KR20110097199A - The virus experiment system for virus filtration efficiency test and the method thereof - Google Patents

Abstract

The present invention relates to an apparatus for evaluating the effectiveness of a filter against a virus and a method for evaluating the effect.
An apparatus for evaluating the effectiveness of a filter against a virus of the present invention is installed in communication with a first aseptic chamber for accommodating viruses from the outside, and in communication with the first aseptic chamber to inhale air from the first aseptic chamber, thereby allowing the first aseptic First detecting means comprising a first collecting portion for detecting the amount of virus in the air of the upper chamber; The virus is installed between the first aseptic chamber of the first detection means and the second aseptic chamber of the second detection means to filter air flowing from the first aseptic chamber to the second aseptic chamber. Filter means for filtering out; And a second aseptic chamber which is installed in communication with the first aseptic chamber through the filter means and allows the air of the first aseptic chamber filtered by the filter means to enter and receive the air, and the air of the second aseptic chamber. And a second detecting means comprising a second collecting portion which collects the virus in the air from the first aseptic chamber through the filter means and detects the amount of virus in the air of the second aseptic chamber.
According to the present invention, it is possible to evaluate the effectiveness of the filter against a variety of viruses and scientifically and easily evaluate the effect of the filter on a living animal by a negative pressure air conditioner, there is a convenient effect.

Description

Evaluation device and evaluation method of filter against virus {THE VIRUS EXPERIMENT SYSTEM FOR VIRUS FILTRATION EFFICIENCY TEST AND THE METHOD THEREOF}

The present invention is to evaluate the effectiveness of the newly developed and manufactured biological filter (Virus filtration efficency: VEF) as well as to evaluate the effectiveness of the filter against a variety of viruses, and to evaluate the effectiveness of the filter scientifically and easily in living animals. The present invention relates to a device for evaluating the effectiveness of a filter against a virus and a method for evaluating the effect thereof.

In recent years, these diseases and prevention have been caused by the development of life-threatening diseases caused by viruses such as H1N1, high pathogenic avian influenza (HPAI), and severe acute respiratory syndrome (SARS). There is a great deal of social interest in.

One of the simplest and most economical methods of removing these viruses is to filter them.

The virus filtration efficency (VFE), which is currently globally recognized, is based on the American Society for Testing and Testing (Military standard MIL-M-36954) and medical mask manufacturing testing standards. materials) The procedure specified in F2101 is followed.

Summarizing the VFE test method, one of the smallest viruses with a diameter of about 27 nm (0.027 μ), an unenveloped icosahedron, phiX174 bacteriophage, is used for filter efficacy evaluation. Using a device designed to collect bacteriophage suspensions at constant pressure and hydraulic pressure using a neicaizer to approx. The VFE test should use a filter stored for at least 4 hours at 21 ± 5 ° C and 5% RH, and the average VFE control should be 2200 ± 500 PFU. In the Anderson sampler, the agar medium in which E. coli was cultured was placed and collected for 12-24 hours at 37 ° C. to measure the filter efficiency by counting the number of plaques due to the growth of the collected bacteriophages.

As a research report on the effectiveness of filters on viruses, Roelants et al. (1968) reported the virus removal efficiency of commercial filters in the air. For this purpose, the experimental system was designed and the results of the experiment were reported.

In addition, reports on virus filtration efficency (VFE) in laboratory animals are very rare, but Burmester et al. (1972) have Marek's, a type of herpesvirus. We report the results of a study evaluating the efficacy of commercially available filters using disease virus (MDV) virus as a change in pathological and serological criteria according to infection status in chickens with very high sensitivity to MDV. have.

However, these reported techniques cannot assess the effectiveness of specific filters against various kinds of viruses, and in particular, cannot evaluate the efficacy on the living organisms most similar to the actual effects of the filters.

P. Roelants, B. Boon and W. Lhoest, Evaluation of a commercial air filter for removal of viruses from the air, 1968, Applied Microbiology, Vol. 16, No. 10, pp. 1465-1467. H. W. Li, C. Y. Wu, F. Tepper, J. H. Lee and C. N. Lee. Removal and retention of viral aerosols by a novel alumina nanofiber filter, 2009, Aerosol Science, Vol. 40, 65-71. B. R. Burmester and R. L. Witter, Efficiency of commercial air filters against Marek's diseases virus, 1972, Applied Microbiology, Vol. 23, No. 3, 505-508.

The present invention is to solve the above problems, it is possible to evaluate the effectiveness of the filter against a variety of viruses and scientifically and easily to evaluate the effect of the filter on the experimental animals in vivo by a negative pressure air conditioner is not only convenient, but also experimental animals The purpose of the present invention is to provide a device for evaluating the effectiveness of a filter against a virus that can perform experiments in accordance with the ethical standards.

In addition, there is also provided an apparatus for evaluating the effectiveness of the filter against viruses that can block the external leakage of microorganisms by the HEPA filter to safely experiment with high-risk viruses.

In addition, it is easy to disinfect by fumigation before and after experiments for various viruses, and also provides an apparatus for evaluating the effectiveness of the filter against viruses that can reduce the experimental error due to contamination.

The apparatus for evaluating the effectiveness of a filter for a virus of the present invention for achieving the above object is installed in communication with a first aseptic chamber and a first aseptic chamber for receiving a virus introduced from the outside from the first aseptic chamber First detection means comprising a first collecting portion which collects viruses in the air by inhaling and detects the amount of viruses in the air of the first aseptic chamber; The virus is installed between the first aseptic chamber of the first detection means and the second aseptic chamber of the second detection means to filter air flowing from the first aseptic chamber to the second aseptic chamber. Filter means for filtering out; And a second aseptic chamber which is installed in communication with the first aseptic chamber through the filter means and allows the air of the first aseptic chamber filtered by the filter means to enter and receive the air, and the air of the second aseptic chamber. Inhalation allows the air of the first aseptic chamber to move to the second aseptic chamber via the filter means, collects the virus in the inhaled air and detects the amount of virus in the air of the second aseptic chamber. It comprises a second detection means consisting of a second collecting unit.

The virus flowing into the first aseptic chamber is characterized in that the sprayed by any one of the injection nozzle or nebulizer.

In addition, the first detection means, the filter means and the second detection means are respectively provided with a virus inlet breaker characterized in that the virus is introduced or blocked.

The first collecting unit may include a first inlet pipe through which the virus in the air is introduced, a first sampler installed in communication with the first inlet pipe, and detecting the amount of virus in the air introduced through the first inlet pipe; It is installed in communication with the first sampler is characterized in that made of a first vacuum pump to suck the virus in the air to the first sampler through the first inlet pipe.

In addition, the second collecting unit is installed in communication with the second inlet pipe through which the filtered air flows through the filter means, and the second inlet pipe detects the amount of unfiltered virus in the air flowing through the second inlet pipe. And a second vacuum pump installed in communication with the second sampler so as to suck viruses in the air into the second sampler through the second inlet pipe.

In addition, after the evaluation of the effectiveness of the filter means is completed, when the evaluation of the effectiveness of the experimental animals with mice, ferrets, etc., the second aseptic chamber and to control the ventilation and air flow inside the second aseptic chamber It is characterized in that the negative pressure air conditioner is further installed in communication.

In addition, a hepa filter is further installed between the second aseptic chamber and the negative pressure air conditioner to prevent external leakage of microorganisms.

In addition, the efficacy evaluation method of the filter against the virus of the present invention can be evaluated by the following method.

A first step of spraying the virus in the interior of the first aseptic chamber in the state of opening only the virus inlet breaker of the first detection means; A second step of detecting a virus in the air of the first aseptic chamber by operating the first collecting unit to suck air from the first aseptic chamber; Close the virus inlet breaker of the first detection means and operate the second collector with the virus inlet breaker of the filter means and the second detection means open, so that the air in the first aseptic chamber passes through the filter means to the second aseptic chamber. A third step of detecting the amount of virus in the air of the second aseptic chamber by collecting the virus in the inhaled air; And a fourth step of evaluating the effectiveness of the filter means by comparing the amount of virus detected by the first detection means with the amount of virus detected by the second detection means.

The present invention by the above solving means is possible to evaluate the effectiveness of the filter against a variety of viruses and scientifically and easily evaluate the effect of the filter on the experimental animals in vivo by a negative pressure air conditioner is not only convenient but also suitable for the ethics of the experimental animals There is an effect that the experiment can be performed as a reference.

In addition, by blocking the external leakage of microorganisms by hepa filter, there is also an effect that can safely experiment with high-risk viruses.

In addition, it is easy to disinfect by fumigation before and after experiments with various viruses, thereby reducing the experimental error due to contamination.

1 is a view showing an apparatus for evaluating the effectiveness of a filter against a virus of the present invention.
2 is a photograph of an apparatus for evaluating the effectiveness of a filter against a virus of the present invention.
Figure 3 is a photograph showing the filter means of the apparatus for evaluating the effectiveness of the filter against the virus of the present invention.
Figure 4 is a block diagram showing a method for evaluating the effectiveness of the filter against the virus of the present invention.
Figure 5 of the gene to be amplified in the process of testing the filter with swine influenza virus using the filter effect evaluation device of the virus of the present invention and the efficiency of the SYBR-Green real-time polymerase chain reaction (RT-PCR method) A graph showing quantitative and Ct values.
6 is a half-step dilution of swine influenza virus using a filter effect evaluation device for the virus of the present invention 2 ⁶ , 2 ⁵ , 2 ⁴ , 2 ³ , 2 ² , 2 ¹ hemagglutination unit (HAU Fig. 11 shows a standard curve prepared by RT-PCR using a virus of Hemagglutination Unit) concentration.
7 is a view showing the weight change of the experimental animal according to the filter means in the apparatus for evaluating the effectiveness of the filter against the virus of the present invention.

1 is a view showing an apparatus for evaluating the effectiveness of the filter against the virus of the present invention, FIG. 2 is a photograph of the apparatus for evaluating the effectiveness of the filter against the virus of the present invention, and FIG. Photo showing the filter means, Figure 4 is a block diagram showing a method for evaluating the effect of the filter against the virus of the present invention, Figure 5 is a filter for testing the effect of swine influenza virus using the apparatus for evaluating the effect of the filter against the virus of the present invention Figure 6 is a graph showing the quantitative and Ct value of the amplified gene in the process of investigating the efficiency by the SYBR-Green real-time polymerase chain reaction (RT-PCR method), Figure 6 is an effect evaluation device of the filter against the virus of the present invention by using a dilution of the swine influenza virus in 1/2 stages ^{26, 25, 24, 23, 22, 21} hemagglutination units (HAU; hemagglutination unit) by the concentration of A diagram showing a standard curve prepared by RT-PCR using the bus, Figure 7 is a view showing the weight change of the animals with and without the filter means in effect evaluation apparatus of the filter for the virus of the present invention.

As shown in the figure, the apparatus for evaluating the effectiveness of the virus of the present invention comprises a first detecting means 100, a filter means 200, and a second detecting means 300.

The first detection means 100 is installed in communication with the first aseptic chamber 110 and the first aseptic chamber 110 for receiving a virus introduced from the outside air from the first aseptic chamber 110 Inhalation of the virus in the air to collect the virus in the air of the first aseptic chamber 110, the first trap 130 is made.

In this case, the virus flowing into the first aseptic chamber 110 may be sprayed by any one of the injection nozzle or the nebulizer 50.

The nebulizer 50 generates the virus solution in a spray state of 0.5 to 5.5 microns by using the 2.5 kg / cm 2 pressure generated in the compressor 70.

The first collecting unit 130 is installed in communication with the first inlet pipe 131 for introducing the virus in the air, the first inlet pipe 131 in the air introduced through the first inlet pipe 131 The first sampler 133 for detecting the amount of virus of the virus and the first sampler 133 is installed in communication with the first sample pipe 131 through the first sampler 133 to suck the virus in the air It consists of a vacuum pump 135.

At this time, the first aseptic chamber 110 and the first inlet pipe to introduce or block the virus flowing into the first inlet pipe 131 from the first aseptic chamber 110 by the first vacuum pump 135 The virus inflow blocker 150 is installed between the 131.

The filter means 200 is installed between the second aseptic chamber 310 of the second detection means 300 in communication with the first aseptic chamber 110 of the first detection means 100 is the first aseptic The air flowing from the upper chamber 110 to the second aseptic chamber 310 is provided to filter the virus.

At this time, the filter means 200 is to replace the filter by blocking the virus flowing into the filter means 200 by installing a virus inlet breaker 250 at both ends in order to replace the filter for a variety of viruses.

The second detecting means 300 is installed in communication with the first aseptic chamber 110 through the filter means 200 and inflows air from the first aseptic chamber 110 filtered by the filter means 200. By sucking the air of the second aseptic chamber 310 and the second aseptic chamber 310 so that the air of the first aseptic chamber 110 passes through the filter means 200. The second collection part 330 is made to be movable to the chamber 310 and to collect the virus in the inhaled air and to detect the amount of virus in the air of the second aseptic chamber 310.

The second collecting unit 330 is installed in communication with the second inlet pipe 331 and the second inlet pipe 331 through which the filtered air flows through the filter means 200, and the second inlet pipe 331. The second sampler 333 for detecting the unfiltered virus amount in the air flowing through the) and the second sampler 333 is installed in communication with the second sampler 333 through the second inlet pipe 331. It consists of a second vacuum pump 335 to inhale viruses in the air.

At this time, the second aseptic chamber 310 and the second inlet pipe to introduce or block the virus flowing into the second inlet pipe 331 from the second aseptic chamber 310 by the second vacuum pump 335. Between the 331, the virus inflow blocker 350 is provided.

As described above, the present invention can evaluate the effect of the filter means 200 by comparing the amount of virus detected by the first detection means 100 with the amount of virus detected by the second detection means 300.

In addition, in the present invention, the negative pressure air conditioner 400 is further installed in communication with the second aseptic chamber 310 to test the effectiveness of the filter against various viruses in the experimental animal which is a living body.

The negative pressure air conditioner 400 is to control the ventilation and air flow inside the second aseptic chamber 310 required for breeding of the experimental animal.

At this time, the condition for selecting a laboratory animal that is a living being is that the virus to be tested must be sensitive to the laboratory animal. Therefore, you should choose a laboratory animal of susceptible breed depending on the type of virus you are testing.

In addition, a hepa filter 410 is further installed between the second aseptic chamber 310 and the negative pressure air conditioner 400 to prevent external leakage of microorganisms.

On the other hand, the apparatus for evaluating the effectiveness of the filter against the virus of the present invention may be tested by the method for evaluating the efficacy of the filter against the virus.

First, the virus spraying step 190 sprays the virus inside the first aseptic chamber 110 while only the virus inlet breaker 150 of the first detection means 100 is opened.

Second, the first detection step 290 operates the first collecting unit 130 to inhale air from the first aseptic chamber 110 to collect viruses in the air and thus to remove the first aseptic chamber 110. Detect viruses in the air

Third, the second detection step 390 is to close the virus inlet breaker 150 of the first detection means 100 and open the virus inlet breakers 250 and 350 of the filter means 200 and the second detection means (300). In this state, the second collecting unit 330 is operated to allow the air of the first aseptic chamber 110 to move to the second aseptic chamber 310 via the filter means 200, and the inhaled air Virus is collected and the amount of virus in the air of the second aseptic chamber 310 is detected.

Fourth, the efficacy evaluation step 490 evaluates the effectiveness of the filter means 200 by comparing the virus amount detected by the first detection means 100 and the virus amount detected by the second detection means 300.

In this case, the efficiency of the filter can be calculated by the same principle as the virus filtration efficiency (VFE) method, and the quantitative difference between the collected viruses can be expressed as% VFE.

The calculation method can be calculated by the difference in plaque number or tissue culture infectious dose (TCID) on tissue culture as follows.

Another method is to use a real-time (RT) polymerase chain reaction (PCR) method to quantitatively amplify a specific gene with a specific primer for a specific virus collected, and The virus efficiency of the filter can be calculated by comparing the same standard curve with the Ct value (cycle threshold value) and estimating the copy of the viral gene and comparing it with the following equation.

Experimental Example 1

In order to amplify the swine influenza virus by the SYBR-Green Real-Time Polymerase Chain Reaction (RT-PCR) method, the previously known M + 25 primers 5'-AGCTGAGTCTTCTAACCGAGGTCG-3 'and M-124 primers 5'-TGCAAAAACATCTTCAAGTCTCTG-3' primers were used. Synthesis (Bioneer, Korea) was used and SYBR-Green RT-PCR was performed.

Viral Gene-spin ^TM in stocks with 2 ⁶ , 2 ⁵ , 2 ⁴ , 2 ³ , 2 ² , 2 ¹ HAU concentrations by diluting half of the stock (2 ⁷ HA titer) porcine influenza virus Virus RNA was extracted using a kit (Intron), and real time RT-PCR was performed on an I cycler IQ (Bio-Rad) PCR machine using a LightCycler RNA Amplification Kit SYBR Geen I (Roche).

The mixture required for the reaction was 0.4 μl of LightCycler RT-PCR Enzyme Mix, 0.4 μl of LightCycler RT-PCR recation Mix SYBR Green I, MgCl ₂ 2.4 μl of stock solution, 2 μl of viral RNA, AIV common primer in Table 1 were used as a forword and reverse primer at 5pm concentration, and 1 μl of each and sterilized tertiary distilled water were added to form 20 μl of the final reaction solution. In addition, the reaction mixture was prepared by removing RNA and adding sterilized tertiary distilled water as a negative control.

The reaction process was reverse transcription at 55 ° C. for 25 minutes, denatured at 95 ° C. for 30 seconds, followed by 45 cycles at 95 ° C. for 5 seconds, 55 ° C. 15 seconds, and 72 ° C. 25 seconds. As a result, if an amplification curve was generated in 35 cycles or more based on the Ct value, the result was judged as a negative signal.

A standard curve was prepared as shown in FIG. 6 using the threshold cycle (Ct, threshold cycle) indicated for each virus concentration. A regression model of y = -4.842x + 36.31 was shown.

Swine influenza virus showed a PR8 strain of 5 × 10 ⁵ PFU / mL titer, and the Ct value of the filter-free control virus was 21.96, followed by filters C, A-1, C-1, and A in order of 25.85, 31.7, 33.12 and 33.47 showed that the filtration capacity of the filter was excellent in the order of A, C-1, A-1 and C. Also, if the value of x is obtained by substituting the y value of Ct at y = -4.842x +36.31 using the standard curve, the amount of detected virus can be calculated in units of hemagglutination (HA). , A-1 and C can be quantified with 2 ^3.0 , 2 ^2.1 , 2 ^1.0 , 2 ^0.7 and 2 ^0.6 HAU / 50 μl, respectively.

Experimental Example 2

Viral infection suppression experiments of in vivo filters using biological animals show the mortality or infection rate of the experimental animals in the same number of controls, i.e. without filters, and the mortality or infection rates of the experimental animals in the presence of the test filter. It can evaluate by comparing.

Viruses such as Ozesky's disease virus or influenza virus that are susceptible to normal ICR or Balb / c mice can be disclosed. That is, certain influenza strains require animals such as ferrets. In other words, prior knowledge of test viruses and susceptible experimental animals is required. The number of experimental animals to be published is set to the minimum number that can be statistically significant for animal welfare.

In this experiment, in the case of comparing the infection control ability of the filter to the actual animal, the experiment equipped with the filter having relatively less virus contamination was performed first, and after disinfection of the device after the first experiment to maintain the objectivity of the experiment results use.

Swine influenza virus published a PR8 week of 5 × 10 ⁵ PFU / mL titers and compared the mean body weight change of the five experimental mice after virus exposure. For weight loss of more than 50%, the mice were euthanized.

As a result of the filter efficacy evaluation using the present invention, the average mouse weight was 53.8 g after the inoculation date, one day after inoculation, two days after inoculation, three days after inoculation, and four days after inoculation, respectively. The average body weight of the unfiltered mice was reduced to 56.4 g, 56.2 g, 51.2 g, 47.6 g and 30.9 g, while increasing to 54.8 g, 55.2 g, 55.4 g and 56.2 g.

When the average weight of the inoculation date was converted into 100% by weight in terms of weight per day, the filtered group increased to 100, 101.9, 102.6, 103.0, and 104.5 as shown in FIG. It was reduced to 99.6, 90.8, 84.4 and 54.8 to evaluate the filter efficacy in vivo.

As can be seen through the experiment, the present invention is not only convenient, but also useful for evaluating the effectiveness of the filter against various viruses, and for evaluating the effect of the filter scientifically and easily on a test animal that is a living body by a negative pressure air conditioner. It is effective to conduct experiments on the basis of proper ethics.

As described above, according to the present invention, it is possible to evaluate the effectiveness of the filter against various viruses and scientifically and easily evaluate the effect of the filter on a living animal by a negative pressure air conditioner. Provide a device for evaluating the effectiveness of a filter against viruses that can perform experiments on appropriate criteria.

In addition, there is also provided an apparatus for evaluating the effectiveness of the filter against viruses that can block the external leakage of microorganisms by the HEPA filter to safely experiment with high-risk viruses.

In addition, it is easy to disinfect by fumigation before and after experiments for various viruses, and also provides an apparatus for evaluating the effectiveness of the filter against viruses that can reduce the experimental error due to contamination.

50: Nebulizer 55: Virus pain
70: compressor 100: first detection means
110: first aseptic chamber 130: first collecting unit
131: first inlet pipe 133: first sampler
135: first vacuum pump 150, 250, 350, 450: virus inlet breaker
190: virus spraying step 200: filter means
290: first detection step 300: second detection means
310: second aseptic chamber 330: second collecting part
331: second inlet pipe 333: second sampler
335: second vacuum pump 390: second detection step
400: negative pressure air conditioner 410: HEPA filter
490: Efficacy evaluation step

Claims (8)

It is installed in communication with the first aseptic chamber 110 and the first aseptic chamber 110 to receive the virus flowing from the outside to collect the virus in the air by sucking the air from the first aseptic chamber 110 First detecting means (100) comprising a first collecting part (130) for detecting the amount of virus in the air of the first aseptic chamber (110);
It is installed between the second aseptic chamber 310 of the second detection means 300 in communication with the first aseptic chamber 110 of the first detection means 100 from the first aseptic chamber 110. Filter means (200) for filtering air flowing into the second aseptic chamber (310) to filter out viruses; And
The second aseptic chamber is installed in communication with the first aseptic chamber 110 through the filter means 200, the second aseptic chamber is introduced to accommodate the air of the first aseptic chamber 110 filtered by the filter means (200) By sucking the air in the chamber 310 and the second aseptic chamber 310, the air in the first aseptic chamber 110 is movable to the second aseptic chamber 310 via the filter means 200. And second detection means (300) comprising a second collecting portion (330) for collecting the virus in the inhaled air and detecting the amount of virus in the air of the second aseptic chamber (310). ,
The amount of virus detected by the first detection means 100 and the amount of virus detected by the second detection means 300 are compared to evaluate the effect of the filter means 200. Efficacy Evaluation Device. The apparatus of claim 1, wherein the virus introduced into the first aseptic chamber is sprayed by either a spray nozzle or a nebulizer. According to claim 1, wherein the first detection means 100, the filter means 200 and the second detection means 300 are virus inlet breakers (150, 250, 350, 450) are respectively installed, characterized in that the virus is introduced or blocked Device for evaluating the effectiveness of a filter against viruses. According to claim 1, wherein the first collecting unit 130 is installed in communication with the first inlet pipe 131 and the first inlet pipe 131 for introducing the virus in the air, the first inlet pipe 131 The first sampler 133 for detecting the amount of virus in the air introduced through the air, and the first sampler 133 is installed in communication with the first sampler 133 through the first inlet pipe 131 Apparatus for evaluating the effect of the filter on the virus, characterized in that consisting of a first vacuum pump 135 for inhaling the virus. According to claim 1, The second collecting unit 330 is installed in communication with the second inlet pipe 331, the second inlet pipe 331 through which the filtered air flows through the filter means 200 The second sampler 333 for detecting the unfiltered virus amount in the air flowing through the second inlet pipe 331 and the second sampler 333 are installed in communication with the second sample pipe 331. The second vacuum pump (335) to suck the virus in the air to the sampler 333, the effect evaluation device for the virus, characterized in that made. According to claim 1, After the evaluation of the effectiveness of the filter means 200, when evaluating the effect on the living animals such as mice, ferrets, etc. Ventilation and air flow inside the second aseptic chamber 310 Efficacy evaluation device of the filter for viruses, characterized in that the negative pressure air conditioner 400 is further installed in communication with the second aseptic chamber 310 to control. According to claim 6, Hepa filter 410 is further installed between the second aseptic chamber 310 and the negative pressure air conditioner 400, the effect of the filter against the virus, characterized in that to prevent the external leakage of microorganisms Evaluation device. A virus spraying step 190 of spraying a virus into the first aseptic chamber 110 in a state in which only the virus inlet breaker 150 of the first detection means 100 is opened;
First detection step of operating the first collecting unit 130 to suck the air from the first aseptic chamber 110 to collect the virus in the air to detect the virus in the air of the first aseptic chamber 110 (290);
Operating the second collecting unit 330 while closing the virus inlet breaker 150 of the first detection means 100 and opening the virus inlet breakers 250 and 350 of the filter means 200 and the second detection means 300. The air of the first aseptic chamber 110 is moved to the second aseptic chamber 310 via the filter means 200, and the virus in the sucked air is collected to collect the second aseptic chamber ( A second detection step 390 of detecting the amount of virus in the air of 310; And
It is characterized in that it consists of a efficacy evaluation step (490) for evaluating the effectiveness of the filter means 200 by comparing the amount of virus detected by the first detection means 100 and the amount of virus detected by the second detection means (300) How to evaluate the effectiveness of a filter against a virus.

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