air microorganism sampling and detecting integrated device, which comprises: the device comprises an air microorganism sampling device, an elution cavity and a chip, wherein the sampling cavity of the air microorganism sampling device is used for containing magnetic beads and lysis binding liquid; the elution cavity is used for containing an elution solution; the sampling cavity, the elution cavity and the chip are sequentially arranged from top to bottom, and the sampling cavity is communicated with the elution cavity through a first channel between the sampling cavity and the elution cavity; the elution cavity is communicated with the sample adding port of the chip through a second channel between the elution cavity and the sample adding port; first passageway with it shifts valve and second to be provided with respectively to move about in the second passageway, first transfer valve can with first passageway seals and is used for with the intracavity of sampling the magnetic bead shifts to the elution intracavity, the second transfer valve can with the second passageway seals and is used for shifting the eluant to in the sample addition port.
Preferably, an open/close valve is provided in each of the air inlet channel and the air outlet channel of the air microorganism sampling device.
Preferably, the first transfer valve comprises a first rotary ball and a first rotary handle, the first rotary ball is arranged in the first channel, the first rotary ball can close the first channel, the surface of the first rotary ball is recessed inwards to form two first transfer grooves, one end of the first rotary handle is fixedly connected with the first rotary ball, the other end of the first rotary handle extends out of the first channel, and the first rotary handle can be driven to rotate by rotating around the axis of the first rotary handle;
the first transfer valve is provided with a first rotation handle and a first rotation ball, the first rotation handle is arranged in the first transfer valve, the first rotation ball is arranged in the first transfer valve, the first rotation handle is arranged in the first transfer valve, the first rotation ball is arranged in the first transfer valve, and the first rotation ball is arranged in the first transfer valve;
the second transfer valve comprises a second rotary ball and a second rotary handle, the second rotary ball is arranged in the second channel and can seal the second channel, the surface of the second rotary ball is inwards recessed to form two second transfer grooves, one end of the second rotary handle is fixedly connected with the second rotary ball, the other end of the second rotary handle extends out of the second channel, and the second rotary handle can be driven to rotate by rotating around the axis of the second rotary handle;
in an initial state, the two first transfer grooves vertically face the sampling cavity and the elution cavity respectively; the two second transfer grooves are respectively vertically towards the elution cavity and the sample adding port.
Preferably, the air microorganism sampling device includes sampling head, sampling pipe and air inlet unit, the cavity that forms in the sampling pipe is promptly the sampling chamber, the sampling head includes lid and intake pipe, set up on the lid with external intercommunication inlet channel with outlet channel, the top of intake pipe fixed and with inlet channel intercommunication, the bottom of intake pipe to the first transfer groove extension of the bottom of sampling pipe.
Preferably, the two open/close valves each include a valve pipe and a knob, one end of the valve pipe is open, the other end of the valve pipe is fixedly connected with the knob, an overflow port is formed in a side wall of the valve pipe, two valve cavities are formed in part of the air inlet channel and part of the air outlet channel respectively, the two open/close valves correspond to the two valve cavities respectively, the two valve pipes are located in the two valve cavities respectively, the two knobs are exposed from the surface of the cover body, the valve pipes can be driven to rotate by rotating the knobs, and the air inlet channel and the air outlet channel can be opened and closed by rotating the valve pipes.
Preferably, the chip is a paper-based microfluidic chip and is made of a transparent material.
Preferably, the sampling tube and the air inlet tube are both made of flexible and compressible materials.
Preferably, the device also comprises a supporting device and a bottom module, wherein the supporting device comprises a supporting frame and a base, and the supporting frame is fixedly arranged on the base; the elution cavity, the first channel and the second channel are formed in the bottom die block, and the bottom die block is fixedly connected with the air microorganism sampling device and the chip;
the bottom module, the air microorganism sampling device and the chip are integrally cuboid, the support frame comprises two support rod groups which are arranged up and down, each support rod group comprises two support rods, the positions, close to the top, of the two opposite side walls of the air microorganism sampling device are respectively provided with a first groove, the two opposite side walls of the bottom module are respectively provided with a second groove, the two support rods in one support rod group are respectively embedded into the two first grooves, and the two support rods in the other support rod group are respectively embedded into the two second grooves; or
The bottom module is of a cylindrical structure, the cover body and the sampling pipe are of a cylindrical structure integrally, an air outlet and an air inlet are formed in the outer side wall of the cover body, elongated pipes are fixedly arranged on the air outlet and the air inlet, and the tail ends of the two elongated pipes extend in the direction away from the cover body along the radial direction of the cover body; the support frame comprises support sleeves which are arranged up and down, the two support sleeves are both of arc structures, the radian of the upper support sleeve is matched with that of the outer wall of the cover body, the radian of the lower support sleeve is matched with that of the outer wall of the bottom die block, the upper support sleeve is abutted against the outer wall of the cover body and/or the outer wall of the sampling tube, and the support sleeves are positioned below the two elongated tubes and abutted against the lower surfaces of the elongated tubes; the supporting sleeve at the lower part is propped against the outer wall of the bottom module.
Preferably, a heating device is arranged in the base, and the chip is placed on the base.
The invention also provides a sampling detection method for carrying out sampling detection by using the air microorganism sampling detection integrated device, which comprises the following steps:
the method comprises the following steps: connecting the air inlet channel with the air inlet device, opening the two opening and closing valves, opening the air inlet device for air sampling, pre-storing the cracking binding solution and the nucleic acid adsorption magnetic beads at the bottom of the sampling tube, and pre-storing the eluent in the elution cavity;
step two: after sampling is finished, the air inlet device is closed, the magnetic rod is inserted into the magnetic rod cavity of the first transfer valve, and magnetic beads at the bottom of the sampling tube are adsorbed to the bottom of the first transfer groove of the first transfer valve;
step three: horizontally placing the air microorganism sampling and detecting integrated device, wherein when the device is horizontally placed, the height of the cracking binding liquid in the sampling pipe is lower than that of the first transfer groove, so that the cracking binding liquid is separated from the magnetic beads;
step four: opening the air inlet device to enable air to enter the sampling tube from the air inlet channel and the air inlet tube and air-dry residual cracking binding solution on the magnetic beads at the bottom of the sampling tube, wherein an outlet of the air inlet tube is over against the first transfer groove at the bottom of the sampling tube;
step five: closing the air inlet device, closing the opening and closing valve, rotating the first transfer valve by 180 degrees to transfer the magnetic beads to the elution cavity, pulling out the magnetic rod, and vertically placing the device again;
step six: the magnetic beads naturally fall into the eluent in the elution cavity, and the elution is carried out on the nucleic acid on the magnetic beads through the shaking device;
step seven: inserting the magnetic rod into the magnetic rod cavity of the first transfer valve, and inverting the device to adsorb the magnetic beads in the elution cavity to the bottom of the first transfer groove of the first transfer valve;
step eight: rightly placing the device, and rotating the second transfer valve for 180 degrees to transfer the eluent to the chip sample addition port and automatically diffuse the eluent into the reaction chamber;
step nine: the device is placed on the heating device for detection reaction and observation of the detection result.
Compared with the prior art, the invention has the following technical effects:
the air microorganism sampling and detecting integrated device and the sampling and detecting method provided by the invention integrate air microorganism sample collection, nucleic acid extraction and detection, collect air microorganisms by directly taking a cracking binding solution as a sampling solution, enrich nucleic acid released by microorganism cracking by utilizing nucleic acid adsorption magnetic beads, and enable eluent obtained by eluting nucleic acid on the magnetic beads to directly enter a detection chip for detection, thereby accelerating the detection process of the air microorganisms; and the tightness of detection reaction can be ensured, aerosol pollution is avoided, and subsequent detection reaction is influenced, so that the device can collect air microorganism samples and extract and detect nucleic acid respectively, is simple to operate, avoids aerosol pollution caused by mixed bacteria pollution or detection reaction caused by complicated operation, and is convenient and rapid.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
Fig. 1 is a schematic structural diagram of an integrated air microorganism sampling and detecting device provided in the first embodiment when no first groove and no second groove are provided and no supporting device is provided;
FIG. 2 is a cross-sectional view of FIG. 1;
FIG. 3 is a schematic structural view of the sampling head and the sampling tube in FIG. 1;
FIG. 4 is a cross-sectional view of the structure of FIG. 3 when open;
FIG. 5 is a cross-sectional view of the structure of FIG. 3 when closed;
FIG. 6 is a schematic view of the first transfer valve of FIG. 1 disposed in the first passage;
FIG. 7 is a cross-sectional view of FIG. 6;
FIG. 8 is a schematic diagram of the chip of FIG. 1;
FIG. 9 is a cross-sectional view of FIG. 8;
FIG. 10 is a schematic view of an integrated apparatus for sampling and detecting microorganisms in air according to an embodiment;
FIG. 11 is a schematic structural view of an integrated air microorganism sampling and detecting device according to a first embodiment, wherein a supporting device is not provided;
FIG. 12 is a schematic view of the integrated air microorganism sampling and detecting device according to the second embodiment without a supporting device;
FIG. 13 is a cross-sectional view of FIG. 12;
FIG. 14 is a schematic view of an integrated apparatus for sampling and detecting microorganisms in air according to a second embodiment;
in the figure: 1. an air microorganism sampling device; 11. an air intake passage; 12. an opening and closing valve; 121. a flow-through port; 13. a cover body; 14. an air outlet channel; 15. an air inlet pipe; 16. a sampling tube; 17. a first groove; 2. a bottom module; 21. an elution chamber; 22. a second groove; 3. a chip; 31. a sample addition port; 32. a reaction chamber; 4. a first transfer valve; 41. a first rotating ball; 42. a magnetic rod cavity; 43. a first transfer slot; 44. a first rotation handle; 5. a second transfer valve; 6. an upper module; 7. a support device; 71. a support bar; 72. a base; 73. a support sleeve; 8. and (5) lengthening the pipe.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. 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 invention aims to provide an air microorganism sampling and detecting integrated device and a sampling and detecting method, which are used for solving the problems in the prior art.
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in further detail below.
Example one
The embodiment provides an air microorganism sampling detection integrated device, as shown in fig. 1-11, including: the device comprises an air microorganism sampling device 1, an elution cavity 21 and a chip 3, wherein a sampling cavity of the air microorganism sampling device 1 is used for containing magnetic beads and lysis binding liquid; the elution cavity 21 is used for containing eluent; the sampling cavity, the elution cavity 21 and the chip 3 are sequentially arranged from top to bottom, and the sampling cavity is communicated with the elution cavity 21 through a first channel between the sampling cavity and the elution cavity 21; the elution chamber 21 and the sample adding port 31 of the chip 3 are communicated through a second channel between the elution chamber 21 and the sample adding port 31; the first channel and the second channel are respectively and movably provided with a first transfer valve 4 and a second transfer valve 5, the first transfer valve 4 can seal the first channel and is used for transferring the magnetic beads in the sampling cavity to the elution cavity 21, and the second transfer valve 5 can seal the second channel and is used for transferring the eluent to the sample port 31.
The air microorganism sampling and detection integrated device provided by the embodiment integrates air microorganism sample collection, nucleic acid extraction and detection, directly collects air microorganisms by taking a lysis binding solution as a sampling solution, enriches nucleic acid released by microorganism lysis by using nucleic acid adsorption magnetic beads, and can directly feed eluent obtained by eluting the nucleic acid on the magnetic beads into a detection chip 3 for detection so as to accelerate the detection process of the air microorganisms; and the airtightness of detection reaction can be ensured, aerosol pollution is avoided, and subsequent detection reaction is influenced, so that the device can collect air microorganism samples, extract nucleic acid and detect the air microorganism samples respectively, is simple to operate, avoids aerosol pollution caused by impurity bacteria pollution or detection reaction due to complex operation, and is convenient and rapid.
In another embodiment, the inlet channel 11 and the outlet channel 14 of the air microorganism sampling device 1 are both provided with the open/close valve 12, and the open/close valve 12 has two functions, one is to facilitate sealing of products in transportation and prevent the sampling cavity from being polluted, and the other is to avoid the need of shaking the device or even inverting the whole device when the device is applied to sampling, elution and nucleic acid detection, and at this time, the liquid in the sampling cavity can be prevented from spilling only by closing the open/close valve 12.
In another embodiment, in order to realize that the first transfer valve 4 can transfer magnetic beads conveniently, the first transfer valve 4 is designed to have a structure that the first transfer valve 4 includes a first rotary ball 41 and a first rotary handle 44, the first rotary ball 41 is disposed in the first channel, the first rotary ball 41 can close the first channel, the surface of the first rotary ball 41 is recessed inwards to form two first transfer grooves 43, one end of the first rotary handle 44 is fixedly connected to the first rotary ball 41, and the other end extends out of the first channel, and rotating the first rotary handle 44 around the axis of the first rotary handle 44 can drive the first rotary ball 41 to rotate;
the first transfer valve 4 is internally provided with a magnetic rod cavity 42, the magnetic rod cavity 42 extends from one end of the first rotation handle 44, which is far away from the first rotation ball 41, to a position between the two first transfer grooves 43 along the direction from the first rotation handle 44 to the first rotation ball 41, and one end of the magnetic rod can extend into the magnetic rod cavity 42 between the two first transfer grooves 43;
in order to realize that the second transfer valve 5 can conveniently transfer the eluent, the second transfer valve 5 is designed into a structure, the second transfer valve 5 comprises a second rotary ball and a second rotary handle, the second rotary ball is arranged in the second channel, the second channel can be sealed by the second rotary ball, the surface of the second rotary ball is recessed inwards to form two second transfer grooves, one end of the second rotary handle is fixedly connected with the second rotary ball, the other end of the second rotary handle extends out of the second channel, and the second rotary handle can be rotated around the axis of the second rotary handle to drive the second rotary ball to rotate;
in the initial state, the two first transfer grooves 43 are respectively vertically oriented to the sampling chamber and the elution chamber 21; the two second transfer wells are vertically oriented toward the elution chamber 21 and the sample addition port 31, respectively.
In another embodiment, the air microorganism sampling apparatus 1 includes a sampling head, the sampling tube 16 and an air inlet device, the air inlet device can adopt a fan, a cavity formed in the sampling tube 16 is a sampling cavity, the sampling head includes a cover 13 and an air inlet tube 15, an air inlet channel 11 and an air outlet channel 14 communicated with the outside are provided on the cover 13, the top end of the air inlet tube 15 is fixed and communicated with the air inlet channel 11, the bottom end of the air inlet tube 15 extends to a first transfer groove 43 at the bottom of the sampling tube 16, so that the air outlet end of the air inlet tube 15 is aligned to the first transfer groove 43, and further, air enters the sampling tube 16 from the air inlet channel 11 and the air inlet tube 15 and air-dries the residual cracking and combining liquid on the magnetic beads at the bottom of the sampling tube 16, so as to avoid the influence on the detection result due to the existence of the residual sampling liquid on the magnetic beads.
In another embodiment, the bottom of the sampling chamber is a cavity with a wide top and a narrow bottom, and the inner side wall is a slope to facilitate the natural flow of the sampling solution or magnetic beads to the bottom of the sampling tube 16, and the bottom of the sampling tube is connected and communicated with the first transfer groove 43.
In another embodiment, the two opening/closing valves 12 each include a valve tube and a knob, one end of the valve tube is open, the other end of the valve tube is fixedly connected to the knob, the side wall of the valve tube is provided with an overflow port 121, a part of the air inlet channel 11 and a part of the air outlet channel 14 form two valve cavities respectively, the two opening/closing valves 12 correspond to the two valve cavities respectively, the two valve tubes are located in the two valve cavities respectively, the two knobs are exposed from the surface of the cover body 13, and the valve tubes can be rotated by a worker, and can be opened and closed by rotating the knobs, so that the air inlet channel 11 and the air outlet channel 14 can be opened and closed, and the opening and closing of the air inlet channel 11 and the air outlet channel 14 can be realized relatively simply.
In another embodiment, the chip 3 is a paper-based microfluidic chip, the chip 3 is made of a transparent material, the eluent can automatically diffuse into each reaction chamber 32 after entering the sample inlet 31 of the paper-based microfluidic chip, the collected sample can be detected by heating the chip 3, the chip 3 is made of a transparent material, a nucleic acid detection reagent with a color or fluorescence indicator can be used as the reaction reagent, and the detection result can be directly judged by observing the color change in the reaction chamber 32.
In another embodiment, the sampling tube 16 and the inlet tube 15 are made of flexible and compressible materials, such as silicone, bellows, etc., to reduce the volume of the device for stacking storage and transportation; the sampling tube 16 portion of the device is compressed and folded when the device is not in use, and the device can be placed on the support means 7 for sampling and testing as described below when the device is in use.
In another embodiment, the integrated device for sampling and detecting air microorganisms further comprises a supporting device 7 and a bottom module 2, wherein the supporting device 7 comprises a supporting frame and a base 72, and the supporting frame is fixedly arranged on the base 72; the elution cavity 21, the first channel and the second channel are all formed in the bottom die block 2, and the bottom die block 2 is fixedly connected with the air microorganism sampling device 1 and the chip 3;
In another embodiment, the bottom module 2 comprises a middle module, an upper module 6 and a lower module, the upper module 6 and the lower module are respectively disposed above and below the middle module and are fixedly connected, the middle module is provided with an elution cavity 21, the upper module 6 is provided with a first channel, and the lower module is provided with a second channel.
In another embodiment, a heating device is disposed in the base 72, and the chip 3 is placed on the base 72, and the heating device is used for heating the chip 3.
In addition, the device has simple structure, and the structures except the air inlet device and the supporting device 7 in the device can be used as disposable consumables, thereby avoiding the cross contamination caused by repeated use.
Example two
As shown in fig. 12 to 14, the difference from the first embodiment is that a bottom mold block 2 in the integrated apparatus for sampling and detecting airborne microorganisms provided in this embodiment is of a cylindrical structure, a cover 13 and a sampling tube 16 are also of a cylindrical structure as a whole, an air outlet and an air inlet are provided on an outer side wall of the cover 13, elongated tubes 8 are fixedly provided on the air outlet and the air inlet, and ends of the two elongated tubes 8 extend in a direction away from the cover 13 along a radial direction of the cover 13; the supporting frame comprises supporting sleeves 73 which are arranged up and down, the two supporting sleeves 73 are both arc-shaped, the radian of the upper supporting sleeve 73 is matched with the radian of the outer wall of the cover body 13, the radian of the lower supporting sleeve 73 is matched with the radian of the outer wall of the bottom die block 2, the upper supporting sleeve 73 is propped against the outer wall of the cover body 13 and/or the sampling tube 16, and the supporting sleeve 73 is positioned below the two elongated tubes 8 and propped against the lower surfaces of the elongated tubes 8; the lower support sleeve 73 rests on the outer wall of the bottom module 2.
EXAMPLE III
The embodiment provides a sampling detection method for sampling detection by using the integrated device for sampling and detecting air microorganisms, which comprises the following steps:
the method comprises the following steps: connecting an air inlet channel 11 with an air inlet device, opening two opening and closing valves 12, starting the air inlet device to sample air, pre-storing a cracking combination liquid and nucleic acid adsorption magnetic beads at the bottom of a sampling tube 16, and pre-storing eluent in an elution cavity 21; the open-close valve 12 defaults to a closed state;
in the method, a lysis binding solution is used as a sampling solution, in the sampling process, air microorganisms are lysed under the action of the lysis binding solution, and nucleic acid samples of the microorganisms are released to the lysis binding solution and are adsorbed on magnetic beads;
step two: after sampling is finished, the air inlet device is closed, the magnetic rod is inserted into the magnetic rod cavity 42 of the first transfer valve 4, and the magnetic beads at the bottom of the sampling tube 16 are adsorbed to the bottom of the first transfer groove 43 of the first transfer valve 4;
step three: horizontally placing the sampling and detection integrated device, wherein when the device is horizontally placed, the height of the cracking binding liquid in the sampling pipe 16 is lower than the heights of the first transfer groove 43 and the two opening and closing valves, so that the cracking binding liquid is separated from the magnetic beads, and meanwhile, the cracking binding liquid is prevented from flowing out of the sampling pipe;
step four: opening the air inlet device, enabling air to enter the sampling tube 16 from the air inlet channel 11 and the air inlet tube 15, and air-drying residual cracking binding solution on magnetic beads at the bottom of the sampling tube 16, wherein the outlet of the air inlet tube 15 is over against the first transfer groove 43 at the bottom of the sampling tube 16;
step five: closing the air inlet device, closing the opening and closing valve 12, rotating the first transfer valve 4 by 180 degrees to transfer the magnetic beads to the elution cavity 21, pulling out the magnetic bar, and vertically placing the device again;
step six: the magnetic beads naturally fall into the eluent in the elution cavity 21, and the nucleic acid on the magnetic beads is eluted by the eluent through a shaking device;
step seven: inserting a magnetic rod into a magnetic rod cavity 42 of the first transfer valve 4, inverting the device to enable the magnetic beads in the elution cavity 21 to be adsorbed to the bottom of a first transfer groove 43 of the first transfer valve 4, and utilizing the magnetism of the magnetic rod to play a role in fixing the magnetic beads so as to enable only eluent to be transferred when a second transfer groove of the second transfer valve 5 rotates;
step eight: rightly placing the device, rotating the second transfer valve 5 by 180 degrees, so that the eluent is transferred to the sample adding port 31 of the chip 3 and automatically diffuses into the reaction chamber 32, and the reagent required by the detection reaction is pre-embedded in the reaction chamber 32;
step nine: the device is placed on a heating device for detection reaction and observation of a detection result.
The principle and the implementation mode of the invention are explained by applying a specific example, and the description of the embodiment is only used for helping to understand the method and the core idea of the invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, the specific embodiments and the application range may be changed. In view of the above, the present disclosure should not be construed as limiting the invention.
Claims (10)
1. The utility model provides an air microorganism sampling detects integrated device which characterized in that: the device comprises an air microorganism sampling device, an elution cavity and a chip, wherein the sampling cavity of the air microorganism sampling device is used for containing magnetic beads and lysis binding liquid; the elution cavity is used for containing an elution solution; the sampling cavity, the elution cavity and the chip are sequentially arranged from top to bottom, and the sampling cavity is communicated with the elution cavity through a first channel between the sampling cavity and the elution cavity; the elution cavity is communicated with the sample adding port of the chip through a second channel between the elution cavity and the sample adding port; first passageway with it shifts valve and second to be provided with respectively to move about in the second passageway, first transfer valve can with first passageway seals and is used for with the intracavity of sampling the magnetic bead shifts to the elution intracavity, the second transfer valve can with the second passageway seals and is used for shifting the eluant to in the sample addition port.
2. The integrated air microorganism sampling and detecting device according to claim 1, characterized in that: and open and close valves are arranged in the air inlet channel and the air outlet channel of the air microorganism sampling device.
3. The integrated air microorganism sampling and detecting device according to claim 1, characterized in that: the first transfer valve comprises a first rotary ball and a first rotary handle, the first rotary ball is arranged in the first channel and can seal the first channel, the surface of the first rotary ball is inwards recessed to form two first transfer grooves, one end of the first rotary handle is fixedly connected with the first rotary ball, the other end of the first rotary handle extends out of the first channel, and the first rotary handle can be driven to rotate by rotating around the axis of the first rotary handle;
the first transfer valve is internally provided with a magnetic rod cavity, the magnetic rod cavity extends to a position between the two first transfer grooves from one end, far away from the first rotating ball, of the first rotating handle along the direction from the first rotating handle to the first rotating ball, and one end of the magnetic rod can extend into the magnetic rod cavity between the two first transfer grooves;
the second transfer valve comprises a second rotary ball and a second rotary handle, the second rotary ball is arranged in the second channel and can seal the second channel, the surface of the second rotary ball is inwards recessed to form two second transfer grooves, one end of the second rotary handle is fixedly connected with the second rotary ball, the other end of the second rotary handle extends out of the second channel, and the second rotary handle can be driven to rotate by rotating around the axis of the second rotary handle;
in an initial state, the two first transfer grooves vertically face the sampling cavity and the elution cavity respectively; the two second transfer grooves are respectively vertically towards the elution cavity and the sample adding port.
4. The integrated air microorganism sampling and detecting device according to claim 2, characterized in that: the air microorganism sampling device comprises a sampling head, a sampling pipe and an air inlet device, wherein a cavity formed in the sampling pipe is the sampling cavity, the sampling head comprises a cover body and an air inlet pipe, the cover body is provided with an air inlet channel and an air outlet channel which are communicated with the outside, the top end of the air inlet pipe is fixed and is communicated with the air inlet channel, and the bottom end of the air inlet pipe extends to a first transfer groove at the bottom of the sampling pipe.
5. The integrated air microorganism sampling and detecting device according to claim 4, characterized in that: two the open and close valve all includes valve pipe and knob, the one end opening of valve pipe, the other end with knob fixed connection, the mouth that overflows has been seted up to the lateral wall of valve pipe, and part inlet channel and part outlet channel form two valve chambeies respectively, two the open and close valve is corresponding to two respectively the valve chamber, two the valve pipe is located two respectively the valve intracavity, two the knob is all followed the surface of lid exposes, rotates the knob can drive the valve pipe rotates, the valve pipe rotates can open and close inlet channel with the outlet channel.
6. The integrated air microorganism sampling and detecting device according to claim 1, characterized in that: the chip is a paper-based micro-fluidic chip and is made of transparent materials.
7. The air microorganism sampling and detection integrated device according to claim 4, characterized in that: the sampling pipe and the air inlet pipe are both made of flexible and compressible materials.
8. The integrated air microorganism sampling and detecting device according to claim 7, characterized in that: the support device comprises a support frame and a base, and the support frame is fixedly arranged on the base; the elution cavity, the first channel and the second channel are formed in the bottom module, and the bottom module is fixedly connected with the air microorganism sampling device and the chip;
the bottom module, the air microorganism sampling device and the chip are integrally cuboid, the support frame comprises two support rod groups which are arranged up and down, each support rod group comprises two support rods, the positions, close to the top, of two opposite side walls of the air microorganism sampling device are respectively provided with a first groove, the two opposite side walls of the bottom module are respectively provided with a second groove, the two support rods in one support rod group are respectively embedded into the two first grooves, and the two support rods in the other support rod group are respectively embedded into the two second grooves; or
The bottom module is of a cylindrical structure, the cover body and the sampling pipe are of a cylindrical structure integrally, an air outlet and an air inlet are formed in the outer side wall of the cover body, lengthened pipes are fixedly arranged on the air outlet and the air inlet, and the tail ends of the two lengthened pipes extend in the direction far away from the cover body along the radial direction of the cover body; the support frame comprises support sleeves which are arranged up and down, the two support sleeves are both of arc structures, the radian of the upper support sleeve is matched with that of the outer wall of the cover body, the radian of the lower support sleeve is matched with that of the outer wall of the bottom die block, the upper support sleeve is abutted against the outer wall of the cover body and/or the outer wall of the sampling tube, and the support sleeves are positioned below the two elongated tubes and abutted against the lower surfaces of the elongated tubes; the supporting sleeve at the lower part is propped against the outer wall of the bottom module.
9. The integrated air microorganism sampling and detecting device according to claim 8, characterized in that: the base is internally provided with a heating device, and the chip is placed on the base.
10. A sampling detection method for sampling detection by using the integrated air microorganism sampling detection device as defined in any one of claims 1 to 9, the method comprising the steps of: an open-close valve is arranged in the air inlet channel and the air outlet channel of the air microorganism sampling device; the first transfer valve comprises a first rotary ball and a first rotary handle, the first rotary ball is arranged in the first channel and can seal the first channel, the surface of the first rotary ball is inwards recessed to form two first transfer grooves, one end of the first rotary handle is fixedly connected with the first rotary ball, the other end of the first rotary handle extends out of the first channel, and the first rotary handle can be driven to rotate by rotating around the axis of the first rotary handle; the first transfer valve is provided with a first rotation handle and a first rotation ball, the first rotation handle is arranged in the first transfer valve, the first rotation ball is arranged in the first transfer valve, the first rotation handle is arranged in the first transfer valve, the first rotation ball is arranged in the first transfer valve, and the first rotation ball is arranged in the first transfer valve; the second transfer valve comprises a second rotary ball and a second rotary handle, the second rotary ball is arranged in the second channel and can seal the second channel, the surface of the second rotary ball is inwards recessed to form two second transfer grooves, one end of the second rotary handle is fixedly connected with the second rotary ball, the other end of the second rotary handle extends out of the second channel, and the second rotary handle can be driven to rotate by rotating around the axis of the second rotary handle; in the initial state, the two first transfer grooves vertically face the sampling cavity and the elution cavity respectively; the two second transfer grooves are respectively vertically towards the elution cavity and the sample adding port; the air microorganism sampling device comprises a sampling head, a sampling pipe and an air inlet device, wherein a cavity formed in the sampling pipe is the sampling cavity, the sampling head comprises a cover body and an air inlet pipe, the cover body is provided with an air inlet channel and an air outlet channel communicated with the outside, the top end of the air inlet pipe is fixed and communicated with the air inlet channel, and the bottom end of the air inlet pipe extends to a first transfer groove at the bottom of the sampling pipe;
the sampling detection method comprises the following steps:
the method comprises the following steps: connecting the air inlet channel with the air inlet device, opening the two opening and closing valves, opening the air inlet device for air sampling, pre-storing the cracking binding solution and the nucleic acid adsorption magnetic beads at the bottom of the sampling tube, and pre-storing the eluent in the elution cavity;
step two: after sampling is finished, the air inlet device is closed, the magnetic rod is inserted into the magnetic rod cavity of the first transfer valve, and magnetic beads at the bottom of the sampling tube are adsorbed to the bottom of the first transfer groove of the first transfer valve;
step three: horizontally placing the air microorganism sampling and detecting integrated device, wherein when the device is horizontally placed, the height of the cracking binding liquid in the sampling pipe is lower than that of the first transfer groove, so that the cracking binding liquid is separated from the magnetic beads;
step four: opening the air inlet device to enable air to enter the sampling tube from the air inlet channel and the air inlet tube and air-dry residual cracking binding solution on the magnetic beads at the bottom of the sampling tube, wherein an outlet of the air inlet tube is over against the first transfer groove at the bottom of the sampling tube;
step five: closing the air inlet device, closing the opening and closing valve, rotating the first transfer valve by 180 degrees to transfer the magnetic beads to the elution cavity, pulling out the magnetic rod, and vertically placing the device again;
step six: the magnetic beads naturally fall into the eluent in the elution cavity, and the elution is carried out on the nucleic acid on the magnetic beads through the shaking device;
step seven: inserting the magnetic rod into the magnetic rod cavity of the first transfer valve, and inverting the device to adsorb the magnetic beads in the elution cavity to the bottom of the first transfer groove of the first transfer valve;
step eight: rightly placing the device, and rotating the second transfer valve for 180 degrees to transfer the eluent to the chip sample addition port and automatically diffuse the eluent into the reaction chamber;
step nine: the device is placed on the heating device for detection reaction and observation of the detection result.
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Application publication date: 20221004 |