JP5552001B2 - Virus collection device and virus inspection system - Google Patents

Description

  The present invention relates to a virus collection device that collects viruses floating in the air, and a virus inspection system that collects viruses floating in the air and detects their presence or absence.

Conventionally, viruses have been confirmed by taking a sample of nasal discharge, sputum, etc. that contain a large amount of virus from a person suspected of having a virus infection, and culturing, separating, and identifying the sample based on the sample. Yes. However, since it takes several days to obtain a result when such a method is used, a simple test kit using immunochromatography has been developed in recent years (see, for example, Patent Document 1). This is to detect the presence of a virus by using an antibody that specifically develops color by an antigen-antibody reaction using a virus as an antigen, and observing the color development caused by contact between the virus as an antigen and the antibody. According to this method, it has become possible to detect a virus within minutes to tens of minutes and effectively utilize it for the treatment of infected patients.
However, the test kit disclosed in Patent Document 1 needs to use a sample collected from a site where a relatively large amount of virus such as nasal discharge or sputum exists.
Here, in order to suppress a virus, particularly a pandemic of a virus infection such as influenza, which has been threatened in recent years, it is desired to collect viruses floating in the air. The test kit disclosed in 1 was not capable of collecting viruses floating in the air.

  In addition, a simple virus detection system using MEMS (Micro Electro Mechanical System) has been proposed (Non-patent Document 1). Even in this detection system, basically, the detection target is a virus with a high concentration. It was premised to be a solution containing, and it was not a technique that could collect viruses floating in the air.

JP 2008-275511 A

Fernando Patolsky, Analytical Chemistry 78, 23, 4260-4269 (2006)

  As described above, the existing technology described above relates to a virus detection system that detects a solution containing a high concentration of virus, and does not relate to a technology that collects viruses floating in the air. . For this reason, these existing technologies have not been able to collect the low-concentration virus floating in the air, which is necessary for preventing the spread of virus infection. Moreover, it has not been a system for analyzing the collected virus quickly and simply on the spot and monitoring the presence of the virus in real time. In other words, for example, a system that can detect the presence of a virus in a room such as a hospital or an elderly facility in real time is desired.

  The present invention has been made in view of the above circumstances, and its purpose is to quickly and easily analyze a virus collection device that can appropriately collect viruses floating in the air and the collected viruses on the spot. Thus, the present invention is to provide a virus inspection system capable of monitoring the presence of viruses in real time.

In order to achieve the above object, the characteristic configuration of the virus collection device of the present invention is:
A main body casing having a cylindrical inner space;
A discharge path forming member that forms an air discharge path for discharging air from the internal space portion on the central axis side to the outside of the apparatus at one end of the main body casing along the central axis;
Suction path formation that forms an air suction path along the inner peripheral surface of the main body casing along the outer peripheral surface of the main body casing at the other end of the main body casing. A member,
A suction / discharge unit that sucks air outside the apparatus through the air suction path, the internal space, and the air discharge path, and discharges the air outside the apparatus;
In the dust processing space between the suction path forming member and the main body casing, the sucked air is swung along the inner peripheral surface of the main body casing and guided from the other end side to the one end side, and the air A cyclone that separates the collection target from the air by centrifugal force generated by the swirling of, and collects the collection target using the one end side as a collection unit,
The collection unit includes an extract introduction unit that introduces an extract that can extract a virus, and an extract extraction unit that derives the extract from a site different from the extraction solution introduction unit of the collection unit. And an extract supply means for supplying the extract to the extract introduction section,
The said collection part exists in the point comprised as an annular groove located in the said one end side of the said main body casing.

Viruses usually float in the air in a state where they are attached to minute water droplets or dust contained in sneezes or the like. Therefore, in the following, a minute water droplet or dust attached with a virus will be described as a virus adhering substance. The weight of the virus-adhering substance is sufficiently larger than that of air because the virus adheres to relatively small water droplets or dust.
According to the above characteristic configuration, the cyclone guides the virus adhering substance as a collection target together with air to the dust treatment space between the suction path forming member and the main casing, and swirls along the inner peripheral surface of the main casing. Let At this time, the virus-adhering substance having a weight sufficiently larger than that of air is subjected to an appropriate centrifugal force by swirling, and is appropriately separated from the air and guided to a collecting portion on one end side of the main body casing.
Furthermore, by introducing an extract that can extract viruses into the collection unit, the virus adhering substances separated from the air are separated and collected by centrifugal force accompanying the swirling, and the extraction is introduced into this part. The virus can be effectively extracted with the extract by contacting with the solution. Thereby, the extraction efficiency of a virus can be improved.
Furthermore, according to the above-described characteristic configuration, the collection part is an annular groove located on one end side of the main body casing, and therefore the direction of swirling of the virus-adhering substance that swirls the annular groove along the inner peripheral surface of the main body casing. It can arrange | position in the state which followed along. As a result, the virus-adhering substance can be efficiently recovered in the annular groove arranged along the direction of rotation of the virus-adhering substance.
As mentioned above, the virus collection apparatus which can collect the virus which floats in the air appropriately was realizable.

Further features of the virus collection device of the present invention are as follows:
A humidifying means for humidifying the air in the vicinity of the inlet of the air suction path is provided.

Some of the viruses floating in the air do not adhere to minute water droplets or dust and exist alone. When such a single virus is sucked into the cyclone of the present invention together with air, it will be swirled in the dust treatment space, but because it is light, it can be guided to the outside of the apparatus together with the air without being separated from the air. Increases nature. Also, some microparticles containing viruses are too small to be separated from the air, and there is a high possibility that these microparticles are also introduced outside the apparatus.
According to the characteristic configuration described above, the humidifying unit can humidify the air near the inlet of the air suction path and include minute water droplets in the air. Thereby, the microparticles containing the virus present alone in the air or the virus that cannot be separated by the cyclone are attached to the minute water droplets, and the virus-adhering substance having a weight sufficiently larger than that of the air can be obtained. Further, the virus adhering substance is appropriately separated from the air and collected by applying an appropriate centrifugal force in the dust treatment space. Therefore, virus collection can be promoted.

Further features of the virus collection device of the present invention are as follows:
The extract introduction part and the extract lead-out part are provided to be opened at the groove bottom of the annular groove,
Both are provided with the central axis of the main body casing interposed therebetween.

  According to the above characteristic configuration, the extraction liquid introduction section and the extraction liquid outlet section are provided to be opened at the groove bottom portion of the annular groove and are provided with the central axis of the main body casing interposed therebetween, so that the airflow swirls Along with this, the extraction liquid flows substantially evenly through the groove on one side and the groove on the other side that connect the extraction liquid introduction part and the extraction liquid lead-out part in the annular groove. As a result, the annular groove is in a state in which the extraction liquid flows through the entire region, and the virus-adhering substance can be appropriately collected by the extraction liquid.

Further features of the virus collection device of the present invention are as follows:
Provided with a substrate solution supply means for supplying a substrate solution containing a chromogenic substrate that develops an enzyme reaction with a virus,
A mixing unit that mixes the substrate solution supplied by the substrate solution supply unit and the extract derived from the extract deriving unit;
It is in the point provided with the sending means which sends out the liquid mixture of the substrate solution mixed with the above-mentioned mixing part, and an extract.

  According to the above characteristic configuration, the mixing unit mixes the extract derived from the extract deriving unit with a substrate solution containing a color developing substrate that reacts with the virus to develop a color, and develops color when the virus is present. A liquid mixture is produced | generated and the sending means can send out the said liquid mixture appropriately.

The characteristic configuration of the virus inspection system of the present invention is as follows:
While equipped with the virus collection device described above,
A receiving unit that receives the mixed solution of the substrate solution and the extract to be sent out by the sending unit, and a fluorescence detector that detects fluorescence by irradiating the mixed solution received in the receiving unit with inspection light; It is in.

According to the above characteristic configuration, the receiving unit receives the mixed solution sent out by the sending unit, and the fluorescence detector irradiates the mixed solution received in the receiving unit with the inspection light, thereby generating fluorescence as a color. It can be detected. As a result, in a state where fluorescence is detected only when the virus is contained in the mixed solution, the collected virus can be analyzed quickly and simply on the spot to detect the presence or absence of the virus floating in the air.
Furthermore, according to the above-described characteristic configuration, it is possible to monitor the presence of the virus continuously and in real time. That is, the extract supply means continuously supplies the extract to the collection part, and the suction / discharge means continuously sucks air, whereby the virus is collected by the extract of the collection part, and the substrate The solution supply means continuously supplies the substrate solution, the mixing unit continuously mixes the extract and the substrate solution and guides it to the receiving unit, and the fluorescence detector is continuously guided to the receiving unit. Moreover, by continuously irradiating the inspection light, it is possible to continuously detect fluorescence as a color.
In addition, the extract is configured to allow continuous flow through the extract introduction unit, the collection unit, the extract extraction unit, the mixing unit, and the fluorescence detector. Thus, the time taken from the collection of the virus to the analysis by the fluorescence detector can be made sufficiently short, and the virus can be detected substantially in real time.
As a result, a virus inspection system that can continuously monitor the presence of viruses in real time has been realized.

It is a schematic block diagram which shows a virus collection apparatus and a virus inspection system. It is BB sectional drawing of FIG. It is a graph which shows the result of the collection test of a virus.

In particular, the present invention continuously and in real time examines a virus collection device capable of appropriately collecting a virus floating in the air A, and an extract V1 containing the virus collected by the virus collection device. It can relate to a virus inspection system. As a virus, in particular, an influenza virus U, which has recently been feared to be a pandemic, can be targeted for collection and detection.
Below, after explaining a virus collection apparatus first, a virus inspection system provided with the said virus collection apparatus is demonstrated.
The following embodiments are described in order to more specifically illustrate the present invention, and various modifications are possible without departing from the spirit of the present invention. The present invention is described below. It is not limited to.

As shown in FIGS. 1 and 2, the virus collection device of the present invention includes a cyclone 40 that sucks air A containing influenza virus U from the outside of the device and separates the influenza virus U from air A.
The cyclone 40 includes a main body casing 11 that forms a cylindrical inner space 10, and one end of the main body casing 11 (the end on the base end side of the arrow Z in FIG. 1). A discharge path forming member 15 that forms an air discharge path 14 for discharging air A from the site to the outside of the apparatus along the central axis L, and the other end of the main body casing 11 (the end on the tip side of the arrow Z in FIG. 1) Thus, an air suction path 16 for sucking air A from the outside of the apparatus into the internal space 10 along the inner peripheral surface 12 of the main body casing 11 is formed along the cylinder outer peripheral surface 13 (shown in FIG. 2) of the main body casing 11. The suction path forming member 17 is provided.
The cyclone 40 further includes a suction motor 29 (an example of a suction / discharge unit) that sucks the air A outside the apparatus through the air suction path 16, the internal space 10, and the air discharge path 14, and discharges the air A to the outside of the apparatus. In the dust processing space M between the suction path forming member 17 and the main body casing 11, the sucked air A is swung along the inner peripheral surface 12 of the main body casing 11, and the one end side (in FIG. 1) The tip of the arrow Z is guided to the other end side, and the collection target is separated from the air A by the centrifugal force generated by the swirling of the air A, and the collection target is collected using the one end side as the collection unit 18. It is configured as follows.

Here, the main body casing 11 of the cyclone 40 has one end provided with the collecting portion 18 directed downward in the vertical direction (base end side of the arrow Z in FIG. 1) and the other end in the vertical direction upward. It is preferable to arrange it toward (the tip side of arrow Z in FIG. 1). Thereby, the extract V1 introduced from the extract introduction part 19 is guided to the lower side by gravity, and thus is held in the collecting part 18 at one end on the lower side of the main casing 11.

  When the suction motor 29 sucks the air A, the influenza virus U is trapped in the collection unit 18 as a collection target separated from the air A in the dust treatment space M as minute water drops or dust in the air A. The influenza virus adhering substance W that is attached is introduced. Here, the collection unit 18 is provided with an extract introduction unit 19 that introduces an extract V1 that can extract the influenza virus U from the influenza virus adhering substance W, and the extract introduction unit 19 of the collection unit 18 Is provided with an extract extracting unit 20 for extracting the extract V1 from different parts. Then, the extract V1 delivered from the extract reservoir 21 by the first delivery pump 22 is guided to the extract introduction unit 19. The extract reservoir 21 and the first delivery pump 22 function as extract supply means.

  With the above-described configuration, in the virus collection device of the present invention, the suction motor 29 sucks the air A so that the influenza virus adhering substance W is swung along the inner peripheral surface 12 of the main body casing 11 of the cyclone 40. The influenza virus adhering substance W is separated from the air A by centrifugal force, and the influenza virus adhering substance W is passed through the collecting part 18 provided on one end side of the cyclone 40 (the base end side of the arrow Z in FIG. 1). It is effectively immersed in the flowing extract V1. Thereby, the collection efficiency of influenza virus U is improved.

  In particular, as shown in FIG. 2, the collection portion 18 is configured as an annular groove 23 in a state along the inner peripheral surface 12 of the main body casing 11 in a plan view of the cyclone 40. Thereby, the flow direction Y of the influenza virus adhering substance W swirling along the inner peripheral surface 12 of the main body casing 11 is substantially matched with the ring shape of the annular groove 23, and the influenza virus adhering substance W is collected in the annular groove 23. be able to. Since the extract V1 flows through this part, the influenza virus adhering substance W is immersed in the extract V1.

  Further, as shown in FIG. 2, each of the extraction liquid introduction section 19 and the extraction liquid outlet section 20 is provided to open to the groove bottom 33 of the annular groove 23 as the collection section 18, and the central axis of the main body casing 11. It is preferable to be provided across L. Thereby, in the plan view of the cyclone 40, the extract V1 starts from the extract introduction part 19 and a part thereof flows through the groove 23a on one side of the annular groove 23, and the remaining part is the groove on the other side. It will be guide | induced to the extraction liquid derivation | leading-out part 20 with the form which flows through 23b. As a result, the extract V1 can be made to exist over substantially the entire area of the annular groove 23.

In the air A, the influenza virus U sometimes floats alone. Since the single influenza virus U is light in weight, an appropriate centrifugal force is not applied to the main body casing 11 of the cyclone 40 and is difficult to be separated from the air A. As a result, it is difficult to collect the single influenza virus U with the cyclone 40.
Therefore, in the cyclone 40 of the present invention, a mist device 26 (an example of a humidifying unit) that humidifies the air A is provided in the vicinity of the suction port 25 of the air suction path 16. In the present embodiment, as shown in FIG. 1, the mist device 26 employs a configuration that can humidify the vicinity of the inlet of the air suction path 16. That is, the mist device 26 is arranged so as to spray the mist N directly in front of the air suction path 16. By spraying the mist N, a single influenza virus U can be attached to the mist N to form an influenza adhering substance W. As a result, influenza virus U can be collected appropriately.

Influenza virus U has inolaminidase in its epidermis, and has the property of releasing inolaminidase from influenza virus U to extract V1 when the influenza virus U dissolves in extract V1 containing a surfactant or the like. Yes.
Therefore, the virus collection device according to the present invention is configured as follows in order to detect the influenza virus U by a virus inspection system to be described later using the properties of the influenza virus U.

That is, the virus collection device is configured to be able to mix the substrate solution V2 for detecting the presence of inolaminidase with the extract V1 derived from the extract derivation unit 20 on the downstream side of the extract derivation unit 20. Has been.
Specifically, as shown in FIG. 1, a substrate solution reservoir 27 (an example of a substrate solution supply unit) that stores a substrate solution V2 containing a chromogenic substrate that develops an enzyme reaction with inolaminidase, and a substrate solution reservoir 27 A second delivery pump 28 (an example of a substrate poultry feeding means) for delivering the substrate solution V2, a substrate solution V2 delivered from the second delivery pump 28, and an extract V1 derived from the extract extractor 20; And a third delivery pump 41 (an example of delivery means) for delivering the extract V1 and the substrate solution V2 to the downstream side.
Thus, when the extract V1 contains the influenza virus U, the mixture of the extract V1 containing the inolaminidase released from the influenza virus U and the substrate solution V2 containing the chromogenic substrate is such that the inolaminidase reacts with the chromogenic substrate. Color. Therefore, if the color of the mixed solution is detected, the presence or absence of the influenza virus U in the extract V1 can be detected.

  Here, as the chromogenic substrate, for example, sialic acid such as 4-methylumbelliferyl α-D inoramic acid ammonium (4-Methylumbelliferyl-N-acetyl-α-D-neuraminic acid, Ammonium salt: 4MU-NANA) In addition, 5-bromoindoxyl 4,7-dimethoxy-N-acetylinolamic acid α-ketoside (5-bromoindoxyl-4,7-dimethyloxy-N-acetylnelactic acid α-) can be used. etc.) can be used. This chromogenic substrate is usually decomposed into an inolaminic acid moiety and a chromogenic product by inoraminidase, and the chromogenic product develops color (light emission in the case of a fluorescent substance), so that the presence of inolaminidase and influenza virus U in the extract V1 is present. Detected.

[Virus inspection system]
In the following, a specific configuration for detecting the presence of the influenza virus U by detecting the fluorescence when the chromogenic substrate emits fluorescence in the virus inspection system provided with the virus collection device will be described.
The virus inspection system of the present invention includes the virus collection device described so far, and also includes a receiving unit 31 that receives the mixed solution of the extract V1 and the substrate solution V2 mixed by the mixing valve 30. In addition to irradiating the test light to the mixed solution received in 31, a fluorescence detector 32 for detecting the fluorescence emitted from the colored product of the chromogenic substrate of the mixed solution is provided.
According to this system, when fluorescence is detected by the fluorescence detector 32, it can be determined that the air A contains the influenza virus U. If no fluorescence is detected, the air A does not contain the influenza virus U. I can judge.

Here, in this virus inspection system, the influenza virus U can be collected and detected continuously and in real time. That is, as shown in FIG. 1, the flow path through which the extract V1 can continuously flow through the extract introduction section 19, the collection section 18, the extract extraction section 20, the mixing valve 30, and the fluorescence detector 32. Is formed.
In the said structure, the 1st delivery pump 22 sends out the extract V1 continuously, the influenza virus U is continuously collected in the collection part 18 with the said extract V1, and the mixing valve 30 is the said extraction. The liquid V1 and the substrate solution V2 are continuously mixed to form a liquid mixture, and the fluorescence detector 32 continuously detects the fluorescence of the liquid mixture and continuously detects the presence or absence of the influenza virus U. .
Furthermore, in the above-described configuration, the extraction liquid introduction unit 19, the collection unit 18, the extraction liquid extraction unit 20, the mixing valve 30, and the fluorescence detector 32 are connected through a continuous flow path. The extract V1 can collect and detect the influenza virus U while flowing through the flow path in a short time of about several tens of seconds to several minutes, and the presence or absence of the influenza virus U can be inspected in substantially real time.

Below, the verification experiment which verified the capture capability of the influenza virus U of the said virus collection apparatus is shown.
[Verification test]
The verification test using the influenza virus U requires considerable equipment and is not realistic. As a model virus that can be used in a normal dusting laboratory, the phage ΦX174 is sprayed and the verification test shown below The virus collection ability was verified by confirming whether or not the phage ΦX174 could be collected using a simple virus collection apparatus.
○ Simple virus collection device for verification test Since this verification test is to confirm the virus collection capability of the virus collection device, in the virus collection device shown in FIG. The cyclone 40 in a state satisfying V1 is prepared, and in the concentration of the model virus, the extraction liquid introduction unit 19 and the extraction liquid extraction unit 20 do not introduce or extract the extraction liquid V1, that is, have been described so far. In the virus collection device, the extract introduction unit 19, the extract solution reservoir 21, the first delivery pump 22, the extract delivery unit 20, the third delivery pump 41, the substrate solution reservoir 27, the second delivery pump 28, and the mixing valve 30. The verification test was carried out under the following conditions using a simple virus collection device for verification test that does not include
○ Gelatin system for subject test As a control test device, a gelatin system having the following sampling unit was used. The sampling unit includes a filter unit to which a gelatin filter (to be described later) can be attached and detached, and an air flow unit having an air pump that allows air A to flow through the filter unit. Thus, when the air pump in the sampling unit is driven, the sampling unit introduces air A from the outside of the sampling unit through the filter unit, and then forms a circulating flow that is returned to the outside of the sampling unit.

= Materials etc. =
○ Model virus phage φX174:
NBRC 103405: Obtained from NBRC

Host E. coli:
NBRC 13898: Obtained from National Institute of Technology and Evaluation (NBRC)

Condensate 702:
Polypeptone: 10g
East Extract: 2g
MgSO ₄ · 7H ₂ O: 1g
An appropriate amount of water is added to the above material and dissolved, and then adjusted to pH 7.0, and water is added to 1L. This is autoclaved.

Agar plate medium (medium 802):
Polypeptone 10g
East Extract 2g
MgSO ₄ · 7H ₂ O 1g
An appropriate amount of water is added to the above material and dissolved, and then adjusted to pH 7.0, and water is added to 1L. After adding 15 g of Agar and sterilizing by autoclave, 10 ml each is aseptically dispensed into a shallow petri dish so that the liquid is evenly distributed in the petri dish and solidified.

○ Filter gelatin filter:
Sartorius, diameter: 9 cm, pore size: 3 μm

Simple virus collector:
DC16 made by DYSON

Sampling unit:
MD8 Airport Sartorius

= Measurement method =
Preparation of host E. coli and phage solution Both host E. coli and phage φX174 were obtained from NBRC (http://www.nbrc.nite.go.jp/). According to the manual attached to the NBRC, the ampule of the host E. coli was opened, and 200 μl of condensate 702 was added. After standing for several minutes, the bottom of the ampoule was tapped and mixed, and several μl was applied to a plurality of agar plate media (medium 802). In addition, tens of μl was added to the condensate 702 and 4 ml previously dispensed in the culture test tube and mixed gently. The plate was placed in an incubator set at 30 ° C., the plate agar plate was allowed to stand, the culture test tube was shaken at 180 rpm, and cultured overnight. Since the growth of E. coli was confirmed in both the agar plate medium and the culture test tube, it was restored by liquid culture, and an appropriate amount of the E. coli solution was planted in fresh condensate 702, and expanded while shaking at 30 ° C. After confirming sufficient growth, sterilized dimethylformamide (DMSO) was added to the E. coli solution to a final concentration of 10% or 20%, and stored at −80 ° C.

  Phage φX174 was prepared by the plate lysate method. As with the host E. coli, the ampule of phage φX174 was opened, 200 μl of condensate 702 was added, and the mixture was allowed to stand for several minutes. After tapping the ampoule bottom and mixing, 50 μl was taken, added to 250 μl of the host E. coli solution that was considered to be in the logarithmic growth phase, mixed well by tapping, and then cultured with shaking (150 rpm) at 37 ° C. for 15 minutes. After mixing with 7 ml of soft agar (0.6% w / v solution) prepared in advance and incubated at 45 ° C, the mixture was spread on an agar plate medium, and after soft agar solidified, it was cultured overnight at 37 ° C. did. After confirming the plaque formation on the soft agar surface, 10 ml of condensate was added to one agar plate medium and pipetted to recover the upper layer liquid. After centrifugation (3000 rpm × 15 minutes), the supernatant was recovered (phage solution). By performing this operation a plurality of times, a phage solution having a sufficient infectivity was prepared and used as a model virus.

Collection of Phage φX174 Using the above model virus, the model virus was sprayed into a dust generation experimental space with a capacity of 24 m ³ , and a model virus collection test using a simple virus collection device and a gelatin system was performed.

A certain amount of model virus (about 1 × 10 ¹⁵ to 1 × 10 ¹⁷ cells / ml × 0.9 ml) infected with E. coli and increased in copy number is sprayed in a sealed dusting laboratory (3.5 L). Spray for 5 minutes). Next, the model virus floating in the dust generation laboratory was collected with a simple virus collector (39.6 m ³ / h × 15 seconds) and a gelatin system (2 m ³ / h × 30 minutes).

  In addition, the environment in the dust generation laboratory when the verification test was performed was in the range of 26 to 30 ° C. and 20 to 40% RH.

○ Model virus recovery by virus collector After the completion of collection, the host virus is infected with the model virus contained in the condensate 702 as the extract V1, and after a certain period of time, plaque formation by the model virus is detected by the plaque assay method. Observed.

○ Model virus recovery by gelatin system After completion of collection, the gelatin filter was removed, and the collected product collected on the filter was recovered by dissolving in the condensate 702 as the extract V1. Furthermore, the model virus contained in the collected product was infected with host E. coli, and after a certain period of time, the plaque formation by the model virus was observed by a plaque assay method.

○ Quantification of collected model viruses (plaque assay method)
The model virus was diluted 10-fold with condensate 702 (450 μl of condensate 702 was added to 50 μl of phage φX174) to prepare a dilution series from 10 ⁻¹ to 10 ⁻¹⁴ . The host Escherichia coli culture solution, which seems to be in the logarithmic growth phase, was dispensed into culture tubes in a volume of 250 μl, and then 100 μl of the model virus dilution series was added and tapped to mix well. Shaking culture was performed at 37 ° C. for 15 minutes. To each test tube after the culture, 7 ml of a 0.6% soft agar solution that had been incubated at 45 ° C. was added, and rapidly stirred by pipetting so as not to foam. Inoculated in advance on an agar plate medium incubated at 37 ° C., smoothed so that the soft agar spread over the entire surface, and solidified at room temperature. After standing in an incubator set at 37 ° C. and culturing for 3 hours (afterwards stored at 4 ° C.), the number of plaques formed was counted. The phage concentration (PFU / ml) was determined from the counted number of plaques. The results are shown in FIG.

○ Virus recovery rate In FIG. 3, the concentration of phage in the stock solution (PFU / ml) is the amount of model virus (approximately 1 × 10 ¹⁵ to 1 × 10 ¹⁷ cells) sprayed in the dusting laboratory under the conditions described above. /Ml×0.9 ml). On the other hand, the phage concentration (PFU / ml) of the recovered solution is the model virus contained in the recovered solution (extract V1) after the model virus is collected in the gelatin system according to the virus collection device and the control test of the present invention. Phage concentration. Here, the virus recovery rate is defined as the phage concentration of the recovered solution divided by the phage concentration of the stock solution. In this case, the model virus recovery rate by the virus trapping device is about 10 ^-12, 10 ^-10 viral recoveries of the control test gelatin system. That is, the recovery rate of the model virus by the virus collector is about 1/100 of the virus recovery rate of the gelatin system of the control test. However, in view of the fact that the suction time of the virus collection device is 15 seconds and is about 1/120 of the suction time of the gelatin system of the control test, which is about 120 times, the model virus recovery rate of the virus collection device Can be said to be at least as high as that using a gelatin system.

○ Recovered amount of model virus Furthermore, in FIG. 3, when focusing on the phage concentration (PFU / ml) of the recovered solution as the recovered amount of the model virus, the virus collector of the present invention has a short suction time of 15 seconds. Nevertheless, a recovery solution with a phage concentration of about 10 ⁵ PFU / ml can be obtained, which is less than the phage concentration of about 10 ⁶ PFU / ml in the recovery solution of the gelatin system of the control test. It was possible to obtain a recovered liquid having a concentration capable of detection.

[Another embodiment]
( 1 ) In the above-described embodiment, an example in which one air suction path 16 is provided has been described. However, a plurality of air suction paths 16 may be provided. At this time, each of the suction path forming members 17 that form the plurality of air suction paths 16 is configured to rotate the air A sucked from the air suction path 16 along the inner peripheral surface 12 of the main body casing 11. It is preferable that it is provided along the cylinder outer peripheral surface 13.

( 2 ) In the above embodiment, the extract introduction part 19 and the extract lead-out part 20 are each provided in the annular groove 23 as the collection part 18, but a plurality of these may be provided separately. You may comprise.

( 3 ) In the above embodiment, an ionizer that neutralizes the influenza virus U, minute water droplets, dust and the like contained in the air A in the atmosphere may be provided in the vicinity of the suction port 25 of the air suction path 16. As a result, the single influenza virus U, minute water droplets, dust, and the like are neutralized and do not repel each other, and the single influenza virus U is adhered to the minute water droplets, dust, etc., and the influenza virus adhering substance W is obtained. it can. As a result, the influenza virus U can be easily collected by the cyclone 40.

  The virus collection device and virus inspection system of the present invention are a virus collection device that can appropriately collect viruses floating in the air, and the collected viruses can be quickly and easily analyzed on the spot to detect the presence of viruses. It can be effectively used as a virus inspection system capable of monitoring the situation in real time.

U: Influenza virus (an example of a virus)
V1: Extraction solution V2: Substrate solution A: Air M: Dust processing space L: Center axis 10: Internal space 11: Main body casing 12: Inner peripheral surface 13 of main body casing: Cylinder outer peripheral surface 14 of main body casing: Air discharge path 15 : Discharge path forming member 16: Air suction path 17: Suction path forming member 18: Collection section 19: Extract liquid introduction section 20: Extract liquid discharge section 21: Extract liquid reservoir (an example of extract liquid supply means)
22: First delivery pump (an example of extract supply means)
23: annular groove (an example of a collecting part)
25: Air suction passage suction port (air suction passage entrance)
26: Mist device (an example of humidifying means)
27: Substrate solution reservoir (an example of substrate solution supply means)
28: Second delivery pump (an example of substrate solution supply means)
29: Suction motor (an example of suction discharge means)
30: Mixing valve (an example of a mixing unit and a delivery means)
31: Receiving part 32: Fluorescence detector 40: Cyclone 41: Third delivery pump (an example of delivery means)

Claims (5)

A main body casing having a cylindrical inner space;
A discharge path forming member that forms an air discharge path for discharging air from the internal space portion on the central axis side to the outside of the apparatus at one end of the main body casing along the central axis;
Suction path formation that forms an air suction path along the inner peripheral surface of the main body casing along the outer peripheral surface of the main body casing at the other end of the main body casing. A member,
A suction / discharge unit that sucks air outside the apparatus through the air suction path, the internal space, and the air discharge path, and discharges the air outside the apparatus;
In the dust processing space between the suction path forming member and the main body casing, the sucked air is swung along the inner peripheral surface of the main body casing and guided from the other end side to the one end side, and the air A cyclone that separates the collection target from the air by centrifugal force generated by the swirling of, and collects the collection target using the one end side as a collection unit,
The collection unit includes an extract introduction unit that introduces an extract that can extract a virus, and an extract extraction unit that derives the extract from a site different from the extraction solution introduction unit of the collection unit. And an extract supply means for supplying the extract to the extract introduction section,
The said collection part is a virus collection apparatus comprised as an annular groove located in the said one end side of the said main body casing.   The virus collection device according to claim 1, further comprising a humidifying unit that humidifies air near an inlet of the air suction path. The extract introduction part and the extract lead-out part are provided to be opened at the groove bottom of the annular groove,
Both are provided with the virus collection apparatus of Claim 1 or 2 provided on both sides of the center axis | shaft of the said main body casing. Provided with a substrate solution supply means for supplying a substrate solution containing a chromogenic substrate that develops an enzyme reaction with a virus,
A mixing unit that mixes the substrate solution supplied by the substrate solution supply unit and the extract derived from the extract deriving unit;
The virus collection device according to any one of claims 1 to 3, further comprising a delivery unit that delivers a mixed solution of the substrate solution and the extract mixed in the mixing unit. While comprising the virus collection device according to claim 4,
A virus comprising a receiving unit that receives a mixed solution of a substrate solution and an extract to be sent out by the sending unit, and a fluorescence detector that detects fluorescence by irradiating the mixed solution received in the receiving unit with test light. Inspection system.

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