CN106967595B - 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
- Publication number
- CN106967595B CN106967595B CN201710346839.4A CN201710346839A CN106967595B CN 106967595 B CN106967595 B CN 106967595B CN 201710346839 A CN201710346839 A CN 201710346839A CN 106967595 B CN106967595 B CN 106967595B
- Authority
- CN
- China
- Prior art keywords
- air
- sample
- sampling
- online
- liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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
-
- 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]
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Pathology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to bacteriums in a kind of air or viral online acquisition and online automatic detection method can rapidly and sensitively detect out pathogenic microorganisms that may be present in air in conjunction with biochip or fluorescent quantitative PCR technique.The device is mainly made of negative pressure air-suction device, air flow meter, adsorption liquid and absorption particle.Utilize air extractor, pass through negative-pressure adsorption air, with constant gas amount per minute, by liquid acquisition area, under the action of absorption particle in solution and solution, collect the virion in air in the liquid of small size, while cracking the microorganism of collection, releasing microbe DNA (RNA) is adsorbed on absorption particle.Then pass through on-line filtration, collect the particle in liquid, with commercially available DNA (RNA) elution DNA (RNA), then microbial biomass is detected using round pcr, the copy number of measurement detects the micro organism quantity in resulting unit air divided by the volume of air of absorption.
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 technique
Air borne is the main path of some pathogenic bacteria or viral transmission, and is most difficult to detect and control in current environment
The approach of system.
It is quick-fried to influence domestic H1N1 and 2013 year by 2009 for the SARS epidemic situation for bringing enormous impact to the whole world from 2003
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 the nasal mucus in dye source (human or animal), phlegm and saliva, pass through airborne droplet and contact transmission.Patient is by coughing, sneezing, very
Into the air that virus is diffused into surrounding with saliva droplet, dust particle etc. by Shi Douhui of speaking up, flows and propagate with air.
Why SARS initial stage causes a large amount of personnel to infect especially medical staff and brings huge sacrifice, except safeguard procedures
Outside reason, the virus being largely because in air is difficult to caused by monitoring and taking precautions against.
Although the more than ten years in SARS epidemic situation past, it is to the tremendous influence of social life bring and social panic mood
Let us is remembered clearly.Even to this day, for harmful microbe early warning in air and monitoring still lack effective tool and
Method.Existing detecting instrument and detection method program are complicated, take a long time, it is difficult to provide epidemic situation scene accurately data letter in time
Breath is to take rapidly the precautionary measures.
With the development of society, the contacts of country variant, 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.Especially within the hospital, cross infection in hospital is always the problem of enabling hospital's headache.
When avian influenza virus prevalence, because government is often in no positive evidence without device for fast detecting
Poultry farms are blindly slaughtered, are all brought about great losses to government and farm.
Summary of the invention
The object of the present invention is to provide bacterium in a kind of air or viral online acquisitions and online automatic detection method, with solution
Must not to bacterium in air or virus carry out early warning or monitoring the problem of, with realize in air bacterium or virus carry out it is fast
The accurate on-site test of speed.
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 device, detection device
And automatic control device, it further include pressurizing device and negative pressure device；The following steps are included:
1) online acquisition and on-line automatic detection device are opened, is wanted according to object to be detected and corresponding PCR kit
It asks, pre-sets PCR amplification program, be inserted into integrated sampling box；
2) sample volume is set, is then turned on the negative pressure device, automatic collection sample is to being arranged sample volume；
3) the setting time is stood, the liquid of sample region is sucked waste liquid bottle, adsorbed DNA or RNA by the starting of institute's negative pressure device
Particle be trapped on the filter medium of the sample region；
4) pressurizing device is opened, DNA or RNA eluent is added in the sample region, described on the particle
Under DNA or RNA elution；
5) by the sample-adding control device, the eluent of DNA described under the elution or RNA is added to the PCR
Amplification device carries out PCR amplification by the PCR amplification program that pre-sets；
6) after the sample after amplification being detected by detection device, output test result.
The sampling integrated sampling box and the sample-adding control device pass through piping connection；The sample-adding control device with
The PCR amplification device cooperation；
The automatic control device respectively with the sample-adding control device, the PCR amplification device, the detection device,
The pressurizing device and negative pressure device electrical connection；
The pressurizing device and the negative pressure device pass through piping connection with the sampling integrated sampling box.
The sampling integrated sampling box includes sampling container, eluent storage container and waste liquid cylinder；Eluent storage is held
Device and the sampling container pass through piping connection；The bottom of the waste liquid cylinder and the sampling container passes through piping connection；
The pressurizing device is provided on pipeline between the sampling container and the eluent storage container；
The sampling container passes through pipeline with the waste liquid cylinder respectively and connect with the negative pressure device；
Porous air distributor and filter are provided in the sampling container；The entrance of the porous air distributor
With the piping connection for connecting the sampling container Yu the negative pressure device, the filter is set to 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 lower than the adsorption liquid
Liquid level；The adsorption liquid is made of microbial lytic liquid and absorption particle；
Eluent is provided in the eluent storage container.
In the adsorption liquid, the volume ratio of the microbial lytic liquid and the absorption particle is 10-20:1.
The pressurizing device is the first peristaltic pump, and the negative pressure device is vacuum pump.
It further include pulling and inserting type interface arrangement, for sampling the fast plugging of integrated sampling box.
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 and the outlet of the check valve pass through piping connection；The check valve
Entrance connect with the outlet at bottom of the sampling container by pipeline.
The PCR amplification device includes sample-adding plate, PCR kit and PCR temperature control equipment, the sample-adding plate with it is described
PCR kit connection, the PCR kit are connect with the PCR temperature control equipment.
The detection device 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 device, 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 automatic control device.
The automatic detection device includes central controller, and the central controller is cyclelog, single-chip microcontroller or work
One of control machine；
The automatic detection device further includes blue tooth interface, wireless WIFI interface, Ethernet interface, USB interface, RS232
One of serial ports or RS485 serial ports or multiple combinations.
The beneficial effects of the present invention are:
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 testing, DNA such as expand and detect online at the components, pass through acquisition air
In microorganism particle, cracked in real time in a liquid, extract DNA (or RNA), after PCR amplification, carried out online real-time
Detection.
The technical program, together in one, can be analyzed respectively the sample of acquisition, and be suitble to by sampling, extraction and detection collection
Different types of microorganisms quantity in analytical unit volumes of air.
The novel air microbe sampling detecting instrument that the technical program is developed, in conjunction with biochip or fluorescent quantitation
Round pcr can rapidly and sensitively detect out pathogenic microorganisms that may be present in air.The device is mainly filled by negative inspiratory pressure
Set (hand-held or vacuum pump), air flow meter, adsorption liquid and absorption particle composition.Using air extractor, pass through negative-pressure adsorption
Air, by liquid acquisition area, under the action of absorption particle in solution and solution, makes sky with constant gas amount per minute
Virion in gas is collected in the liquid of small size, while cracking the microorganism of collection, and releasing microbe DNA (RNA) inhales
It is attached on absorption particle.Then by on-line filtration, the particle in liquid is collected, with commercially available DNA (RNA) elution
DNA (RNA) then detects microbial biomass using round pcr, and the copy number of measurement is divided by the volume of air of absorption, i.e. detection institute
The micro organism quantity in unit air obtained.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of apparatus of the present invention；
Fig. 2 is present invention sampling integrated sampling box operation schematic diagram.
Description of symbols
1 vacuum pump, 2 first solenoid 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 solenoid valves, 11 waste liquid cylinders, 12 excitation light emission devices, 13 fluorescent acceptors, 14
Interactive system, 15 sample-adding plates, 16PCR temperature control equipment, 17 filters.
Specific embodiment
Carry out the technical solution that the present invention will be described in detail by the following examples, embodiment below is merely exemplary, only
It can be used to explanation and illustration technical solution of the present invention, and be not to be construed as the limitation to technical solution of the present invention.
The present invention develops novel air microbe sampling detecting instrument, in conjunction with biochip or quantitative fluorescent PCR skill
Art can rapidly and sensitively detect out pathogenic microorganisms that may be present in air.The device is mainly by negative pressure air-suction device (hand
Hold formula or vacuum pump), air flow meter, adsorption liquid and absorption particle composition.Using air extractor, by negative-pressure adsorption air,
With constant gas amount per minute, by liquid acquisition area, under the action of absorption particle in solution and solution, make in air
Virion is collected in the liquid of small size, while cracking the microorganism of collection, and releasing microbe DNA (RNA) is adsorbed on
On attached particle.Then by on-line filtration, the particle in liquid is collected, with commercially available DNA (RNA) elution DNA
(RNA), microbial biomass is then detected using round pcr, the copy number of measurement detects resulting divided by the volume of air of absorption
Micro organism quantity in unit air.
Current such sampler in the market is only applicable to the air microbe sampling of high concentration, for the disease compared with low-density
Poison can not trap piconavirus particle, therefore due to the limitation of the factors such as residence time of the air in the liquid of smaller size smaller
Sensitivity is not high, while can not on-line determination.The present invention is collected by sampling, extraction and detection together in one, can be by acquisition
Sample is analyzed respectively, and is suitble 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 institute
Show, including sampling integrated sampling box, sample-adding control device, PCR amplification device, detection device and automatic control device, also wraps
Include the pressurizing device and negative pressure device matched with sampling integrated sampling box.
Sampling integrated sampling box and sample-adding control device pass through piping connection；It is loaded control device and PCR amplification device
Cooperation；The sampling integrated sampling box of the application is inserted by pulling and inserting type interface arrangement for sampling the quick of integrated sampling box
It dials.
Automatic control device is filled with sample-adding control device, PCR amplification device, detection device, pressurizing device and negative pressure respectively
Set electrical connection；Automatic detection device includes central controller, and central controller is in cyclelog, single-chip microcontroller or industrial personal computer
One kind is realized by programming from air sampling to detecting that microbe species and concentration are performed fully automatic.
Automatic detection device further includes blue tooth interface, wireless WIFI interface, Ethernet interface, USB interface, RS232 serial ports
Or one of RS485 serial ports or multiple combinations, pass through wired or wireless way and host computer or mobile terminal communication, realizes pair
The long-range monitoring of the present apparatus, testing staff can obtain testing result without going deep into epidemic-stricken area, it is infected to avoid testing staff
Risk.Meanwhile these interfaces can be communicated with other equipment such as host computer, mobile phone etc., set testing result can by other
Standby application, automatic detection device can also be controlled by other equipment, can also realize the remote transmission to detection data or cloud
Management and the long-range control to automatic detection device.
Pressurizing device and negative pressure device pass through piping connection with sampling integrated sampling box.
Pressurizing device is the first peristaltic pump 4；Negative pressure device is vacuum pump 1 in this application.
Sampling integrated sampling box includes sampling container 3, eluent storage container 5 and waste liquid cylinder 11；Eluent storage is held
Device and sampling container pass through piping connection；The bottom of waste liquid cylinder and sampling container passes through piping connection.In sampling integrated sampling
In box, liquid flows through pressure control, controls gas flow direction by check valve on pipeline, holds to control sampling
Pressure in device, eluent storage container or waste liquid cylinder realizes the directed flow of liquid (adsorption liquid or eluent).
Pressurizing device is provided on pipeline between sampling container and eluent storage container；
Sampling container 3 passes through pipeline with waste liquid cylinder respectively and connect with negative pressure device；In this application, negative pressure device is vacuum
Pump 1, is provided with the first solenoid valve 2 between vacuum pump and sampling container, and the second electricity is provided between vacuum pump and waste liquid cylinder
Magnet valve 10；First solenoid valve is electrically connected with automatic control device with second solenoid valve, passes through automatic control device control first
Solenoid valve and second solenoid valve open or close.
Porous air distributor and filter 17 are provided in sampling container；The entrance of porous air distributor and connection
The piping connection of sampling container and negative pressure device, filter are set at the outlet at bottom of sampling container, are set in sampling container
It is equipped with adsorption liquid 7, the gas outlet of porous air distributor is lower 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 air-flow is too big in order to prevent, and liquid is gone out in direct short-circuit,
Multiple pores air that will carry disease germs is scattered in minute bubbles, increases the contact area of carry disease germs air and adsorption liquid, improves in absorption air
The efficiency of microorganism.
Adsorption liquid is made of microbial lytic liquid and absorption particle.
Eluent 6 is provided in 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 absorption particle is 10-20:1.
Adsorbing particle is that DNA adsorbs particle or RNA adsorbs particle.
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 configuration method is, by NaCl, KCl,
Na2HPO4And KH2PO4, it is dissolved in distilled water, adjusts the pH value of solution to 7.4 with HCl, finally plus distilled water constant volume, 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
HCl adjusts the pH value of solution to 7.4, and finally plus distilled water is settled to 1L.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 forms: it is 10-30 milligrams straight that particle diameter distribution is added in 1ml PBS solution
Diameter is the chitin nanometer of 10-200nm.
In this application, as formed using specific adsorption liquid, addition particle diameter distribution is 10-30. milli in 1ml PBS solution
Gram diameter is that the surface of 10-200nm has the nanoparticle of Antibody of Influenza modification.
In this application, DNA adsorb particle or RNA absorption particle all can be commercially available absorption resin, such as the raw work in Shanghai
Or the DNA (RNA) of Beijing Quan Shijindeng company production adsorbs resin.
Being loaded control device includes the second peristaltic pump 8 and check valve 9；The outlet of second peristaltic pump and PCR amplification device phase
Corresponding, the outlet of the entrance and check valve of the second peristaltic pump passes through piping connection；The entrance of check valve is held by pipeline and sampling
The outlet at bottom of device connects.The application sample-adding plate sample application zone, be loaded using the second peristaltic pump, using drop recorder into
The metering of row sample-adding amount.
PCR amplification device includes sample-adding plate 15, PCR kit and PCR temperature control equipment 16, is loaded plate and PCR reagent
Box connection, PCR kit are connect with PCR temperature control equipment.Adding liquid paraffin carries out reaction solution close in PCR kit
Envelope.The PCR kit of the application is commercially available PCR kit.
Detection device includes excitation light emission device 12, fluorescent acceptor 13 and interactive system 14；Excitation light emission device
It is relatively arranged on the detection zone of PCR amplification device with fluorescent acceptor, fluorescent emitters and fluorescent acceptor are and human-computer dialogue
System electrical connection, interactive system are electrically connected with automatic control device.Interactive system can also set related inspection simultaneously
Survey parameter and equipment maintenance and management.
Specific working principle is:
According to test object and PCR kit requirement, PCR amplification program, insertion sampling integrated sampling box are set；Setting
Good sampling volume is then turned on device, and automatic control device can control vacuum pump startup, realizes automatic sampling to setting volume；
After standing 2 minutes, automatic control device control vacuum pump is again started up, and is made for negative pressure in waste liquid cylinder, will be in sampling container
Liquid be drawn into waste liquid cylinder, adsorbed the particle cutoff of DNA (RNA) on the filter medium in sampling container；It automatically controls
Device controls the first wriggling pump startup, pressurizes for 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 in advance added in commercially available DNA (RNA) test sample PCR kit, and preset according to the requirement of PCR kit
Program carries out PCR amplification；After the detection device detection that sample after amplification passes through detection zone, exports result and give automatic control dress
It sets, obtains the type letter concentration of microorganism, and will test result and host computer or movement are passed to eventually by wired or wireless communication
End.
In this application, all breakdown actions control each device by automatic control device and are automatically performed, motion parts
It is controlled and is completed by automatic control device using stepper motor.
The application provides bacterium or viral online acquisition and online automatic detection method in a kind of air,
Use online acquisition and on-line automatic detection device, comprising 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 correspondence
PCR kit requirement, pre-set PCR amplification program, be inserted into integrated sampling box, in this application, all consumptive materials are equal
Need aseptic process.
2) sample volume is set, is then turned on negative pressure device, in the present embodiment, negative pressure device is vacuum pump, is adopted automatically
Collection sample is to being arranged sample volume.
3) the setting time is stood, selects stand 2 minutes in the present embodiment, institute's negative pressure device starting, by the liquid of sample region
Body sucks waste liquid bottle, and the particle for having adsorbed DNA or RNA is trapped on the filter medium of sample region.
4) pressurizing device is opened, in the present embodiment, pressurizing device is the first peristaltic pump, and DNA or RNA is added in sample region
Eluent, by particle DNA or RNA elution under.
5) pass through sample-adding control device, the eluent that will elute lower DNA or RNA is added to PCR amplification device by presetting
It sets PCR amplification program and carries out PCR amplification.
6) after being detected the sample after amplification by detection device, output test result obtains the type of microorganism
And concentration.
Embodiment 1
Particle is adsorbed in the liquid addition commercially available DNA of 1ml (RNA) in liquid acquisition area, respectively with the NaOH of 0.01mol/L and
HCl adjusts the pH to 6-8 of solution, carries out the acquisition experiment of influenza virus in air.Mouse lung adapted strain influenza A virus is passed through
After chicken embryo rejuvenation, it is configured to 100000 viruses/ml solution with the phosphate solution of 0.1mol/L, then to volume about 50m3's
In confined space, virus is sprayed in air with air sparger, specific virion number subtracts 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 3:1,
0.1m is acquired in different ways3Influenza virus, carry out sample detecting experiment, every time three batches, experimental result in two times
(table 1).PCR kit is detected using commercially available H1N1 quantification kit.
The collection result of virion in 1 air of table
Note: sample mode, after spraying, at once with line acquisition and on-line automatic detection device respectively in four corners in room
It is sampled simultaneously at the liftoff 1.6m in middle position, then calculates average value, 1 virus is 1 copy in table 1.
Embodiment 2
Particle is adsorbed in the liquid addition commercially available DNA of 1ml (RNA) in liquid acquisition area, respectively with the NaOH of 0.01mol/L and
HCl adjusts the pH to 6-8 of solution, carries out the acquisition experiment of Escherichia coli in air.Bacillus coli DH 5 alpha is used into conventional method
Culture to concentration is 5 × 106Afterwards, it is configured to 100000 cells/ml solution with the phosphate solution of 0.1mol/L, then to body
Product about 50m3Confined space in, spray virus in air with air sparger, specific microorganism population is according to spraying
(the microbial cell concentration in air is about 1000/m for the reduction amount calculating of liquid in device3).The liquid bulk in liquid acquisition area
Product 15ml, ratio of height to diameter 3:1 acquire 0.1m in different ways3Escherichia coli, in two times carry out sample detecting experiment, often
Secondary three batches, experimental result (table 2).It is detected using commercially available colibacillus PCR immue quantitative detection reagent box.
The collection result of Escherichia coli in 2 air of table
Note: note: sample mode, it is liftoff in four corners in room and middle position respectively with sampler at once after spraying
It is sampled simultaneously at 1.6m, then calculates average value.1 coliform is 1 pfu in table 2.
The above is only the descriptions of the preferred embodiment of the present invention, it is noted that due to the finiteness of literal expression, and
Objectively there is unlimited specific structure, for those skilled in the art, is not departing from the principle of the invention
Under the premise of, several improvements and modifications can also be made, these modifications and embellishments should also be considered as the scope of protection of the present invention.
Claims (9)
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 device, detection device and from
Dynamic control device, further includes pressurizing device and negative pressure device；The following steps are included:
1) online acquisition and on-line automatic detection device are opened, according to object to be detected and the requirement of corresponding PCR kit, in advance
PCR amplification program is set, integrated sampling box is inserted into；
2) sample volume is set, is then turned on the negative pressure device, automatic collection sample is to being arranged sample volume；
3) the setting time is stood, the liquid of sample region is sucked waste liquid bottle, adsorbed of DNA or RNA by the starting of institute's negative pressure device
Grain is trapped on the filter medium of the sample region；
4) open the pressurizing device, DNA or RNA eluent be added in the sample region, by the particle the DNA or
Under the RNA elution；
5) by the sample-adding control device, the eluent of DNA described under the elution or RNA is added to the PCR amplification
Device carries out PCR amplification by the PCR amplification program that pre-sets；
6) after the sample after amplification being detected by detection device, output test result；
The sampling integrated sampling box includes sampling container, eluent storage container and waste liquid cylinder；Eluent storage container with
The sampling container passes through piping connection；The bottom of the waste liquid cylinder and the sampling container passes through piping connection；
The pressurizing device is provided on pipeline between the sampling container and the eluent storage container；
The sampling container passes through pipeline with the waste liquid cylinder respectively and connect with the negative pressure device；
Porous air distributor and filter are provided in the sampling container；The entrance of the porous air distributor and company
The piping connection of the sampling container Yu the negative pressure device is connect, the filter is set to 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 lower than the liquid of the adsorption liquid
Face；The adsorption liquid is made of microbial lytic liquid and absorption particle；
Eluent is provided in the eluent storage container.
2. bacterium or viral online acquisition and online automatic detection method, feature exist in air according to claim 1
In: the sampling integrated sampling box and the sample-adding control device pass through piping connection；The sample-adding control device with it is described
The cooperation of PCR amplification device；
The automatic control device respectively with the sample-adding control device, the PCR amplification device, the detection device, described
Pressurizing device and negative pressure device electrical connection；
The pressurizing device and the negative pressure device pass through piping connection with the sampling integrated sampling box.
3. bacterium or viral online acquisition and online automatic detection method, feature exist in air according to claim 1
In: in the adsorption liquid, the volume ratio of the microbial lytic liquid and the absorption particle is 10-20:1.
4. bacterium or viral online acquisition and online automatic detection method, feature exist in air according to claim 1
In: the pressurizing device is the first peristaltic pump, and the negative pressure device is vacuum pump.
5. bacterium or viral online acquisition and online automatic detection method, feature exist in air according to claim 1
In: it further include pulling and inserting type interface arrangement, for sampling the fast plugging of integrated sampling box.
6. bacterium or viral online acquisition and online automatic detection method, feature exist in air according to claim 1
In: the sample-adding control device includes the second peristaltic pump and check valve；The outlet of second peristaltic pump and the PCR amplification fill
Set corresponding, the entrance of second peristaltic pump and the outlet of the check valve pass through piping connection；The entrance of the check valve
It is connect by pipeline with the outlet at bottom of the sampling container.
7. bacterium or viral online acquisition and online automatic detection method, feature exist in air according to claim 1
In: the PCR amplification device includes sample-adding plate, PCR kit and PCR temperature control equipment, and the sample-adding plate and the PCR are tried
The connection of agent box, the PCR kit are connect with the PCR temperature control equipment.
8. bacterium or viral online acquisition and online automatic detection method, feature exist in air according to claim 1
In: the detection device 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 device, 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 automatic control device.
9. bacterium or viral online acquisition and online automatic detection method, feature exist in air according to claim 1
In: the automatic detection device includes central controller, and the central controller is in cyclelog, single-chip microcontroller or industrial personal computer
One kind；
The automatic detection device further includes blue tooth interface, wireless WIFI interface, Ethernet interface, USB interface, RS232 serial ports
Or one of RS485 serial ports or multiple combinations.
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 CN106967595A (en) | 2017-07-21 |
| CN106967595B true CN106967595B (en) | 2019-11-29 |
Family
ID=59326610
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710346839.4A Active CN106967595B (en) | 2017-05-16 | 2017-05-16 | Bacterium or viral online acquisition and online automatic detection method in a kind of air |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN106967595B (en) |
| WO (1) | WO2018210128A1 (en) |
Families Citing this family (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106967595B (en) * | 2017-05-16 | 2019-11-29 | 上海德具生物科技有限公司 | Bacterium or viral online acquisition and online automatic detection method in a kind of air |
| 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 |
| TWI756746B (en) * | 2020-07-21 | 2022-03-01 | 威盛電子股份有限公司 | Virus collection assembly, virus storage device and virus collection method |
| CN111811078B (en) * | 2020-07-22 | 2022-03-11 | 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 |
| CN111650173B (en) * | 2020-07-22 | 2022-03-15 | 广州建设工程质量安全检测中心有限公司 | Air virus monitoring devices |
| WO2022024035A1 (en) * | 2020-07-29 | 2022-02-03 | Istituto Nazionale Di Astrofisica | Apparatus and method for sampling and detecting a pathogen in air |
| CN112557364B (en) * | 2020-12-11 | 2023-03-07 | 天津市职业大学 | Intelligent indoor air quality virus detection system and detection method |
| FR3123725A1 (en) | 2021-06-04 | 2022-12-09 | Valérie COMPAN | MOBILE AND AUTONOMOUS DEVICE AND METHOD FOR DETECTING ANALYTES IN THE AIR |
| CN114058498B (en) * | 2022-01-11 | 2022-04-19 | 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 |
| CN115902088B (en) * | 2023-03-09 | 2023-05-16 | 合肥泷睿机器人有限责任公司 | Nanometer titanium dioxide photocatalysis screen virus killing performance testing device |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102559486A (en) * | 2011-12-22 | 2012-07-11 | 中国人民解放军军事医学科学院微生物流行病研究所 | Full-automatic air microbe sampler |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1208113C (en) * | 2003-05-01 | 2005-06-29 | 东南大学 | Device for collecting micro-substance in air and collection method thereof |
| ITTO20120320A1 (en) * | 2012-04-12 | 2013-10-13 | St Microelectronics Srl | DEVICE AND METHOD FOR THE PREPARATION OF BIOLOGICAL SAMPLES, IN PARTICULAR FOR THE EXTRACTION OF DNA, AND THE LOADING IN DRAINAGE FOR THE NEXT EXECUTION OF PCR |
| US9677975B2 (en) * | 2014-10-31 | 2017-06-13 | General Electric Company | Systems and methods for aseptic sampling |
| CN104568680A (en) * | 2015-01-14 | 2015-04-29 | 浙江大学 | Community monitoring method of microorganism carried by air particulate matters with multiple particle sizes |
| CN106967595B (en) * | 2017-05-16 | 2019-11-29 | 上海德具生物科技有限公司 | Bacterium or viral online acquisition and online automatic detection method in a kind of air |
-
2017
- 2017-05-16 CN CN201710346839.4A patent/CN106967595B/en active Active
-
2018
- 2018-05-02 WO PCT/CN2018/085293 patent/WO2018210128A1/en active Application Filing
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102559486A (en) * | 2011-12-22 | 2012-07-11 | 中国人民解放军军事医学科学院微生物流行病研究所 | Full-automatic air microbe sampler |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2018210128A1 (en) | 2018-11-22 |
| CN106967595A (en) | 2017-07-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106967595B (en) | Bacterium or viral online acquisition and online automatic detection method in a kind of air | |
| CN107118961B (en) | Online collection and online automatic detection device for bacteria or viruses in air | |
| TWI833984B (en) | Triggered sampling systems and methods | |
| Hermann et al. | Optimization of a sampling system for recovery and detection of airborne porcine reproductive and respiratory syndrome virus and swine influenza virus | |
| KR20230005825A (en) | Diagnosis of respiratory diseases using exhaled breath and aerosol analysis | |
| Pyankov et al. | Inactivation of airborne influenza virus in the ambient air | |
| CN111323511B (en) | Rapid detection kit and method for inactivating new coronavirus | |
| CN102888397A (en) | Kit using magnetic bead to extract whole blood genomic DNA and use of kit | |
| Agranovski et al. | New personal sampler for viable airborne viruses: feasibility study | |
| CN102323115A (en) | Air sampler | |
| Cho et al. | Continuous surveillance of bioaerosols on-site using an automated bioaerosol-monitoring system | |
| CN104560796B (en) | It is a kind of to be resistant to and the grignard Serratieae of enriching heavy metal and its application | |
| US20220034763A1 (en) | Platform for sampling viral particles on surfaces, porous structures, and in the air | |
| CN104483339A (en) | On-line analyzer and analysis method of mercury in flue gas based on wet enrichment | |
| CN107118951B (en) | Sampling integration sampling box structure | |
| CN104977388A (en) | Detection system for detecting purifying rate of air purifier and air purifying material, and detection method thereof | |
| CN105368949A (en) | Detection method of city wastewater treatment plant/station microbial aerosol | |
| CN105203510B (en) | A kind of microorganism in food rapid detection method | |
| CN204101457U (en) | A kind of reaction unit detecting microorganism for ATP bioluminescence method | |
| WO2023116477A1 (en) | Method for detecting bacterial filtration efficiency of mask | |
| CN102134554A (en) | Portable biologic sampler | |
| CN205352790U (en) | Atmospheric sampling device for environment measuring | |
| CN113249203A (en) | Air detector and using method thereof | |
| CN204789523U (en) | A detecting system for being directed at purifying rate of air purifier and air purification material detects | |
| CN108949896A (en) | A kind of fluorescence labeling method of microbes in air and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |