CN114459981A - Method for detecting bacteria filtering efficiency of mask - Google Patents
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- CN114459981A CN114459981A CN202111579682.2A CN202111579682A CN114459981A CN 114459981 A CN114459981 A CN 114459981A CN 202111579682 A CN202111579682 A CN 202111579682A CN 114459981 A CN114459981 A CN 114459981A
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- 238000001914 filtration Methods 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 21
- 241000894006 Bacteria Species 0.000 title claims abstract description 15
- 230000001580 bacterial effect Effects 0.000 claims abstract description 58
- 239000001888 Peptone Substances 0.000 claims abstract description 27
- 108010080698 Peptones Proteins 0.000 claims abstract description 27
- 239000000443 aerosol Substances 0.000 claims abstract description 27
- 235000019319 peptone Nutrition 0.000 claims abstract description 27
- 238000012360 testing method Methods 0.000 claims abstract description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000000725 suspension Substances 0.000 claims abstract description 12
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 10
- 238000005507 spraying Methods 0.000 claims abstract description 10
- 230000035515 penetration Effects 0.000 claims abstract description 8
- 239000013641 positive control Substances 0.000 claims abstract description 8
- 230000000149 penetrating effect Effects 0.000 claims abstract description 3
- 239000002245 particle Substances 0.000 claims description 7
- 108010050327 trypticase-soy broth Proteins 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims 1
- 238000007865 diluting Methods 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 4
- 238000002360 preparation method Methods 0.000 abstract description 2
- 238000003908 quality control method Methods 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000007689 inspection Methods 0.000 description 3
- 230000001143 conditioned effect Effects 0.000 description 2
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000006150 trypticase soy agar Substances 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000003616 anti-epidemic effect Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000006199 nebulizer Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- 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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/06—Quantitative determination
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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
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/14—Streptococcus; Staphylococcus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1006—Investigating individual particles for cytology
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/305—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F)
- G01N2333/31—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F) from Staphylococcus (G)
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
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Abstract
The invention relates to the technical field of mask detection, and discloses a method for detecting the bacterial filtration efficiency of a mask. The invention comprises the following steps: (1) sample pretreatment; (2) preparation of bacterial suspensions for testing: (3) atomizing the staphylococcus aureus bacterial suspension into aerosol to be sprayed out, vertically spraying the aerosol through a pipeline onto a mask fixed on a sampler at the flow rate of 1mL/min for 1min, stopping spraying, spraying sterile air for 1min, enabling the aerosol to enter an impact type liquid collector through the mask, collecting the aerosol penetrating through a sample by using 20mL sterile peptone water for each collector, and controlling the gas flow rate passing through the sampler to be at the condition of 28.3L/min; (4) counting the peptone water in the collector by using an electronic bacteria counter in a sterile operation manner to respectively obtain the bacterial number in the peptone water with the mask sample, namely a bacterial penetration value, and the bacterial number in the peptone water without the mask, namely a positive control value; (5) and calculating the result according to a formula.
Description
Technical Field
The invention belongs to the technical field of mask detection, and particularly relates to a method for detecting mask bacterial filtration efficiency.
Background
Medical masks and surgical masks play an important role in clinical and epidemic prevention, are used for covering the mouth, nose and lower collar of a user, and provide a physical barrier for preventing pathogen microorganisms, body fluids, particulate matters and the like from directly permeating. The bacterial filtration efficiency is an important index of medical masks and surgical masks, and is concerned in the inspection and detection of the medical masks and surgical masks. Bacterial filtration efficiency; BFE refers to the percentage of bacteria-containing suspended particles that were filtered out by the mask material at the specified flow rate. The epidemic situation outbreak, gauze mask has become the effective sharp ware of anti-epidemic situation at the epidemic prevention initial stage, and bacterial filtration efficiency is as the important index of gauze mask protection level, and is receiving great attention. The existing biological testing method for the filtration efficiency of bacteria comprises the following steps: will be configured as 5 x 105The staphylococcus aureus with the concentration of CFU/mL passes through a test system, is atomized and sprayed onto a mouth mask fixed on a sampler under the condition that the gas flow rate is 28.3L/min, and the aerosol is collected by using a six-grade Anderson sampler provided with a trypsin soybean agar plate. The collected positive, negative and sample agar plates were incubated (48. + -.4) at (37. + -.2) ℃ for h, and then colony forming units (positive wells) formed by bacterial particle aerosol were counted and converted to the number of possible impinging EUT particles using a positive well conversion table. The converted values are used to determine the average level of aerosol of bacterial particles delivered to the test sample. And finally, calculating the bacterial filtration efficiency of the mask by using the converted data through a formula.
The existing mask bacterial filtration efficiency testing method has the following defects:
1. the time required for inspection is long. At least 2 days are required from the start of the test to the outcome, which greatly affects the speed of the test. Even if the culture time is changed from 48 hours to 24 hours by the EU standard, the time is still long compared with other inspection items of the mask.
2. The anderson sampler is prone to errors. Because the Anderson sampler has wall loss, and the granule slips and is smashed the circumstances such as from the collection face, above all can all produce the error to the experimental result, influence the reliability of experimental result.
3. The sampling steps are complicated, and more agar plates are needed. The installation and the dismantlement of collector all need be gone on many times, have increased operating personnel's intensity of labour and test cost.
The calculation is cumbersome. In order to calculate the bacterial filtration efficiency, raw data must be subjected to data conversion by using a positive well conversion table, and each culture medium plate is counted, so that the data amount is large.
Disclosure of Invention
The invention aims to solve the problems and provides a novel method for detecting the bacterial filtering efficiency of a mask, which has the following characteristics: the time cost is greatly saved, and the reliability of experimental data is improved.
In order to achieve the purpose, the invention provides the following technical scheme: a method for detecting the bacterial filtration efficiency of a mask, comprising the steps of:
(1) pre-treating the sample in an environment with the temperature of 21 +/-5 ℃ and the relative humidity of 85 +/-5% for at least 4 h;
(2) staphylococcus aureus CMCC (B)26003 was inoculated into an appropriate amount of trypticase soy peptone broth, cultured with shaking at 37. + -. 2 ℃ for 24. + -.2 h, and then diluted with 1.5% peptone to a bacterial suspension at a concentration of 5X 105 CFU/mL;
(3) atomizing the staphylococcus aureus bacterial suspension obtained in the step (2), wherein the average particle diameter (MPS) of the sprayed bacterial aerosol is 3.0 +/-0.3 mu m; the geometric standard deviation of the distribution of the bacterial aerosol is not more than 1.5, the aerosol is vertically sprayed to a mask fixed on a sampler at the flow rate of 1mL/min, the spraying is stopped for 1min, the spraying is stopped, the sterile air is sprayed for 1min, the aerosol enters an impact liquid collector through the mask, each collector collects the aerosol penetrating through a sample by using 20mL sterile peptone water, and the gas flow rate passing through the sampler is controlled under the condition of 28.3L/min, and three parallel tests are carried out totally;
(4) counting the peptone water in the collector by using an electronic bacteria counter in a sterile operation manner to respectively obtain the bacterial number in the peptone water with the mask sample, namely a bacterial penetration value, and the bacterial number in the peptone water without the mask, namely a positive control value;
(5) calculating the result according to the formula:
in the formula:
t: value of bacterial penetration
C: positive control value mean.
Compared with the prior art, the invention has the following beneficial effects:
1. the method can save the time of bacterial culture, shorten the experiment time from more than 48 hours to several hours, and greatly save the time cost.
2. The method can reduce the system error caused by depending on the Anderson sampler and increase the reliability of experimental data.
3. The method can reduce the workload of testers, simplify the work flow and operation, improve the reliability of the test result and enhance the authenticity of data.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. 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 invention.
1. The reagents used were: trypticase soy agar medium (TSA); trypticase Soy peptone broth (TSB); peptone water; staphylococcus aureus CMCC (B) 26003.
2. Sample pretreatment: the samples were pre-conditioned for at least 4 hours prior to testing in an environment having a temperature of (21. + -. 5 ℃) and a relative humidity of (85. + -. 5%).
3. Preparation of bacterial suspensions for testing: staphylococcus aureus CMCC (B)26003 was inoculated into an appropriate amount of trypticase Soytone broth, cultured with shaking at 37. + -. 2 ℃ for 24. + -.2 h, and then diluted to 5X 10 with 1.5% peptone5CFU/mL concentration of bacterial suspension.
4. Atomizing staphylococcus aureus bacterial suspension, wherein the average particle diameter (MPS) of the sprayed bacterial aerosol is 3.0 +/-0.3 mu m; the geometric standard deviation of the distribution of the bacterial aerosol is not more than 1.5, the bacterial aerosol is vertically sprayed to a mouth mask fixed on a sampler at the flow rate of 1mL/min for 1min, then the spraying is stopped, and the sterile air is sprayed for 1 min. The aerosol passed through the mask into impact liquid collectors, each collector collecting the sample-penetrated aerosol with 20mL sterile peptone water, the gas flow rate through the sampler was controlled at 28.3L/min. A total of three replicates were performed.
5. After the test is finished, the peptone water in the collector is counted by an electronic bacteria counter in an aseptic operation, and the bacterial number in the peptone water with the mask sample, namely the bacterial penetration value, and the bacterial number in the peptone water without the mask, namely the positive control value, are respectively obtained.
6. Calculating the result according to the formula:
in the formula:
t: value of bacterial penetration
C: mean value of positive control values
Example (b):
3 masks were taken and the samples were pre-conditioned for 4 hours at 21 ℃ and 85% relative humidity prior to testing.
Staphylococcus aureus CMCC (B)26003 was inoculated into 50mL trypticase Soytone broth and shake-cultured at 37 ℃After 24h, the culture was diluted to 5X 10 with 1.5% peptone5CFU/mL concentration of bacterial suspension.
The test system is not placed in an inlet cover, is connected with an impact type liquid collector, controls the flow rate of gas passing through the sampler to be 28.3L/min, sets the time for conveying the bacterial suspension to the sprayer to be 1min, sets the air pressure and the running time of the sampler to be 2min, runs for three times, and balances the test system.
And starting a test system without putting a sample, collecting the bacterial aerosol into a sterilized impact type liquid collector containing 20mL of sterile peptone water as positive quality control, counting the individual quantity of bacteria by using a bacterial counter for the peptone water in the collector, determining the aerosol flow, wherein the aerosol flow is (2200 +/-500) CFU, and adjusting the concentration of the bacterial liquid and the gas flow rate according to the situation.
After the positive quality control test is finished, a new sterilized impact type liquid collector containing 20mL of sterile peptone water is put in, the test sample is clamped at the upper end of the sampler, and the tested surface faces upwards. Sampling was performed according to the procedure described above. After a batch of test samples is tested, the positive quality control is tested again. A total of three trials were performed.
After completion of the test of one batch of test samples, a 2min sample of air in the aerosol chamber was then collected as a negative quality control, during which time the bacterial suspension could not be delivered to the nebulizer.
After the test is finished, the peptone water in the collector is counted by a bacteria counter through aseptic operation, and the bacterial number in the peptone water with the mask sample, namely the bacterial penetration value, and the bacterial number in the peptone water without the mask, namely the positive control value, are respectively obtained.
And calculating the bacterial filtration efficiency of each mask by using a formula.
The bacteria filtration efficiency of the mask batch detected by the method is 99.8%, the bacteria filtration efficiency of the mask batch detected by the original method is 99.9%, and the detection result has no obvious difference.
The bacteria counter may be a cell flow counter or an electronic bacteria counter, or other instrumentation that may be used to count individual bacteria in a liquid.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (5)
1. A method for detecting the bacterial filtration efficiency of a mask, comprising the steps of:
(1) pre-treating the sample in an environment with the temperature of 16-26 ℃ and the relative humidity of 80-90% for at least 4 h;
(2) inoculating Staphylococcus aureus CMCC (B)26003 into trypticase Soytone liquid medium, culturing at 35-39 deg.C for 22-26 hr under shaking, and diluting the culture with 1.5% peptone to 5 × 105Bacterial suspension with the concentration of CFU/mL;
(3) atomizing the staphylococcus aureus bacterial suspension obtained in the step (2) into aerosol to be sprayed out, vertically spraying the aerosol to a mask fixed on a sampler at the flow rate of 1mL/min, spraying for 1min, stopping spraying, spraying sterile air for 1min, enabling the aerosol to enter an impact type liquid collector through the mask, collecting the aerosol penetrating through a sample by using 20mL sterile peptone water for each collector, and controlling the flow rate of gas passing through the sampler to be at the condition of 28.3L/min;
(4) counting the peptone water in the collector by using an electronic bacteria counter in a sterile operation manner to respectively obtain the bacterial number in the peptone water with the mask sample, namely a bacterial penetration value, and the bacterial number in the peptone water without the mask, namely a positive control value;
(5) calculating the result according to the formula:
in the formula:
t: value of bacterial penetration
C: positive control value mean.
2. The method for detecting the bacterial filtering efficiency of the mask according to claim 1, wherein in the step (2), the mean particle diameter (MPS) of the bacterial aerosol is 3.0 ± 0.3 μm, and the geometric standard deviation of the distribution of the bacterial aerosol is not more than 1.5.
3. The method for detecting the bacterial filtration efficiency of the mask according to claim 1, wherein said step (2) requires three parallel tests.
4. The method for detecting the bacterial filtration efficiency of the mask according to claim 1, wherein in the step (1), the temperature is 21 ℃ and the humidity is 85%.
5. The method for detecting the bacterial filtration efficiency of the mask according to claim 1, wherein in the step (2), the culture is performed at 37 ℃ for 24 hours under shaking.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111579682.2A CN114459981A (en) | 2021-12-22 | 2021-12-22 | Method for detecting bacteria filtering efficiency of mask |
| PCT/CN2022/138248 WO2023116477A1 (en) | 2021-12-22 | 2022-12-10 | Method for detecting bacterial filtration efficiency of mask |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN202111579682.2A CN114459981A (en) | 2021-12-22 | 2021-12-22 | Method for detecting bacteria filtering efficiency of mask |
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| CN114459981A true CN114459981A (en) | 2022-05-10 |
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| CN202111579682.2A Pending CN114459981A (en) | 2021-12-22 | 2021-12-22 | Method for detecting bacteria filtering efficiency of mask |
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| WO (1) | WO2023116477A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023116477A1 (en) * | 2021-12-22 | 2023-06-29 | 河北省药品医疗器械检验研究院(河北省化妆品检验研究中心) | Method for detecting bacterial filtration efficiency of mask |
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- 2021-12-22 CN CN202111579682.2A patent/CN114459981A/en active Pending
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2022
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