CN109406215A - A kind of water body environment DNA intelligent acquisition device and acquisition method - Google Patents
A kind of water body environment DNA intelligent acquisition device and acquisition method Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 46
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- 230000008569 process Effects 0.000 claims abstract description 18
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 claims description 33
- 239000012528 membrane Substances 0.000 claims description 25
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/10—Devices for withdrawing samples in the liquid or fluent state
- G01N1/14—Suction devices, e.g. pumps; Ejector devices
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- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/10—Devices for withdrawing samples in the liquid or fluent state
- G01N1/14—Suction devices, e.g. pumps; Ejector devices
- G01N2001/1418—Depression, aspiration
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Abstract
The invention discloses a kind of water body environment DNA intelligent acquisition device and acquisition methods, belong to environment DNA detection technique field.Water body environment DNA intelligent acquisition device and acquisition method provided by the present invention; integrate intelligent control, sample acquisition, the emptying of residual night and pipeline blockage decompression protection function; both the acquisition of aqueous bio sample had been can be carried out; it can be carried out water sampling again; biological sample sampling process efficiency can effectively be improved; drainage amount calculates accuracy, ensure that the timeliness of sample, so that the environment of detection water sample is closer to natural environment；Have the advantages that be not easy to plug, is simple structure and reasonable design, easily fabricated.
Description
Technical field
The invention belongs to environment DNA detection technique fields, more specifically to a kind of water body environment DNA intelligent acquisition
Device and acquisition method.
Background technique
Species distribution, species richness situation are all the main problem of Freshwater ecosystems Diversity Study, these
Play the role of for implementation bio-diversity conservation measure vital.DNA is obtained from species viability environment reflects species
Distribution, the information such as abundance are a kind of non-intrudings, quick species detection means.With the fast development of high throughput sequencing technologies,
Diversity Study method based on environment DNA and Protocols in Molecular Biology (the macro bar codes technique of eDNA) starts more and more
Be applied in Ecology health assessment, significantly improve our level of understandings to ecosystem species diversity.Such as
The researchers such as Ficetda (Species detection using environmental DNA from water samples
[J] Biol Lett, 2008,4 (4): 432-435) environment DNA detection method is just once successfully utilized, it has detected in natural pond
Invade species Rana castes beiana shaw there are situations.
It is well known that the error of analytic process can be gradually decreased by technological progress, but sampling technique is very one section long
Progress is insufficient in period, therefore the overall error for analyzing measurement final result often comes from sampling process.With traditional biological multiplicity
The sampling mode of the acquisition bion sample of sex investigation is different, and the macro bar codes technique of DNA needs in quick obtaining receiving water body
Total environment DNA therefore to obtain correct, reliable analysis as a result, the representativeness of institute's sample thief, accuracy and complete
Property become water environment DNA monitoring matter of utmost importance.The correct selection method of sampling and device, improve sampling technique, for improving
Research and application quality is extremely important., whereas if sampling fault, subsequent analytic process will lose meaning, or even cause
Huge waste.
Traditional sample collection method is subsequent to be carried out dehydrating in the lab to obtain environment based on manually acquiring
DNA acquires sample, such as Chinese utility model patent, Authorization Notice No.: CN203811429U, authorized announcement date:
2014.09.3, a kind of netting gear for the qualitative sample acquisition of planktonic organism in torrent water body is disclosed, including bag-shaped dictyosome,
The front end opening end of dictyosome is equipped with valve, and the middle part of dictyosome is equipped with hollow pipe collar, and dictyosome is divided into funnelform net by pipe collar
Body and positioned at body rear and the net tripe that is connected with body, can be widely used for planktonic organism sample acquisition field.But this method
With following obvious shortcoming: (1) manual operation, inefficiency: big by constraints such as water body bank, ships, sample range is limited, sample
Product are representative poor；And too many human and material resources are spent, working efficiency is low；(2) sampling location is difficult to accurately control: by water currents,
Plankton net easily drifts out the influence of many factors such as sample point and ship when sampling, can not carry out ship for certain waters
Only sample；(3) the calculating error of drainage amount is larger: due to the variation of plankton net sampling depth, being discharged and is grown according to wirerope
There are large errors for the drainage amount that degree, sampling angle and sampling time estimate, and with the calculated plant of swimming of the drainage amount
Object biological density is not inconsistent with actual conditions；(4) environmental water sample easily changes, and measures this period from analysis is collected
It is interior, due to the change of environmental condition, the influence of the biological metabolic activity in water body, chemical action and physical action,
The variation that can cause acquired sample causes the variation of environment DNA even to be degraded, and then influences the verification and measurement ratio of species, meanwhile,
Also the testing results such as species distribution be will affect.And these variations are cracking, therefore, it is necessary to take necessary safeguard measure pair
Sample is saved.Although the macro bar codes technique of eDNA greatly improves environmental monitoring as emerging environmental monitoring means
Efficiency and accuracy, but traditional biological sampling method and the demand for being unsatisfactory for this novel monitoring technology, also limit this
Large-scale promotion application of the method in environment friendly system.Therefore, when carrying out water body environment sampling, the guarantee and control of sampling quality are carried out
System is very important.Such as Chinese utility model patent, the applying date: 2016.05.25, application number: 201620486713.8,
A kind of water body environment DNA sampling apparatus is disclosed, including shield, pumping rod, water pumper and the pumping conduit being attached thereto, is taken out
The other end of waterpipe is equipped with filter paper, and filter paper is fixed by fixed ring, and fixed ring further includes multiple for covering the fan of filter paper
Leaf, multiple flabellums cooperatively form circle, one of angle of flabellum and retainer plate axis connection, and the edge of flabellum also passes through connection
Pipe is connected to shield；The shield includes the necklace and movable branching rod of bottom, and necklace is mounted on pumping conduit, activity branch
Bar is connect with strut, the collection in worksite work of the achievable water body environment DNA of the utility model, without subsequent heavy filtering behaviour
Make, it is time saving and energy saving, but still have, quantitative collection can not be carried out, drainage amount error is big, and especially equipment is in use process
In, it is easy to happen obstruction, gently then influences normal use, it is heavy then damage pipeline
In addition, it is worth noting that, the investigation of species richness is based on environment DNA concentration, and environment DNA concentration is by DNA's
The influence of release and degradation process, while environment DNA extracting method and water sample volume also will affect environment DNA concentration.eDNA
Macro bar codes technique be mainly pass through obtain receiving water body in planktonic organism (including plant, animal body, microbial body and
Various sizes of organism), based on suspended particulate substance in water sample etc., to detect environment DNA, therefore, if thinking most true
React the DNA in corresponding water body environment, it is necessary to guarantee the real effectiveness of acquired sample.
For above-mentioned variation is reduced to minimum level, certain safeguard measure can be generally taken sample, for example taken cold
Hiding, the measure of freezing or addition stabilizer etc.；Then, shorten haulage time, as early as possible analysis measurement as much as possible.Such as China
Patent of invention, authorized announcement date 2018.07.17,105636937 B of Authorization Notice No. CN are disclosed a kind of for saving biology
The stabilizer of sample, in use, stabilizer is mixed according to a certain ratio with biological sample, it can be non-frozen using the stabilizer
Under the conditions of keep the integrity and stability of biological sample for a long time；But still have stabilizer interference monitoring item be added
The risk of range estimation.
Therefore, environment DNA sample gathering apparatus is developed, existing sampling technique is improved, improves sampling efficiency, reduction sampled
Influence disturbing factor in journey reduces environment DNA extracting method and water sample volume to the influence factor of environment DNA, for mentioning
The research and application quality of high water body environment species richness just seems and its important.
Summary of the invention
1. to solve the problems, such as
For existing water body environment biological sample sampling process inefficiency, drainage amount calculating error is big, raw after acquisition
Object sample easily changes and existing sample devices is easily because of the problem of blocking, causing conduit failure, and the present invention provides a kind of water
Body environment DNA intelligent acquisition device and acquisition method, which can make portable quantitative enrichment and sampling uses, and can also cooperate nothing
The vehicle equipments such as people's ship or unmanned plane use, and can greatly reduce artificial intensity, improve sample accuracy and representativeness.
2. technical solution
To solve the above-mentioned problems, the technical solution adopted in the present invention is as follows:
A kind of water body environment DNA intelligent acquisition device, including intelligent control mechanism and sampling main body mechanism, described adopts
Sample main body mechanism includes sample collection unit, pressure release unit and water emptying unit；
The sample collection unit includes water pump, collector and flowmeter, described water pump one end and outside body of water
Connection, the other end are connected with the water inlet of collector；The flowmeter is connected with the water outlet of collector；
The described water emptying unit includes admission line and vacuum pump, described admission line one end and collector into
The mouth of a river is connected, and the other end is communicated with outside atmosphere；Described vacuum pump one end is connected with flowmeter, the other end and external rings
Border is connected；
The pressure release unit includes relief valve, and the relief valve is arranged between water pump and collector, and with outside
Atmosphere；
The intelligent control mechanism includes master control borad and the GPS positioning block being connected with master control borad, the master control
Plate empties opening or closing for unit to control acquisition unit and water, and receives the location information of GPS positioning block.
Preferably, the sampling main body mechanism further includes triple valve, the first import of the triple valve and external water
Body connection；The outlet of the triple valve is connected with water pump；Second import of the triple valve and water emptying unit into
Feed channel is connected.
Preferably the sample collection unit further includes coarse filter, solenoid valve II and check valve；The thick mistake
Filter is arranged on the first inlet pipeline of triple valve, and the pipeline between the first import of coarse filter and triple valve is equipped with electricity
Magnet valve II；The check valve is in shape in parallel with vacuum pump, is connected in pipeline.
Preferably, water emptying unit further includes that solenoid valve I described in solenoid valve I is located on admission line.
Preferably, the collector is that enrichment collector is used for water sample biological concentration, is set in the enrichment collector
It is equipped with filter membrane；The collector may be receiving flask, for acquiring water sample.
Preferably, the enrichment collector includes upper pedestal, middle pedestal and lower base；The upper pedestal and middle seat
Body is connected together to form upper chamber；The middle pedestal and lower base is connected together to form lower chambers, installs between upper and lower chambers
There is filter membrane.
The middle pedestal quantity can be one or more, be connected together between adjacent middle pedestal, and be equipped with difference
The filter membrane in aperture；The pore size filter of the filter membrane is 0.2~200 micron.
When there is multiple filter membranes, the aperture of the filter membrane is successively successively decreased from big to small on water inlet direction.
Preferably, the intelligent control mechanism further include the power supply system being connected respectively with master control borad, display screen with
And manual control panel；The real time execution parameter of the display screen display device；The manual control panel is used to carry out parameter to master control borad
It is default；
The intelligent control mechanism further includes wireless controlled making sheet and the remote controler with wireless controlled making sheet wireless connection, institute
The wireless controlled making sheet stated is connected with master control borad and GPS positioning block simultaneously, and the remote controler is by wireless controlled making sheet to master
Control the instruction of plate input control.
The master control borad is connect with power supply system, display screen and manual control panel；Manual control panel setup parameter can be read in real time, counted
Calculation is converted into control system respectively and such as pumps and valve movement and operating status；Operating status is shown back through display unit simultaneously
Show.
Preferably, the water body environment DNA intelligent acquisition device, can be switched by multiple electromagnetic pipelines, implement multitube
Road water sample biological concentration or water sampling..
A kind of water body environment DNA intelligent acquisition method, includes the following steps:
A. a kind of above-mentioned water body environment DNA acquisition device is built；
B. parameter is default: sampling parameter when carrying out device operation to master control borad using manual control panel is default；
C. sample acquires: after acquisition device is placed into collecting location, activation system, master control borad is according to order parameter, hair
It instructs out, control sampling unit work, solenoid valve II and water pump are opened, and under the driving of water pump, water sample enters collector, carries out
Sample；Water emptying unit and pressure release unit are in off working state at this time；Aqueous bio example enrichment in water sample is to richness
Collect in collector；Extra water body flows through flowmeter, by check valve discharger, mistake inside the real-time accumulated reading device of flowmeter
Water, and signal is delivered to master control borad, when water amount is consistent with preset flow parameter, acquisition unit work terminates；Master control
Plate controls water and empties cell operation, empties to the water of enriched sample, and after the completion of water emptying, the acquisition of aqueous bio sample is completed；
D. water empties: after acquisition unit works, master control borad controls water and empties cell operation, and vacuum pump is opened, to
Residual liquid in emptying enrichment collector, after the completion of water emptying, a course of work terminates, can be by replacement collector
The next sub-sampling of row；
E. pressure release: if the blockage phenomenon in sample collection process, it is excessive that it will cause loine pressures, at this point, as managed
Road pressure is greater than the critical pressure value of relief valve, and relief valve will automatically turn on, and forms liquid communication pipeline, adjusts loine pressure,
Device is effectively protected.
The working principle of apparatus of the present invention is as follows:
Parameter is default: using manual control panel to water body total flow parameter V1, the water emptying parameters such as runing time are configured simultaneously
It is transferred on main control module；
Sample acquisition: after acquisition device reaches collecting location, start command is exported using remote control device, is received by wireless signal
It sends out device to issue, the wireless communication module in wireless controlled making sheet receives signal and feeds back to master control borad, and master control borad controls solenoid valve II
It is opened with water pump, under the driving of water pump, water body enters pipeline by coarse filter, by solenoid valve II, triple valve and water
Pump enters collector, and the biological sample in water body is enriched on filter membrane, and extra residual night flows through flowmeter, is discharged by check valve
Device；During this, water amount t inside the real-time accumulated reading device device of flowmeter2, and signal is delivered to master control borad, when
Water amount V2With total flow parameter V1When consistent, solenoid valve II and water pump are closed in master control borad output signal, control；
Water emptying: then, solenoid valve I is opened in master control borad output signal, control, and starts vacuum pump, carries out delay operation
Emptying, vacuum pump negative-pressure adsorption effect under, outside atmosphere is inhaled into via air inlet pipe, successively by solenoid valve I, water pump,
Collector, flowmeter and vacuum pump, last discharger empty the residual liquid in collector, and water empties runing time
After arrival, system is automatically stopped；At this point, a sample collecting work process terminates, taken next time by replacing collector
Sample；
Pressure release: if the blockage phenomenon in sample collection process, pipeline is unsmooth, and it is excessive that it will cause loine pressures, this
When, the critical pressure value of (such as loine pressure is greater than relief valve), relief valve will automatically turn on, and form the runner pipe of liquid outlet
Road adjusts loine pressure, device is effectively protected；
In device operational process, master control borad reads setup parameter and is converted into signal, the movement of real-time control pipeline components
And operating status, while the parameter read being calculated compared with setting value, it is shown back through display device, is intelligence
The core processing unit of control；Power supply system is preset to provide electric energy, manual control panel to carry out parameter to master control borad, wireless controlled
Making sheet shows the real time execution parameter of screen display equipment for the signal transmitting effect between master control borad and remote controler；GPS is fixed
Position plate is used to positioning in real time, and feeds back to master control borad and remote controler, is shown on display screen and remote controller displays screen.
3. beneficial effect
Compared with the prior art, the invention has the benefit that
(1) water body environment DNA intelligent acquisition device and acquisition method provided by the present invention, collection intelligent control, sample are adopted
Integrate, residual night emptying and pipeline blockage decompression protection function, have the effect that
1. can realize accurate sampling according to parameter preset automatic running by Intelligent Single-Chip Based process control, be equipped with
GPS system may be implemented precise positioning sampling high degree of automation, avoid personnel from going to sampling area by power ship, sample
Efficiency is fast, and saves human and material resources cost；
2. flowmeter is placed in after collector in sample collection process, can real-time accumulated reading water amount, reach default
After water amount parameters, it is automatically stopped sampling, realizes to the accurate control of sampling water, it is larger to overcome prior art drainage amount error
The problem of, accurate quantification enrichment and the sampling functions sample of water environment DNA are really realized, the meter of planktonic organism density is improved
Precision is calculated, samples taken more closing to reality situation is made, is conducive to the accuracy and representativeness that improve sample；
3. after the completion of sample acquisition, the residual solution being enriched in collector can be emptied to the maximum extent by emptying unit by water
Body avoids, when remaining aqueous exists, due to the change of environmental condition, giving birth to from collecting in analysis measurement this period
The influence of metabolic activity of the object in water body, chemical action and physical action, the variation of caused environment DNA is even
It is degradation, and then influences the verification and measurement ratio of species, ensure that the timeliness of sample to greatest extent, so that the environment of detection water sample is more
Close to natural environment；
4. water body environment DNA acquisition device provided by inventing, by the combination of positive press filtration and negative-pressure adsorption, really
Realize live in-situ enrichment；
5. it is acquired in the prior art in aqueous bio sample, the limitation for more being equipped with pore size filter because collecting filter device,
It is easy to generating device clogging, at this point, such as cannot timely be handled, it will damage even damage equipment；The present invention
Provided water body environment DNA acquisition device is equipped with decompression function, if the blockage phenomenon in sample collection process, is caused
Loine pressure is greater than the critical pressure value of relief valve, and relief valve will automatically turn on, and forms liquid communication pipeline, adjusts pipeline pressure
Device is effectively protected in power.
(2) water body environment DNA intelligent acquisition device and acquisition method provided by the present invention, enrichment collector are equipped with multilayer
Grade filter membrane, biological water body example is directly anchored on filter membrane, and active section is completely in vacuum state, a large amount of without acquisition
Water sample takes back laboratory, carries out the separation of later period aqueous bio sample, avoiding later period water body example separation operation process can energy band
The pollution come, is in aqueous bio sample as far as possible in the reset condition of acquisition, the confidence level of sample treatment is increased
Add；
Enrichment collector adds multiple groups multi-layer filter membrane simultaneously, and filter membrane is according to direction from inlet water to outlet water, aperture
Successively successively decrease from big to small, the filtering of different size grain size category may be implemented, and water sample can be made to pass through collection to the maximum extent
Device improves working efficiency, avoids blocking.
(3) water body environment DNA intelligent acquisition device and acquisition method provided by the present invention are furnished with wireless remote control device,
It can be used for the sampling of fixed or long-distance remote control, high degree of automation is easy to use, and sampling efficiency is high, good reliability.
Detailed description of the invention
Fig. 1 is the overall structure main view of apparatus of the present invention；
Fig. 2 is the remote controller structure schematic diagram of apparatus of the present invention；
Fig. 3 is the overall structure diagram for being enriched with collector；
Fig. 4 is the middle base structure schematic diagram for being enriched with collector；
Fig. 5 is the receiving flask schematic diagram of sample collection device；
In figure: 1, cabinet；2, solenoid valve I；3, solenoid valve II；4, collector；410, upper pedestal；420, middle pedestal；421,
Boss；422, water hole；423, groove；430, lower base；440, filter membrane；450, water inlet；460, water outlet；5, flow
Meter；6, vacuum pump；7, water inlet pipe；8, coarse filter；9, check valve；10, master control borad；11, manual control panel；111, manual buttons；12,
GPS positioning block；13, wireless controlled making sheet；14, antenna；15, power supply system；16, display screen；17, remote control device；171, wireless communication
Number transceiver；172, remote control display screen；173, button is manipulated；18, water pump；19, relief valve；20, triple valve.
Specific embodiment
The present invention is further described below combined with specific embodiments below.
It should be noted that it can directly on the other element when element is referred to as " being fixedly arranged on " another element
Or there may also be elements placed in the middle；When an element is considered as " connection " another element, it, which can be, is directly connected to
To another element or it may be simultaneously present centering elements；Term as used herein " vertical ", " horizontal ", " left side ",
" right side " and similar statement are for illustrative purposes only.
Unless otherwise defined, all technical and scientific terms used herein and belong to technical field of the invention
The normally understood meaning of technical staff is identical；Term as used herein in the specification of the present invention is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention；Term " and or " used herein includes one or more phases
Any and all combinations of the listed item of pass.
Embodiment 1
As shown in Figure 1, a kind of water body environment DNA intelligent acquisition device, the intelligence including cabinet 1 and inside cabinet 1
Can control mechanism with sampling main body mechanism；The sampling main body mechanism includes sample collection unit, pressure release unit, water emptying
Unit and triple valve 20；
Intelligent control mechanism includes master control borad 10 and the GPS positioning block 12 being connected with master control borad 10 in the present embodiment,
Power supply system 15, display screen 16 and the manual control panel 11 being connected with master control borad 10 are wirelessly connected distant with wireless controlled making sheet 13
Control device 17；The master control borad 10 empties opening or closing for unit to control acquisition unit and water, and receives GPS positioning
The location information of block 12.The manual control panel 11 is preset by manual buttons 111 to carry out parameter to master control borad 10, described
Display screen 16 shows the operating parameter of master control borad 10；The manual control panel 11 is used to carry out parameter to master control borad 10 to preset, described
Remote controler 17 instructed remote controler 17 to realize remote real time remote control 10 input control of master control borad by wireless controlled making sheet 13
The start and stop of equipment.
Further, the remote control device in the present embodiment, be equipped with wireless signal transceiver 171, remote control display screen 172 and
Button 173 is manipulated, by the manipulation input instruction of button 173, instruction sends wireless controlled making sheet to by wireless signal transceiver 171
13, remote control display screen 172 is synchronous with display screen 16, the current operating parameter of display device.
Sample collection unit in the present embodiment includes water pump 18, collector 4, flowmeter 5, coarse filter 8, solenoid valve II
3 (solenoid valve II 3 in the present embodiment is normally closed solenoid valve: model: De Lixi 2W series voltage: 24 volts) and check valve 9, institute
The coarse filter 8 stated is arranged on the first inlet pipeline of triple valve 20, and between 20 first import of coarse filter 8 and triple valve
Connecting pipe be equipped with solenoid valve II 3；The outlet of the triple valve 20 is connected with one end of water pump 18；Water pump 18 it is another
One end is connected with the water inlet 450 of collector 4, and described 5 one end of flowmeter is connected with the water outlet 460 of collector 4；Institute
The master control borad 10 stated is connect with water pump 18 and solenoid valve II 3, to control the open and close of the two；When remote controler 17 passes through
After wireless controlled making sheet 13 inputs enabled instruction to master control borad 10, master control borad 10 controls water pump 18 and solenoid valve II 3 is opened, in water
Under the driving of pump 18, water body enters pipeline by coarse filter 8, enters by solenoid valve II 3, triple valve 20 and water pump 18 and receives
Storage 4, the biological sample in water body are enriched on filter membrane 440, and extra residual night flows through flowmeter, pass through the discharge dress of check valve 9
It sets；During this, water amount t inside the real-time accumulated reading device of flowmeter 52, and signal is delivered to master control borad 10, serve as water
Measure V2With total flow parameter V1When consistent, solenoid valve II 3 and water pump 18 are closed in master control borad output signal, control.
Water emptying unit in the present embodiment includes admission line, I 2 (the present embodiment of solenoid valve on admission line
In solenoid valve I 2 be normally closed solenoid valve: model: De Lixi 2W series voltage: 24 volts) and vacuum pump 6, the air inlet pipe
Road one end is communicated with outside atmosphere, and the other end is connected by the second import of triple valve 20 with the water inlet 450 of collector 4；
Described 6 one end of vacuum pump is connected with flowmeter 5, and the other end is connected with external environment；The vacuum pump 6 and check valve
9, in shape in parallel, are connected in pipeline；Master control borad 10 is connected with vacuum pump 6 and solenoid valve I 2, to control the unlatching of the two
With closing；After sampling, solenoid valve I 2 is opened in 10 output signal of master control borad, control, and starts vacuum pump 6, is delayed
Operation emptying, vacuum pump 6 negative-pressure adsorption effect under, outside atmosphere is inhaled into via air inlet pipe, successively by solenoid valve I 2,
Water pump 18, collector 4, flowmeter 5 and vacuum pump 6, last discharger empty the residual liquid in collector 4, water row
After the dry running time reaches, system is automatically stopped；At this point, a sample collecting work process terminates, by replacement collector 4 into
The next sub-sampling of row；
Pressure release unit in the present embodiment includes relief valve 19, and relief valve 19 is arranged between water pump 18 and collector 4, and
It is connected to outside atmosphere；Usually relief valve 19 is in normally off, if the blockage phenomenon in sample collection process, pipeline is not
Freely, it is excessive that it will cause loine pressures, at this point, as loine pressure be greater than relief valve 19) critical pressure value, relief valve 19 incite somebody to action from
It is dynamic to open, the flow pipe of liquid outlet is formed, loine pressure is adjusted, device is effectively protected；
Collector can be enrichment collector in the present embodiment, be enriched with for water sample biological sample, main to collect life of swimming
Suspended particulate substance etc. in object, water sample；The enrichment collector includes upper pedestal 410, middle pedestal 420 and lower base 430；
Water inlet 450 is located at 410 top of upper pedestal, and the lower part of upper pedestal 410 is cavity, and cavity lateral wall is equipped with boss；In described
Pedestal 420 divides for upper cavity and lower chamber, and upper cavity inner wall is equipped with groove 423, and lower chamber lateral wall is equipped with boss 421, up and down
Water hole 422 is equipped among cavity；The lower base bottom is equipped with water outlet 460, and cavity inner wall is equipped with boss；Upper pedestal
410 are connected together to form upper chamber by its cavity lateral wall boss and middle 420 upper cavity inboard wall groove 423 of pedestal, described
Middle pedestal 420 is connected together to form cavity of resorption by the groove of its lower chamber lateral wall boss 421 and 430 cavity inner wall of lower base
Room；Between the middle pedestal 420 and upper pedestal 410, between middle pedestal 420 and lower base 430, be mounted on sealing ring and
Filter membrane 440；Middle 420 quantity of pedestal can increase and decrease on demand, be set as one or more, engage between adjacent middle pedestal 420
Connection, in and the filter membrane 440 of sealing ring and different pore size is installed；The pore size filter of the filter membrane 440 be 0.2~
200 microns；The filter membrane 440 on the arragement direction of water inlet 450 to water outlet 460, successively pass from big to small by aperture
Subtract.
After water body flows through enrichment collector, the biological sample in water body is enriched on filter membrane 440, after the completion of enrichment, is beaten
Collector 4 is opened, presss from both sides out filter membrane 440 with aseptic nipper, is fitted into preprepared sterile centrifugation tube (centrifugation pipe volume 2-10
Milliliter), (stored frozen and room temperature storage may be selected) in storage.It is worth noting that the collector 4 in the present embodiment can also
To select receiving flask, water sample collection, for the extraction of nucleic acid, the analysis including DNA, RNA and environmental contaminants are carried out.
The invention device cooperates remote control device such as unmanned boat, unmanned plane in use, its Function is especially prominent, especially logical
Crossing GPS positioning block 12 can real-time positioning device.Device remote control is arrived by carrier (unmanned boat unmanned plane) in some specific environments
Behind collection point, because being artificially difficult to view its operating status and specific location at a distance.The GPS positioning that the device passes through setting
Function, energy real-time confirmation position, while passing through the operation of wireless module energy real-time control apparatus and display state), and by distant
Control device 17 realizes the start and stop of remote real time remote control equipment.
Certainly, above description is not limitation of the present invention, within the essential scope of the present invention, the variation made, remodeling,
Addition or replacement, all should belong to protection scope of the present invention.
Embodiment 2
One of the present embodiment water body environment DNA intellegent sampling method specific steps are as follows:
A. a kind of a kind of water body environment DNA acquisition device as described in claim 1~8 is built；
B. parameter is default: sampling parameter when carrying out device operation to master control borad 10 using manual control panel 11 is default；
C. sample acquires: after acquisition device is placed into collecting location, activation system, master control borad 10 according to order parameter,
Instruction, control sampling unit work are issued, solenoid valve II 3 and water pump 18 are opened, and under the driving of water pump 18, water sample enters collection
Device 4 carries out sample；Water emptying unit and pressure release unit are in off working state at this time；Aqueous bio sample in water sample
It is enriched in enrichment collector；Extra water body flows through flowmeter 5, passes through 9 discharger of check valve, the real-time accumulated reading of flowmeter 5
Water amount inside device is taken, and signal is delivered to master control borad 10, when water amount is consistent with preset flow parameter, acquisition unit
Work terminates；Master control borad 10 controls water and empties cell operation, empties to the water of enriched sample, after the completion of water emptying, aqueous bio
Sample acquisition is completed；
D. water empties: after acquisition unit works, master control borad 10 controls water and empties cell operation, and vacuum pump 6 is opened, and uses
To empty the residual liquid in enrichment collector, after the completion of water emptying, a course of work terminates, and is by replacing collector 4
It can carry out next sub-sampling；
E. pressure release: if the blockage phenomenon in sample collection process, it is excessive that it will cause loine pressures, at this point, as managed
Road pressure is greater than the critical pressure value of relief valve 19, and relief valve 19 will automatically turn on, and forms liquid communication pipeline, adjusts pipeline pressure
Device is effectively protected in power.
The method of sampling in the present embodiment, sampling is accurate, and high degree of automation, sampling efficiency is fast, save human and material resources at
This.In collection process, can real-time accumulated reading water amount, after reaching said preset amount of water parameter, be automatically stopped sampling, realize to taking
The accurate control of sample water overcomes the larger problem of prior art drainage amount error, really realizes the essence of water environment DNA
It determines amount enrichment and sampling functions sample, improves the computational accuracy of planktonic organism density, make samples taken more closing to reality feelings
Condition is conducive to the accuracy and representativeness that improve sample；
After the completion of sample acquisition, by water air emptying function, residual liquid can be emptied to the maximum extent, avoided from collecting
It is biological new old in water body due to the change of environmental condition when remaining aqueous exists in analysis measurement this period
The variation of the influence of metabolic activity, chemical action and physical action, caused environment DNA is even degraded, and then is influenced
The verification and measurement ratio of species ensure that the timeliness of sample to greatest extent, so that the environment of detection water sample is closer to natural environment.
Certain embodiment of the invention above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
Limitations on the scope of the patent of the present invention therefore cannot be interpreted as；It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to guarantor of the invention
Protect range；Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (9)
1. a kind of water body environment DNA intelligent acquisition device, it is characterised in that: including intelligent control mechanism and sampling main body machine
Structure, the sampling main body mechanism include sample collection unit, pressure release unit and water emptying unit；
The sample collection unit includes water pump (18), collector (4) and flowmeter (5), the water pump (18) one end
It is connect with outside body of water, the other end is connected with the water inlet (450) of collector (4)；The flowmeter (5) and collector
(4) water outlet (460) is connected；
The described water emptying unit includes admission line and vacuum pump (6), described admission line one end and collector (4)
Water inlet (450) is connected, and the other end is communicated with outside atmosphere；Described vacuum pump (6) one end is connected with flowmeter (5),
The other end is connected with external environment；
The pressure release unit includes relief valve (19), the relief valve (19) setting water pump (18) and collector (4) it
Between, and be connected to outside atmosphere；
The intelligent control mechanism includes master control borad (10) and the GPS positioning block (12) being connected with master control borad (10), institute
The master control borad (10) stated empties opening or closing for unit to control acquisition unit and water, and receives GPS positioning block (12)
Location information.
2. a kind of water body environment DNA intelligent acquisition device according to claim 1, it is characterised in that: the sampling master
Body mechanism further includes triple valve (20), and the first import of the triple valve (20) is connect with outside body of water；The triple valve
(20) outlet is connected with water pump (18)；The admission line phase of second import of the triple valve (20) and water emptying unit
Connection.
3. a kind of water body environment DNA intelligent acquisition device according to claim 1, it is characterised in that: the sample is adopted
Collecting unit further includes coarse filter (8), solenoid valve II (3) and check valve (9)；The coarse filter (8) is arranged in threeway
On first inlet pipeline of valve (20), and the pipeline between (20) first import of coarse filter (8) and triple valve is equipped with electromagnetism
Valve II (3)；The check valve (9) is in shape in parallel with vacuum pump (6), is connected in pipeline.
4. a kind of water body environment DNA intelligent acquisition device according to claim 1, it is characterised in that: the water emptying
Unit further includes solenoid valve I (2), and the solenoid valve I (2) is located on admission line.
5. a kind of water body environment DNA intelligent acquisition device according to claim 1, it is characterised in that: the collector
(4) it is enrichment collector, is used for water sample biological concentration, is provided with filter membrane (440) in the enrichment collector；The receipts
Storage (4) or receiving flask, for acquiring water sample.
6. a kind of water body environment DNA intelligent acquisition device according to claim 5, it is characterised in that: the enrichment is received
Storage includes upper pedestal (410), middle pedestal (420) and lower base (430)；The upper pedestal (410) and middle pedestal (420)
It is connected together to form upper chamber, the middle pedestal (420) and lower base (430) are connected together to form lower chambers, upper and lower chambers
Between filter membrane (440) are installed.
7. a kind of water body environment DNA intelligent acquisition device according to claim 6, it is characterised in that: the enrichment is received
Middle pedestal (420) quantity of storage can be one or more, be connected together between adjacent middle pedestal (420), and be equipped with not
With the filter membrane (440) in aperture；The pore size filter of the filter membrane (440) is 0.2~200 micron.
When there is multiple filter membranes (440), the aperture of the filter membrane (440) is successively passed from big to small on water inlet direction
Subtract.
8. a kind of water body environment DNA intelligent acquisition device described in -7 according to claim 1, it is characterised in that: the intelligence
Control mechanism further includes the power supply system (15) being connected respectively with master control borad (10), display screen (16) and manual control panel (11)；
The real time execution parameter of described display screen (16) display device；The manual control panel (11) to master control borad (10) for being joined
Number is default；
The intelligent control mechanism further includes wireless controlled making sheet (13) and the remote control with wireless controlled making sheet (13) wireless connection
Device (17), the wireless controlled making sheet (13) are connected with master control borad (10) and GPS positioning block (12) simultaneously, the remote control
Device (17) instructs master control borad (10) input control by wireless controlled making sheet (13).
9. a kind of water body environment DNA intelligent acquisition method, includes the following steps:
A. a kind of any water body environment DNA acquisition device according to claims 1-8 is built；
B. parameter is default: sampling parameter when using manual control panel (11) master control borad (10) are carried out with device operation is default；
C. sample acquires: after acquisition device is placed into collecting location, activation system, master control borad (10) is according to order parameter, hair
It instructs out, control sampling unit work, solenoid valve II (3) and water pump (18) are opened, and under the driving of water pump (18), water sample enters
Collector (4) carries out sample；Water emptying unit and pressure release unit are in off working state at this time；Water body in water sample is raw
In object example enrichment to enrichment collector；Extra water body flows through flowmeter (5), passes through check valve (9) discharger, flowmeter
(5) water amount inside real-time accumulated reading device, and signal is delivered to master control borad (10), when water amount and preset flow parameter
When consistent, acquisition unit work terminates；Master control borad (10) controls water and empties cell operation, empties to the water of enriched sample, water row
After the completion of sky, the acquisition of aqueous bio sample is completed；
D. water empties: after acquisition unit works, master control borad (10) controls water and empties cell operation, and vacuum pump (6) is opened, and uses
To empty the residual liquid in enrichment collector, after the completion of water emptying, a course of work terminates, by replacing collector (4)
It can carry out next sub-sampling；
E. pressure release: if the blockage phenomenon in sample collection process, it is excessive that it will cause loine pressures, at this point, such as pipeline pressure
Power is greater than the critical pressure value of relief valve (19), and relief valve (19) will automatically turn on, and forms liquid communication pipeline, adjusts pipeline pressure
Device is effectively protected in power.
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| CN114058485B (en) * | 2021-11-01 | 2023-06-16 | 烟台大学 | Environmental DNA collection device in ocean water |
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| CN114518257A (en) * | 2022-01-12 | 2022-05-20 | 中国科学院水生生物研究所 | Intelligent acquisition method of environmental DNA |
| CN114577524A (en) * | 2022-02-21 | 2022-06-03 | 中国科学院水生生物研究所 | Cartridge magazine type filter membrane replacing device and system based on automatic return for environmental DNA sampling |
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| CN116982602A (en) * | 2023-09-18 | 2023-11-03 | 淮北师范大学 | Basin biodiversity sample collection box |
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