CN106967595A - Bacterium or viral online acquisition and online automatic detection method in a kind of air - Google Patents
Bacterium or viral online acquisition and online automatic detection method in a kind of air Download PDFInfo
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- CN106967595A CN106967595A CN201710346839.4A CN201710346839A CN106967595A CN 106967595 A CN106967595 A CN 106967595A CN 201710346839 A CN201710346839 A CN 201710346839A CN 106967595 A CN106967595 A CN 106967595A
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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
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
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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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
Abstract
The present invention relates to bacterium in a kind of air or viral online acquisition and online automatic detection method, with reference to biochip or fluorescent quantitative PCR technique, pathogenic microorganisms that may be present in air can be enzyme rapidly and sensitively detected.The device is mainly made up of negative pressure air-suction device, mass air flow sensor, adsorption liquid and granule for absorbing.Utilize air extractor, pass through negative-pressure adsorption air, with constant gas amount per minute, by liquid acquisition area, in the presence of granule for absorbing in solution and solution, the virion in air is set to collect in the liquid of small size, while cracking the microorganism collected, releasing microbe DNA (RNA), is adsorbed on granule for absorbing.Then on-line filtration is passed through, collect the particle in liquid, with commercially available DNA (RNA) elution DNA (RNA), then microbial biomass is detected using round pcr, micro organism quantity in the copy number of measure divided by the volume of air of absorption, i.e. unit air obtained by detection.
Description
Technical field
The invention belongs to technical field of microbial detection, particularly relate in a kind of air bacterium or viral online acquisition and
Line automatic testing method.
Background technology
Air borne is the main path of some pathogenic bacteria or viral transmission, is also most to be difficult to detect and control in current environment
The approach of system.
The SARS epidemic situations of enormous impact were brought to the whole world from 2003, H1N1 and 2013 year to the influence country in 2009 is quick-fried
The bird flu epidemic situation of hair, being all can be by airborne viral disease.By taking SARS virus as an example：Virus is primarily present in biography
In nasal mucus, phlegm and the saliva in dye source (human or animal), pass through air transmitted by droplets and direct contact.Patient by coughing, sneezing, very
To speaking up, virus is diffused into the air of surrounding by Shi Douhui with the saliva spittle, dust particle etc., is propagated with air flow.
Why SARS causes a large amount of personnel's infection particularly medical personnel to bring huge sacrifice initial stage, except safeguard procedures
Outside reason, the virus being largely because in air is difficult to monitor and takes precautions against what is caused.
Although the more than ten years in SARS epidemic situations past, tremendous influence and the panic mood of society that it brings to social life
Let us is remembered clearly.Even to this day, for harmful microbe early warning in air with monitoring still lack effective instrument and
Method.Existing detecting instrument and detection method program are complicated, take longer, it is difficult to provide epidemic situation scene accurately data letter in time
Cease to take the precautionary measures rapidly.
With the development of society, country variant, the contacts of regional people are more and more frequent, therefore public transport, friendship
The crowded closed areas such as logical website, school, hospital, market, office building have become pathophorous main in air
Place.Particularly within the hospital, cross infection in hospital is always the problem of making hospital's headache.
When avian influenza virus is popular, because without device for fast detecting, government is often in the case of no positive evidence
Poultry farms are blindly slaughtered, all brought about great losses to government and plant.
The content of the invention
It is an object of the invention to provide bacterium in a kind of air or viral online acquisition and online automatic detection method, to solve
The problem of early warning or monitoring must not being carried out to bacterium in air or virus, it is fast to the bacterium in air or virus progress to realize
Fast accurately Site Detection.
The present invention is achieved by the following technical solutions：
Bacterium or viral online acquisition and online automatic detection method in a kind of air,
Use online acquisition and on-line automatic detection device, including sample-adding control device, PCR amplification devices, detection means
And automaton, in addition to pressue device and negative pressure device；Comprise the following steps：
1) online acquisition and on-line automatic detection device are opened, will according to object to be detected and corresponding PCR kit
Ask, pre-set PCR amplification programs, be inserted into integrated sampling box；
2) sample volume is set, the negative pressure device is then turned on, automatic data collection sample is to setting sample volume；
3) the setting time is stood, institute's negative pressure device starts, the liquid of sample region is sucked into waste liquid bottle, DNA or RNA has been adsorbed
Particle be trapped within the filter medium of the sample region；
4) pressue device is opened, DNA or RNA eluents are added in the sample region, described on the particle
Under DNA or described RNA elutions；
5) by the sample-adding control device, the eluent of the DNA or described RNA under elution is added to the PCR
Amplification device enters performing PCR amplification by the PCR amplification programs that pre-set；
6) after the sample after amplification is detected by detection means, testing result is exported.
The sampling integrated sampling box is connected with the sample-adding control device by pipeline；It is described sample-adding control device with
The PCR amplification devices coordinate；
The automaton respectively with it is described sample-adding control device, the PCR amplification devices, the detection means,
The pressue device and negative pressure device electrical connection；
The pressue device is connected with the negative pressure device with the sampling integrated sampling box by pipeline.
The sampling integrated sampling box includes sampling container, eluent storage container and waste liquid cylinder；Eluent storage is held
Device is connected with the sampling container by pipeline；The waste liquid cylinder is connected with the bottom of the sampling container by pipeline；
The pressue device is provided with pipeline between the sampling container and the eluent storage container；
The sampling container is connected by pipeline with the negative pressure device respectively with the waste liquid cylinder；
Porous air distributor and filter are provided with the sampling container；The entrance of the porous air distributor
It is connected with being connected the sampling container with the pipeline of the negative pressure device, the filter is arranged at the bottom of the sampling container
Exit, is provided with adsorption liquid in the sampling container, and the gas outlet of the porous air distributor is less than the adsorption liquid
Liquid level；The adsorption liquid is made up of microbial lytic liquid and granule for absorbing；
Eluent is provided with the eluent storage container.
In the adsorption liquid, the volume ratio of the microbial lytic liquid and the granule for absorbing is 10-20：1.
The pressue device is the first peristaltic pump, and the negative pressure device is vavuum pump.
Also include pulling and inserting type interface arrangement, the fast plugging for integrated sampling box of sampling.
The sample-adding control device includes the second peristaltic pump and check valve；The outlet of second peristaltic pump and the PCR
Amplification device is corresponding, and the entrance of second peristaltic pump is connected with the outlet of the check valve by pipeline；The check valve
Entrance be connected by pipeline with the outlet at bottom of the sampling container.
The PCR amplification devices include sample-adding plate, PCR kit and PCR temperature control equipments, the sample-adding plate with it is described
PCR kit is connected, and the PCR kit is connected with the PCR temperature control equipments.
The detection means includes excitation light emission device, fluorescent acceptor and interactive system；The excitation light emission
Device and the fluorescent acceptor are relatively arranged on the detection zone of the PCR amplification devices, the excitation light emission device with it is described glimmering
Optical receiver is electrically connected with the interactive system, and the interactive system is electrically connected with the automaton.
The automatic detection device includes central controller, and the central controller is cyclelog, single-chip microcomputer or work
One kind in control machine；
The automatic detection device also includes blue tooth interface, wireless WIFI interface, Ethernet interface, USB interface, RS232
One or more combinations in serial ports or RS485 serial ports.
The beneficial effects of the invention are as follows：
The bacterium of the technical program or viral online acquisition and online automatic detection method, including negative pressure ventilation, microorganism
Online acquisition, air microbe absorption, DNA or RNA On-line testings, DNA such as expand and detected online at the part, by gathering air
In microorganism particle, cracked in real time in a liquid, extract DNA (or RNA), after being expanded by PCR, carried out online real-time
Detection.
The technical program, together in one, can be analyzed the sample of collection respectively, and be adapted to by sampling, extraction and detection collection
Different types of microorganisms quantity in analytical unit volumes of air.
The new air microbe sampling detecting instrument that the technical program is developed, with reference to biochip or fluorescent quantitation
Round pcr, can enzyme rapidly and sensitively detect pathogenic microorganisms that may be present in air.The device is mainly filled by negative inspiratory pressure
Put (hand-held or vavuum pump), mass air flow sensor, adsorption liquid and granule for absorbing composition.Using air extractor, pass through negative-pressure adsorption
Air, with constant gas amount per minute, by liquid acquisition area, in the presence of the granule for absorbing in solution and solution, makes sky
Virion in gas is collected in the liquid of small size, while cracking the microorganism collected, releasing microbe DNA (RNA) inhales
It is attached on granule for absorbing.Then by on-line filtration, the particle in liquid is collected, with commercially available DNA (RNA) elution
DNA (RNA), then detects microbial biomass, the copy number of measure divided by the volume of air of absorption using round pcr, that is, detects institute
Micro organism quantity in the unit air obtained.
Brief description of the drawings
Fig. 1 is the structural representation of apparatus of the present invention；
Fig. 2 is present invention sampling integrated sampling box operating diagram.
Description of reference numerals
1 vavuum pump, 2 first magnetic valves, 3 sampling containers, 4 first peristaltic pumps, 5 eluent storage containers, 6 eluents, 7 inhale
Attached liquid, 8 second peristaltic pumps, 9 check valves, 10 second magnetic valves, 11 waste liquid cylinders, 12 excitation light emission devices, 13 fluorescent acceptors, 14
Interactive system, 15 sample-adding plates, 16PCR temperature control equipments, 17 filters.
Embodiment
Describe technical scheme in detail by the following examples, following embodiment be only it is exemplary, only
Explanation and illustration technical scheme can be used for, and be not to be construed as the limitation to technical solution of the present invention.
The present invention develops new air microbe sampling detecting instrument, with reference to biochip or quantitative fluorescent PCR skill
Art, can enzyme rapidly and sensitively detect pathogenic microorganisms that may be present in air.The device is main by negative pressure air-suction device (hand
Hold formula or vavuum pump), mass air flow sensor, adsorption liquid and granule for absorbing composition.Using air extractor, by negative-pressure adsorption air,
With constant gas amount per minute, by liquid acquisition area, in the presence of the granule for absorbing in solution and solution, make in air
Virion is collected in the liquid of small size, while cracking the microorganism collected, releasing microbe DNA (RNA), absorption is being inhaled
On attached particle.Then by on-line filtration, the particle in liquid is collected, with commercially available DNA (RNA) elutions DNA
(RNA) microbial biomass, the copy number of measure divided by the volume of air of absorption then, are detected using round pcr, i.e., obtained by detection
Micro organism quantity in unit air.
Such sampler of current in the market is only applicable to the air microbe sampling of high concentration, for the disease compared with low-density
Poison, due to the limitation of the factors such as residence time of the air in the liquid of smaller size smaller, it is impossible to trap piconavirus particle, therefore
Sensitivity is not high, while can not on-line determination.The present invention is collected together in one by sampling, extraction and detection, can be by collection
Sample is analyzed respectively, and is adapted to different types of microorganisms quantity in analytical unit volumes of air.
The application provides bacterium or viral online acquisition and on-line automatic detection device in a kind of air, such as Fig. 1 and Fig. 2 institutes
Show, including sampling integrated sampling box, sample-adding control device, PCR amplification devices, detection means and automaton, also wrap
Include the pressue device and negative pressure device being engaged with sampling integrated sampling box.
Sampling integrated sampling box is connected with sample-adding control device by pipeline；It is loaded control device and PCR amplification devices
Coordinate；The sampling integrated sampling box of the application is by pulling and inserting type interface arrangement, and quick for integrated sampling box of sampling inserts
Dial.
Automaton is filled with sample-adding control device, PCR amplification devices, detection means, pressue device and negative pressure respectively
Put electrical connection；Automatic detection device includes central controller, and central controller is in cyclelog, single-chip microcomputer or industrial computer
One kind, by programming realization from air sampling to detecting that microbe species and concentration is performed fully automatic.
Automatic detection device also includes blue tooth interface, wireless WIFI interface, Ethernet interface, USB interface, RS232 serial ports
Or one or more combinations in RS485 serial ports, pass through wired or wireless way and host computer or mobile terminal communication, realization pair
The remote monitoring of the present apparatus, testing staff need not go deep into epidemic-stricken area and just can obtain testing result, it is to avoid testing staff is infected
Risk.Meanwhile, these interfaces can be communicated with miscellaneous equipment such as host computer, mobile phone etc., testing result is set by other
Standby application, automatic detection device can also be controlled by miscellaneous equipment, can also realize the remote transmission to detecting data or high in the clouds
Manage and to the remote control of automatic detection device.
Pressue device is connected with negative pressure device with sampling integrated sampling box by pipeline.
Pressue device is the first peristaltic pump 4；Negative pressure device is vavuum pump 1 in this application.
Integrated sampling box of sampling includes sampling container 3, eluent storage container 5 and waste liquid cylinder 11；Eluent storage is held
Device is connected with sampling container by pipeline；The bottom of waste liquid cylinder and sampling container is connected by pipeline.In sampling integrated sampling
In box, flowing through on air pressure control, pipeline by check valve control gas flow direction for liquid is held so as to control to sample
Pressure in device, eluent storage container or waste liquid cylinder, realizes the directed flow of liquid (adsorption liquid or eluent).
Pressue device is provided with pipeline between sampling container and eluent storage container；
Sampling container 3 is connected by pipeline with negative pressure device respectively with waste liquid cylinder；In this application, negative pressure device is vacuum
Pump 1, is provided with the first magnetic valve 2 between vavuum pump and sampling container, and the second electricity is provided between vavuum pump and waste liquid cylinder
Magnet valve 10；First magnetic valve is electrically connected with the second magnetic valve with automaton, and first is controlled by automaton
Magnetic valve and the second magnetic valve are turned on and off.
Porous air distributor and filter 17 are provided with sampling container；The entrance of porous air distributor and connection
The pipeline connection of sampling container and negative pressure device, filter is arranged at the outlet at bottom of sampling container, set in sampling container
Adsorption liquid 7 is equipped with, the gas outlet of porous air distributor is less than the liquid level of adsorption liquid；The gas outlet of porous air distributor passes through
The air that carries disease germs of suction is distributed in adsorption liquid by multiple pore modes, and in order to prevent air-flow too big, liquid is gone out in direct short-circuit,
Multiple pores air that will carry disease germs is scattered in minute bubbles, and increase is carried disease germs the contact area of air and adsorption liquid, is improved in absorption air
The efficiency of microorganism.
Adsorption liquid is made up of microbial lytic liquid and granule for absorbing.
Eluent 6 is provided with eluent storage container 5, in this application, eluent is DNA (RNA) eluent,
For commercially available DNA (RNA) eluent.
In adsorption liquid, the volume ratio of microbial lytic liquid and granule for absorbing is 10-20：1.
Granule for absorbing is DNA granule for absorbing or RNA granule for absorbing.
Ratio of height to diameter of the adsorption liquid in sampling container is 6：1-3：1.
In this application, microbial lytic liquid uses phosphate buffer PBS, and collocation method is, by NaCl, KCl,
Na2HPO4And KH2PO4, it is dissolved in distilled water, the pH value for adjusting solution with HCl finally adds distilled water constant volume to 7.4, and
Steam sterilization 20 minutes under 121 DEG C of high pressures, are stored in room temperature or 4 DEG C of refrigerators.
For example, 8g NaCl, 0.2g KCl, 1.44g Na2HPO4With 0.24g KH2PO4, it is dissolved in 800ml distilled water, uses
The pH value of HCl regulation solution finally adds distilled water to be settled to 1L to 7.4.Steam sterilization 20 minutes under 121 DEG C of high pressures, is protected
It is stored in room temperature or 4 DEG C of refrigerators.
In this application, broad-spectrum high efficacy adsorption liquid is constituted：It is 10-30 milligrams straight that particle diameter distribution is added in 1ml PBS solutions
Footpath is 10-200nm chitin nanometer.
In this application, as constituted using specific adsorption liquid, addition particle diameter distribution is 10-30. millis in 1ml PBS solutions
Gram a diameter of 10-200nm surface carries the nano-particle that Antibody of Influenza is modified.
In this application, DNA granule for absorbing or RNA granule for absorbing can be commercially available polymeric adsorbent, such as Shanghai life work
Or DNA (RNA) polymeric adsorbent of Beijing Quan Shijindeng companies production.
Being loaded control device includes the second peristaltic pump 8 and check valve 9；The outlet of second peristaltic pump and PCR amplification device phases
Correspondence, the outlet of the entrance and check valve of the second peristaltic pump is connected by pipeline；The entrance of check valve is held by pipeline with sampling
The outlet at bottom connection of device.In the sample application zone of the sample-adding plate of the application, it is loaded, is entered using drop recorder using the second peristaltic pump
The metering of row sample-adding amount.
PCR amplification devices include sample-adding plate 15, PCR kit and PCR temperature control equipments 16, sample-adding plate and PCR reagent
Box is connected, and PCR kit is connected with PCR temperature control equipments.Adding liquid paraffin carries out close to reaction solution in PCR kit
Envelope.The PCR kit of the application is commercially available PCR kit.
Detection means includes excitation light emission device 12, fluorescent acceptor 13 and interactive system 14；Excitation light emission device
Be relatively arranged on the detection zone of PCR amplification devices with fluorescent acceptor, fluorescent emitters with fluorescent acceptor and human-computer dialogue
System is electrically connected, and interactive system is electrically connected with automaton.Interactive system can also set related inspection simultaneously
Survey parameter and equipment maintenance and management.
Specifically operation principle is：
According to detection object and PCR kit requirement, PCR amplification programs, insertion sampling integrated sampling box are set；Set
Good sampling volume, is then turned on device, and automaton can control vacuum pump startup, realizes automatic sampling to setting volume；
After standing 2 minutes, automaton control vavuum pump is again started up, and it is negative pressure to make in waste liquid cylinder, by sampling container
Liquid be drawn into waste liquid cylinder, adsorbed DNA (RNA) particle cutoff on the filter medium in sampling container；Automatically control
Device controls the first wriggling pump startup, is the pressurization of eluent storage container, DNA (RNA) eluent is pressed into sampling container,
Under the DNA (RNA) adsorbed on bright attached particle is eluted, the washing DNA (RNA) under eluting by check valve and the second peristaltic pump
De- liquid is added to be detected in sample P CR kits added with commercially available DNA (RNA) in advance, and default according to the requirement of PCR kit
Program enters performing PCR amplification；Sample after amplification passes through after the detection means detection of detection zone, and output result is automatically controlled dress
Put, obtain the species letter concentration of microorganism, and testing result is passed into host computer or mobile whole by wired or wireless communication
End.
In this application, all breakdown actions control each device to be automatically performed by automaton, motion parts
Controlled to complete by automaton using stepper motor.
The application provides bacterium or viral online acquisition and online automatic detection method in a kind of air,
Using online acquisition and on-line automatic detection device, comprise the following steps：
1) online acquisition and on-line automatic detection device are opened, preheating is booted up, according to object to be detected and correspondingly
PCR kit requirement, pre-set PCR amplification programs, be inserted into integrated sampling box, in this application, all consumptive materials are equal
Need aseptic process.
2) sample volume is set, negative pressure device is then turned on, in the present embodiment, negative pressure device is vavuum pump, is adopted automatically
Collection sample is to setting sample volume.
3) the setting time is stood, in the present embodiment from standing 2 minutes, institute's negative pressure device starts, by the liquid of sample region
Body sucks waste liquid bottle, and the particle for having adsorbed DNA or RNA is trapped within the filter medium of sample region.
4) pressue device is opened, in the present embodiment, pressue device is the first peristaltic pump, and DNA or RNA is added in sample region
Eluent, by under the DNA on particle or RNA elutions.
5) by being loaded control device, the eluent that lower DNA or RNA will be eluted be added to PCR amplification devices pass through it is default
Put PCR amplification programs and enter performing PCR amplification.
6) after the sample after amplification is detected by detection means, testing result is exported, the species of microorganism is obtained
And concentration.
Embodiment 1
Liquid addition 1ml commercially available DNA (RNA) granule for absorbing in liquid acquisition area, respectively with 0.01mol/L NaOH and
HCl adjusts the pH to 6-8 of solution, carries out the collection experiment of influenza virus in air.By mouse lung adapted strain influenza A virus warp
After chicken embryo rejuvenation, 100000 viruses/ml solution is configured to 0.1mol/L phosphate solution, then to volume about 50m3's
In confined space, virus is sprayed in atmosphere with air sparger, specific virion is several to be subtracted according to liquid in sprayer
(virion in air is about 1000/m for a small amount of calculating3).The liquid volume 15ml in liquid acquisition area, ratio of height to diameter is 3:1,
0.1m is gathered in different ways3Influenza virus, sample detecting experiment, every time three batches, experimental result are carried out in two times
(table 1).PCR kit is detected using commercially available H1N1 quantification kits.
The collection result of virion in the air of table 1
Note：Sample mode, after spraying, at once with line collection and on-line automatic detection device respectively in four corners in room
With sampling simultaneously at the liftoff 1.6m in centre position, average value is then calculated, 1 virus is 1 copy in table 1.
Embodiment 2
Liquid addition 1ml commercially available DNA (RNA) granule for absorbing in liquid acquisition area, respectively with 0.01mol/L NaOH and
HCl adjusts the pH to 6-8 of solution, carries out the collection experiment of Escherichia coli in air.Bacillus coli DH 5 alpha is used into conventional method
Culture to concentration is 5 × 106Afterwards, 100000 cells/ml solution is configured to 0.1mol/L phosphate solution, then to body
Product about 50m3Confined space in, spray virus in atmosphere with air sparger, specific microorganism population is according to spraying
(the microbial cell concentration in air is about 1000/m for the decrement calculating of liquid in device3).The liquid bulk in liquid acquisition area
Product 15ml, ratio of height to diameter is 3:1,0.1m is gathered in different ways3Escherichia coli, in two times carry out sample detecting experiment, often
Secondary three batches, experimental result (table 2).Detected using commercially available colibacillus PCR immue quantitative detection reagent box.
The collection result of Escherichia coli in the air of table 2
Note：Note:Sample mode, it is liftoff in four corners in room and centre position respectively with sampler at once after spraying
Sampled simultaneously at 1.6m, then calculate average value.1 coliform is 1 pfu in table 2.
It the above is only the description of the preferred embodiment of the present invention, it is noted that due to the finiteness of literal expression, and
Objectively there is unlimited concrete structure, for those skilled in the art, do not departing from the principle of the invention
On the premise of, some improvements and modifications can also be made, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (10)
1. bacterium or viral online acquisition and online automatic detection method in a kind of air, it is characterised in that：
Using online acquisition and on-line automatic detection device, including sample-adding control device, PCR amplification devices, detection means and from
Dynamic control device, in addition to pressue device and negative pressure device；Comprise the following steps：
1) online acquisition and on-line automatic detection device are opened, is required according to object to be detected and corresponding PCR kit, in advance
PCR amplification programs are set, integrated sampling box is inserted into；
2) sample volume is set, the negative pressure device is then turned on, automatic data collection sample is to setting sample volume；
3) the setting time is stood, institute's negative pressure device starts, the liquid of sample region is sucked into waste liquid bottle, DNA or RNA has been adsorbed
Grain is trapped within the filter medium of the sample region；
4) open the pressue device, DNA or RNA eluents added in the sample region, by the DNA on the particle or
Under the RNA elutions；
5) by the sample-adding control device, the eluent of the DNA or described RNA under elution is added to the PCR and expanded
Device enters performing PCR amplification by the PCR amplification programs that pre-set；
6) after the sample after amplification is detected by detection means, testing result is exported.
2. bacterium or viral online acquisition and online automatic detection method in air according to claim 1, its feature exist
In：The sampling integrated sampling box is connected with the sample-adding control device by pipeline；It is described sample-adding control device with it is described
PCR amplification devices coordinate；
The automaton respectively with the sample-adding control device, the PCR amplification devices, the detection means, described
Pressue device and negative pressure device electrical connection；
The pressue device is connected with the negative pressure device with the sampling integrated sampling box by pipeline.
3. bacterium or viral online acquisition and online automatic detection method, its feature in air according to claim 1 or 2
It is：The sampling integrated sampling box includes sampling container, eluent storage container and waste liquid cylinder；Eluent storage container with
The sampling container is connected by pipeline；The waste liquid cylinder is connected with the bottom of the sampling container by pipeline；
The pressue device is provided with pipeline between the sampling container and the eluent storage container；
The sampling container is connected by pipeline with the negative pressure device respectively with the waste liquid cylinder；
Porous air distributor and filter are provided with the sampling container；The entrance of the porous air distributor and company
The pipeline that the sampling container is connect with the negative pressure device is connected, and the filter is arranged at the outlet at bottom of the sampling container
Place, is provided with adsorption liquid in the sampling container, and the gas outlet of the porous air distributor is less than the liquid of the adsorption liquid
Face；The adsorption liquid is made up of microbial lytic liquid and granule for absorbing；
Eluent is provided with the eluent storage container.
4. bacterium or viral online acquisition and online automatic detection method in air according to claim 3, its feature exist
In：In the adsorption liquid, the volume ratio of the microbial lytic liquid and the granule for absorbing is 10-20：1.
5. bacterium or viral online acquisition and online automatic detection method in air according to claim 1, its feature exist
In：The pressue device is the first peristaltic pump, and the negative pressure device is vavuum pump.
6. bacterium or viral online acquisition and online automatic detection method in air according to claim 3, its feature exist
In：Also include pulling and inserting type interface arrangement, the fast plugging for integrated sampling box of sampling.
7. bacterium or viral online acquisition and online automatic detection method in air according to claim 3, its feature exist
In：The sample-adding control device includes the second peristaltic pump and check valve；The outlet of second peristaltic pump is expanded with the PCR and filled
Put corresponding, the entrance of second peristaltic pump is connected with the outlet of the check valve by pipeline；The entrance of the check valve
It is connected by pipeline with the outlet at bottom of the sampling container.
8. bacterium or viral online acquisition and online automatic detection method in air according to claim 3, its feature exist
In：The PCR amplification devices include sample-adding plate, PCR kit and PCR temperature control equipments, and the sample-adding plate is tried with the PCR
Agent box is connected, and the PCR kit is connected with the PCR temperature control equipments.
9. bacterium or viral online acquisition and online automatic detection method in air according to claim 1, its feature exist
In：The detection means includes excitation light emission device, fluorescent acceptor and interactive system；The excitation light emission device and institute
State the detection zone that fluorescent acceptor is relatively arranged on the PCR amplification devices, the excitation light emission device and the fluorescence reception
Device is electrically connected with the interactive system, and the interactive system is electrically connected with the automaton.
10. bacterium or viral online acquisition and online automatic detection method in air according to claim 1, its feature exist
In：The automatic detection device includes central controller, and the central controller is in cyclelog, single-chip microcomputer or industrial computer
One kind；
The automatic detection device also includes blue tooth interface, wireless WIFI interface, Ethernet interface, USB interface, RS232 serial ports
Or one or more combinations in RS485 serial ports.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710346839.4A CN106967595B (en) | 2017-05-16 | 2017-05-16 | Bacterium or viral online acquisition and online automatic detection method in a kind of air |
| PCT/CN2018/085293 WO2018210128A1 (en) | 2017-05-16 | 2018-05-02 | Method for on-line collection and on-line automatic detection of bacteria or viruses in air |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710346839.4A CN106967595B (en) | 2017-05-16 | 2017-05-16 | Bacterium or viral online acquisition and online automatic detection method in a kind of air |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106967595A true CN106967595A (en) | 2017-07-21 |
| CN106967595B CN106967595B (en) | 2019-11-29 |
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| WO2018210128A1 (en) * | 2017-05-16 | 2018-11-22 | 上海德具生物科技有限公司 | Method for on-line collection and on-line automatic detection of bacteria or viruses in air |
| CN111650173A (en) * | 2020-07-22 | 2020-09-11 | 广州建设工程质量安全检测中心有限公司 | Air virus monitoring devices |
| CN111811078A (en) * | 2020-07-22 | 2020-10-23 | 广州建设工程质量安全检测中心有限公司 | Air conditioner ventilation system's virus monitoring system |
| CN113249203A (en) * | 2020-02-12 | 2021-08-13 | 苏州百源基因技术有限公司 | Air detector and using method thereof |
| CN114058498A (en) * | 2022-01-11 | 2022-02-18 | 至美时代生物智能科技(北京)有限公司 | Full-automatic air microorganism detection system and detection method |
| CN114594208A (en) * | 2022-03-07 | 2022-06-07 | 谱瑞前海(深圳)智能科技有限公司 | Real-time detection device for virus spread to epidemic disease in air environment |
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| CN114058498A (en) * | 2022-01-11 | 2022-02-18 | 至美时代生物智能科技(北京)有限公司 | Full-automatic air microorganism detection system and detection method |
| CN114594208A (en) * | 2022-03-07 | 2022-06-07 | 谱瑞前海(深圳)智能科技有限公司 | Real-time detection device for virus spread to epidemic disease in air environment |
| EP4290210A1 (en) | 2022-06-08 | 2023-12-13 | Fundació Institut de Ciències Fotòniques | A device, a method, and a computer program, for detecting airborne particulate matter in aerosols |
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| CN106967595B (en) | 2019-11-29 |
| WO2018210128A1 (en) | 2018-11-22 |
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