WO2018210127A1 - 一种空气中细菌或病毒在线采集及在线自动检测装置 - Google Patents
一种空气中细菌或病毒在线采集及在线自动检测装置 Download PDFInfo
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- WO2018210127A1 WO2018210127A1 PCT/CN2018/085292 CN2018085292W WO2018210127A1 WO 2018210127 A1 WO2018210127 A1 WO 2018210127A1 CN 2018085292 W CN2018085292 W CN 2018085292W WO 2018210127 A1 WO2018210127 A1 WO 2018210127A1
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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]
Definitions
- the invention belongs to the technical field of microbial detection, in particular to an online collection and online automatic detecting device for bacteria or viruses in the air.
- Airborne transmission is the main route for some pathogens or viruses to spread, and it is the most difficult way to detect and control in the current environment.
- the SARS virus is mainly found in the nose, sputum and saliva of the source of infection (human or animal) and is transmitted by air droplets and contact. When a patient coughs, sneezes, or even speaks loudly, the virus spreads to the surrounding air with saliva droplets, dust particles, etc., and spreads with the air.
- the object of the present invention is to provide an online collection and online automatic detection device for bacteria or viruses in the air to solve the problem that the bacteria or virus in the air cannot be early warning or monitored, so as to realize the rapid and accurate detection of bacteria or viruses in the air. On-site inspection.
- An online collection and online automatic detection device for bacteria or viruses in the air comprising a sampling integrated sampling box, a sample loading control device, a PCR amplification device, a detection device and an automatic control device, and a sampling and integration sampling box Cooperating pressurizing device and vacuum device;
- the sampling integrated sampling box is connected to the sample loading control device through a pipeline; the sample loading control device is matched with the PCR amplification device;
- the automatic control device is electrically connected to the sample loading control device, the PCR amplification device, the detection device, the pressing device, and the negative pressure device, respectively;
- the pressing device and the negative pressure device are both connected to the sampling integrated sampling box through a pipeline.
- the sampling integrated sampling box includes a sampling container, an eluent storage container and a waste liquid tank; the eluent storage container is connected to the sampling container through a pipeline; the waste liquid tank and the bottom of the sampling container pass through the tube Road connection
- sampling container and the waste liquid tank are respectively connected to the negative pressure device through a pipeline;
- a porous air distributor and a filter are disposed in the sampling container; an inlet of the porous air distributor is connected to a pipeline connecting the sampling container and the negative pressure device, and the filter is disposed on the sampling At the bottom outlet of the container, an adsorption liquid is disposed in the sampling container, and an air outlet of the porous air distributor is lower than a liquid surface of the adsorption liquid; the adsorption liquid is composed of a microorganism lysate and adsorbed particles;
- An eluent is disposed in the eluent storage container.
- the volume ratio of the microorganism lysate to the adsorbed particles is 10-20:1.
- the pressurizing device is a first peristaltic pump.
- It also includes a plug-in interface device for sampling the quick insertion of the integrated sampling box.
- the sample loading control device includes a second peristaltic pump and a one-way valve; an outlet of the second peristaltic pump corresponds to the PCR amplification device, an inlet of the second peristaltic pump and an outlet of the one-way valve Connected by a line; the inlet of the one-way valve is connected to the bottom outlet of the sampling container through a line.
- the PCR amplification device includes a sample plate, a PCR kit, and a PCR temperature control device.
- the sample plate is connected to the PCR kit, and the PCR kit is connected to the PCR temperature control device.
- the detecting device includes an excitation light emitter and a fluorescence receiver; the excitation light emitter is disposed opposite to the fluorescence receiver in a detection area of the PCR amplification device.
- a human-machine dialog system the fluorescent transmitter and the fluorescent receiver being electrically coupled to the human-machine dialog system, the human-machine dialog system being electrically coupled to the automatic control device.
- the automatic detecting device includes a central controller, and the central controller is one of a program controller, a single chip microcomputer, or an industrial computer;
- the automatic detecting device further comprises one or more combinations of a Bluetooth interface, a wireless WIFI interface, an Ethernet interface, a USB interface, an RS232 serial port or an RS485 serial port.
- the invention provides an online collection and online automatic detection device for bacteria or viruses, including negative pressure pumping, on-line microbial collection, air microbial adsorption, DNA or RNA online extraction, DNA online amplification and detection, etc., by collecting microorganisms in the air.
- the particles are lysed in real time in a liquid, extracted DNA (or RNA), amplified by PCR, and subjected to online real-time detection.
- the technical scheme can analyze the collected samples separately by sampling, extracting and detecting, and is suitable for determining the number of different kinds of microorganisms in a unit volume of air.
- the new air microbial sampling and detecting instrument developed by the technical solution, combined with biochip or real-time PCR technology, can quickly and sensitively detect pathogenic microorganisms that may exist in the air.
- the device is mainly composed of a negative pressure suction device (handheld or vacuum pump), an air flow meter, an adsorption liquid and adsorbed particles.
- a suction device adsorbing air through a vacuum, at a constant gas flow rate per minute, through the liquid collection zone, under the action of adsorbed particles in the solution and the solution, the virions in the air are collected in a small volume of liquid, while The collected microorganisms are lysed to release microbial DNA (RNA) and adsorbed on the adsorbed particles.
- RNA microbial DNA
- RNA DNA
- RNA DNA
- RNA DNA
- Figure 1 is a schematic view showing the structure of the device of the present invention.
- FIG. 2 is a schematic view showing the operation of the sampling integrated sampling box of the present invention.
- the invention develops a novel air microbial sampling and detecting instrument, and combines biochip or real-time PCR technology to quickly and sensitively detect pathogenic microorganisms which may exist in the air.
- the device is mainly composed of a negative pressure suction device (handheld or vacuum pump), an air flow meter, an adsorption liquid and adsorbed particles.
- a suction device adsorbing air through a vacuum, at a constant gas flow rate per minute, through the liquid collection zone, under the action of adsorbed particles in the solution and the solution, the virions in the air are collected in a small volume of liquid, while The collected microorganisms are lysed to release microbial DNA (RNA) and adsorbed on the adsorbed particles.
- RNA microbial DNA
- RNA DNA
- RNA DNA
- RNA DNA
- the invention integrates sampling, extraction and detection into one body, can separately analyze the collected samples, and is suitable for determining the number of different living microorganisms in a unit volume of air.
- the present application provides an online collection and online automatic detection device for bacteria or viruses in the air, as shown in FIG. 1 and FIG. 2, including a sampling integrated sampling box, a sample loading control device, a PCR amplification device, a detection device and an automatic control device. It also includes a pressurizing device and a vacuum device matched with the sampling integrated sampling box.
- sampling integrated sampling box and the sample loading control device are connected by a pipeline; the sample loading control device is matched with the PCR amplification device; the sampling integrated sampling box of the present application is used for sampling the integrated sampling box by the plug-in interface device. Plug in.
- the automatic control device is respectively electrically connected with the sample control device, the PCR amplification device, the detection device, the pressing device and the negative pressure device;
- the automatic detection device comprises a central controller, and the central controller is a program controller, a single chip microcomputer or an industrial computer One is to fully automate the process from sampling air to detecting the type and concentration of microorganisms.
- the automatic detecting device further comprises one or a combination of a Bluetooth interface, a wireless WIFI interface, an Ethernet interface, a USB interface, an RS232 serial port or an RS485 serial port, and communicates with the host computer or the mobile terminal through a wired or wireless manner to implement the device.
- Remote monitoring allows the tester to obtain test results without going deep into the affected area, thus avoiding the risk of the tester being infected.
- these interfaces can communicate with other devices such as the top computer, mobile phone, etc., so that the detection result can be applied by other devices, the automatic detection device can also be controlled by other devices, and can also realize remote transmission or cloud management of detection data and Remote control of the automatic detection device.
- Both the pressurizing device and the negative pressure device are connected to the sampling integrated sampling box through a pipeline.
- the pressurizing device is the first peristaltic pump 4; in the present application, the vacuum device is the vacuum pump 1.
- the sampling integrated sampling box comprises a sampling container 3, an eluent storage container 5 and a waste liquid tank 11; the eluent storage container and the sampling container are connected by a pipeline; the waste liquid tank and the bottom of the sampling container are connected by a pipeline.
- the flow of the liquid is controlled by the air pressure, and the flow direction of the gas is controlled by the check valve on the pipeline, thereby controlling the pressure in the sampling container, the eluent storage container or the waste liquid tank to realize the liquid (adsorbed liquid or Directional flow of the eluent).
- a pressurizing device is disposed on the pipeline between the sampling container and the eluent storage container;
- the sampling container 3 and the waste liquid tank are respectively connected to the vacuum device through a pipeline; in the present application, the vacuum device is a vacuum pump 1, and a first electromagnetic valve 2 is disposed between the vacuum pump and the sampling container, in the vacuum pump and the waste liquid cylinder A second solenoid valve 10 is disposed therebetween; both the first solenoid valve and the second solenoid valve are electrically connected to the automatic control device, and the first solenoid valve and the second solenoid valve are controlled to be turned on or off by the automatic control device.
- a porous air distributor and a filter 17 are disposed in the sampling container; the inlet of the porous air distributor is connected to a pipeline connecting the sampling container and the negative pressure device, and the filter is disposed at the bottom outlet of the sampling container, and is disposed in the sampling container There is an adsorption liquid 7, the outlet port of the porous air distributor is lower than the liquid level of the adsorption liquid; the air outlet of the porous air distributor distributes the drawn air into the adsorption liquid through a plurality of pores, in order to prevent the airflow from being too large The direct short circuit rushes out the liquid, and the plurality of pores distribute the carrier air into small bubbles, increasing the contact area between the carrier air and the adsorption liquid, and improving the efficiency of adsorbing microorganisms in the air.
- the adsorption solution 7 is composed of a microorganism lysate and adsorbed particles.
- the eluent 6 is provided in the eluent storage container 5.
- the eluate is a DNA (RNA) eluate, and both are commercially available DNA (RNA) eluents.
- the volume ratio of the microorganism lysate to the adsorbed particles is 10-20:1.
- the adsorbed particles are DNA adsorbing particles or RNA adsorbing particles.
- the height-to-diameter ratio of the adsorbent in the sampling container is 3:1-6:1.
- the microbial lysate is phosphate buffered PBS by dissolving NaCl, KCl, Na 2 HPO 4 and KH 2 PO 4 in distilled water, and adjusting the pH of the solution to 7.4 with HCl. Add distilled water to make up to volume, and steam sterilize at 121 ° C for 20 minutes under high pressure, and store in room temperature or 4 ° C refrigerator.
- a broad-spectrum high-efficiency adsorption liquid composition is added to a 1 ml PBS solution to which a chitosan nanoparticle having a particle size distribution of 10 to 30 mg and a diameter of 10 to 200 nm is added.
- a particle size distribution of 10-30. mg of a diameter of 10-200 nm is added to a 1 ml PBS solution with influenza virus antibody-modified nanoparticles.
- the DNA adsorption particles or the RNA adsorption particles may each be a commercially available adsorption resin such as a DNA (RNA) adsorption resin produced by a company such as Shanghai Shenggong or Beijing Quanjin.
- a DNA (RNA) adsorption resin produced by a company such as Shanghai Shenggong or Beijing Quanjin.
- the sample loading control device comprises a second peristaltic pump 8 and a check valve 9; the outlet of the second peristaltic pump corresponds to the PCR amplification device, and the inlet of the second peristaltic pump and the outlet of the check valve are connected by a pipeline; The inlet is connected to the bottom outlet of the sampling vessel via a line.
- the second peristaltic pump is used for the sample application, and the metering amount is measured by the dripper.
- the PCR amplification device comprises a sample plate 15, a PCR kit and a PCR temperature control device 16.
- the sample plate is connected to the PCR kit, and the PCR kit is connected to the PCR temperature control device.
- the reaction solution was sealed by adding liquid paraffin to the PCR kit.
- the PCR kit of the present application is a commercially available PCR kit.
- the detecting device comprises an excitation light emitter 12, a fluorescence receiver 13 and a human-machine dialogue system 14; the excitation light emitter and the fluorescence receiver are disposed opposite to the detection area of the PCR amplification device, and the fluorescence emitter and the fluorescence receiver are both connected to the human-machine
- the dialogue system is electrically connected, and the human-machine dialogue system is electrically connected to the automatic control device.
- the human-machine dialogue system can also set relevant detection parameters and equipment maintenance management.
- the automatic control device will control the vacuum pump to start, realize automatic sampling to the set volume; After 2 minutes, the automatic control device controls the vacuum pump to start again, so that the waste liquid cylinder is under negative pressure, and the liquid in the sampling container is sucked into the waste liquid tank, and the particles adsorbed with DNA (RNA) are cut off on the filter medium in the sampling container.
- the automatic control device controls the first peristaltic pump to start, pressurizes the eluent storage container, and hydraulically elutes the DNA (RNA) into the sampling container, and elutes the adsorbed DNA (RNA) on the exposed particles to pass through
- the check valve and the second peristaltic pump add the eluted DNA (RNA) eluate to a PCR kit pre-loaded with a commercially available DNA (RNA) detection sample, and preset according to the requirements of the PCR kit.
- PCR amplification is performed; after the amplified sample is detected by the detection device of the detection area, the result is output to an automatic control device to obtain the type and concentration of the microorganism, and the detection result is transmitted through wired or wireless communication.
- PC or a mobile terminal PC or a mobile terminal.
- all the opening actions are automatically completed by the automatic control device, and the moving part is controlled by the automatic control device using the stepping motor.
Abstract
Description
Claims (10)
- 一种空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:包括采样一体化采样盒、加样控制装置、PCR扩增装置、检测装置及自动控制装置,还包括与所述采样一体化采样盒相配合的加压装置及负压装置;所述采样一体化采样盒与所述加样控制装置通过管路连接;所述加样控制装置与所述PCR扩增装置配合;所述自动控制装置分别与所述加样控制装置、所述PCR扩增装置、所述检测装置、所述加压装置及所述负压装置电连接;所述加压装置与所述负压装置均与所述采样一体化采样盒通过管路连接。
- 根据权利要求1所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:所述采样一体化采样盒包括采样容器、洗脱液存储容器及废液缸;洗脱液存储容器与所述采样容器通过管路连接;所述废液缸与所述采样容器的底部通过管路连接;在所述采样容器与所述洗脱液存储容器之间的管路上设置有所述加压装置;所述采样容器与所述废液缸分别通过管路与所述负压装置连接;在所述采样容器内设置有多孔空气分布器及过滤器;所述多孔空气分布器的入口与连接所述采样容器与所述负压装置的管路连接,所述过滤器设置于所述采样容器的底部出口处,在所述采样容器内设置有吸附液,所述多孔空气分布器的出气口低于所述吸附液的液面;所述吸附液由微生物裂解液及吸附粒子组成;在所述洗脱液存储容器内设置有洗脱液。
- 根据权利要求2所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:在所述吸附液中,所述微生物裂解液与所述吸附粒子的体积比为10-20:1。
- 根据权利要求1所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:所述加压装置为第一蠕动泵。
- 根据权利要求1所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:还包括插拨式接口装置,用于采样一体化采样盒的快速插拨。
- 根据权利要求2所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:所述加样控制装置包括第二蠕动泵及单向阀;所述第二蠕动泵的出口与所述PCR扩增装置相对应,所述第二蠕动泵的入口与所述单向阀的出口通过管路连接;所述单向阀的入口通过管路与所述采样容器的底部出口连接。
- 根据权利要求1所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:所述PCR扩增装置包括加样板、PCR试剂盒及PCR温度控制装置,所述加样板与所述PCR试剂盒连通,所述PCR试剂盒与所述PCR温度控制装置连接。
- 根据权利要求1所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:所述检测装置包括激发光发射器及荧光接收器;所述激发光发射器与所述荧光接收器相对设置于所述PCR扩增装置的检测区。
- 根据权利要求8所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:还包括人机对话系统,所述荧光发射器与所述荧光接收器均与所述人机对话系统电连接,所述人机对话系统与所述自动控制装置电连接。
- 根据权利要求1所述的空气中细菌或病毒在线采集及在线自动检测装置,其特征在于:所述自动检测装置包括中央控制器,所述中央控制器为程序控制器、单片机或工控机中的一种;所述自动检测装置还包括蓝牙接口、无线WIFI接口、以太网接口、USB接口、RS232串口或RS485串口中的一种或多种组合。
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CN107118961B (zh) * | 2017-05-16 | 2020-06-16 | 上海德具生物科技有限公司 | 一种空气中细菌或病毒在线采集及在线自动检测装置 |
CN108239599B (zh) * | 2018-03-26 | 2019-12-13 | 胥振国 | 一种空气细菌采样、培养及检测一体化装置 |
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EP3887051A2 (en) * | 2018-11-30 | 2021-10-06 | Illumina, Inc. | Temperature-controllable reagent cartridge and temperature control system for the same |
CN110229751A (zh) * | 2019-07-17 | 2019-09-13 | 中国人民解放军军事科学院军事医学研究院 | mRNA制备系统及制备方法 |
CN111607483B (zh) * | 2020-05-07 | 2023-09-15 | 广州再生医学与健康广东省实验室 | 体外诊断仪及提取装置 |
CN112221544B (zh) * | 2020-09-27 | 2022-06-17 | 北京理工大学重庆创新中心 | 一种集采样与检测一体化的微流控芯片 |
CN114058498B (zh) * | 2022-01-11 | 2022-04-19 | 至美时代生物智能科技(北京)有限公司 | 一种全自动空气微生物检测系统及检测方法 |
CN114594208A (zh) * | 2022-03-07 | 2022-06-07 | 谱瑞前海(深圳)智能科技有限公司 | 一种在空气环境中对流行病传播病毒的实时检测装置 |
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