JP2004130196A - Method for decomposing and disinfecting virus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for decomposing and disinfecting a virus contained in water thinly, and an apparatus therefor. <P>SOLUTION: In a preferable mode, SRSV contained in sewage discharge water thinly is decomposed and disinfected by preparing an ultraviolet ray emitting device using a medium pressure mercury lamp as an ultraviolet ray source and a photocatalyst for generating radicals upon the reception of the irradiation with ultraviolet rays and exposing SRSV to both of ultraviolet rays emitted from the ultraviolet ray emitting device and radicals generated from the photocatalyst to decompose the virus. An apparatus, which decomposes and disinfects the virus contained in water thinly, includes the ultraviolet ray emitting device using the medium pressure mercury lamp as the ultraviolet ray source and the photocatalyst for generating radicals upon the reception of the irradiation with ultraviolet rays device using the medium pressure mercury lamp as the ultraviolet ray source and the photocatalyst for generating radicals upon the reception of the irradiation with ultraviolet rays. <P>COPYRIGHT: (C)2004,JPO

Description

【００５２】
図５に、ＰＣＲ分析の標的としたＳＲＳＶのＲＮＡポリメラーゼ遺伝子についてＰＣＲにより増幅されるＤＮＡ断片を模式的に示す。ＳＲＳＶはｇｅｎｏｔｙｐｅ　Ｉとｇｅｎｏｔｙｐｅ　ＩＩに大別されるため、いずれの遺伝子型についても検出できるＰＣＲプライマー・セットを使用した。図５中、Ｇ１、Ｇ２、ＳＭ３１は、ＲＴ−ＰＣＲ用プライマーを示し、ＮＩ、Ｅ３は、ｎｅｓｔｅｄ　ＰＣＲ用プライマーを示す。
【００５３】
結果：図６に、ｎｅｓｔｅｄ　ＰＣＲ産物の電気泳動写真を示す。実験原水（対照）、及びＩＩＩ系（紫外線（低圧））水について、ＳＲＳＶ　ｇｅｎｏｔｙｐｅ　１及び２の共通断片に相当する約１１０塩基対のＰＣＲ増幅ＤＮＡ断片が明確に観察された。ＩＩ系（紫外線（中圧））水については、上記約１１０塩基対断片が僅かに観察された。一方、Ｉ系（光触媒／紫外線（中圧））水については、上記約１１０塩基対断片は、全く観察されなかった。したがって、紫外線中圧水銀ランプと光触媒を併用することにより、下水処理（放流）水中に希薄に含まれるＳＲＳＶを完全に分解消毒することが分かる。
【００５４】
【発明の効果】
紫外線中圧水銀ランプと光触媒を併用することにより、中圧紫外線による直接的なウイルス不活化に加え、中圧紫外線が光触媒を活性化して生じるラジカルの効果によってウイルス粒子を分解して、下水処理（放流）水中に希薄に含まれるウイルスを分解消毒することができる。
【００５５】
【配列表】

【図面の簡単な説明】
【図１】本発明に係る紫外線／光触媒装置の概略図。
【図２】本発明に係る実験装置のフロー、及び実験装置の主な仕様を示す図。
【図３】ウイルス検査フロー図。
【図４】各実験系の図面に代わる写真。
【図５】ＰＣＲ分析の標的遺伝子に関する模式図。
【図６】各実験系水及び実験原水（対照）についてのｎｅｓｔｅｄ　ＰＣＲの電気泳動写真。
【図７】本発明に係る紫外線中圧水銀ランプと光触媒との併用効果の概説図。[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a method and an apparatus for eliminating and poisoning a virus diluted in water. More particularly, the present invention relates to a method and an apparatus for removing and poisoning SRSV contained in sewage treatment (discharge) water in combination with an ultraviolet and medium pressure mercury lamp and a photocatalyst.
[0002]
[Prior art]
As a pathogen of food poisoning caused by seafood and drinking water, a microspherical virus (hereinafter, referred to as SRSV (Small Round Structured Virus)) has been raised as a problem. In particular, if SRSV is detected in cultured oysters, the commercial value of raw oysters will be significantly reduced, causing considerable damage to the local economy where fishing is the main industry.
[0003]
SRSV is not believed to grow in the body of fish and shellfish. Therefore, for example, the fact that SRSV is detected in the body of an oyster means that the oyster enters the growing area of the oyster via sewage, etc., and is concentrated and retained in the oyster body as a result of oyster feeding behavior. I do. Therefore, a technology for removing SRSV in sewage discharge water, which is a source of SRSV pollution, is keenly desired.
[0004]
It is generally said that viruses are difficult to inactivate by chlorination. It is also reported that SRSV cannot be inactivated at a chlorine concentration of 10 mg / L. On the other hand, ultraviolet (UV) disinfection using a low-pressure mercury lamp is generally considered to be effective because it damages the gene of a virus photochemically. There is a report that it cannot be distinguished from DNA by the PCR method (see Non-Patent Document 7 described below). On the other hand, there is also a report that it is difficult for bacteria to recover light when a medium-pressure mercury lamp is used as an ultraviolet light source (see Non-Patent Document 10 described below). In any case, to date, there has been no report examining the effect of UV disinfection on SRSV.
[0005]
As a sanitary inspection of seafood and drinking water, a method of checking for the presence or absence of virus contamination by an RT-PCR method or the like is employed. However, as described above, in the RT-PCR method, regardless of whether or not the virus is inactivated, a positive determination is made that the inactivated virus is present in the test sample. Therefore, there is a demand for a technique for completely decomposing virus particles to such an extent that it cannot be detected even by RT-PCR or the like, thereby disinfecting a virus diluted in sewage discharge water.
[0006]
As described below, the present inventors used a UV-pressure medium-pressure mercury lamp and a photocatalyst in addition to direct virus inactivation by the medium-pressure UV, and radicals generated by the medium-pressure UV activation of the photocatalyst. A method and an apparatus have been developed for decomposing virus particles by the effect of (1) and eliminating and poisoning the virus diluted in sewage treatment (discharge) water. In the development of the method and the device, it is necessary to establish in advance a test method to confirm whether the virus contained in water is diluted and poisoned in order to evaluate the performance of the method and the device. Was. Therefore, the present invention has been completed only after such an inspection method has been established.
[0007]
Hereinafter, a description will be given of a test method for confirming whether or not a virus diluted in water is detoxified, and a prior art relating to the virus detoxification method and apparatus according to the present invention.
[0008]
In general, there are many reports on the detection of viruses in fish and shellfish containing high concentrations of viruses.However, viruses in very dilute water, especially in sewage effluent containing large amounts of contaminants, have been reported. There are few reports on the detection method, and there is no report on SRSV.
[0009]
In Non-Patent Document 1 listed below, DEAE-cellulose, which is an anion exchanger, is directly added to sample water (0.03%) to adsorb viruses in water, and then cellulose is flocculated with an anionic organic flocculant. A method is described in which the aggregate is collected with a nylon mesh, and the virus is induced with a beef extract solution. The virus used in Non-Patent Document 1 is attenuated poliovirus type 1 (Lsc, 2ab strain). Since the above method is a batch process, it is not suitable for treating a large amount of water. Further, it is necessary to appropriately examine the optimal injection amount of the flocculant in accordance with the test water.
[0010]
Non-Patent Document 2 listed below describes a method for concentrating enterovirus in 100 to 1000 L of sample water using a special glass wool, oiled sodatic glass glass (Saint Gobain R.725). The glass wool filter recovered 62-75% of some enteroviruses (laboratory or wild strain) and rotavirus SA11 added to tap water. The above method recovered 62% and 57% of poliovirus added to river water and treated wastewater, respectively. The above method has the advantage that it is not necessary to change the pH of the treated water because no ion exchanger is used, but has the disadvantage that the treatment flow rate is small and that the special glass wool is difficult to obtain.
[0011]
Non-Patent Document 3 listed below discloses a method for simultaneously concentrating enterovirus, hepatitis E virus, and rotavirus from a drinking water sample by passing through a filtration column filled with granular activated carbon (GAC). Urea-arginine phosphate buffer (UAPB) is used as eluent at pH 9.0 for effective desorption and elution from GAC. By further concentrating the virus with magnesium chloride, nucleic acid extraction, cDNA synthesis, and synthesis using primer sets specific to each virus become possible. The efficiency of the method in a 100 L water sample inoculated with poliovirus-1 was 74% in the activated carbon based UAPB-RT-PCR. However, the applicability of the above method to SRSV is unknown.
[0012]
Patent Document 1 listed below describes a virus concentration material having an antibody against a virus on the surface, and a virus concentration method using the material. However, in the above method, it is necessary to prepare an antibody for each virus species to be separated and concentrated, and it is not suitable for treating a large amount of water.
[0013]
Patent Document 2 listed below discloses a method for capturing a virus from a fluid containing a virus, in which a polymer having a monomer having an anionic group in a molecule as one component is retained on at least a part of a substrate. A virus capturing method characterized by passing a fluid containing the virus through the virus capturing body thus formed and adsorbing the virus to the virus capturing body is described. The above-mentioned method aims at increasing the capture rate of poliovirus, which was difficult to capture by the conventional method, and the capture rate of poliovirus is improved to 90% or more in the pH range of 4 to 9. However, the applicability of the above method to SRSV is unknown.
[0014]
Non-Patent Document 4 listed below discloses that coliphages in drinking water and wastewater samples are adsorbed onto a positively charged membrane filter, eluted with a urea-arginine phosphate buffer (UAPB), reconcentrated, and reconcentrated. E. coli C (ATCC 13706). The membrane filter-based UAPB method for enrichment and detection of coliphage is comparable to the beef extract and reconcentration procedure described in the Standard Methods for the Examination of Water and Wastewater and coliphage detection. In particular, when the phage titer is very low, the method described in Non-Patent Literature 4 uses the UAPB from a positively charged membrane filter to test the entire amount of the concentrated sample, and thus the method described in the above-mentioned standard method is used. Gives high coliphage recovery. However, the method described in Non-Patent Document 4 has a disadvantage that the filter is easily clogged by a suspended component in the test water, and it is unknown whether the method can be applied to SRSV.
[0015]
Non-Patent Document 5 listed below describes a nested reverse transcriptase PCR assay for the detection of SRSV in Molluscan Shellfish containing environmentally contaminated oysters.
[0016]
Non-Patent Document 6, listed below, states that SRSV does not multiply or produce toxins in food (fish and shellfish), and that food simply acts as a vehicle for passive transport of SRSV; It is a review on SRSV and the like, describing that it is a major contamination route of fish and shellfishes living along the coast.
[0017]
Non-Patent Document 7 listed below describes that E. coli RNA phage Qβ is inactivated by UV irradiation, and the methods of RT-PCR and nested PCR are described. However, it is described that the inactivated virus is detected as positive in the RT-PCR method.
[0018]
Non-Patent Document 8 listed below describes that an anion exchange fiber was added to a sample collected from a natural water body, aggregated and precipitated, and the precipitate was separated to concentrate the virus.
[0019]
Non-Patent Document 9 listed below is a review on SRSV and the like. It also states that there is no SRSV activity test method.
[0020]
Non-Patent Document 10 listed below describes that when a medium-pressure mercury lamp is used as an ultraviolet light source, light recovery of bacteria hardly occurs.
[0021]
Patent Document 3 listed below describes an apparatus for removing turbidity, bacteria, viruses, and trihalomethane precursors in raw water by membrane separation and ozone treatment. The virus concentrate that has been subjected to solid-liquid separation is to be discarded, but there remains a problem in its treatment method.
[0022]
Patent Literature 4 listed below describes a small and lightweight water purification device using a hollow fiber formed of a reverse osmosis membrane. Since the small-sized water purification device is small in size, it does not have a high processing capacity, and there remains a problem in a method of disposing of the separated virus concentrate.
[0023]
Patent Literature 5 listed below describes a method for removing minute substances such as viruses by passing an electric current between the electrodes to denature the micro-harmful substances including viruses or to adsorb them to the electrodes to make them harmless. I have. Even if the virus can be adsorbed to the electrode, it is unknown whether the virus has been damaged and the growth function has been lost or has simply been adsorbed, and there remains a problem in a method of disposing of the adsorbed virus concentrate.
[0024]
Patent Literature 6 listed below describes a method of removing viruses by predating viruses on ciliates that live in a filter tank. The above removal method is a biological treatment, and there is no guarantee that a high removal is always possible.
[0025]
Patent Literature 7 listed below describes a water purification method using ozone oxidation, in which turbidity by membrane filtration and promotion of an ozone oxidation reaction by a catalyst are combined. Ozone treatment may be able to inactivate the virus, but it is unclear whether the virus can be degraded.
[0026]
Patent Literature 8 listed below describes a method of dispersing a photocatalyst in blood or the like infected with a virus, and irradiating ultraviolet rays to reduce the infectivity. The ultraviolet light used is near-ultraviolet light with a wavelength of about 350 nm from black light, and the photocatalyst used is described as titanium dioxide. However, no examples showing how much the infectious titer of the virus was actually reduced are described. Also, there is no description as to whether the virus can be degraded. Since the target is blood, the applicability to wastewater treatment technology remains to be solved in terms of treatment efficiency and the like because the scale of the apparatus is significantly different.
[0027]
Patent Document 9 listed below discloses a device in which a photocatalyst contacts a material held on a substrate surface with blood or a blood product, and irradiates the photocatalyst on this device with light to remove contaminating viruses and the like. A system for reducing infectivity is described. Photocatalyst used _{is, TiO 2, ZnO, SrTiO 3} , Fe 2 O 3, CdS, CdSe, WO 3, FeTiO 3, GaP, GaAs, RuO 2, MoS 3, LaRhO 3, CdFeO 3, BiO 3, MoS 2 , In ₂ O ₃ , CdO, and SnO ₂ . The light irradiation device used is a 100 W xenon lamp. However, no examples showing how much the infectious titer of the virus was actually reduced are described. Also, there is no description as to whether the virus can be degraded. Since the target is blood, the applicability to wastewater treatment technology remains to be solved in terms of treatment efficiency and the like because the scale of the apparatus is significantly different.
[0028]
[Non-patent document 1]
Yano et al., "Examination of virus testing method in water by cellulose adsorption / aggregation method," Water Works Association Magazine, Vol. 60, No. 5 (No. 680), pp. 10-23 (1991).
[Non-patent document 2]
Ph. Vilagin et al. , "Glass Wool for Virus Concentration at Ambient Water pH Level", Wat. Sci. Tech. Vol. 27, No. 3-4 pp. 299-306 (1993)
[Non-Patent Document 3]
N. Jothikumar et al. , "A simple device for the concentration and detection of enterovirus, hepatitis E virus and rotavirus from water samples by reverse transcription-polymerase chain reation", Journal of Virological Methods 55 (1995) 401-415
[Non-patent document 4]
N. Jothikumar et al. , "Elution and reconcentration of coliphages in water from positively charged membrane filters with urea-arginine phosphate buffer".
[Non-Patent Document 5]
J. Green, et al. , "A nested reverse transcriptase PCR assay for detection of small round-structured virtues in environmentally related annually related overseas marketing. 64, No. 3, Mar, 1998, p. 858-863
[Non-Patent Document 6]
Hazle Appleton, "Control of food-born viruses", British Medical Bulletin 2000; 56 (No. 1), 172-183.
[Non-Patent Document 7]
Katayama et al. (1997) Quantification of RNA phage inactivated by UV irradiation by RT-PCR, Journal of Environmental Engineering, 34: 83-91.
[Non-Patent Document 8]
Tomoko Kitahashi et al. (1999) Detection of HAV, SRSV, and Astrovirus Genes from Native Oysters along the Coast of Chiba City, Journal of Infectious Diseases, 73 (6): 559-564.
[Non-Patent Document 9]
D. Lees, "Virus and bivalve shellfish", International Journal of Food Microbiology 59 (2000) 81-116.
[Non-Patent Document 10]
Kumiko Oguma et al., Evaluation of light recovery characteristics of medium-pressure ultraviolet lamp by quantification of pyrimidine dimer, 36th Annual Meeting of Japan Society on Water Environment, p390, August 21, 2002 [Patent Document 1]
Japanese Patent Application Laid-Open No. 2002-78485 [Patent Document 2]
JP-A-5-84071 [Patent Document 3]
JP-A-6-328069 [Patent Document 4]
JP-A-10-137754 [Patent Document 5]
Japanese Patent Application Laid-Open No. H5-169064 [Patent Document 6]
Japanese Patent Application Laid-Open No. Hei 10-137783 [Patent Document 7]
Japanese Patent Application Laid-Open No. H10-113659 [Patent Document 8]
JP-A-8-23970 [Patent Document 9]
JP 2000-41667 A
[Problems to be solved by the invention]
As described above, in the RT-PCR method, a positive determination is made that the inactivated virus is present in the test sample regardless of whether the virus is inactivated. Therefore, there is a demand for a technique for completely decomposing virus particles to such an extent that it cannot be detected even by the RT-PCR method or the like, thereby disinfecting the virus diluted in the sewage discharge water.
[0030]
[Means for Solving the Problems]
As a result of repeated experiments to solve the above-mentioned problems, the present inventors have found that, by using a UV-Medium pressure mercury lamp and a photocatalyst in combination, in addition to direct virus inactivation by the medium-pressure UV light, The present invention has been completed by developing a method and an apparatus for decomposing virus particles by the effect of radicals generated by activation and for eliminating and poisoning the virus diluted in sewage treatment (discharge) water.
[0031]
That is, according to one aspect of the present invention, there is provided a method for eliminating and poisoning a virus diluted in water, comprising the following steps:
An ultraviolet ray generator using a medium pressure mercury lamp as an ultraviolet ray source, and a photocatalyst which generates radicals upon irradiation with the ultraviolet ray are prepared;
The water is exposed to both the ultraviolet light generated from the ultraviolet light generator and the radical generated from the photocatalyst to decompose the virus,
There is provided the method comprising:
[0032]
The water can be sewage (treatment) effluent. Before exposing the water to the ultraviolet rays or radicals, the turbid matter may be removed in advance by pre-filtration.
[0033]
The virus can be SRSV. The exposing is for a period of time sufficient to damage the DNA of the virus and to degrade the envelope of the virus. The degradation is degradation of damaged DNA to the extent that DNA specific to the virus cannot be detected by PCR.
[0034]
In another aspect of the present invention, there is provided a device for removing and poisoning a virus diluted in water, comprising:
An ultraviolet ray generator using a medium pressure mercury lamp as an ultraviolet ray source; and a photocatalyst that generates radicals upon irradiation with the ultraviolet ray,
There is provided the device comprising: The water can be sewage (treatment) effluent. Further, the virus may be SRSV.
[0035]
As shown in Example 2 below, a nested PCR detects a fragment of about 110 base pairs derived from SRSV only by ultraviolet light (low pressure) and ultraviolet light (medium pressure). On the other hand, according to the present invention, the combined use of the ultraviolet / medium pressure mercury lamp and the photocatalyst makes it impossible to detect a fragment of about 110 base pairs derived from SRSV in the nested PCR. This means that the SRSV target gene is fragmented as a fragment shorter than about 110 base pairs in length according to the method of the present invention. While not wishing to be bound by any particular theory, the combined use of a UV-Medium pressure mercury lamp and a photocatalyst allows direct virus inactivation by the medium-pressure UV light, as well as the hydroxyl generated by the medium-pressure UV light activating the photocatalyst. It is thought that radicals and superoxide ions decompose the envelope of the virus, and accordingly, DNA damaged by medium-pressure ultraviolet rays is further decomposed. This is outlined in FIG.
[0036]
The present invention will be described in detail with reference to the following examples and the accompanying drawings, which should not be construed as limiting the scope of the present invention.
[0037]
【Example】
Example 1: Experimental apparatus and method Ultraviolet / photocatalyst according to the present invention was installed in a sewage treatment plant in a certain town in Iwate prefecture in order to verify the effect of eliminating SRSV in sewage discharge water by ultraviolet / photocatalyst. A demonstration experiment was conducted on the effect of the method and apparatus for removing and poisoning SRSV components according to the present invention in winter with the apparatus installed.
[0038]
FIG. 1 shows a schematic diagram of an ultraviolet / photocatalytic device according to the present invention.
[0039]
A photocatalyst having a SUS corrugated foil surface coated with titanium by ion plating and having a cylindrical shape was inserted around the ultraviolet and medium pressure mercury lamp.
[0040]
As Comparative Example 1, a medium-pressure mercury lamp without a photocatalyst was used, and as Comparative Example 2, a low-pressure mercury lamp without a photocatalyst was used.
[0041]
FIG. 2 shows a flow of the experimental apparatus and main specifications of the experimental apparatus.
[0042]
Biologically treated water at a sewage treatment plant (before chlorination) is used as experimental raw water, and is used as an I system (photocatalyst / ultraviolet light (medium pressure)), a II system (ultraviolet light (medium pressure)), and a III system (ultraviolet light (low pressure)). ), And the water after passing through each system and the experimental raw water (control) were sampled, and the presence or absence of SRSV was confirmed according to the virus test flow shown in FIG. The lower part of FIG. 2 shows the main specifications of the experimental apparatus of each system described above.
[0043]
FIG. 4 shows a photograph replacing the drawing of each system.
[0044]
As described above, FIG. 3 shows a schematic diagram of a test flow of a virus diluted in a large amount of water. For example, a sample water (sample) from which contaminants have been removed in advance by a means such as pre-filtration is passed through a 50 mL column filled with DEAE cellulose at a rate of, for example, 10 L / hour (concentration step). The virus adsorbed on DEAE is eluted with a small amount, for example, 60 mL of an alkaline solution (elution step). The virus is precipitated and separated from the eluate using polyethylene glycol (sedimentation separation step), RNA is extracted from the precipitate (RNA extraction step), and this is amplified by RT-PCR and / or nested PCR. The DNA fragment thus obtained is observed on an electrophoretic photograph. By performing nested PCR (nested PCR) after RT-PCR (reverse transcription PCR), a large amount of virus-specific fragments can be amplified if there is a small amount of viral RNA.
[0045]
Example 2: Assay of SRSV in system I (photocatalyst / ultraviolet (medium pressure)) water, system II (ultraviolet (medium pressure)) water, system III (ultraviolet (low pressure)) water, and experimental raw water (control) < About 10 L of each title water was collected and stored refrigerated.
[0046]
In the pre-filtration, the water was passed through a filter (No. 5A) to remove dust, and then passed through a DEAE column (10 L / hour). The DEAE column eluate was assayed for the presence of SRSV (see FIG. 3).
[0047]
The PCR reaction and electrophoresis conditions used are shown below:
(1) RNA Extraction The eluate of the DEAE column was reconcentrated with polyethylene glycol to reconcentrate the virus. 140 µL of each precipitate solution was extracted with QIAGEN's QIAamp viral RNA MINI Kit to extract RNA to obtain 60 µL of RNA solution. Was.
(2) RT-PCR reaction TaKaRa RNA LA PCR Kit (AMV) ver. The RT-PCR reaction was performed using 1.1.
[0048]
i) RT reaction reaction solution composition (20 μL): as in the Kit instruction manual, with the following changes:
RNA solution volume used: 5 μL
Using primers: _{N 9} that comes in the Kit, and SM31
Reaction conditions: 30 ° C: 10 minutes → 42 ° C: 30 minutes → 99 ° C: 5 minutes → 5 ° C: ∞
ii) PCR reaction solution composition: as per the Kit instructions above, with the following changes:
RNA solution volume used: 50 μL (of which 20 μL is an RT reaction solution)
Mg concentration: about 1.5 to 2 mM
Primers used: G1 and G2 (and SM31 derived from the above RT reaction solution)
Reaction conditions: # 1 = 94 ° C: 2 minutes # 2 = 95 ° C: 1 minute → 40 ° C: 1 minute → 72 ° C: 1 minute (50 cycles)
# 3 = 72 ° C: 10 minutes → 4 ° C: ∞
(3) Nested PCR reaction solution composition (50 μL): as in the Kit instruction manual, with the following changes:
5 μL of an RT-PCR reaction solution was used as a template.
[0049]
Primers used: NI and E3
Reaction conditions: # 1 = 94 ° C: 2 minutes # 2 = 95 ° C: 1 minute → 40 ° C: 1 minute → 72 ° C: 1 minute (30 cycles)
# 3 = 72 ° C: 10 minutes → 4 ° C: ∞
(4) Confirmation of PCR-amplified DNA fragment by electrophoresis By agarose electrophoresis (3.0% by mass NuSieve 3: 1 agarose / 0.5 × TBE buffer system), an RT-PCR reaction solution and a nested PCR reaction solution were used, respectively. 10 μL was run. The agarose gel after electrophoresis was stained with a fluorescent dye, ethidium bromide, and visualized on a transilluminator to observe the intensity of the DNA band.
[0050]
Primers used for the PCR reaction were prepared according to Non-Patent Document 5. The primers used have the base sequences shown in Table 1 below:
[0051]
[Table 1]

[0052]
FIG. 5 schematically shows a DNA fragment amplified by PCR for the SRSV RNA polymerase gene targeted for PCR analysis. Since SRSV is roughly classified into genotype I and genotype II, a PCR primer set that can detect any genotype was used. In FIG. 5, G1, G2, and SM31 indicate RT-PCR primers, and NI and E3 indicate nested PCR primers.
[0053]
Results : FIG. 6 shows an electrophoresis photograph of the nested PCR product. For experimental raw water (control) and III-system (ultraviolet (low pressure)) water, a PCR amplified DNA fragment of about 110 base pairs corresponding to the common fragment of SRSV genotypes 1 and 2 was clearly observed. With respect to type II (ultraviolet (medium pressure)) water, the above approximately 110 base pair fragment was slightly observed. On the other hand, for I-system (photocatalyst / ultraviolet (medium pressure)) water, the above-mentioned approximately 110 base pair fragment was not observed at all. Therefore, it can be seen that the combined use of the ultraviolet- and medium-pressure mercury lamp and the photocatalyst completely eliminates and poisons SRSV that is rarely contained in the sewage treatment (discharge) water.
[0054]
【The invention's effect】
By using a UV-Medium pressure mercury lamp and a photocatalyst together, in addition to direct virus inactivation by the medium-pressure UV, virus particles are decomposed by the effect of radicals generated when the medium-pressure UV activates the photocatalyst, and sewage treatment ( (Discharge) It is possible to dissolve and poison viruses that are rarely contained in water.
[0055]
[Sequence list]

[Brief description of the drawings]
FIG. 1 is a schematic diagram of an ultraviolet / photocatalytic device according to the present invention.
FIG. 2 is a diagram showing a flow of an experimental apparatus according to the present invention and main specifications of the experimental apparatus.
FIG. 3 is a flowchart of a virus test.
FIG. 4 is a photograph replacing a drawing of each experimental system.
FIG. 5 is a schematic diagram of a target gene for PCR analysis.
FIG. 6 is an electrophoresis photograph of nested PCR for each experimental system water and experimental raw water (control).
FIG. 7 is a schematic diagram showing the combined effect of the ultraviolet and medium pressure mercury lamp and the photocatalyst according to the present invention.

Claims (9)

A method for removing and poisoning a virus diluted in water, comprising the following steps:
An ultraviolet ray generator using a medium pressure mercury lamp as an ultraviolet ray source, and a photocatalyst which generates radicals upon irradiation with the ultraviolet ray are prepared;
The water is exposed to both the ultraviolet light generated from the ultraviolet light generator and the radical generated from the photocatalyst to decompose the virus,
The method comprising: The method of claim 1, wherein the water is sewage (treatment) effluent. The method according to claim 1, wherein the turbid matter is previously removed by pre-filtration before exposing the water to the ultraviolet rays or radicals. 2. The method according to claim 1, wherein said virus is SRSV. The method of claim 1, wherein the exposing occurs for a period of time sufficient to damage DNA of the virus and to degrade the envelope of the virus. The method according to claim 1, wherein the degradation is degradation of damaged DNA to a degree that PCR cannot detect DNA specific to the virus. A poisoning device for the elimination of viruses diluted in water, comprising:
An ultraviolet ray generator using a medium pressure mercury lamp as an ultraviolet ray source; and a photocatalyst that generates radicals upon irradiation with the ultraviolet ray,
The device comprising: The apparatus of claim 7, wherein the water is sewage (treatment) effluent. The device according to claim 7, wherein the virus is SRSV.

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