CN114045216B - Air sampling bottle, air sampling system and air sampling method - Google Patents
Air sampling bottle, air sampling system and air sampling method Download PDFInfo
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- CN114045216B CN114045216B CN202210023859.9A CN202210023859A CN114045216B CN 114045216 B CN114045216 B CN 114045216B CN 202210023859 A CN202210023859 A CN 202210023859A CN 114045216 B CN114045216 B CN 114045216B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/06—Magnetic means
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
Abstract
The invention discloses an air sampling bottle, an air sampling system and an air sampling method, and relates to the technical field of microbial detection. The invention integrates air microorganism sampling and nucleic acid extraction, collects the air microorganism sample, simultaneously cracks the air microorganism, and utilizes the magnetic beads to adsorb the nucleic acid released by microorganism cracking, thereby realizing the enrichment of the microorganism nucleic acid, and the eluent obtained by eluting the nucleic acid on the magnetic beads can be directly used for detection, thereby accelerating the detection process of the air microorganism.
Description
Technical Field
The invention relates to the technical field of microorganism detection, in particular to an air sampling bottle, an air sampling system and an air sampling 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 kind 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 various, wherein the impact type air sampling bottle collects microorganism particles in the air into sampling liquid in a jet airflow mode. In the sampling process, after the sampling liquid is added into the air sampling bottle, the air extraction power is started, air enters from the inlet of the air sampling bottle, microbial particles in the air impact the sampling liquid of the air sampling bottle, and the microbial particles are captured due to the adhesion of the liquid.
Common impact type air sampling bottle simple structure, the air microorganism sample of collection generally can not directly be used for detecting, but need with air microorganism sample, sampling solution, can obtain detectable microorganism sample after carrying out operations such as extra cultivation or enrichment, nucleic acid extraction, complex operation, required instrument and equipment are many, consuming time is long, and introduce non-sampling bacterial contamination easily in follow-up operation, be unfavorable for the short-term test to air microorganism.
Disclosure of Invention
The invention aims to provide an air sampling bottle, an air sampling system and an air sampling method, which are used for solving the problems in the prior art.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides an air sampling bottle which comprises a bottle body and a support, wherein the support is arranged on the outer side of the bottle body and connected with the bottle body, an electromagnet is arranged on the support, an air inlet, an air exhaust port, a lysate inlet, an eluent inlet, a washing liquid inlet and an eluent outlet are formed in the upper end of the bottle body, the air inlet is used for collecting air microorganism samples, the air exhaust port is used for extracting air in the bottle body, the lysate inlet is used for introducing lysate and magnetic beads, the magnetic beads can be attracted by the electromagnet, the eluent inlet is used for introducing eluent, the washing liquid inlet is used for introducing washing liquid, the eluent outlet is used for discharging the eluent, a waste liquid outlet is formed in the lower end of the bottle body, and the waste liquid outlet is used for discharging waste liquid and magnetic beads.
Preferably, the support includes the support body, the upper end of support body is provided with solid fixed ring, gu fixed ring cover is established the outside of bottle upper end bottleneck, be provided with annular electro-magnet on the support body, the bottom of support body is provided with the fixed block, be provided with the clearing hole on the fixed block, the waste liquid export is provided with the waste liquid discharge pipe, the waste liquid discharge pipe passes the clearing hole.
Preferably, the bottom of the bottle body is in an inverted cone shape, and the waste liquid outlet is located at the center of the bottom of the bottle body.
Preferably, the air sampling bottle further comprises an air inlet pipe, an air exhaust pipe, a lysate inlet pipe, an eluent inlet pipe, a washing liquid inlet pipe and an eluent outlet pipe, one end of the air inlet pipe is the air inlet, the other end of the air inlet pipe extends into the bottle body, one end of the air exhaust pipe is the air exhaust opening, the other end of the air exhaust pipe extends into the bottle body and is positioned above the liquid level of the bottle body, one end of the lysate inlet pipe is the lysate inlet, the other end of the lysate inlet pipe extends into the bottle body, one end of the eluent inlet pipe is the eluent inlet, the other end of the eluent inlet pipe extends into the bottle body, one end of the washing liquid inlet pipe is the washing liquid inlet, the other end of the washing liquid inlet pipe extends into the bottle body, one end of the eluent outlet pipe is the eluent outlet, and the other end of the eluent outlet pipe extends into the bottle body and is positioned at the bottom of the bottle body.
The invention also provides an air sampling system, which comprises a lysate storage bottle, an eluent storage bottle, a washing liquid storage bottle, a waste liquid bottle, a sample tube, a vacuum pump and the air sampling bottle; the lysate storage bottle is used for storing lysate, the lysate storage bottle is communicated with the lysate inlet of the air sampling bottle through a first pipeline, and a magnetic bead storage structure is arranged on the first pipeline and used for storing magnetic beads; the eluent storage bottle is used for storing eluent and is communicated with the eluent inlet of the air sampling bottle through a second pipeline; the washing liquid storage bottle is used for storing washing liquid, and the washing liquid storage bottle is communicated with the washing liquid inlet of the air sampling bottle through a third pipeline; the waste liquid bottle is used for storing waste liquid, and is communicated with the waste liquid outlet of the air sampling bottle through a fourth pipeline; the sample tube is used for storing a sample to be detected, and the sample tube is communicated with the eluent outlet of the air sampling bottle through a fifth pipeline; the vacuum pump is communicated with the air pumping port of the air sampling bottle through a sixth pipeline.
Preferably, the vacuum pump is communicated with the sample tube through a seventh pipeline, and a valve is arranged on the seventh pipeline.
Preferably, valves are arranged on the first pipeline, the second pipeline, the third pipeline, the fourth pipeline and the sixth pipeline.
Preferably, the magnetic bead storage structure includes a storage structure body and a turntable, the turntable is connected with the storage structure body in a rotating mode, at least one groove is formed in the turntable, a magnetic bead storage cavity is formed in the storage structure body and used for storing magnetic beads, an outlet is formed in the magnetic bead storage cavity, when the turntable rotates to the groove and the outlet is communicated, the magnetic beads in the magnetic bead storage cavity can enter the groove, and when the turntable continues to rotate to the groove and the first pipeline is communicated, the magnetic beads enter the first pipeline.
The invention also provides an air sampling method adopting the air sampling system, which comprises the following steps:
step one, adding magnetic beads into a first pipeline by a magnetic bead storage structure;
opening a valve on a sixth pipeline and a valve on the first pipeline, starting a vacuum pump, sucking the lysate in the lysate storage bottle into the air sampling bottle, and enabling the magnetic beads in the first pipeline to flow into the air sampling bottle along with the lysate in the process that the lysate flows into the air sampling bottle from the lysate storage bottle;
step three, closing a valve on the first pipeline, opening a valve on the air inlet pipe, carrying out air sampling, cracking an air microorganism sample under the action of a cracking solution in the sampling process, and releasing and adsorbing nucleic acid onto magnetic beads;
step four, after sampling is finished, the vacuum pump is turned off, the valve on the sixth pipeline is turned off, the electromagnet is electrified, and the magnetic beads are adsorbed to the side wall of the air sampling bottle;
opening a valve on a waste liquid discharge pipe, and enabling the lysate to flow into a waste liquid bottle;
step six, closing a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, powering off an electromagnet, opening a valve on a sixth pipeline and a valve on a third pipeline, starting a vacuum pump, sucking the cleaning solution into the air sampling bottle from the cleaning solution storage bottle, and cleaning nucleic acid;
step seven, the vacuum pump is turned off, the valve on the sixth pipeline and the valve on the third pipeline are turned off, the electromagnet is electrified, and the magnetic beads are adsorbed to the side wall of the air sampling bottle;
step eight, opening a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, and enabling the washing liquid to flow into a waste liquid bottle;
step nine, closing a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, powering off an electromagnet, opening a valve on a sixth pipeline and a valve on a second pipeline, starting a vacuum pump, sucking eluent from an eluent storage bottle into an air sampling bottle, and eluting nucleic acid from magnetic beads;
step ten, turning off the vacuum pump, turning off a valve on the sixth pipeline and a valve on the second pipeline, electrifying the electromagnet, and adsorbing the magnetic beads to the side wall of the air sampling bottle;
step eleven, opening a valve on the seventh pipeline and a valve on the air inlet pipe, starting a vacuum pump, and sucking eluent containing the air microorganism nucleic acid sample into a sample tube, wherein the air microorganism nucleic acid sample in the sample tube can be used for detecting air microorganisms;
step twelve, closing a valve on the seventh pipeline and a valve on the air inlet pipe, powering off the electromagnet, opening a valve on the second pipeline, sucking the eluent into the air sampling bottle by the vacuum pump, washing the air sampling bottle, and simultaneously suspending magnetic beads in the eluent;
step thirteen, the vacuum pump is closed, the valve on the second pipeline is closed, the valve on the air inlet pipe and the valve on the waste liquid discharge pipe are opened, and the magnetic beads flow into the waste liquid bottle along with the flushing liquid;
step fourteen, closing a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, opening a valve on a second pipeline, starting a vacuum pump, sucking eluent into the air sampling bottle by the vacuum pump, and flushing the air sampling bottle again;
and step fifteen, closing the vacuum pump, closing a valve on the second pipeline, opening a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, enabling the washed eluent to flow into the waste liquid bottle, and closing the valve on the air inlet pipe and the valve on the waste liquid discharge pipe.
Compared with the prior art, the invention has the following technical effects:
the invention integrates air microorganism sampling and nucleic acid extraction, directly takes lysis solution as sampling solution to collect air microorganisms, collects air microorganism samples and simultaneously lyses the air microorganisms, and utilizes magnetic beads to adsorb nucleic acid released by microbial lysis, thereby realizing the enrichment of microbial nucleic acid, and eluent obtained by eluting nucleic acid on the magnetic beads can be directly used for detection, thereby accelerating the detection process of the air microorganisms; in addition, the sampling system needs few instruments and equipment, can realize full-automatic operation through a central controller, and is simple and convenient to operate and low in cost.
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 view of an air sampling bottle of the present invention;
FIG. 2 is a schematic view of a bottle of the present invention;
FIG. 3 is a schematic view of an air inlet tube, an air exhaust tube, a lysate inlet tube, an eluent inlet tube, a washing liquid inlet tube and an eluent outlet tube according to the present invention;
FIG. 4 is a front view of the stand of the present invention;
FIG. 5 is a top view of the bracket of the present invention;
FIG. 6 is a schematic view of an air sampling system of the present invention;
FIG. 7 is a schematic diagram of a magnetic bead storage structure according to the present invention;
FIG. 8 is a schematic diagram illustrating an operation process of a magnetic bead storage structure according to the present invention;
wherein: 100-air sampling bottle, 200-air sampling system, 1-bottle body, 2-bracket, 3-electromagnet, 4-air inlet, 5-air extraction opening, 6-lysate inlet, 7-eluent inlet, 8-washing liquid inlet, 9-eluent outlet, 10-waste liquid outlet, 11-bracket body, 12-fixing ring, 13-fixing block, 14-through hole, 15-waste liquid discharge pipe, 16-air inlet pipe, 17-air extraction pipe, 18-lysate inlet pipe, 19-eluent inlet pipe, 20-washing liquid inlet pipe, 21-eluent outlet pipe, 22-lysate storage bottle, 23-eluent storage bottle, 24-washing liquid storage bottle, 25-waste liquid bottle, 26-sample pipe, 27-vacuum pump, 28-a first pipeline, 29-a second pipeline, 30-a third pipeline, 31-a fourth pipeline, 32-a fifth pipeline, 33-a sixth pipeline, 34-a seventh pipeline, 35-a valve, 36-a magnetic bead storage structure, 37-a storage structure body, 38-a turntable, 39-a magnetic bead storage cavity, 40-a groove, 41-a magnetic bead, 42-a bottle stopper and 43-a bottle neck.
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 obtained by a person skilled in the art without inventive effort based on the embodiments of the present invention, are within the scope of the present invention.
The invention aims to provide an air sampling bottle, an air sampling system and an air sampling 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
As shown in fig. 1-5: the embodiment provides an air sampling bottle 100, which comprises a bottle body 1 and a support 2, the support 2 is arranged outside the bottle body 1 and connected with the bottle body 1, the support 2 is provided with an electromagnet 3, the upper end of the bottle body 1 is provided with an air inlet 4, an air suction opening 5, a lysate inlet 6, an eluent inlet 7, a washing liquid inlet 8 and an eluent outlet 9, the air inlet 4 is used for collecting air microorganism samples, the air suction opening 5 is used for extracting air in the bottle body 1 to form negative pressure in the bottle body 1, the lysate inlet 6 is used for introducing lysate and magnetic beads 41, the magnetic beads 41 can adsorb nucleic acid of the air microorganisms, the magnetic beads 41 can enter the air sampling bottle 100 together with the lysate when the lysate enters the air sampling bottle 100, the magnetic beads 41 can be attracted by the electromagnet 3 after being electrified, the eluent inlet 7 is used for introducing eluent, the washing liquid is introduced into the washing liquid inlet 8, after the elution of the nucleic acid on the magnetic beads 41 by the eluent, the eluent can be guided out through an eluent outlet 9, the eluent can be directly used for detection, a waste liquid outlet 10 is formed in the lower end of the bottle body 1, and the waste liquid outlet 10 is used for discharging waste liquid and the magnetic beads 41.
In this embodiment, support 2 includes support body 11, support body 11's upper end is provided with solid fixed ring 12, gu fixed ring 12 cover establishes the outside at bottleneck 43, gu fixed ring 12's diameter is greater than the diameter of bottleneck 43 and is less than the diameter of bottle 1 to fix support 2's upper end in bottleneck 43 department, be provided with annular electro-magnet 3 on the support body 11, when electro-magnet 3 circular telegram, this electro-magnet 3 has magnetism, can attract magnetic bead 41, when electro-magnet 3 outage, this electro-magnet 3 loses magnetism. The bottom of the support body 11 is provided with a fixed block 13, the fixed block 13 is provided with a through hole 14, the waste liquid outlet 10 is provided with a waste liquid discharge pipe 15, the waste liquid discharge pipe 15 passes through the through hole 14, and the waste liquid discharge pipe 15 is provided with a valve 35 which can be used for controlling the discharge of waste liquid and magnetic beads 41 in the bottle body 1.
In this embodiment, the bottom of the bottle body 1 is an inverted cone, and the waste liquid outlet 10 is located at the center of the bottom of the bottle body 1, that is, the bottom of the bottle body 1 is inclined by a certain angle (low center and high all around) from the center to the four circumferential directions, so that the liquid in the bottle body 1 flows out from the waste liquid outlet 10, and the residue of the liquid in the bottle body 1 is reduced.
In this embodiment, the air sampling bottle 100 further includes an air inlet tube 16, an air exhaust tube 17, a lysate inlet tube 18, an eluent inlet tube 19, a washing liquid inlet tube 20 and an eluent outlet tube 21, the air inlet tube 16, the air exhaust tube 17, the lysate inlet tube 18, the eluent inlet tube 19, the washing liquid inlet tube 20 and the eluent outlet tube 21 are fixed by a bottle stopper 42 at the bottle mouth of the bottle body 1, one end of the air inlet tube 16 is an air inlet 4, the other end of the air inlet tube 16 extends into the bottle body 1, one end of the air exhaust tube 17 is an air exhaust opening 5, the other end of the air exhaust tube 17 extends into the bottle body 1 and is located above the liquid level of the bottle body 1, one end of the lysate inlet tube 18 is a lysate inlet 6, the other end of the lysate inlet tube 18 extends into the bottle body 1, one end of the eluent inlet tube 19 is an eluent inlet 7, the other end of the eluent inlet tube 19 extends into the bottle body 1, one end of the washing liquid inlet tube 20 is a washing liquid inlet 8, the other end of the washing liquid inlet pipe 20 extends into the bottle body 1, one end of the eluent outlet pipe 21 is an eluent outlet 9, and the other end of the eluent outlet pipe 21 extends into the bottle body 1 and is positioned at the bottom of the bottle body 1.
The air sampling bottle 100 of this embodiment has contained the collection of air microorganism sample and has extracted required structure with nucleic acid, utilizes this air sampling bottle 100, can accomplish collection and nucleic acid extraction to air microorganism sample, easy operation, and the nucleic acid extract that obtains can directly be used for detecting, avoids the miscellaneous fungus pollution that loaded down with trivial details operation introduced, convenient and fast.
Example two
As shown in fig. 6 to 8, the present embodiment provides an air sampling system 200, which includes a lysis solution storage bottle 22, an eluent storage bottle 23, a washing solution storage bottle 24, a waste solution bottle 25, a sample tube 26, a vacuum pump 27, and the air sampling bottle 100 of the first embodiment; the lysate storage bottle 22 is used for storing lysate, the lysate storage bottle 22 is communicated with the lysate inlet 6 of the air sampling bottle 100 through a first pipeline 28, a magnetic bead storage structure 36 is arranged on the first pipeline 28, and the magnetic bead storage structure 36 is used for storing magnetic beads 41; the eluent storage bottle 23 is used for storing eluent, and the eluent storage bottle 23 is communicated with the eluent inlet 7 of the air sampling bottle 100 through a second pipeline 29; the cleaning solution storage bottle 24 is used for storing cleaning solution, and the cleaning solution storage bottle 24 is communicated with the cleaning solution inlet 8 of the air sampling bottle 100 through a third pipeline 30; the waste liquid bottle 25 is used for storing waste liquid, the waste liquid bottle 25 is communicated with the waste liquid outlet 10 of the air sampling bottle 100 through a fourth pipeline 31, and a valve 35 is arranged on the fourth pipeline 31; the sample tube 26 is used for storing a sample to be detected, and the sample tube 26 is communicated with the eluent outlet 9 of the air sampling bottle 100 through a fifth pipeline 32; the vacuum pump 27 is in communication with the suction port 5 of the air sampling bottle 100 through a sixth line 33.
In this embodiment, the vacuum pump 27 is communicated with the sample tube 26 through a seventh pipeline 34, and the seventh pipeline 34 is provided with a valve 35. Valves 35 are provided on the first line 28, on the second line 29, on the third line 30 and on the sixth line 33. The valve 35 prevents airborne microorganisms from entering the lysate storage bottle 22, the eluent storage bottle 23, the wash solution storage bottle 24 and the vacuum pump 27.
In this embodiment, the magnetic bead storage structure 36 includes storage structure body 37 and carousel 38, carousel 38 rotates with storage structure body 37 to be connected, at least one recess 40 has been seted up on carousel 38, this embodiment includes the recess 40 that two symmetries set up, two recesses 40 are the same in size and shape, can transfer the magnetic bead 41 ration in the magnetic bead storage structure 36 to first pipeline 28, be provided with magnetic bead storage chamber 39 on the storage structure body 37, magnetic bead storage chamber 39 is used for storing magnetic bead 41, magnetic bead storage chamber 39 has seted up the export, when carousel 38 rotates to recess 40 and export intercommunication, magnetic bead 41 in the magnetic bead storage chamber 39 can get into in the recess 40, when carousel 38 continues to rotate to recess 40 and first pipeline 28 intercommunication, magnetic bead 41 gets into in the first pipeline 28.
Specifically, fig. 8 is a schematic diagram of a working flow of the magnetic bead storage structure 36, in a default state, two grooves 40 of the turntable 38 are in a vertical direction, when the turntable 38 is in operation, the turntable 38 rotates according to the sequence in fig. 8 (in the figure, the rotation direction of the turntable 38 is clockwise, and may also rotate counterclockwise, and the achieved effect is the same), along with the rotation of the turntable 38, the magnetic beads 41 in the grooves 40 are transferred out of the magnetic bead 41 storage cavity, and when the grooves 40 of the turntable 38 are in the vertical direction again, the transferred magnetic beads 41 fall into the first pipeline 28. Wherein, the volume of the groove 40 is fixed, so as to ensure the volume of the magnetic beads 41 transferred each time is consistent.
The air sampling system 200 of this embodiment collects microorganism and extracts in an organic whole with nucleic acid, lysate storage bottle 22, eluant storage bottle 23, the washing liquid storage bottle 24 and the magnetic bead store and contain lysate, eluant, washing liquid and magnetic bead 41 respectively in the structure 36, except air sampling bottle 100, other storage bottles and storage chamber are acquiescent with atmosphere intercommunication, the position of intercommunication can set up the filter membrane, prevent that air microorganism from getting into corresponding bottle, in the cavity, valve 35 in the system is acquiescent for the closed condition.
The air sampling system of this embodiment includes a central controller, and each valve 35, the vacuum pump 27 and the magnetic bead storage structure 36 are all connected to the central controller, and the central controller can automatically control the opening and closing of each valve 35, the vacuum pump 27 and the operation of the magnetic bead storage structure 36, thereby realizing the full-automatic operation of this system. The control process of the central controller is prior art.
EXAMPLE III
The present embodiment provides a method for sampling air by using the air sampling system 200 of the second embodiment, which includes the following steps:
step one, the magnetic bead storage structure 36 adds the magnetic bead 41 into the first pipeline 28;
step two, opening a valve 35 on the sixth pipeline 33 and a valve 35 on the first pipeline 28, starting the vacuum pump 27, sucking the lysate in the lysate storage bottle 22 into the air sampling bottle 100, and in the process that the lysate flows into the air sampling bottle 100 from the lysate storage bottle 22, allowing the magnetic beads 41 in the first pipeline 28 to flow into the air sampling bottle 100 along with the lysate;
step three, closing the valve 35 on the first pipeline 28, opening the valve 35 on the air inlet pipe 16, sampling air, cracking the air microorganism sample under the action of a cracking solution in the sampling process, and releasing and adsorbing nucleic acid onto the magnetic beads 41;
step four, after the sampling is finished, the vacuum pump 27 is turned off, the valve 35 on the sixth pipeline 33 is turned off, the electromagnet 3 is powered on, and the magnetic beads 41 are adsorbed to the side wall of the air sampling bottle 100;
step five, opening a valve 35 on the waste liquid discharge pipe 15, and enabling the lysate to flow into a waste liquid bottle 25;
step six, closing the valve 35 on the air inlet pipe 16 and the valve 35 on the waste liquid discharge pipe 15, powering off the electromagnet 3, opening the valve 35 on the sixth pipeline 33 and the valve 35 on the third pipeline 30, starting the vacuum pump 27, sucking the cleaning solution from the cleaning solution storage bottle 24 into the air sampling bottle 100, and washing the nucleic acid;
step seven, the vacuum pump 27 is turned off, the valve 35 on the sixth pipeline 33 and the valve 35 on the third pipeline 30 are turned off, the electromagnet 3 is powered on, and the magnetic beads 41 are adsorbed to the side wall of the air sampling bottle 100;
step eight, opening a valve 35 on the air inlet pipe 16 and a valve 35 on the waste liquid discharge pipe 15, and enabling the washing liquid to flow into the waste liquid bottle 25;
step nine, closing the valve 35 on the air inlet pipe 16 and the valve 35 on the waste liquid discharge pipe 15, powering off the electromagnet 3, opening the valve 35 on the sixth pipeline 33 and the valve 35 on the second pipeline 29, starting the vacuum pump 27, sucking the eluent from the eluent storage bottle 23 into the air sampling bottle 100, and eluting the nucleic acid from the magnetic beads 41;
step ten, turning off the vacuum pump 27, turning off the valve 35 on the sixth pipeline 33 and the valve 35 on the second pipeline 29, electrifying the electromagnet 3, and adsorbing the magnetic beads 41 to the side wall of the air sampling bottle 100;
step eleven, opening a valve 35 on the seventh pipeline 34 and a valve 35 on the air inlet pipe 16, starting the vacuum pump 27, sucking the eluent containing the air microorganism nucleic acid sample into the sample pipe 26, wherein the air microorganism nucleic acid sample in the sample pipe 26 can be used for detecting the air microorganism;
step twelve, closing the valve 35 on the seventh pipeline 34 and the valve 35 on the air inlet pipe 16, powering off the electromagnet 3, opening the valve 35 on the second pipeline 29, sucking the eluent into the air sampling bottle 100 by the vacuum pump 27, washing the air sampling bottle 100, and suspending the magnetic beads 41 in the eluent;
step thirteen, the vacuum pump 27 is closed, the valve 35 on the second pipeline 29 is closed, the valve 35 on the air inlet pipe 16 and the valve 35 on the waste liquid discharge pipe 15 are opened, and the magnetic beads 41 flow into the waste liquid bottle 25 along with the flushing liquid;
step fourteen, closing the valve 35 on the air inlet pipe 16 and the valve 35 on the waste liquid discharge pipe 15, opening the valve 35 on the second pipeline 29, starting the vacuum pump 27, sucking the eluent into the air sampling bottle 100 by the vacuum pump 27, and flushing the air sampling bottle 100 again;
step fifteen, the vacuum pump 27 is closed, the valve 35 on the second pipeline 29 is closed, the valve 35 on the air inlet pipe 16 and the valve 35 on the waste liquid discharge pipe 15 are opened, the washed eluent flows into the waste liquid bottle 25, and the valve 35 on the air inlet pipe 16 and the valve 35 on the waste liquid discharge pipe 15 are closed.
The principle and the implementation mode of the present invention are explained by applying specific examples in the present specification, and the above descriptions of the examples are only used to help understanding the method and the core idea of the present 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 (9)
1. An air sampling bottle which is characterized in that: the device comprises a bottle body and a support, the support sets up the outside of bottle and with the bottle is connected, be provided with the electro-magnet on the support, the upper end of bottle is provided with air inlet, extraction opening, lysate import, eluent import, washing liquid import and eluent export, the air inlet is used for gathering air microorganism sample, the extraction opening is used for the extraction air in the bottle, the lysate import is used for letting in lysate and magnetic bead, the magnetic bead can be by the electro-magnet attraction, the eluent import is used for letting in the eluent, the washing liquid import lets in the washing liquid, the eluent export is used for discharging the eluent, the waste liquid export has been seted up to the lower extreme of bottle, the waste liquid export is used for discharging waste liquid and magnetic bead.
2. The air sampling bottle of claim 1, wherein: the support includes the support body, the upper end of support body is provided with solid fixed ring, gu fixed ring cover is established the outside of bottle upper end bottleneck, be provided with annular electro-magnet on the support body, the bottom of support body is provided with the fixed block, be provided with the clearing hole on the fixed block, the waste liquid export is provided with the waste liquid discharge pipe, the waste liquid discharge pipe passes the clearing hole.
3. The air sampling bottle of claim 1, wherein: the bottle bottom of the bottle body is in an inverted cone shape, and the waste liquid outlet is positioned in the center of the bottle bottom of the bottle body.
4. The air sampling bottle of claim 1, wherein: the air sampling bottle also comprises an air inlet pipe, an air exhaust pipe, a lysate inlet pipe, an eluent inlet pipe, a washing liquid inlet pipe and an eluent outlet pipe, one end of the air inlet pipe is the air inlet, the other end of the air inlet pipe extends into the bottle body, one end of the air exhaust pipe is the air exhaust opening, the other end of the air exhaust pipe extends into the bottle body and is positioned above the liquid level of the bottle body, one end of the lysate inlet pipe is the lysate inlet, the other end of the lysate inlet pipe extends into the bottle body, one end of the eluent inlet pipe is the eluent inlet, the other end of the eluent inlet pipe extends into the bottle body, one end of the washing liquid inlet pipe is the washing liquid inlet, the other end of the washing liquid inlet pipe extends into the bottle body, one end of the eluent outlet pipe is the eluent outlet, and the other end of the eluent outlet pipe extends into the bottle body and is positioned at the bottom of the bottle body.
5. An air sampling system, characterized by: comprising a lysate storage bottle, an eluent storage bottle, a washing solution storage bottle, a waste liquid bottle, a sample tube, a vacuum pump and an air sampling bottle according to any one of claims 1-4; the lysate storage bottle is used for storing lysate, the lysate storage bottle is communicated with the lysate inlet of the air sampling bottle through a first pipeline, and a magnetic bead storage structure is arranged on the first pipeline and used for storing magnetic beads; the eluent storage bottle is used for storing eluent and is communicated with the eluent inlet of the air sampling bottle through a second pipeline; the washing liquid storage bottle is used for storing washing liquid, and the washing liquid storage bottle is communicated with the washing liquid inlet of the air sampling bottle through a third pipeline; the waste liquid bottle is used for storing waste liquid, and is communicated with the waste liquid outlet of the air sampling bottle through a fourth pipeline; the sample tube is used for storing a sample to be detected, and the sample tube is communicated with the eluent outlet of the air sampling bottle through a fifth pipeline; the vacuum pump is communicated with the air pumping port of the air sampling bottle through a sixth pipeline.
6. The air sampling system of claim 5, wherein: the vacuum pump is communicated with the sample tube through a seventh pipeline, and a valve is arranged on the seventh pipeline.
7. The air sampling system of claim 5, wherein: valves are arranged on the first pipeline, the second pipeline, the third pipeline, the fourth pipeline and the sixth pipeline.
8. The air sampling system of claim 5, wherein: the magnetic bead storage structure comprises a storage structure body and a rotary table, the rotary table is connected with the storage structure body in a rotating mode, at least one groove is formed in the rotary table, a magnetic bead storage cavity is formed in the storage structure body and used for storing magnetic beads, an outlet is formed in the magnetic bead storage cavity, when the rotary table rotates to the groove and the outlet is communicated, the magnetic beads in the magnetic bead storage cavity can enter the groove, and when the rotary table continues to rotate to the groove and the first pipeline is communicated, the magnetic beads enter the first pipeline.
9. An air sampling method using the air sampling system according to any one of claims 5 to 8, characterized in that: the method comprises the following steps:
step one, adding magnetic beads into a first pipeline by a magnetic bead storage structure;
opening a valve on a sixth pipeline and a valve on the first pipeline, starting a vacuum pump, sucking the lysate in the lysate storage bottle into the air sampling bottle, and enabling the magnetic beads in the first pipeline to flow into the air sampling bottle along with the lysate in the process that the lysate flows into the air sampling bottle from the lysate storage bottle;
step three, closing a valve on the first pipeline, opening a valve on the air inlet pipe, carrying out air sampling, cracking an air microorganism sample under the action of a cracking solution in the sampling process, and releasing and adsorbing nucleic acid onto magnetic beads;
step four, after sampling is finished, the vacuum pump is turned off, the valve on the sixth pipeline is turned off, the electromagnet is electrified, and the magnetic beads are adsorbed to the side wall of the air sampling bottle;
opening a valve on a waste liquid discharge pipe, and enabling the lysate to flow into a waste liquid bottle;
step six, closing a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, powering off an electromagnet, opening a valve on a sixth pipeline and a valve on a third pipeline, starting a vacuum pump, sucking the cleaning solution into the air sampling bottle from the cleaning solution storage bottle, and cleaning nucleic acid;
step seven, the vacuum pump is turned off, the valve on the sixth pipeline and the valve on the third pipeline are turned off, the electromagnet is electrified, and the magnetic beads are adsorbed to the side wall of the air sampling bottle;
step eight, opening a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, and enabling the washing liquid to flow into a waste liquid bottle;
step nine, closing a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, powering off an electromagnet, opening a valve on a sixth pipeline and a valve on a second pipeline, starting a vacuum pump, sucking eluent from an eluent storage bottle into an air sampling bottle, and eluting nucleic acid from magnetic beads;
step ten, turning off the vacuum pump, turning off a valve on the sixth pipeline and a valve on the second pipeline, electrifying the electromagnet, and adsorbing the magnetic beads to the side wall of the air sampling bottle;
step eleven, opening a valve on the seventh pipeline and a valve on the air inlet pipe, starting a vacuum pump, and sucking eluent containing the air microorganism nucleic acid sample into a sample tube, wherein the air microorganism nucleic acid sample in the sample tube can be used for detecting air microorganisms;
step twelve, closing a valve on the seventh pipeline and a valve on the air inlet pipe, powering off the electromagnet, opening a valve on the second pipeline, sucking the eluent into the air sampling bottle by the vacuum pump, washing the air sampling bottle, and simultaneously suspending magnetic beads in the eluent;
step thirteen, the vacuum pump is closed, the valve on the second pipeline is closed, the valve on the air inlet pipe and the valve on the waste liquid discharge pipe are opened, and the magnetic beads flow into the waste liquid bottle along with the flushing liquid;
step fourteen, closing a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, opening a valve on a second pipeline, starting a vacuum pump, sucking eluent into the air sampling bottle by the vacuum pump, and flushing the air sampling bottle again;
and step fifteen, closing the vacuum pump, closing a valve on the second pipeline, opening a valve on the air inlet pipe and a valve on the waste liquid discharge pipe, enabling the washed eluent to flow into the waste liquid bottle, and closing the valve on the air inlet pipe and the valve on the waste liquid discharge pipe.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114774270A (en) * | 2022-06-22 | 2022-07-22 | 至美时代生物智能科技(北京)有限公司 | Automatic sampling device and sampling method |
| CN114774264B (en) * | 2022-06-22 | 2022-09-09 | 至美时代生物智能科技(北京)有限公司 | Automatic air microorganism detection system |
| CN115141739A (en) * | 2022-09-06 | 2022-10-04 | 至美时代生物智能科技(北京)有限公司 | Air microorganism sampling and detection integrated device and sampling and detection method |
| CN115639030B (en) * | 2022-11-28 | 2023-06-23 | 北京慧荣和科技有限公司 | Aerosol collecting and washing integrated machine and unmanned transportation equipment |
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