Method for inactivating virus and application thereof

Abstract

The invention provides a method for inactivating a virus. The method comprises the steps that electromagnetic waves are adopted to radiate a sample, the radiation power density of the electromagnetic waves is 100-120 milliwatts per square centimeter, and the radiation frequency of the electromagnetic waves is 2.8-3.0 gigahertz; the sample comprises coronavirus, and the pH value of the sample is 7-9. The method disclosed by the invention can be used for remarkably improving the virus inactivation effect of electromagnetic wave radiation and is used for inactivating coronavirus; and the method is suitable for virus inactivation at normal temperature, and is beneficial to virus inactivation in daily work and life scenes. The method and the virus inactivation device can be further used for killing coronavirus, and are suitable for popularization and application in closed or semi-closed spaces such as scientific research laboratories, hospitals, shopping malls and vehicles.

Description

Technical Field

[0001] The present invention relates to the field of virus inactivation, and in particular to a method for inactivating viruses by using electromagnetic waves and an application thereof. Background Art

[0002] Pathogenic virus infection has long been an important public health safety issue facing the world. The demand for efficient and safe methods to inactivate pathogenic viruses is even more urgent.

[0003] Traditional technical methods for inactivating pathogenic viruses have certain effects, but there are still certain limitations; electromagnetic waves have the characteristics of strong penetration, physical resonance, and no pollution. Their ability to inactivate pathogenic viruses has gradually attracted attention, and their basic and applied research in the field of pathogenic virus inactivation has been continuously explored and developed. Electromagnetic wave radiation is a type of non-ionizing radiation, which is widely used to inactivate microorganisms in liquids and on the surfaces of objects. Some researchers have used electromagnetic waves to conduct experimental studies on the killing and destructive effects of respiratory, intestinal, arboviruses and hepatitis viruses, which can be used for the disinfection of hospital infectious wards and related medical supplies, and have achieved good results. However, there are currently no reports on the application conditions of electromagnetic wave radiation for virus inactivation.

[0004] Therefore, there is an urgent need to develop application conditions and methods for electromagnetic wave radiation to effectively inactivate viruses. Summary of the invention

[0005] The present invention aims to solve at least one of the technical problems existing in the prior art to a certain extent. To this end, the present invention provides a method and device for inactivating viruses, which can significantly improve the effect of inactivating viruses by electromagnetic wave radiation, and can be used for inactivating coronaviruses; and the method is suitable for inactivating viruses at room temperature, which is conducive to inactivating viruses in daily work and life scenarios. The method of the present invention and the device for inactivating viruses can be further used for the disinfection of coronaviruses, and are suitable for promotion and application in closed or semi-enclosed spaces such as scientific research laboratories, hospitals, shopping malls, and transportation vehicles.

[0006] It should be stated that the present invention is completed based on the following work of the inventor:

[0007] In the process of studying the inactivation effect of electromagnetic wave radiation on coronavirus, the inventors found that electromagnetic wave exposure with an electromagnetic radiation power density of 100 to 120 milliwatts per square centimeter and a radiation frequency of 2.8 to 3.0 gigahertz can effectively reduce the activity of coronaviruses (such as mouse hepatitis virus MHV-A59). Based on a large number of screening and optimization work on inactivation virus conditions, it was found that the pH value of the new coronavirus sample can affect the effect of electromagnetic radiation on virus inactivation. Beyond the inventors' expectations, the combination of sample pH values ​​of 7 to 9 and the above-mentioned electromagnetic radiation conditions showed a synergistic inactivation effect on coronaviruses (such as mouse hepatitis virus MHV-A59). The method of the present invention significantly improves the effect of electromagnetic wave radiation inactivating viruses and can be used for the inactivation of coronaviruses.

[0008] Therefore, in the first aspect of the present invention, the present invention provides a method for inactivating viruses. The method comprises:

[0009] The sample is irradiated with electromagnetic waves, wherein the radiation power density of the electromagnetic waves is 100 to 120 milliwatts per square centimeter, and the radiation frequency of the electromagnetic waves is 2.8 to 3.0 gigahertz;

[0010] The sample includes coronavirus, and the pH value of the sample is 7-9.

[0011] It should be noted that the samples of the present invention are selected from samples whose pH is adjustable, or samples whose coronaviruses can be placed in the expected pH range by alternative means. Exemplarily, the "alternative means" can be spraying a PBS buffer with a pH value of 7 to 9 on the surface of a solid sample; or dissolving a solid sample (such as a soluble inorganic salt) in a solution with a pH value of 7 to 9; or adjusting the pH value of a liquid, suspension, semi-solid, etc. sample by an acid-base reagent to be within the range of 7 to 9.

[0012] In the experiment, the inventors adjusted the power density and frequency of the electromagnetic wave radiation to the above range and then irradiated the coronavirus sample with electromagnetic waves, and creatively adjusted the pH of the coronavirus sample to 7-9. The two factors of electromagnetic wave radiation conditions and sample pH synergistically enhanced the effect of inactivating viruses, and exerted a stronger virus killing effect. Therefore, compared with electromagnetic wave radiation alone, the effect of inactivating viruses in the method of the present invention is significantly enhanced, and can be used for inactivating coronaviruses. Based on the method of the present invention and its working principle, a device for inactivating viruses can be further developed.

[0013] According to an embodiment of the present invention, the radiation power density of the electromagnetic wave is 100 milliwatts per square centimeter, and the radiation frequency of the electromagnetic wave is 2.8 gigahertz.

[0014] According to an embodiment of the present invention, the temperature of the sample is room temperature.

[0015] It should be noted that the normal temperature described in the present invention specifically refers to a temperature of 15 to 45°C. According to an embodiment of the present invention, the inventor creatively designed an experiment and determined that when the temperature of the above-mentioned virus sample is controlled within this range, the method of the present invention has a better virus inactivation effect. Therefore, the method of the present invention has low requirements for temperature control of the virus sample and is easy to achieve, and is more suitable for promotion and application in daily life scenarios. According to an embodiment of the present invention, the temperature is 15 to 42.5°C.

[0016] According to an embodiment of the present invention, the temperature is 15-32.5°C.

[0017] According to an embodiment of the present invention, the temperature is 20-37.5°C.

[0018] According to an embodiment of the present invention, the temperature is 25-42.5°C.

[0019] According to an embodiment of the present invention, under the above-mentioned electromagnetic wave radiation conditions, adjusting the pH value of the sample to 7-9 can significantly enhance the effect of electromagnetic wave radiation inactivating viruses; the two factors affecting the electromagnetic wave virus inactivation effect, the electromagnetic wave radiation conditions and the sample pH, synergize and enhance the effect, and are statistically significant.

[0020] According to an embodiment of the present invention, the radiation duration of the electromagnetic wave is not less than 15 minutes.

[0021] According to an embodiment of the present invention, the radiation duration of the electromagnetic wave is not less than 30 minutes.

[0022] The longer the electromagnetic wave irradiation time is, the stronger the virus inactivation effect is. In a specific implementation scheme, the electromagnetic wave radiation time can be adjusted according to the properties of the virus to obtain the expected virus inactivation effect.

[0023] According to an embodiment of the present invention, the pH value of the sample is 7.5-8.5.

[0024] According to an embodiment of the present invention, the pH value of the sample is 8.

[0025] According to an embodiment of the present invention, the pH of the virus sample to be inactivated can be appropriately adjusted according to the radiation power density and radiation frequency of the electromagnetic wave and the properties of the virus, thereby obtaining the expected enhanced virus inactivation effect.

[0026] According to an embodiment of the present invention, the sample is selected from an aerosol sample, a liquid sample, a suspension sample or a semi-solid sample.

[0027] According to an embodiment of the present invention, the sample is a suspension sample.

[0028] According to an embodiment of the present invention, the pH value of the suspension sample is 7.5-8.5.

[0029] According to an embodiment of the present invention, the pH value of the suspension sample is 8.

[0030] According to an embodiment of the present invention, the pH of a sample including coronavirus is appropriately adjusted, thereby obtaining the expected enhanced virus inactivation effect.

[0031] According to an embodiment of the present invention, the radiation is performed at room temperature.

[0032] According to an embodiment of the present invention, the radiation is performed at 15-40°C.

[0033] According to an embodiment of the present invention, the radiation is performed at 15-25°C.

[0034] Temperature also affects the effect of electromagnetic wave radiation inactivating viruses. According to an embodiment of the present invention, the method of the present invention is suitable for virus inactivation at room temperature, which is beneficial to the inactivation of viruses in daily work and life scenarios.

[0035] According to an embodiment of the present invention, the coronavirus is a mammalian coronavirus.

[0036] According to an embodiment of the present invention, the mammalian coronavirus is selected from at least one of mouse hepatitis virus, human coronavirus HCoV-229E, human coronavirus HCoV-OC43, human coronavirus HCoV-HKUI, human coronavirus HCoV-NL63, human coronavirus MERS-CoV, human coronavirus SARS-CoV and human coronavirus SARS-CoV-2.

[0037] According to an embodiment of the present invention, the mouse hepatitis virus is selected from at least one of MHV-1, MHV-2, MHV-3, MHV-4 (JHM), MHV-A59 and MHV-S.

[0038] According to an embodiment of the present invention, the mouse hepatitis virus is MHV-A59.

[0039] According to an embodiment of the present invention, the mammalian coronavirus is selected from at least one of SARS-CoV-2, SARS-CoV and MERS-CoV or their model viruses.

[0040] It should be noted that the model viruses of coronaviruses such as SARS-CoV-2, SARS-CoV and MERS-CoV of the present invention can replace the above coronaviruses under laboratory conditions to evaluate and verify the inactivation effect of the virus inactivation method of the present invention on the above coronaviruses. Exemplarily, the "model virus" can be mouse hepatitis virus MHV, and more specifically, the "model virus" can be mouse hepatitis virus MHV-A59. MHV is a model virus commonly used in coronavirus research, and the electromagnetic wave radiation method that has an inactivation effect on it has the same or similar inactivation effect on other coronaviruses, especially coronaviruses that have serological cross-reactivity with certain strains of other mammalian coronaviruses of the genus MHV.

[0041] In a second aspect of the present invention, a device for inactivating viruses is provided. The device for inactivating viruses comprises:

[0042] An electromagnetic wave transmitting module, used for irradiating the sample;

[0043] The control module is used to control the electromagnetic wave transmitting module, the control module is connected to the electromagnetic wave transmitting module, and the control module is controlled based on the above method. Thus, the method of inactivating viruses of the present invention is promoted and applied in closed or semi-closed spaces such as scientific research laboratories, hospitals, shopping malls, and transportation vehicles.

[0044] In a third aspect of the present invention, a computer-readable storage medium is provided. The storage medium stores a program, and the program can be executed by a processor to implement the aforementioned method or to implement the virus inactivation function of the aforementioned device. Thus, the method for inactivating viruses of the present invention is automated and intelligently promoted and applied in closed or semi-closed spaces such as scientific research laboratories, hospitals, shopping malls, and transportation vehicles.

[0045] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0047] Figure 1 This is a morphological structure diagram of the mouse hepatitis virus (MHV-A59) used in Example 1 of the present invention observed under a transmission electron microscope;

[0048] Figure 2 This is a morphological picture of 17CL-1 cells required for culturing viruses in Example 1 of the present invention observed under a fluorescent inverted phase contrast microscope;

[0049] Figure 3 This is a graph showing the results of investigating the changes in virus titer of a virus sample after exposure to electromagnetic waves for 0, 15, and 30 minutes in Example 2 of the present invention;

[0050] Figure 4 This is a graph showing the results of investigating the changes in pH values ​​of two groups of virus suspension samples, the pH 2 group and the pH 2+ electromagnetic wave exposure group, before and after electromagnetic wave irradiation in Example 3 of the present invention;

[0051] Figure 5 This is a graph showing the results of the investigation of pH value changes of two groups of virus suspension samples, the pH 4 group and the pH 4+ electromagnetic wave exposure group, before and after electromagnetic wave irradiation in Example 3 of the present invention;

[0052] Figure 6 This is a graph showing the results of the investigation of pH value changes of two groups of virus suspension samples, the pH 8 group and the pH 8+electromagnetic wave exposure group, before and after electromagnetic wave irradiation in Example 3 of the present invention;

[0053] Figure 7 This is a graph showing the results of the investigation of pH value changes of two groups of virus suspension samples, the pH 12 group and the pH 12+electromagnetic wave exposure group, before and after electromagnetic wave irradiation in Example 3 of the present invention;

[0054] Figure 8 This is a graph showing the results of investigating the changes in virus titer after the inactivation treatment of three groups of virus suspension samples, namely, the pH 2 group, the electromagnetic wave exposure group, and the pH 2+ electromagnetic wave exposure group in Example 3 of the present invention;

[0055] Fig. 9 This is a graph showing the results of investigating the changes in virus titer after the inactivation treatment of three groups of virus suspension samples, namely, the pH 4 group, the electromagnetic wave exposure group, and the pH 4+electromagnetic wave exposure group in Example 3 of the present invention;

[0056] Fig.10 This is a graph showing the results of investigating the changes in virus titer after the inactivation treatment of three groups of virus suspension samples, namely, the pH 8 group, the electromagnetic wave exposure group, and the pH 8+electromagnetic wave exposure group in Example 3 of the present invention;

[0057] Fig.11 This is a graph showing the results of investigating the changes in virus titer after the inactivation treatment of three groups of virus suspension samples, namely, the pH 12 group, the electromagnetic wave exposure group, and the pH 12+electromagnetic wave exposure group in Example 3 of the present invention;

[0058] Fig.12 This is a real-time temperature change diagram of the virus simulation culture medium within 30 minutes of electromagnetic wave exposure in Example 4 of the present invention;

[0059] Fig.13It is a graph showing the virus titer at room temperature (control) in Example 4 of the present invention, and the results of the investigation of the virus titer change under electromagnetic wave exposure to room temperature, ice, and dry ice conditions, wherein A is the investigation result of the virus titer change of the same virus sample after being exposed to electromagnetic waves at room temperature and only placed at room temperature for the same time (control), and B is the investigation result of the virus titer change of the same virus sample after being exposed to electromagnetic waves under different temperature conditions such as room temperature, ice, and dry ice. DETAILED DESCRIPTION

[0060] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

[0061] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. Further, in the description of the present invention, unless otherwise specified, the meaning of "plurality" is two or more.

[0062] It should be noted that the present invention includes samples of coronaviruses, and the pH thereof can be adjusted or by alternative means, so that the coronavirus in the sample is within the expected pH range. "Alternative means" can be, for example, spraying a PBS buffer with a pH value of 7 to 9 on the surface of a solid sample; or dissolving a solid sample (such as a soluble inorganic salt) in a solution with a pH value of 7 to 9; or adjusting the pH value of a liquid, suspension, semi-solid, etc. sample by a strong acid or strong base reagent to make it within the range of 7 to 9.

[0063] Terms and Definitions

[0064] In this article, the term "acidity" refers to the strength of the acidity or alkalinity of a solution, and is represented by the symbol pH. pH is equivalent to "acidity value" and "hydrogen ion concentration index", which is a scale of hydrogen ion activity in a solution, that is, a measure of the acidity or alkalinity of a solution in the usual sense. In conjunction with the context of this article, virus suspensions of different pH values ​​represent virus suspensions of different pH values. For example, a virus suspension of pH 2 represents a virus suspension of pH 2.

[0065] As used herein, the term "electromagnetic waves" refers to a propagating form of energy resulting from the interaction of electric and magnetic fields.

[0066] In this article, the term "electromagnetic wave exposure" refers to the presence of an organism or biological sample in an electromagnetic radiation field, generally referring to the exposure of the human body to an electromagnetic wave radiation field. Combined with the context of this article, it is equivalent to "electromagnetic wave irradiation", "electromagnetic wave radiation", and "electromagnetic radiation". Electromagnetic radiation usually refers to all electromagnetic waves with electromagnetic radiation characteristics. Non-ionizing radiation refers to radio waves, microwaves, infrared rays, visible light, and ultraviolet rays. X-rays and gamma rays are usually considered to be radioactive radiation, called ionizing radiation (Ge Debiao, Yan Yubo. Electromagnetic Wave Time Domain Finite Difference Method. "Xidian University Press", 2002).

[0067] In this article, the term "hertz" is abbreviated as Hz, which is a physical unit of measurement for frequency, referring to the number of periodic fluctuations that occur per second. The main units of measurement for high frequencies are kHz (i.e. 1000Hz), MHz (i.e. 1000kHz), GHz (i.e. 1000MHz), and THz (i.e. 1000GHz). In this article, the term "gigahertz" is equivalent to "GHz" and "gigahertz".

[0068] As used herein, the term "normal temperature" is defined as a temperature between 15 and 45°C.

[0069] In this article, the term "coronavirus" (CoV) refers to a class of enveloped single-stranded positive-strand RNA viruses that can infect humans and a variety of animals, have respiratory, gastrointestinal and nervous system tropism, can cause serious diseases in livestock and companion animals (such as pigs, cattle, chickens, dogs, cats), and can cause humans to suffer from diseases ranging from the common cold to severe acute respiratory syndrome. Coronaviruses that can infect humans include, but are not limited to, human coronavirus 229E (HCoV-229E), NL63 (HCoV-NL63), HKU1 (HCoV-HKU1), OC43 (HCoV-OC43) that cause upper respiratory tract infection symptoms, severe acute respiratory syndrome coronavirus (SARS-CoV) and new coronavirus (SARS-CoV-2), and Middle East respiratory syndrome coronavirus (MERS-CoV). Coronaviruses that can infect animals include, but are not limited to, porcine transmissible gastroenteritis virus (TGEV), porcine delta coronavirus (PDC), porcine hemagglutinating encephalomyelitis virus (PHEV), canine respiratory coronavirus (CrCoV), mouse hepatitis virus, and feline coronavirus (FCoV).

[0070] In this article, the term "mouse hepatitis virus (MHV)" is classified as belonging to the Coronaviridae family and the Coronavirus genus. It is one of the most important viruses that infect mice. Infected mice are distributed throughout the world, but in most cases they are latently infected, subclinically infected, or chronically infected. Under the induction of certain stress factors, the body's resistance is reduced, causing acute illness and death in mice, mainly hepatitis and encephalitis. The main strains of mouse hepatitis virus are MHV-1, MHV-2, MHV-3, MHV-4 (JHM), MHV-A59, and MHV-S. These strains are antigenically similar. MHV has serological cross-reactivity with some strains of other mammalian coronaviruses in this genus, and is often used as a model for studying other coronaviruses ("Experimental Animal Science Course", edited by Hu Jianhua, Yao Ming, and Cui Shufang). MHV is also a commonly used model virus for coronavirus research. It belongs to the 2a subgroup of the β coronavirus genus together with SARS-CoV-2, SARS-CoV, and MERS-CoV. It does not infect humans and is relatively safe. MHV virus particles are spherical or polymorphic, with a diameter of 60 to 160 nm. They have typical rod-shaped protrusions or abrasive particles that are about 20 nm long and 7 nm wide at the top; and are usually inactivated at 56°C for 30 minutes. They have a high degree of structural and thermal sensitivity similarity with SARS-CoV-2 and are the preferred alternative for SARS-CoV-2.

[0071] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be purchased commercially.

[0072] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

[0073] In order to facilitate the investigation of the inactivation effect of the virus inactivation method of the present invention on the above-mentioned coronaviruses, unless otherwise specified, MHV-A59 is selected as a model virus in the embodiments of the present invention to investigate the inactivation effect of the virus inactivation method of the present invention on coronaviruses such as SARS-CoV-2, SARS-CoV and MERS-CoV.

[0074] Example 1: Mouse hepatitis virus (MHV-A59) culture

[0075] 1.1 Culture of mouse hepatitis virus (MHV-A59): Figure 1 ) solution was added to the confluent monolayer or confluent 17CL-1 cells ( Figure 2 ) in a cell culture bottle, adsorb for 2 hours, discard the virus solution, add a medium containing 2% fetal bovine serum to culture the virus, and collect the virus solution after about 48 hours when 2 / 3 of the cells show cytopathic effects. The culture solution containing the virus and host cells is placed at -20°C and repeatedly frozen and thawed 3 times to break the host cells, centrifuged at 3000 rpm for 15 minutes, filtered through a 0.22 micron filter, collected in 1.8 ml cryotubes, and stored in a -80°C ultra-low temperature refrigerator for later use.

[0076] Example 2: Detection of mouse hepatitis virus activity after electromagnetic wave exposure

[0077] 2.1 Virus grouping and electromagnetic wave exposure: Take out the frozen viruses from the -80℃ ultra-low temperature refrigerator, mix all the viruses thoroughly with a vortex oscillator, take 1 ml of the virus stock solution and place it in a 35 mm dish, prepare a total of 15 samples, divided into 0, 15, and 30 minute groups, with 5 replicates in each group. The average power density of the electromagnetic wave radiation source is 100 milliwatts per square centimeter, and the radiation frequency is 2.8 gigahertz. The above three groups of samples are irradiated for 0, 15 and 30 minutes respectively. All virus samples are stored in a 4℃ refrigerator when not irradiated with electromagnetic waves.

[0078] 2.2 Determination of the titer of mouse hepatitis virus (MHV-A59): The virus stock solution was diluted 10 times with a culture medium containing 5% fetal bovine serum. A 96-well cell culture plate containing a monolayer of 17CL-1 cells was taken. Four replicate wells were made for each dilution and a negative control was set up. The plates were placed in a 37°C, 5% carbon dioxide incubator for continuous culture. After 24 hours, the plates were taken out and observed under a microscope. The cytopathic conditions were recorded well by well, and the virus titer of the virus samples was calculated.

[0079] The virus titer was calculated (Reed-Muench method) as the half-cell infection dose. When the cell culture medium containing mouse hepatitis virus (MHV-A59) was continuously passaged on cells for three times without finding cytopathic effects, it was considered to be completely inactivated.

[0080] Logarithm of the 50% cell infection dose = Logarithm of the dilution of the group with a lesion rate higher than 50% + distance ratio

[0081] Wherein, distance ratio = (lesion rate of group above 50% - 50) / (lesion rate of group above 50% - lesion rate of group below 50%)

[0082] The changes in virus titer of each group of samples after 0, 15 and 30 minutes of electromagnetic wave radiation are as follows Figure 3 shown.

[0083] The results showed that the virus titer of the 30-minute group samples was significantly reduced compared with the 0-minute group (P<0.05). The above experimental results show that the electromagnetic wave exposure with an average power density of 100 milliwatts per square centimeter and a radiation frequency of 2.8 gigahertz and a radiation duration of not less than 30 minutes can effectively reduce the activity of mouse hepatitis virus.

[0084] To facilitate the study of the effect of different pH values ​​on the inactivation of viruses by electromagnetic wave radiation, unless otherwise specified, the following embodiments of the present invention adopt the following electromagnetic wave radiation conditions: an average power density of 100 milliwatts per square centimeter and a radiation frequency of 2.8 gigahertz.

[0085] Example 3: Detection of virus activity of mouse hepatitis virus suspensions with different pH values ​​after exposure to electromagnetic waves

[0086] 3.1 Preparation of virus suspensions with different pH values

[0087] 3.1.1 Preparation of virus suspension at pH 2: Take 1 ml of virus suspension and prepare pH 2 virus suspension with 3% hydrogen chloride solution for preparing samples of pH 2 group (2.02±0.04) and pH 2+electromagnetic wave exposure group (1.9±0.00), with 5 replicates in each group. At the same time, set up a blank control group and an electromagnetic wave exposure group, and replace the hydrogen chloride solution with sterile deionized water, with 5 replicates in each group.

[0088] 3.1.2 Preparation of virus suspension at pH 4: Take 1 ml of virus suspension and use 0.1 mol / L citric acid solution to prepare virus suspension at pH 4, which is used to prepare samples of pH 4 group (4.08±0.04) and pH 4+electromagnetic wave exposure group (4.1±0.00), with 5 replicates in each group. At the same time, a blank control group and an electromagnetic wave exposure group were set up, and sterile deionized water was used instead of citric acid solution, with 5 replicates in each group.

[0089] 3.1.3 Preparation of virus suspension at pH 8: Take 1 ml of virus suspension and use 0.2 mol / L sodium dihydrogen phosphate solution to prepare pH 8 virus suspension for the preparation of pH 8 group (7.96±0.05) and pH 8+electromagnetic wave exposure group (7.98±0.04) samples, with 5 replicates in each group. At the same time, set up a blank control group and an electromagnetic wave exposure group, and replace the sodium dihydrogen phosphate solution with sterile deionized water, with 5 replicates in each group.

[0090] 3.1.4 Preparation of virus suspension at pH 12: Take 1 ml of virus suspension and use 0.5 mol / L sodium hydroxide solution to prepare virus suspension at pH 12 for the preparation of samples of pH 12 group (11.92±0.11) and pH 12+electromagnetic wave exposure group (11.96±0.04), with 5 replicates in each group. At the same time, set up a blank control group and an electromagnetic wave exposure group, and replace the sodium hydroxide solution with sterile deionized water, with 5 replicates in each group.

[0091] After preparation, each group of virus samples under the above different pH conditions were placed in a 35 mm dish, stored in a 4°C refrigerator for 24 hours, and then exposed to electromagnetic waves.

[0092] 3.2 Virus grouping and electromagnetic wave exposure: The viruses in each group were uniformly irradiated using an electromagnetic wave radiation source with an average power density of 100 milliwatts per square centimeter for 30 minutes. The viruses in the blank control group and the pH group were placed in the same position on the irradiation table without being irradiated.

[0093] After the radiation, the pH values ​​of the virus suspensions in all pH groups (pH 2, 4, 8, 12) and pH+EMW exposure groups were tested, and the changes in the pH values ​​of the virus suspensions before and after radiation were investigated (the results are shown in Table 2). Figure 4 , 5 , 6, and 7). The experimental results showed that there was no significant difference in the pH values ​​of virus suspensions in different pH groups and in the pH values ​​of virus suspensions in the same group before and after radiation (P>0.05), and the pH of the virus solution was stable within 24 hours. Therefore, the results of each experimental group were credible and comparable. Finally, the virus suspensions in all pH gradients (pH2, 4, 8, 12) and pH+electromagnetic wave exposure groups were buffered to neutral (pH 7).

[0094] 3.3 Mouse hepatitis virus titer determination: Same as Example 2.

[0095] The experimental results are as follows Figures 8 to 11 shown.

[0096] Changes in virus titer at pH 2 Figure 8 As shown. Compared with the blank control group, the viral activity in the pH 2 group, the electromagnetic wave exposure group, and the pH2+electromagnetic wave exposure group was significantly reduced (P<0.001). Compared with the pH 2 group and the electromagnetic wave exposure group, the viral activity in the pH 2+electromagnetic wave exposure group was significantly reduced (P<0.001), and when the pH value of the virus environment was 2 and the electromagnetic wave was exposed for 30 minutes, the virus was completely inactivated. However, according to statistical analysis, pH 2 and electromagnetic wave exposure for 30 minutes did not show a synergistic effect in inactivating mouse hepatitis virus (P>0.05).

[0097] At pH 4, the virus titer changes as Fig. 9As shown. Compared with the blank control group, the viral activity of the electromagnetic wave exposure group and the pH 4+ electromagnetic wave exposure group was significantly reduced (P<0.001), and there was no significant change in the viral activity of the pH 4 group. Compared with the pH 4 group, the viral activity of the electromagnetic wave exposure group and the pH 4+ electromagnetic wave exposure group was significantly reduced (P<0.001). Compared with the electromagnetic wave exposure group, there was no significant change in the viral activity of the pH4+ electromagnetic wave exposure group. According to statistical analysis, pH 4 and electromagnetic wave exposure for 30 minutes did not show a synergistic effect in inactivating mouse hepatitis virus (P>0.05).

[0098] The changes of virus titer at pH 8 are as follows Fig.10 As shown. Compared with the blank control group, the viral activity of the electromagnetic wave exposure group and the pH 8+electromagnetic wave exposure group was significantly reduced (P<0.001), and there was no significant change in the viral activity of the pH 8 group. Compared with the pH 8 group, the viral activity of the electromagnetic wave exposure group and the pH 8+electromagnetic wave exposure group was significantly reduced (P<0.001). Compared with the electromagnetic wave exposure group, the viral activity of the pH8+electromagnetic wave exposure group was significantly reduced (P<0.001). Contrary to the inventor's expectations, after statistical analysis, pH 8 and electromagnetic wave exposure for 30 minutes showed a synergistic effect in inactivating mouse hepatitis virus (P<0.001).

[0099] Virus titer changes at pH 12 Fig.11 As shown. Compared with the blank control group, the viral activity in the pH 12 group, the electromagnetic wave exposure group, and the pH 12 + electromagnetic wave exposure group was significantly reduced (P<0.001). Compared with the electromagnetic wave exposure group, the viral activity in the pH 12 + electromagnetic wave exposure group was significantly reduced (P<0.001). And the viruses in the pH 12 group and the pH 12 + electromagnetic wave exposure group were completely inactivated. According to statistical analysis, pH 12 and electromagnetic wave exposure for 30 minutes did not show a synergistic effect in inactivating mouse hepatitis virus (P>0.05).

[0100] The above experimental results show that under electromagnetic wave radiation conditions that can inactivate mouse hepatitis virus (for example, the radiation power density of the electromagnetic wave is 100-120 milliwatts per square centimeter, and the radiation frequency of the electromagnetic wave is 2.8-3.0 gigahertz), adjusting the pH value of the virus sample to 7-9 can significantly enhance the effect of electromagnetic wave radiation inactivating the virus; the two factors that affect the electromagnetic wave virus inactivation effect, the electromagnetic wave radiation conditions and the virus sample pH, work synergistically and are statistically significant.

[0101] Example 4: Detection of mouse hepatitis virus (MHV-A59) activity after electromagnetic wave exposure (different temperatures)

[0102] 4.1 Virus grouping: The viruses were divided into normal temperature group (20.0℃), electromagnetic wave + normal temperature group (20.0℃), electromagnetic wave + ice group (3.5℃), and electromagnetic wave + dry ice group (0℃), with 5 replicates in each group.

[0103] 4.1.1 Normal temperature group: Place 1 ml of virus stock solution in a 35 mm dish.

[0104] 4.1.2 Electromagnetic wave + normal temperature group: 1 ml of virus stock solution was placed in a 35 mm dish and exposed to electromagnetic waves at normal temperature.

[0105] 4.1.3 Electromagnetic wave + ice group: Take 1000g of ice and spread it on the bottom of the foam box. Place a 35mm dish containing 1ml of virus stock solution on top of the ice and wait until its temperature stabilizes at 3.5℃.

[0106] 4.1.4 Electromagnetic wave + dry ice group: Take 500g of dry ice and spread it on the bottom of the foam box. Place a 35mm dish containing 1ml of virus stock solution on top of the dry ice and wait until its temperature stabilizes at 0℃.

[0107] 4.2 Electromagnetic wave exposure: Each group of virus samples was placed on an irradiation table and irradiated uniformly using an electromagnetic wave radiation source with an average power density of 100 milliwatts per square centimeter for 30 minutes.

[0108] 4.3 Real-time temperature measurement with optical fiber: Take 1 ml of deionized water and put it into a 35 mm dish to simulate virus culture medium. Drill a hole with a diameter of 1 mm 5 mm above the edge of the dish cover. Place it in the irradiation room to equilibrate the room temperature 4 hours in advance. Insert the optical fiber probe along the hole and connect the power supply for real-time temperature measurement. The temperature change is as follows: Fig.12 As shown, during the 30-minute electromagnetic wave exposure, the temperature rose by 17.51°C. The temperature of the electromagnetic wave + ice group was 3.5°C before exposure and 4.5°C after exposure, with a temperature rise of 1°C. This temperature rise had no significant effect on the activity of the virus in a short period of time.

[0109] 4.4 Mouse hepatitis virus titer determination: Same as Example 2. The results are as follows Fig.13 As shown, compared with the normal temperature group, the virus activity in the electromagnetic wave + normal temperature group was significantly reduced (P<0.001). Compared with the electromagnetic wave + normal temperature group, after ice and dry ice cooling treatment, electromagnetic waves had no obvious effect on the activity of the virus, indicating that electromagnetic waves are suitable for virus inactivation at normal temperature, and temperature plays an important role in the process of electromagnetic wave virus inactivation.

[0110] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0111] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A method for inactivating viruses, characterized in that: include: The sample is irradiated with electromagnetic waves, wherein the radiation power density of the electromagnetic waves is 100 to 120 milliwatts per square centimeter, and the radiation frequency of the electromagnetic waves is 2.8 to 3.0 gigahertz; The sample includes coronavirus, and the pH value of the sample is 7-9.

2. The method according to claim 1, characterized in that The radiation power density of the electromagnetic wave is 100 milliwatts per square centimeter, and the radiation frequency of the electromagnetic wave is 2.8 gigahertz; Optionally, the temperature of the sample is room temperature; Optionally, the temperature is 15 to 42.5°C; Optionally, the temperature is 15 to 32.5°C; Optionally, the temperature is 20 to 37.5°C; Optionally, the temperature is 25-42.5°C.

3. The method according to claim 1, characterized in that The radiation duration of the electromagnetic waves is not less than 15 minutes; Optionally, the radiation duration of the electromagnetic waves is not less than 30 minutes.

4. The method according to claim 1, characterized in that: The pH value of the sample is 7.5-8.5; Optionally, the pH of the sample is 8.

5. The method according to claim 1, characterized in that: The sample is selected from an aerosol sample, a liquid sample, a suspension sample or a semi-solid sample; Optionally, the sample is a suspension sample; Optionally, the pH value of the suspension sample is 7.5 to 8.5; Optionally, the pH value of the suspension sample is 8.

6. The method according to claim 1, characterized in that The radiation is carried out at room temperature; Optionally, the irradiation is performed at 15-40°C; Optionally, the irradiation is performed at 15-25°C.

7. The method according to claim 1, characterized in that The coronavirus is a mammalian coronavirus.

8. The method according to claim 7, characterized in that The mammalian coronavirus is selected from at least one of mouse hepatitis virus, human coronavirus HCoV-229E, human coronavirus HCoV-OC43, human coronavirus HCoV-HKUI, human coronavirus HCoV-NL63, human coronavirus MERS-CoV, human coronavirus SARS-CoV and human coronavirus SARS-CoV-2; Optionally, the mouse hepatitis virus is selected from at least one of MHV-1, MHV-2, MHV-3, MHV-4 (JHM), MHV-A59 and MHV-S; Optionally, the mouse hepatitis virus is MHV-A59; Optionally, the mammalian coronavirus is selected from at least one of SARS-CoV-2, SARS-CoV and MERS-CoV or a model virus thereof.

9. A device for inactivating viruses, characterized in that: include: An electromagnetic wave transmitting module, used for irradiating the sample; A control module is used to control the electromagnetic wave transmitting module, the control module is connected to the electromagnetic wave transmitting module, and the control module is controlled based on the method described in any one of claims 1 to 8.

10. A computer-readable storage medium, characterized in that: The storage medium stores a program, which can be executed by a processor to implement the method described in any one of claims 1 to 8 or to implement the virus inactivation function of the device described in claim 9.

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