CN112710516A - Microorganism collector, microorganism sampling system and sampling method - Google Patents
Microorganism collector, microorganism sampling system and sampling method Download PDFInfo
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- 244000005700 microbiome Species 0.000 title claims abstract 44
- 238000005070 sampling Methods 0.000 title claims abstract 16
- 238000000034 method Methods 0.000 title claims abstract 7
- 230000002068 genetic effect Effects 0.000 claims abstract 8
- 239000007788 liquid Substances 0.000 claims 7
- 238000009630 liquid culture Methods 0.000 claims 3
- 239000002609 medium Substances 0.000 claims 3
- 238000000605 extraction Methods 0.000 claims 2
- 239000001963 growth medium Substances 0.000 claims 2
- 238000001514 detection method Methods 0.000 claims 1
- 230000000813 microbial effect Effects 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract 1
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- 238000007689 inspection Methods 0.000 abstract 1
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- 238000004659 sterilization and disinfection Methods 0.000 abstract 1
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/22—Devices for withdrawing samples in the gaseous state
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Abstract
本申请涉及一种微生物采集器、微生物采样系统及采样方法。微生物采集器包括：采集罐，内部为空腔，采集罐包括进气口和出气口，空腔内部设置微生物过滤网，进气口和出气口分别位于微生物过滤网的两侧；采样头，连接采集罐的进气口，采样头用于采集洁净区回风口过滤网上的微生物和/或微生物遗传信息；气泵，连接采集罐的出气口。本申请的微生物采集器在回风口处进行采样，检测结果可用于洁净环境监测、消毒控制和药品监管部门的监督检查。
The present application relates to a microorganism collector, a microorganism sampling system and a sampling method. The microorganism collector includes: a collection tank with a cavity inside, the collection tank includes an air inlet and an air outlet, a microorganism filter screen is arranged inside the cavity, and the air inlet and the air outlet are respectively located on both sides of the microorganism filter screen; the sampling head, connected to The air inlet of the collection tank, the sampling head is used to collect microorganisms and/or the genetic information of microorganisms on the filter net of the return air outlet of the clean area; the air pump is connected to the air outlet of the collection tank. The microorganism collector of the present application conducts sampling at the return air outlet, and the test results can be used for clean environment monitoring, disinfection control and supervision and inspection by drug regulatory authorities.
Description
Technical Field
The application relates to the field of microorganism sampling, in particular to a microorganism collector, a microorganism sampling system and a microorganism sampling method.
Background
Environmental monitoring is crucial to drug production, and critical area microorganism monitoring, contaminated microorganism identification and strain library establishment are important, and can be used for guiding disinfection, process monitoring, risk assessment, contaminated product tracing and the like.
Because of the large general area of the purification space, the contaminated microorganisms conform to the statistical Poisson distribution and may also show overproof or outbreak within a certain time or space. And (3) single or multiple verification detections are carried out, the types and the quantity of extracted microorganisms are extremely limited, and the effect on pollution risk assessment of a clean area is small. At present, the problem of how to effectively analyze the microorganism population which may appear in a clean area in a period of time through one-time detection is needed to be solved.
Disclosure of Invention
Based on the problems of the background art, the application provides a microorganism collector, a microorganism sampling system and a sampling method, wherein the collection of microorganisms is carried out at an air return opening, and the support is provided for the analysis of the microorganisms.
One embodiment of the present application provides a microorganism collector, comprising: the collection tank is internally provided with a cavity, the collection tank comprises an air inlet and an air outlet, a microorganism filter screen is arranged in the cavity, and the air inlet and the air outlet are respectively positioned at two sides of the microorganism filter screen; the sampling head is connected with the air inlet of the collection tank, and is used for collecting microorganisms and/or microorganism genetic information on a filter screen of an air return opening of a clean area and sending the microorganisms and/or microorganism genetic information into the collection tank through air; and the air pump is connected with the air outlet of the collection tank.
According to some embodiments of the present application, the microorganism collector further comprises a liquid medium tank, the liquid medium tank comprising a liquid outlet; the collection tank comprises a liquid inlet, and the liquid outlet of the liquid culture medium tank is connected with the liquid inlet of the collection tank through a control valve.
According to some embodiments of the application, the outer wall of the liquid medium pot is connected to the outer wall of the collection pot by means of fixing rods.
According to some embodiments of the present application, an extraction port is provided on the collection tank, the extraction port being configured to extract microorganisms and/or microorganism genetic information on the microorganism filter screen.
According to some embodiments of the application, a gas control switch is arranged on a connecting pipeline between the gas outlet of the collection tank and the gas pump.
One embodiment of the present application provides a microbial sampling system comprising: a microorganism collector as described above; the microorganism collector is arranged on the shell; the control module is arranged on the shell, and the air pump is connected with the control module; and the power supply module is arranged on the shell and is connected with the control module.
According to some embodiments of the present application, microorganism sampling system still include the return air inlet filter screen, the return air inlet filter screen sets up in clean area's return air inlet department, the return air inlet filter screen includes: the electrostatic screen comprises a first screen layer, a second screen layer and an electrostatic screen layer, wherein the electrostatic screen layer is arranged between the first screen layer and the second screen layer.
One embodiment of the present application provides a method of sampling microorganisms, comprising: placing a sampling head at an air return opening of a clean area, and collecting microorganisms and/or microorganism genetic information attached to a filter screen of the air return opening; the sampled gas enters an acquisition tank and is discharged out of the acquisition tank after passing through a microorganism filter screen, and the microorganism filter screen adsorbs microorganisms and/or microorganism genetic information in the sampled gas; and extracting the microorganisms and/or the genetic information of the microorganisms in the collection tank for analysis, and determining the species and the flora structure of the microorganisms.
According to some embodiments of the application, the extracting the microorganisms in the collection tank for analysis comprises: and injecting a liquid culture medium into the collection tank, and extracting a liquid culture in the collection tank for analysis after culture.
According to some embodiments of the application, the extracting the microorganisms in the collection tank for analysis comprises: and extracting the microorganism and/or microorganism genetic information on the microorganism filter screen through an extraction port.
According to some embodiments of the present application, a plurality of sampling points are disposed within the clean zone.
According to some embodiments of the present application, a plurality of sampling locations are provided at a return air opening.
According to the microorganism collector, the microorganism sampling system and the microorganism sampling method, microorganisms attached to the filter screen are collected at the air return inlet, the microorganism pollution condition of a clean area can be accurately provided through the structural analysis of the flora at the air return inlet, data support is provided for risk assessment, pollution product traceability and the like, powerful technical support is provided for supervision, and a guiding effect is achieved on an enterprise self-construction strain bank.
Drawings
In order to more clearly illustrate the technical solutions of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for a person skilled in the art to obtain other drawings based on these drawings without exceeding the protection scope of the present application.
FIG. 1 is a schematic view of a microorganism collector according to an embodiment of the present application;
FIG. 2 is a schematic diagram of a sampling head according to an embodiment of the present application;
FIG. 3 is a schematic view of another microorganism collector according to an embodiment of the present disclosure;
FIG. 4 is a schematic view of a microbial sampling system according to an embodiment of the present application;
FIG. 5 is a schematic view of a filter screen of an air return inlet according to an embodiment of the present application;
FIG. 6 is a schematic diagram of a sampling point according to an embodiment of the present application;
FIG. 7 is a schematic view of a sampling location of an air return vent according to an embodiment of the present disclosure;
FIG. 8 is a schematic view of another sampling position of the return air inlet in the embodiment of the present application.
Detailed Description
The technical solutions of the present application will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present application, and it is obvious that the described embodiments are some, not all, of the embodiments of the present application. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
Example 1
As shown in fig. 1, an embodiment of the present application provides a microorganism collector 100. The microorganism collector 100 includes: collection jar 1, sampling head 2 and air pump 3. Sampling head 2 is used for gathering gas, and the gas of gathering gets into collection tank 1, and the staff draws the microorganism by gathering in 1 and analyzes, and air pump 3 provides power for gaseous flow.
The collection tank 1 is a closed cavity, and the collection tank 1 comprises an air inlet 11 and an air outlet 12. A microorganism filter screen 13 is arranged in the cavity, and the microorganism filter screen 13 is used for adsorbing microorganisms and/or microorganism genetic information in gas. The genetic information of the microorganism includes DNA or RNA of the microorganism, etc. The material of the microorganism filter screen 13 of this embodiment is selected from the existing materials (i.e., polypropylene and glass fiber are mixed, and the aperture 20-50 um. is that the air inlet 11 and the air outlet 12 are respectively located at the two sides of the microorganism filter screen 13, so that the air flowing in the collection tank 1 must pass through the microorganism filter screen 13, and the microorganism filter screen 13 is convenient to adsorb microorganisms in the air. in this embodiment, the air inlet 11 is located at the bottom of the collection tank 1, and the air outlet 12 is located at the top of the collection tank 1.
The sampling head 2 is connected with the air inlet 11 of the collection tank 1. The sampling head 2 is used for collecting microorganisms and/or microorganism genetic information on a return air filter screen of a clean area and sending the microorganisms and/or microorganism genetic information into the collection tank 1 through gas. The sampling head 2 of the present embodiment is connected to the air inlet 11 of the collection tank 1 through a hose.
The return air inlet is used as the only outlet of all circulating air in the clean area, and almost all microorganisms which may float in the air of the clean area are enriched. Even if the microorganism dies, complete genetic information is left. Therefore, the information such as microorganism species, dominant species and the like can be analyzed by collecting the microorganism and/or microorganism genetic information enriched in the return air inlet and then carrying out gene amplification by using common primers such as 16S, ITS and the like, or by adopting methods such as metagenome sequencing, trace gene analysis and the like, so that the reasonable judgment on the microorganism pollution condition in a clean area can be conveniently made.
For example: the staphylococcus is rich, so the risk of human pollution is high; if micrococcus are abundant, the operation efficiency of the purification system may be reduced; the spore is abundant, so that the environmental humidity control is problematic; the water system may be contaminated if the negative bacilli are abundant; the spores are abundant, the disinfection and cleaning scheme needs to be adjusted, and the pollution risk of non-terminal sterilization products is increased.
The air pump 3 is connected with the air outlet 12 of the collecting tank 1. Air pump 3 provides power for gaseous flow, and air pump 3 of this embodiment passes through hose connection collection jar 1, realizes air pump 3 and collection jar 1's flexonics.
The utility model provides a microorganism collector 100 passes through sampling head 2 and gathers gas in return air inlet department, and gaseous lets in collection jar 1 back, through microorganism or thing biological genetic information in the microorganism filter screen 13 adsorbed gas, through return air inlet fungus crowd structural analysis, can be more accurate provide the microbial contamination condition in clean district, for risk assessment and pollution product traceability etc. provide data support, provide powerful technical support for the supervision, can play the guide effect to the enterprise from building the bacterial seed storehouse.
As shown in fig. 2, in an alternative scheme, the shape of the sampling head 2 is matched with the air return opening, and different sampling heads can be selected according to the height, depth, gap size and the like of the air return opening. For example, the first sampling head 2a has a long holding part, the second sampling head 2b has a trumpet-shaped end part, and the third sampling head 2c has an L-shaped delivery pipe, and those skilled in the art can select other sampling heads 2 according to the requirements.
According to an alternative embodiment of the present application, the microorganism collector 100 further comprises a liquid culture medium pot 4. The liquid culture medium tank 4 comprises a liquid outlet 41, the collection tank 1 comprises a liquid inlet 14, and the liquid outlet 41 of the liquid culture medium tank is connected with the liquid inlet 14 of the collection tank through a control valve 42. The liquid medium tank 4 stores a liquid medium for liquid culture, and when the microorganisms in the collection tank 1 need to be extracted after passing through culture, the control valve 42 is opened to transfer the liquid medium in the liquid medium tank 4 to the collection tank 1 to culture the microorganisms. When the microorganism harvesting apparatus 100 is used, the liquid medium tank 4 and the harvesting tank 1 may be horizontally placed, the liquid medium tank 4 may be positioned above the harvesting tank 1, and the liquid medium in the liquid medium tank 4 may flow into the harvesting tank 1 by gravity after the control valve 42 is opened.
After the culture is completed according to the culture conditions, the liquid culture in the collection tank 1 can be extracted by using an inoculating needle, streaked or coated, a single colony in the culture is separated, and identification and analysis of the strain are carried out by adopting a biochemical or molecular biological method. The classification sequencing method can also be used for classifying and sequencing the microorganisms in the culture and determining the species of the collected microorganisms.
In an alternative solution, the outer wall of the liquid culture medium pot 4 is connected to the outer wall of the collection pot 1 by means of fixing rods 5. The fixed connection of the liquid culture medium pot 4 and the collection pot 1 is achieved by means of a fixing rod 5. In this embodiment, the number of the fixing rods 5 is two, and one of the fixing rods 5 is a hollow tube for connecting the liquid outlet 41 of the liquid medium tank and the liquid inlet 14 of the collection tank. A control valve 42 is arranged on the hollow pipe, and after the control valve 42 is opened, the liquid culture medium in the liquid culture medium tank 4 enters the collection tank 1 through the hollow pipe.
As shown in fig. 3, according to an alternative embodiment of the present invention, the collection tank 1 is provided with an extraction port 15. When the microorganism is extracted by a non-culture method, the microorganism in the collection tank can be extracted without culturing. After the microorganism filter screen 13 finishes the adsorption of the microorganism and/or the microorganism genetic information, the extraction port 15 is opened, and the microorganism and/or the microorganism genetic information on the microorganism filter screen 13 is extracted through the extraction port 15.
After extracting the microorganism or microorganism genetic information from the microorganism filter screen 13, the macro-histology data is obtained by constructing a sequencing library and performing high-throughput sequencing.
After obtaining the microbiome data, firstly, the quality control is carried out, including removing primers and joints which are artificially added in the sequencing and library building processes, low-quality sequences generated in the sequencing process and the like. Host sequences are removed by alignment with the host genome. The obtained pure sequences are then aligned to a reference database or a de novo assembled reference gene set, quantified as a signature table, which can be classified into species or functional genome composition tables according to the type of sequence annotation. And analyzing the distribution and abundance of the microbial species contained in the environmental sample according to the obtained metagenome data identification result.
According to an optional technical scheme of the application, an air control switch 31 is arranged on a connecting pipeline between the air outlet 12 of the collection tank and the air pump 3. After the collection tank 1 finishes the collection of microorganisms, the gas control switch 31 is used for closing the connecting pipeline between the gas outlet 12 of the collection tank and the gas pump 3, so that the interference of external gas on the microorganisms in the collection tank 1 is avoided. If the connecting pipeline between the air outlet 12 of the collecting tank and the air pump 3 is a hose, the air control switch 31 can select a hose clamp. Optionally, an air filter screen is arranged on a connecting pipeline between the air outlet 12 of the collection tank and the air pump 3 to prevent impurities from entering the air pump 3 and affecting the operation of the air pump 3.
Example 2
As shown in fig. 4, the present embodiment provides a microorganism sampling system. A microbiological sampling system comprising: the microorganism collector 100, the housing 200, the control module 300 and the power module 400 as described above. The housing 200 protects the microorganism collector 100, the control module 300 and the power module 400.
The housing 200 is provided with a card slot 201. The microorganism collector 100 is disposed on the housing 200. Specifically, the collection tank 1 is detachably disposed in the clamping groove 201 through a buckle, and the air pump 3 is disposed in the housing 200. When the microorganism and/or the information of the microorganism need to be extracted from the collection tank 1, the collection tank 1 is detached from the housing 200, so that the extraction is convenient.
The control module 300 is disposed on the housing 200, and the air pump 3 is connected to the control module 300. The control module 300 controls the operation of the air pump 3. Optionally, the control module 300 is a touch screen controller, which facilitates the operation of the operator. The control module 300 may also control the control valve 42 and the gas control switch 31. The power module 400 is disposed on the housing 200, and the power module 400 is connected to the control module 300. Alternatively, the power module 400 is a rechargeable battery.
When the microorganism sampling system is used, the collection tank 1 is arranged in the clamping groove 201, the sampling head 2 is placed at the air return opening, the air pump 3 is started through the control module 300, the gas at the air return opening enters the collection tank 1, and microorganisms and microorganism genetic information in the gas are adsorbed by the microorganism filter screen; after the specified sampling time or area is completed, the air pump 3 stops working. The collection tank 1 is removed from the housing 200, and the microorganisms are cultured and then extracted, or the microorganisms and/or the genetic information of the microorganisms are extracted in a non-culture manner, as required.
As shown in fig. 5, according to an alternative embodiment of the present application, the microorganism sampling system further includes a return air inlet filter screen 500, and the return air inlet filter screen 500 is disposed at a return air inlet of the clean area. The return air inlet filter screen 500 includes: a first screen layer 501, a second screen layer 502, and an electrostatic screen layer 503. The electrostatic mesh layer 503 is disposed between the first mesh layer 501 and the second mesh layer 502. The electrostatic net layer 503 is made of aluminum alloy and is connected with the whole air return system through a conducting wire. After the air return system operates, the electrostatic net layer generates static electricity to effectively adsorb microorganisms. The first filter screen layer 501 and the second filter screen layer 502 are made of: diatomite (6-10%), flax fiber (20-30%), polypropylene fiber (40-50%), glycol (5-10%) and polylactic acid (10-20%).
Optionally, the return air inlet filter screen 500 is fixed in the support frame, and the first filter screen layer 501, the second filter screen layer 502 and the electrostatic screen layer 503 can be pulled out from the support frame independently, so that the surface sampling of the microorganism collector 100 is facilitated. The return air inlet filter screen 500 is arranged, so that the adsorption effect on microorganisms is greatly increased under the condition of not influencing return air.
The embodiment provides a microorganism sampling method, which comprises the following steps:
s1, placing the sampling head at the air return opening of the clean area, starting the air pump, and sampling the microorganisms and/or microorganism genetic information attached to the air return opening filter screen;
s2, enabling the sampled gas to enter an acquisition tank, discharging the gas out of the acquisition tank after passing through a microorganism filter screen, and adsorbing microorganisms and/or microorganism genetic information in the sampled gas by the microorganism filter screen;
s3, extracting the microorganisms in the collection tank for analysis, and determining the types of the microorganisms.
According to an optional technical scheme of the present application, after the microorganism filtering net finishes the adsorption of the microorganism and/or the genetic information of the microorganism, step S3 includes injecting a liquid culture medium into the collection tank, finishing the culture according to the preset conditions, and extracting the liquid culture in the collection tank for analysis.
According to an alternative embodiment of the present application, after the microorganism filtering net completes the adsorption of the microorganism and/or the genetic information of the microorganism, step S3 includes extracting the microorganism on the microorganism filtering net through the extraction port.
The analysis of the microorganisms and/or the genetic information of the microorganisms comprises the steps of adopting a biochemical or molecular biology method and carrying out gene sequencing to obtain information such as microorganism types, population distribution, variation trend, abundance and the like.
As shown in fig. 6, according to an alternative solution of the present application, a plurality of sampling points are disposed in the clean zone, so that the sampling result is more accurate. Setting a sampling point every 50 square meters, setting only one sampling point M1 when the square meters is less than 50 square meters, and increasing the sampling points M2, M3, M4, M2 and M6 according to the area of a clean zone. In this embodiment, the quantity of sampling point is six at most, avoids staff intensity of labour too big when satisfying the sampling precision.
As shown in fig. 7 and 8, a plurality of sampling positions are provided at a return air opening. Fig. 7 is a return-air inlet, and fig. 8 is a louver-type return-air inlet. In this embodiment, at least five sampling positions, respectively the sampling position A, B, C, D, E, are set at the air return opening. Five sampling positions can be uniformly distributed at the air return inlet, so that the sampling accuracy is improved.
The microorganism collector, the microorganism sampling system and the sampling method collect environmental microorganisms enriched in a clean area such as a purification workshop and a return air inlet. After sampling is finished, the information such as species, population distribution, change trend and the like of microorganisms is detected by adopting a culture or genetic information analysis method, and the method is used for clean environment monitoring, disinfection control and supervision and inspection of a drug supervision department.
The embodiments of the present application are described in detail above. The principle and the implementation of the present application are explained herein by applying specific examples, and the above description of the embodiments is only used to help understand the technical solutions and the core ideas of the present application. Therefore, the person skilled in the art should, according to the idea of the present application, change or modify the embodiments and applications of the present application based on the scope of protection of the present application. In view of the above, the description should not be taken as limiting the application.
Claims (12)
1. A microorganism collector, comprising:
the collection tank is internally provided with a cavity, the collection tank comprises an air inlet and an air outlet, a microorganism filter screen is arranged in the cavity, and the air inlet and the air outlet are respectively positioned at two sides of the microorganism filter screen;
the sampling head is connected with the air inlet of the collection tank, and is used for collecting microorganisms and/or microorganism genetic information on a return air inlet filter screen of a clean area and sending the microorganisms and/or microorganism genetic information into the collection tank through air;
and the air pump is connected with the air outlet of the collection tank.
2. The microorganism collector of claim 1 further comprising a liquid medium tank, the liquid medium tank comprising a liquid outlet; the collection tank comprises a liquid inlet, and the liquid outlet of the liquid culture medium tank is connected with the liquid inlet of the collection tank through a control valve.
3. The microorganism collector of claim 2 wherein the outer wall of the liquid medium pot is connected to the outer wall of the collection pot by a fixing rod.
4. The microorganism collector of claim 1, wherein the collection tank is provided with an extraction port for extracting microorganisms and/or microorganism genetic information on the microorganism filter screen.
5. The microorganism collector according to claim 1, wherein a gas control switch is arranged on a connecting pipeline between the gas outlet of the collection tank and the gas pump.
6. A microbial sampling system, comprising:
a microorganism collector as claimed in any one of claims 1 to 5;
the microorganism collector is arranged on the shell;
the control module is arranged on the shell, and the air pump is connected with the control module;
and the power supply module is arranged on the shell and is connected with the control module.
7. The microorganism sampling system according to claim 6, further comprising a return air inlet filter screen, the return air inlet filter screen being disposed at a return air inlet of the clean zone, the return air inlet filter screen comprising: the electrostatic screen comprises a first screen layer, a second screen layer and an electrostatic screen layer, wherein the electrostatic screen layer is arranged between the first screen layer and the second screen layer.
8. A method of sampling microorganisms, comprising:
placing a sampling head at an air return opening of a clean area, and collecting microorganisms and/or microorganism genetic information attached to a filter screen of the air return opening;
the sampled gas enters an acquisition tank and is discharged out of the acquisition tank after passing through a microorganism filter screen, and the microorganism filter screen adsorbs microorganisms and/or microorganism genetic information in the sampled gas;
and extracting the microorganisms and/or the genetic information of the microorganisms in the collection tank for detection and analysis, and determining the types of the microorganisms.
9. The method of claim 8, wherein said extracting microorganisms in said collection tank for analysis comprises: and injecting a liquid culture medium into the collection tank, and extracting a liquid culture in the collection tank for analysis after culture.
10. The method of claim 8, wherein said extracting microorganisms in said collection tank for analysis comprises: and extracting the microorganism and/or microorganism genetic information on the microorganism filter screen through an extraction port.
11. The microorganism sampling method according to claim 8, wherein a plurality of return air inlet sampling points are provided in the clean zone.
12. The method for sampling microorganisms of claim 8, wherein a plurality of sampling positions are provided at an air return opening.
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Application publication date: 20210427 |