CN114112461B - Virus removal testing device and method for water purifier - Google Patents
Virus removal testing device and method for water purifier Download PDFInfo
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- CN114112461B CN114112461B CN202111323539.7A CN202111323539A CN114112461B CN 114112461 B CN114112461 B CN 114112461B CN 202111323539 A CN202111323539 A CN 202111323539A CN 114112461 B CN114112461 B CN 114112461B
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 223
- 241000700605 Viruses Species 0.000 title claims abstract description 159
- 238000012360 testing method Methods 0.000 title claims abstract description 78
- 238000000034 method Methods 0.000 title claims description 18
- 238000005070 sampling Methods 0.000 claims abstract description 79
- 230000007246 mechanism Effects 0.000 claims abstract description 16
- 239000003085 diluting agent Substances 0.000 claims description 39
- 239000006916 nutrient agar Substances 0.000 claims description 36
- 239000001963 growth medium Substances 0.000 claims description 33
- 239000011550 stock solution Substances 0.000 claims description 24
- 239000007787 solid Substances 0.000 claims description 21
- 239000000725 suspension Substances 0.000 claims description 21
- 238000002156 mixing Methods 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 11
- 230000001954 sterilising effect Effects 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 9
- 238000010790 dilution Methods 0.000 claims description 8
- 239000012895 dilution Substances 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 8
- 239000002609 medium Substances 0.000 claims description 7
- 238000004659 sterilization and disinfection Methods 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 238000007865 diluting Methods 0.000 claims description 4
- 235000015097 nutrients Nutrition 0.000 claims description 4
- 238000009631 Broth culture Methods 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims description 3
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- 230000008014 freezing Effects 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 238000004064 recycling Methods 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 2
- 238000007747 plating Methods 0.000 claims description 2
- 238000011084 recovery Methods 0.000 abstract description 3
- 241000588724 Escherichia coli Species 0.000 description 7
- 238000002372 labelling Methods 0.000 description 7
- 239000003651 drinking water Substances 0.000 description 5
- 235000020188 drinking water Nutrition 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 238000001223 reverse osmosis Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000011990 functional testing Methods 0.000 description 1
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- 239000012535 impurity Substances 0.000 description 1
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- 230000004048 modification Effects 0.000 description 1
- 239000002957 persistent organic pollutant Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000007655 standard test method Methods 0.000 description 1
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- 238000006467 substitution reaction Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
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- 238000000108 ultra-filtration Methods 0.000 description 1
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01M—TESTING STATIC OR DYNAMIC BALANCE OF MACHINES OR STRUCTURES; TESTING OF STRUCTURES OR APPARATUS, NOT OTHERWISE PROVIDED FOR
- G01M99/00—Subject matter not provided for in other groups of this subclass
- G01M99/008—Subject matter not provided for in other groups of this subclass by doing functionality tests
-
- 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
Abstract
The invention discloses a virus removal testing device of a water purifier, which comprises a pressure conveying mechanism, a first pressure gauge, a shunt tube, a first sampling tube, a plurality of fourth valves, a plurality of second pressure gauges, a plurality of water purifiers, a plurality of flow meters, a plurality of second sampling tubes, a plurality of second drain valves and a controller. The invention realizes stable recovery of virus in water and automatic control.
Description
Technical Field
The invention relates to the technical field of water purification, in particular to a virus removal testing device and method of a water purifier.
Background
The water purifier is also called a water purifier and a water quality processor, and is water treatment equipment for performing deep filtration and purification treatment on water according to the use requirement of water.
In recent years, as the living standard of people is continuously improved, the importance of the sanitary and safety problem of drinking water is increasingly increased. The water purifier can effectively remove bacteria, viruses, heavy metals, organic pollutants and other impurities in water in a manner of ultrafiltration or reverse osmosis and the like, and the water purifier can be directly drunk only by being connected with tap water in the use process, so that the sanitation and safety of drinking water are ensured while convenience is brought. Therefore, the types and the number of water purifiers on the market are increasing. At present, the evaluation of the functional test of the water purifier in the water purifier industry is mainly based on the standards of sanitary safety and function evaluation Specification of Water quality processors for Drinking Water, general Water quality processor (2001), sanitary safety and function evaluation Specification of Water quality processors for Drinking Water, reverse osmosis Water quality processor (2001) and Standard test method for Drinking Water issued by the Ministry of health. The standard focuses on the functional verification of the water purifier, and the removal effect of the water purifier on bacteria, heavy metals and organic matters is detected. The standard has no clear requirement on the testing device, the standard can not evaluate the virus removal of the water purifier, the current standard has no corresponding detection method for the virus in water, and the water purifier has no missing testing device for the virus in water.
Disclosure of Invention
The invention aims to overcome the defects in the prior art, and provides a virus removal testing device and method for a water purifier.
The aim of the invention is achieved by the following technical scheme: the utility model provides a virus removal testing arrangement of purifier, includes pressure conveying mechanism, first manometer, shunt tubes, first sampling tube, a plurality of fourth valve, a plurality of second manometer, a plurality of purifier, a plurality of flowmeter, a plurality of second sampling tube, a plurality of second drain valve and controller, pressure conveying mechanism is connected with the house steward of shunt tubes, the branch tube of shunt tubes is connected with a plurality of fourth valve respectively, first manometer and first sampling tube are installed in proper order in the house steward of shunt tubes, and a plurality of fourth valve is connected with a plurality of purifier through a plurality of first water pipes, and a plurality of second manometer are installed in a plurality of first water pipes, and a plurality of purifier are connected with a plurality of second drain valve through a plurality of second water pipes, a plurality of flowmeter and a plurality of second sampling tube are installed in a plurality of second water pipes in proper order, the controller is connected with pressure conveying mechanism, first manometer, second manometer and flowmeter respectively.
More preferably, the pressure conveying mechanism comprises a water supply pump, a one-way water stop valve, a first valve, an ultraviolet sterilization lamp, a standard adding water tank, a stirrer, a first drain valve, a second valve, a booster pump, a liquid level meter and a third valve, wherein the water supply pump, the one-way water stop valve, the first valve, the ultraviolet sterilization lamp, the standard adding water tank, the second valve, the booster pump and the third valve are sequentially connected, the third valve is connected with a main pipe of the shunt pipe, the stirrer, the liquid level meter and the first drain valve are all arranged in the standard adding water tank, and the stirrer, the liquid level meter, the water supply pump and the booster pump are all connected with the controller.
A virus removal test method of a water purifier comprises the following steps:
s1, preparing virus stock solution through viruses and hosts;
s2, determining the titer of the virus stock;
s3, adding the virus stock solution into the virus removal testing device of the water purifier according to any one of claims 1-2 for testing to obtain a control group water sample and a test group water sample;
s4, determining the titer of the water sample of the control group and the water sample of the test group to obtain a result of the control group and a result of the test group;
s5, calculating the results of the control group and the test group through formulas to obtain the virus removal rate and the virus removal value.
More preferably, the step S1 includes the following steps:
s11, selecting proper viruses and hosts;
s12, placing nutrient agar into a culture medium plate to obtain a nutrient agar culture medium plate, inoculating the host into the nutrient agar culture medium plate, culturing for 24+/-1 h at the temperature of 37+/-1 ℃ to obtain single colonies, inoculating the single colonies into a nutrient broth culture medium, and oscillating for 4-6h at the temperature of 35+/-1 ℃;
s13, pouring the nutrient agar into a first culture dish, and obtaining a solid nutrient agar culture dish after the nutrient agar is solidified;
s14, mixing the host suspension and the virus suspension, and standing for 10-20min at the temperature of 35+/-1 ℃ to obtain a mixed suspension of the host and the virus;
s15, uniformly mixing the mixed suspension and a semi-solid culture medium, pouring the mixed suspension and the semi-solid culture medium into the solid nutrient agar culture dish in the step S13, culturing for 17-19h at the temperature of 35+/-1 ℃, recycling the semi-solid culture medium into a sterile bag by adopting a sterile coating rod, centrifuging the sterile bag, and standing at the ambient temperature of 35+/-1 ℃ to obtain a virus suspension;
s16, transferring the virus suspension into a first centrifuge tube for centrifugal treatment to obtain supernatant, and transferring the supernatant into a second centrifuge tube for secondary centrifugal treatment;
s17, repeating the step S16 twice;
and S18, filtering the supernatant in the step S17 through a filter membrane to obtain a virus stock solution, and freezing or refrigerating the virus stock solution.
More preferably, the step S2 includes the following steps:
s21, diluting the virus stock solution to obtain a first virus diluent;
s22, mixing the first virus diluent and the host according to a certain concentration ratio, and standing for 10-20min at the ambient temperature of 35+/-1 ℃ to obtain a mixed solution of the virus and the host;
s23, uniformly mixing the mixed solution and the semi-solid culture medium containing agar, pouring the mixed solution and the semi-solid culture medium into a prepared nutrient agar culture medium plate, and standing for 16-20h at the ambient temperature of 35+/-1 ℃ to obtain a plurality of nutrient agar culture medium plates with virus spots;
s24, observing the number of virus spots in a plurality of nutrient agar medium plates, selecting the nutrient agar medium plates according to the number of the virus spots for counting, and calculating by a formula to obtain the titer value of the virus.
More preferably, the number of viral plaques in step S24 is 30-150.
More preferably, the calculation formula in the step S24 is as follows
More preferably, the step S3 includes the following steps:
s31, dividing the test into multiple stages according to the nominal rated water purifying amount of the water purifier;
s32, adding sterile pure water into a standard adding water tank of the virus removal testing device, and then adding virus stock solution for dilution to obtain second virus diluent;
s33, introducing the second virus diluent into the virus removal testing device, and sampling through a first sampling tube of the virus removal testing device at the beginning of water introduction to obtain a control group water sample, and sampling through a second sampling tube of the virus removal testing device to obtain a test group water sample;
s34, sampling at the first sampling tube to obtain another water sample of the control group, sampling at the second sampling tube to obtain another water sample of the test group, and cleaning the residual virus diluent in the virus removal testing device;
s35, repeating the steps S31 and S34 to obtain a plurality of water samples of the control group and a plurality of water samples of the test group.
More preferably, the formula of the virus removal rate in step S5 is as follows
More preferably, the calculation formula of the virus removal value in the step S5 is m=lg (V a )-lg(V b )。
Compared with the prior art, the invention has the following advantages:
1. according to the invention, through the pressure conveying mechanism, the first pressure gauge, the shunt tubes, the first sampling tubes, the fourth valves, the second pressure gauges, the water purifiers, the flow meters, the second sampling tubes, the second drain valves and the controller, stable recovery of viruses in water is realized, and automatic control can be realized.
2. The method for detecting the virus removal test of the water purifier can evaluate the virus removal standard of the water purifier, and has good repeatability of the virus recovery rate.
Drawings
FIG. 1 is a schematic diagram showing connection of a virus removal testing device of a water purifier according to the present invention;
the reference numerals for the various parts in the drawings: 1-a pressure delivery mechanism; 2-a water supply pump; 3-a one-way water stop valve; 4-a first valve; 5-ultraviolet sterilizing lamp; 6-adding a standard water tank; 7-a stirrer; 8. a second drain valve; 9-a second valve; 10-a variable-frequency booster pump; 11-a third valve; 12-a first pressure gauge; 13-a first sampling tube; 14-fourth valve; 15-a second pressure gauge; 16-a water purifier; 17-high precision flowmeter; 18-a second sampling tube; 19-a first drain valve; 20-shunt.
Detailed Description
The present invention will be described in further detail with reference to the drawings and specific examples, which are not to be construed as limiting the embodiments of the present invention.
As shown in fig. 1, a virus removal testing device of a water purifier 16 comprises a pressure conveying mechanism 1, 1 first pressure gauge 12, 1 first sampling tube 13, 1 shunt tube 20, 2 fourth valves 14, 2 second pressure gauges 15, 2 water purifiers 16, 2 flow meters 17, 2 second sampling tubes 18, 2 second drain valves 8 and a controller, wherein the pressure conveying mechanism 1 is connected with a main pipe of the shunt tube 20, the first pressure gauge 12 and the first sampling tube 13 are sequentially installed on the main pipe, a switch is arranged on the first sampling tube 13, 2 branch tubes of the shunt tube 20 are respectively connected with water inlet ends of the 2 first water pipes through 2 fourth valves 14, 2 second pressure gauges 15 are installed on the 2 first water pipes, water outlet ends of the 2 shunt tubes are respectively connected with water inlets of the 2 water purifiers 16, water outlets of the 2 water purifiers 16 are respectively connected with the second drain valves 8 through the second water pipes, the 2 flow meters 17 and the 2 second sampling tubes 18 are sequentially installed on the second water pipes, and the controller is respectively connected with the pressure conveying mechanism 1, the first pressure gauge 15 and the second pressure gauge 17.
The pressure delivery mechanism 1 is used to provide a second virus diluent (i.e., phage diluent) at a pressure that allows sterile pure water to be mixed with the virus stock. The shunt tube is used for enabling the second virus diluent to evenly flow in a shunt way; the first pressure gauge 12 is used for displaying the total pipe water pressure of the shunt pipe and feeding back to the controller; the second pressure gauge 15 is used for displaying the water pressure of the branch pipe of the shunt pipe and feeding back to the controller; the fourth valve 14 is used for controlling the second virus diluent to enter the water purifier 16; the water purifier 16 serves as a test sample and has a function of filtering the second virus diluent; the flowmeter 17 is a high-precision flowmeter 17, and is used for calculating the flow rate of the second virus diluent and feeding back the flow rate to the controller; the first sampling tube 13 and the second sampling tube 18 are used for sampling by a tester; the second drain valve 8 is used to control the second viral diluent drain. The controller is a PLC controller, can receive signals of the high-precision flowmeter 17, the first pressure gauge 12, the second pressure gauge 15 and the liquid level gauge, and can control the stirrer 7, the variable-frequency booster pump 10 and the water supply pump 2.
The pressure conveying mechanism 1 comprises a water supply pump 2, a one-way water stop valve 3, a first valve 4, an ultraviolet sterilization lamp 5, a standard adding water tank 6, a stirrer 7, a first drain valve 19, a second valve 9, a booster pump 10, a liquid level meter and a third valve 11, wherein the water supply pump 2, the one-way water stop valve 3, the first valve 4, the ultraviolet sterilization lamp 5, the standard adding water tank 6, the second valve 9, the booster pump 10 and the third valve 11 are sequentially connected, the third valve 11 is connected with a main pipe of a shunt pipe 20, the stirrer 7 and the liquid level meter are all installed in the standard adding water tank 6, the first drain valve 19 is installed at the bottom of the standard adding water tank 6, and the stirrer 7, the liquid level meter, the water supply pump 2 and the booster pump 10 are all connected with a controller.
The water supply pump 2 provides power for the flow of pure water and conveys the sterile pure water to the marking water tank 6; the one-way water stop valve 3 can prevent the sterile pure water from flowing back; the first valve 4 can control the passage of the shunt pipe; the ultraviolet sterilizing lamp 5 is used for sterilizing the sterile pure water and preventing bacteria from affecting the quality of the pure water; the marking water tank 6 is used for storing sterile pure water; the stirrer 7 allows sterile pure water and virus stock to be fully mixed; the first drain valve 19 is used for conveniently draining the remaining second virus diluent in the labeling water tank 6; the second valve 9 is used for controlling the passages of the standard adding water tank 6 and the booster pump 10; the booster pump 10 is a variable-frequency booster pump 10, provides power for the transmission of the second virus diluent, and can automatically control the water pressure according to the data of the first pressure gauge 12 and the second pressure gauge 15; the liquid level meter is used for measuring the height of the second virus diluent in the standard adding water tank 6 and feeding back to the PLC; the third valve 11 is used to control the passage of the booster pump 10 and the shunt pipe.
Description of device use: because the test time of the test is long, viruses (namely phage) can be continuously killed in the test process, the test is carried out according to the rated total water purifying amount (2 m) of the specification of the water purifier 16 3 For example) into 4 phases of 5 sampling nodes, each phase of 0.5m 3 The 5 sampling nodes are respectively the beginning (1 st time) sampling, the end (2 nd time) sampling, the end (3 rd time) sampling, the end (4 th time) sampling and the end (5 th time) sampling of the section (2/4 th time) sampling of the water passing through 0/4. Sampling the water sample collected in the first sampling tube 13 and the second sampling tube 18 of the water purifier 16 which is just filled with water at the beginning (1 st time) of 0/4 water filling; the 1/4 end (2 nd time) sampling is that the water purifying amount of the 1 st stage water purifier 16 reaches 0.5m 3 At the time of sampling water samples collected in the first sampling tube 13 and the second sampling tube 18; the end (3 rd time) of 2/4 segments is sampled to 1.0m for the water purifier 16 3 At the time of sampling water samples collected in the first sampling tube 13 and the second sampling tube 18; the 3/4-section last (4 th time) sampling is that the water purifier 16 reaches 1.5m 3 In the first sampling tube 13 and the second sampling tube 18Collecting a water sample; the end of 4/4 segments (5 th sampling) is that the water purifier 16 reaches 2m 3 At this time, the water sample collected in the first sampling tube 13 and the second sampling tube 18. The total water purification amount per stage of the high-precision flowmeter 17 was set to 0.5m 3 1m is pumped by a water supply pump 2 3 Injecting sterile pure water into the labeling water tank 6, adding virus stock solution into the labeling water tank 6, and adjusting to obtain second virus diluent with concentration of 10 5 PFU/mL～10 6 PFU/mL. After the preparation of the virus diluent is completed, the second valve 9 and the third valve 11 are opened, the variable-frequency booster pump 10 controls the in-tube pressure of the second virus diluent within the range of (0.24+/-0.02) MPa, the virus diluent starts to circulate in the pipeline, the fourth valve 14 is opened to enable the virus diluent to pass through the two water purifiers 16, the purified water outlet ends of the two water purifiers 16 are respectively connected with the two high-precision flow meters 17, then the virus diluent passes through the two second sampling pipes 18, and finally is discharged through the second drain valve 8. When the second virus diluent flows out of the water purifier 16, the first sampling tube 13 and the second sampling tube 18 are sampled for the 1 st time, the sampling at the beginning (the 1 st time) of 0/4 water passing is completed, and when the flow calculated by the high-precision flowmeter 17 reaches 0.5m 3 And then sampling the water from the first sampling tube 13 and the second sampling tube 18 at the end of 1/4 section (the 2 nd time), and controlling the water purifier 16 and the variable-frequency booster pump 10 to stop operating by the controller to prompt that the stage 1 test is finished. At this time, phage titration was performed on 4 water samples (including the 1 st and 2 nd sampling water samples), respectively, and the phage removal rate by the water purifier 16 was calculated by the formula. Then, the first drain valve 19 at the bottom of the labeling water tank 6 is opened, and the virus diluent in the labeling water tank 6 is discharged and cleaned, so that the test of the stage 1 is completed. Then adding the virus stock solution and sterile pure water into the labeling water tank 6 again, and adjusting to obtain second virus diluent with concentration of 10 5 PFU/mL～10 6 PFU/mL, according to the test mode of the 1 st stage, sampling and titration are carried out at the end of 2/4 sections (3 rd time), the end of 3/4 sections (4 th time) and the end of 4/4 sections (5 th time), and phage removal rate is calculated through the titrated water sample result.
A virus removal test method of a water purifier 16, comprising the steps of:
s1, preparing virus stock solution through viruses and hosts;
s11, selecting proper viruses and hosts, wherein the viruses are phage: phi-X174 (ATCC 13706-B1) and Escherichia coli (ATCC 13706) as the host;
s12, placing nutrient agar into a culture medium plate to obtain a nutrient agar culture medium plate, inoculating escherichia coli into the nutrient agar, culturing for 24+/-1 h at the temperature of 37+/-1 ℃, obtaining single bacterial colony, inoculating the single bacterial colony into a nutrient broth culture medium, and oscillating for 4-6h at the rotating speed of 100r/min at the temperature of 35+/-1 ℃;
s13, pouring the nutrient agar into a first culture dish, and obtaining a solid nutrient agar culture dish after the nutrient agar is solidified;
s14, 5mL is concentrated to 10 8 CFU/mL～10 9 CFU/mL of host (E.coli) suspension and 5mL of the solution at a concentration of 10 5 PFU/mL～10 6 Mixing PFU/mL virus (namely phage) suspension (according to concentration ratio of Escherichia coli: phage=1000:1), standing at 35+ -1deg.C for 10-20min to obtain host and virus mixed suspension;
s15, adding 10mL of mixed suspension into a semi-solid culture medium, uniformly mixing, pouring the mixed suspension onto a solid nutrient agar culture dish in the step S13, culturing for 17-19h at the temperature of 35+/-1 ℃, recycling the semi-solid culture medium into a sterile bag by adopting a sterile coating rod, centrifuging the sterile bag (260 r/min for 2 min), and standing at the ambient temperature of 35+/-1 ℃ to obtain a virus suspension;
s16, transferring the virus suspension into a 50mL first centrifuge tube for centrifugation (3500 r/min for 10 min) to obtain a supernatant, and transferring the supernatant into a 50mL second centrifuge tube for secondary centrifugation;
s17, repeating the method of the step S16 for 2 times;
s18, filtering the supernatant obtained in the step S17 through a filter membrane with the diameter of 0.22 mu m to obtain a virus stock solution (namely phage stock solution), and freezing or refrigerating the virus stock solution for storage.
S2, determining the titer of the virus stock solution;
s21, diluting the virus stock solution according to a ten-fold dilution method to release a first virus dilution solution; namely, 100. Mu.l of virus stock solution is added into 900. Mu.l of nutrient broth medium to be 10 -1 PFU/ml, diluted 10 in turn -2 PFU/ml、10 -3 PFU/ml、10 -4 PFU/ml、10 -5 PFU/ml、10 -6 PFU/ml。
S22, 1ml is concentrated to 10 5 ～10 6 PFU/ml phage (i.e., first virus dilution) and 1ml at a concentration of 10 8 ～10 9 Mixing CFU/ml of Escherichia coli (namely host) according to a certain concentration ratio (Escherichia coli: phage=1000:1), and standing at 35+ -1deg.C for 10-20min to obtain a mixed solution of virus and host;
s23, adding 10mL of semi-solid culture medium containing 0.5% of agar into the mixed solution, fully and uniformly mixing, respectively pouring the semi-solid culture medium into a plurality of prepared nutrient agar culture medium plates, and standing for 16-20h at the ambient temperature of 35+/-1 ℃ to obtain a plurality of nutrient agar culture medium plates for generating virus spots;
s24, observing the number of plaques in a plurality of nutrient agar medium plates, selecting a proper nutrient agar medium plate according to the number of plaques to count (30-150 plaques/dish), and calculating according to a formula to obtain the titer value (PFU/ml) of the virus (ATCC 13706-B1).
The calculation formula in step S24 is
Wherein: c = average number of plaques grown on plates at a certain dilution; v = volume of diluent used when plating the plate; m=dilution fold.
S3, adding the virus stock solution into a virus removal testing device of a water purifier for testing to obtain 5 water samples of a control group and 10 water samples of a test group;
s31, according to the nominal rated water purifying amount (total 2m 3 ) The whole marking test process is divided into four stages (each stage is 0.5m 3 )；
S32, adding sterile pure water into a standard adding water tank 6 of a virus removal testing device of a water purifier, and then adding a virus stock solution (namely phage stock solution) for dilution to obtain a second virus diluent (namely phage diluent), wherein the concentration of the second virus diluent is 10 5 PFU/mL～10 6 PFU/mL；
S33, lifting the hydraulic pressure of the second virus diluent to 0.22-0.26Mpa through a booster pump 10 of the virus removal testing device, starting testing the second virus diluent in the virus removal testing device, taking a water sample of a control group through a first sampling tube 13 and obtaining water samples of 2 test groups through a second sampling tube 18 at the beginning of 0/4 water supply, wherein the sampling amount of each time is 100mL, and storing the water samples in different sterilization hard glass bottles;
s34, when the high-precision flowmeter reaches 0.5m 3 In the process, 1 control group water sample is obtained by sampling in the first sampling tube 13, and is 1 control group water sample; respectively obtaining water samples of 1/4 section of end in 2 second sampling tubes 18, wherein the water samples are water samples of 2 test groups; the sampling amount of each time is 100mL, the sample is stored in different sterilized hard glass bottles, and the second virus diluent in the virus removal testing device is cleaned;
s35, repeating the steps S32 and S34, sampling at the end of 1/4 section to obtain 1 control group water sample and 2 test group water samples, obtaining 1 control group water sample and 2 test group water samples at the end of 2/4 section, obtaining 1 control group water sample and 2 test group water samples at the end of 3/4 section, obtaining 1 control group water sample and 2 test group water samples at the end of 4/4 section, and obtaining 5 control group water samples and 10 control group water samples in total.
S4, determining the titer of the 5 control group water samples and the 10 test group water samples to obtain a control group result and a test group result;
diluting 5 comparison group water samples and 10 test group water samples respectively by 10 times with sterile pure water, and then preparing the water samples according to escherichia coli: uniformly mixing phage with the concentration ratio of 1000:1, standing for 10-20min at the temperature of 35+/-1 ℃ to obtain water sample diluent; after standing, adding 10ml of semi-solid culture medium of 0.5% agar, mixing, pouring into a prepared nutrient agar culture medium plate, and standing at 35+ -1deg.CCulturing for 16-20h; plaques in the nutrient agar plates were observed, and the nutrient agar plates with the appropriate number of plaques were selected for counting ((30-150 plaques) per dish) and calculated by the calculation formula to give the titer value (PFU/ml) of the virus (ATCC 13706-B1) as follows
S5, obtaining the virus removal rate and the virus removal value through a calculation formula according to the results of the control group and the test group.
The virus removal rate and the virus removal logarithm of the water purifier 16 are calculated as follows:
M＝lg(V a )-lg(V b )
wherein: r is R V : virus removal rate,%; m: virus log values are removed; v (V) a : concentration of phage recovered in control group, PFU/mL; v (V) b : phage concentration, PFU/mL of the test group.
In this embodiment, the labeling process of the water purifier 16 is divided into four sections according to the rated total water purifying amount, the virus removal rate of the water purifier 16 is calculated by measuring the phage titer before and after the virus diluent enters and exits the water purifier 16, the performance of the water purifier 16 is represented by the virus removal rate, and the virus removal rate result of the water purifier 16 is 99.98%, which indicates that the water purifier 16 can effectively remove viruses in water and has a good virus removal effect.
The above embodiments are preferred examples of the present invention, and the present invention is not limited thereto, and any other modifications or equivalent substitutions made without departing from the technical aspects of the present invention are included in the scope of the present invention.
Claims (10)
1. The utility model provides a virus removal testing arrangement of purifier, its characterized in that includes pressure conveying mechanism, first manometer, shunt tubes, first sampling tube, a plurality of fourth valve, a plurality of second manometer, a plurality of purifier, a plurality of flowmeter, a plurality of second sampling tube, a plurality of second drain valve and controller, pressure conveying mechanism is connected with the house steward of shunt tubes, the branch tube of shunt tubes is connected with a plurality of fourth valve respectively, first manometer and first sampling tube are installed in the house steward of shunt tubes in proper order, a plurality of fourth valve is connected with a plurality of purifier through a plurality of first water pipes, a plurality of second manometer are installed in a plurality of first water pipes, a plurality of purifier is connected with a plurality of second drain valves through a plurality of second water pipes, a plurality of flowmeter and a plurality of second sampling tube are installed in a plurality of second water pipes in proper order, the controller is connected with pressure conveying mechanism, first manometer, second manometer and flowmeter respectively.
2. The virus removal testing device of a water purifier according to claim 1, wherein the pressure conveying mechanism comprises a water supply pump, a one-way water stop valve, a first valve, an ultraviolet sterilization lamp, a standard adding water tank, a stirrer, a first drain valve, a second valve, a booster pump, a liquid level meter and a third valve, the water supply pump, the one-way water stop valve, the first valve, the ultraviolet sterilization lamp, the standard adding water tank, the second valve, the booster pump and the third valve are sequentially connected, the third valve is connected with a main pipe of the shunt pipe, the stirrer, the liquid level meter and the first drain valve are all installed in the standard adding water tank, and the stirrer, the liquid level meter, the water supply pump and the booster pump are all connected with the controller.
3. The virus removal testing method of the water purifier is characterized by comprising the following steps of:
s1, preparing virus stock solution through viruses and hosts;
s2, determining the titer of the virus stock;
s3, adding the virus stock solution into the virus removal testing device of the water purifier according to any one of claims 1-2 for testing to obtain a control group water sample and a test group water sample;
s4, determining the titer of the water sample of the control group and the water sample of the test group to obtain a result of the control group and a result of the test group;
s5, calculating the results of the control group and the test group through formulas to obtain the virus removal rate and the virus removal value.
4. The method for testing virus removal of a water purifier according to claim 3, wherein the step S1 comprises the steps of:
s11, selecting proper viruses and hosts;
s12, placing nutrient agar into a culture medium plate to obtain a nutrient agar culture medium plate, inoculating the host into the nutrient agar culture medium plate, culturing for 24+/-1 h at the temperature of 37+/-1 ℃ to obtain single colonies, inoculating the single colonies into a nutrient broth culture medium, and oscillating for 4-6h at the temperature of 35+/-1 ℃;
s13, pouring the nutrient agar into a first culture dish, and obtaining a solid nutrient agar culture dish after the nutrient agar is solidified;
s14, mixing the host suspension and the virus suspension, and standing for 10-20min at the temperature of 35+/-1 ℃ to obtain a mixed suspension of the host and the virus;
s15, uniformly mixing the mixed suspension and a semi-solid culture medium, pouring the mixed suspension and the semi-solid culture medium into the solid nutrient agar culture dish in the step S13, culturing for 17-19h at the temperature of 35+/-1 ℃, recycling the semi-solid culture medium into a sterile bag by adopting a sterile coating rod, centrifuging the sterile bag, and standing at the ambient temperature of 35+/-1 ℃ to obtain a virus suspension;
s16, transferring the virus suspension into a first centrifuge tube for centrifugal treatment to obtain supernatant, and transferring the supernatant into a second centrifuge tube for secondary centrifugal treatment;
s17, repeating the step S16 twice;
and S18, filtering the supernatant in the step S17 through a filter membrane to obtain a virus stock solution, and freezing or refrigerating the virus stock solution.
5. The method for testing virus removal of a water purifier according to claim 3, wherein the step S2 comprises the steps of:
s21, diluting the virus stock solution to obtain a first virus diluent;
s22, mixing the first virus diluent and the host according to a certain concentration ratio, and standing for 10-20min at the ambient temperature of 35+/-1 ℃ to obtain a mixed solution of the virus and the host;
s23, uniformly mixing the mixed solution and the semi-solid culture medium containing agar, pouring the mixed solution and the semi-solid culture medium into a prepared nutrient agar culture medium plate, and standing for 16-20h at the ambient temperature of 35+/-1 ℃ to obtain a plurality of nutrient agar culture medium plates with virus spots;
s24, observing the number of virus spots in a plurality of nutrient agar medium plates, selecting the nutrient agar medium plates according to the number of the virus spots for counting, and calculating by a formula to obtain the titer value of the virus.
6. The method according to claim 5, wherein the number of virus spots in step S24 is 30-150.
7. The method according to claim 5, wherein the calculation formula in the step S24 is a virus titerThe number of plaques is the average number of plaques, the volume of the diluent used in plating the plate is V, and the dilution is M.
8. The method for testing virus removal of a water purifier according to claim 3, wherein the step S3 comprises the steps of:
s31, dividing the test into multiple stages according to the nominal rated water purifying amount of the water purifier;
s32, adding sterile pure water into a standard adding water tank of the virus removal testing device, and then adding virus stock solution for dilution to obtain second virus diluent;
s33, introducing the second virus diluent into the virus removal testing device, and sampling through a first sampling tube of the virus removal testing device at the beginning of water introduction to obtain a control group water sample, and sampling through a second sampling tube of the virus removal testing device to obtain a test group water sample;
s34, sampling at the first sampling tube to obtain another water sample of the control group, sampling at the second sampling tube to obtain another water sample of the test group, and cleaning the residual virus diluent in the virus removal testing device;
s35, repeating the steps S31 and S34 to obtain a plurality of water samples of the control group and a plurality of water samples of the test group.
9. The method for testing virus removal of a water purifier according to claim 3, wherein the formula of the virus removal rate in step S5 isThe V is a Phage concentration recovered for control group, V b Phage concentration for the test group.
10. The method according to claim 3, wherein the formula of the virus removal value in step S5 is m=lg (V a )-lg(V b ) The V is a Phage concentration recovered for control group, V b Phage concentration for the test group.
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