Method for evaluating microorganisms

Abstract

To provide a microorganism evaluation method capable of collecting microorganisms in a gas and reducing the time required for evaluating the collected microorganisms.SOLUTION: The microbe evaluation method includes a first sampling step (step S101) and a second sampling step (step S103) of collecting microbes contained in air in an object space, and an evaluation step (step S104) of evaluating a change in the state of the collected microbes 1. In the first sampling step (step S101) and the second sampling step (step S103), a collecting liquid containing a labeling material capable of labeling the microorganisms to be collected is used as the collecting liquid. In the evaluation step (step S104), the first labeling result based on the first analyte sample by the first sampling step (step S101) and the second labeling result based on the second analyte sample by the second sampling step (step S103) are compared to evaluate a change in the state of microorganisms in the object space.SELECTED DRAWING: Figure 6

Description

[Technical Field]

[0001] The present disclosure relates to a microbial evaluation method in which microorganisms in a gas are collected, labeled, and evaluated. [Background technology]

[0002] Conventionally, there are known devices and methods for sampling fine particles in a gas using a device that utilizes the inertia or centrifugal force of the fine particles (see, for example, Patent Document 1). More specifically, Patent Document 1 discloses a device and method that generates an electrostatic force using a rotating cylindrical electrode to which a collection liquid is supplied, thereby separating objects to be collected, such as fine particles, from the air and collecting them in the liquid. In addition, when the sampled fine particles contain or may contain microorganisms, a method is known in which the microorganisms are labeled with a labeling substance and detected (see, for example, Patent Document 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2021 / 153155 [Patent Document 2] Japanese Patent Application Laid-Open No. 2006-238779 Summary of the Invention [Problem to be solved by the invention]

[0004] In conventional methods, microorganisms are captured using a microbial sampling device and then labeled for detection. This means that it takes time to visualize the captured microorganisms, making it difficult to quickly evaluate the microorganisms when verifying the effectiveness of space purification using hypochlorous acid gas, etc.

[0005] Therefore, the present disclosure has been made in consideration of the above-mentioned conventional problems, and its purpose is to provide a microbial evaluation method that can capture microorganisms in gas and reduce the time required to evaluate the captured microorganisms. [Means for solving the problem]

[0006] A microbial evaluation method according to the present disclosure uses a cylindrical first electrode that is open at both ends in its axial direction, and a second electrode that extends in the axial direction of the first electrode and is disposed within the first electrode at a distance from the inner surface of the first electrode, to capture and evaluate microorganisms contained in air that is taken in from a target space and circulating within the interior of the target space. The method includes a supply step of supplying a capture liquid into the first electrode and allowing the capture liquid to accumulate on a portion of the inner surface of the first electrode in a direction around the axial center; and a step of rotating the first electrode around a rotation axis that extends in the axial direction of the first electrode and passes through the first electrode while rotating the first electrode. The method includes a first sampling step of applying a voltage between the first electrode and the second electrode to trap microorganisms contained in air circulating inside the target space in a trapping liquid, a recovery step of recovering the trapping liquid from which the microorganisms have been trapped in the trapping step, and recovering the microorganisms as a first specimen sample containing the microorganisms, a second sampling step of recovering a second specimen sample containing the microorganisms by carrying out the supplying step, the trapping step, and the recovery step, and an evaluation step of evaluating the state of the microorganisms trapped in the first and second sampling steps. The trapping step uses a trapping liquid containing a labeling substance capable of labeling the microorganisms to be trapped, and the evaluation step compares a first labeling result based on the first specimen sample with a second labeling result based on the second specimen sample to evaluate a change in the state of the microorganisms in the target space. [Effects of the Invention]

[0007] According to the present disclosure, it is possible to provide a microbial evaluation method that can capture microorganisms in gas and reduce the time required to evaluate the captured microorganisms. [Brief explanation of the drawings]

[0008] [Figure 1] FIG. 1 is a perspective view showing the appearance of a microorganism sampling device used in a microorganism evaluation method according to this embodiment. [Figure 2]FIG. 2 is a side view showing the appearance of the microorganism sampling device of FIG. [Figure 3] FIG. 3 is a cross-sectional view taken along line III-III in FIG. [Figure 4] FIG. 4 is an end view taken along line IV-IV in FIG. [Figure 5] FIG. 5 is a schematic diagram showing a microorganism treatment apparatus used in the microorganism evaluation method according to this embodiment. [Figure 6] FIG. 6 is a flowchart showing the microbial evaluation method according to this embodiment. [Figure 7] FIG. 7 is a flowchart showing detailed steps in the first sampling step and the second sampling step. [Figure 8] FIG. 8 is a cross-sectional view taken along line III-III in FIG. 1, and is an explanatory diagram for explaining an example of the operation performed by the microorganism sampling device in FIG. [Figure 9] FIG. 9 is a diagram showing a detailed flow of the microbial treatment step. DETAILED DESCRIPTION OF THE INVENTION

[0009] Hereinafter, embodiments of the present disclosure will be described with reference to the drawings.

[0010] However, the microbial evaluation method according to the present disclosure is not intended to be limited to the embodiments described below or the configurations shown in the drawings, and also includes configurations equivalent thereto.

[0011] The embodiments described below are all comprehensive or specific examples. The numerical values, shapes, materials, components, the arrangement and connection of the components, steps, and the order of steps shown in the following embodiments are merely examples and are not intended to limit the scope of the claims. Furthermore, the drawings are not necessarily strict illustrations. In the drawings, substantially identical components are designated by the same reference numerals, and duplicated descriptions may be omitted or simplified.

[0012] In the following, terms indicating the relationship between elements, such as parallel and perpendicular, terms indicating the shape of elements, such as cylindrical, and numerical ranges do not only represent the strict meaning, but also include a substantially equivalent range, for example, a difference of about a few percent.

[0013] In the following figures, the X-axis and Y-axis are axes that are perpendicular to each other on a horizontal plane. The Z-axis is an axis that is perpendicular to the horizontal plane. On the Z-axis, the positive direction represents a vertical upward direction, and the negative direction represents a vertical downward direction.

[0014] The microbial evaluation method according to this embodiment involves capturing microorganisms contained in the air of a target space, labeling the captured microorganisms with a target substance, detecting the labeled microorganisms, and evaluating changes in the state of the microorganisms.

[0015] In this specification, the term "target space" refers to an indoor or outdoor space that is partitioned by a roof, wall, partition, or the like, and prevents air from flowing in or out. More specifically, the indoor space refers to, for example, a hospital, office, underground warehouse, house, indoor smoking area, or the like, where frequent ventilation is difficult. Alternatively, for example, a closed space created for experimental purposes may be used as the indoor space. The outdoor space refers to, for example, an outdoor smoking area, or the like.

[0016] As used herein, the term "microorganisms" includes bacteria, archaea, fungi, viruses, and aerosols thereof.

[0017] In this specification, the term "change in the state of microorganisms" includes an increase or decrease in the number of microorganisms or a change in the activity state of microorganisms.

[0018] In this specification, "evaluation" means, when the object of evaluation is an increase or decrease in microorganisms, checking the increase or decrease in the number of microorganisms by comparing the number of microorganisms detected in the first sampling step described below with the number of microorganisms detected in the second sampling step. When the object of evaluation is a change in the activity state of microorganisms, it means comparing the detection results of the activity state of microorganisms in the first sampling step described below with the detection results of the activity state of microorganisms in the second sampling step, and comparing the increase or decrease in activity using images, or calculating and comparing numerical values ​​indicating activity.

[0019] The microbial evaluation method according to this embodiment will be described in detail later, but as shown in FIG. 6, which will be described later, includes a first sampling step (step S101), a space purification step (step S102), a second sampling step (step S103), and an evaluation step (step S104).

[0020] The first sampling step (step S101) and the second sampling step (step S103) each include a step of capturing microorganisms 1 contained in the air in the target space R, a step of processing the captured microorganisms 1, and a step of detecting the processed microorganisms 1. Each step included in the first sampling step (step S101) and the second sampling step (step S103) is performed using a different device.

[0021] In the step of collecting the microorganisms 1, the collection of the microorganisms 1 is carried out using a microorganism sampling device 10 (see FIG. 1, which will be described later). In the step of processing the collected microorganisms 1, the processing of the microorganisms 1 is carried out using a microorganism processing device 90 (see FIG. 5, which will be described later). In the step of detecting the processed microorganisms 1, the detection of the microorganisms 1 is carried out using a microorganism detection device (not shown).

[0022] (Microbial sampling device) First, a microorganism sampling device 10 used to perform the steps of capturing microorganisms 1 contained in the air in the target space R, which are included in each of the first sampling step (step S101) and the second sampling step (step S103), will be described with reference to Figs. 1 to 4. Fig. 1 is a perspective view showing the exterior of the microorganism sampling device 10. Fig. 2 is a side view showing the exterior of the microorganism sampling device 10 of Fig. 1. Fig. 3 is a cross-sectional view taken along line III-III in Fig. 1, showing the internal structure of the microorganism sampling device 10 of Fig. 1. Fig. 4 is an end view taken along line IV-IV in Fig. 1.

[0023] As shown in Figures 1 to 4, the microbial sampling device 10 is a device that samples microorganisms 1 in a collection liquid. Specifically, the microbial sampling device 10 is a device that samples microorganisms 1 in a collection liquid 68 (described below) by capturing microorganisms 1 in the gas in the collection liquid 68. The microbial sampling device 10 includes a duct 12, a first bearing seal 14, a second bearing seal 16, a first flange member 18, a second flange member 20, a first electrode 22, a second electrode 24, a voltage application unit 26, a supply unit 28, a recovery unit 30, and a drive unit 32.

[0024] The microorganism sampling device 10 is configured by surrounding a rotating first electrode 22 and a second electrode 24 disposed at the center of the first electrode 22 with a duct 12, a first flange member 18, and a second flange member 20. A gas such as air is passed through the microorganism sampling device 10. That is, the gas such as air is passed through the microorganism sampling device 10 from the target space R in a direction that passes through the microorganism sampling device 10 (the direction shown by arrow A in FIG. 2). Each component of the microorganism sampling device 10 will now be described.

[0025] The duct 12 is cylindrical and rotatably supports the first electrode 22 inside the duct 12. The duct 12 has a main body 42, a first support portion 44, and a second support portion 46. The main body 42, the first support portion 44, and the second support portion 46 are insulating.

[0026] The main body 42 is cylindrical, and one end and the other end in the axial direction of the main body 42 are open. The first support portion 44 protrudes radially outward from one end of the main body 42 in the axial direction of the main body 42 and is formed integrally with the main body 42. The first support portion 44 is recessed radially outward from the main body 42 and is generally U-shaped (see FIG. 3). The first support portion 44 is annular when viewed from the axial direction of the main body 42. A first bearing seal 14 is disposed inside the first support portion 44. The first bearing seal 14 seals the gap between the first support portion 44 and a first outer flange portion 58 (described below) to prevent gas from leaking between the first support portion 44 and a first outer flange portion 58. The first support portion 44 rotatably supports the first electrode 22 via the first bearing seal 14. The second support portion 46 protrudes radially outward from the other end of the main body 42 in the axial direction and is formed integrally with the main body 42. The second support portion 46 is recessed radially outward from the main body 42 and is generally U-shaped (see FIG. 3). The second support portion 46 is annular when viewed from the axial direction of the main body 42. A second bearing seal 16 is disposed inside the second support portion 46. The second bearing seal 16 seals the gap between the second support portion 46 and a second outer flange portion 60 (described below) to prevent gas from leaking between the second support portion 46 and the second outer flange portion 60. The second support portion 46 rotatably supports the first electrode 22 via the second bearing seal 16.

[0027] The first flange member 18 is cylindrical and connected to the duct 12. The first flange member 18 has a main body 48 and a flange 50.

[0028] The main body 48 is cylindrical, and one end and the other end in the axial direction of the main body 48 are open. The flange 50 protrudes radially outward from one end of the main body 48 in the axial direction of the main body 48 and is formed integrally with the main body 48. The flange 50 is annular when viewed in the axial direction of the main body 48. The other end in the axial direction of the main body 48 is connected to one end in the axial direction of the duct 12.

[0029] The second flange member 20 is cylindrical and connected to the duct 12. The second flange member 20 has a main body 52 and a flange .

[0030] The main body 52 is cylindrical, and one end and the other end in the axial direction of the main body 52 are open. One end in the axial direction of the main body 52 is connected to the other end in the axial direction of the duct 12. The flange 54 protrudes radially outward from the other end in the axial direction of the main body 52 and is formed integrally with the main body 52. ​​The flange 54 is annular when viewed in the axial direction of the main body 52.

[0031] The first electrode 22 is cylindrical, and both axial ends of the first electrode 22 are open. The first electrode 22 is connected to ground via a second electric wire 76 (described later) or the like. The first electrode 22 has a main body 56, a first outer collar portion 58, a second outer collar portion 60, a first inner collar portion 62, and a second inner collar portion 64. For example, the main body 56, the first outer collar portion 58, the second outer collar portion 60, the first inner collar portion 62, and the second inner collar portion 64 are formed using stainless steel such as SUS (Steel Special Use Stainless).

[0032] The main body 56 is cylindrical, and one end and the other end in the axial direction of the main body 56 are open. The axial direction of the main body 56 is the direction in which the axis B of the main body 56 extends (the X-axis direction). The main body 56 has external teeth (not shown) on its outer circumferential surface that mesh with external teeth (not shown) of a gear 86 (described below). The inner surface 66 of the main body 56 may be subjected to a hydrophilic treatment. The hydrophilic treatment here is a treatment that processes the inner surface 66 into a finely uneven shape. For example, the hydrophilic treatment is performed by plasma treatment. Alternatively, the hydrophilic treatment is performed by alkali treatment using potassium hydroxide (KOH). Furthermore, an adhesion-inhibiting member that inhibits adhesion of microorganisms may be attached to the inner surface 66 of the main body 56. For example, the adhesion-inhibiting member is a blocking agent such as skim milk, bovine serum albumin (BSA), or polyethylene glycol (PEG). Here, "adhesion" of microorganisms includes both a state in which the microorganisms are attached to the inner surface 66 of the main body 56 and a state in which the microorganisms form a membrane-like structure on the inner surface 66 of the main body 56 in a biofilm-like state.

[0033] Although details will be described later, in this embodiment, when sampling is performed, the inner surface 66 of the first electrode 22 is hydrophilized using a hydrophilization treatment liquid 67 containing an amphipathic polymer (described later), and then a collection liquid 68 that does not contain an amphipathic polymer is supplied into the electrode as a sampling solution, so that the amphipathic polymer contained in the hydrophilization treatment liquid 67 comes into contact with and adheres to the entire inner surface of the electrode, initializing the hydrophilic state of the inner surface of the electrode, and then the microparticles are sampled by the collection liquid 68. Here, initializing the hydrophilic state refers to restoring the surface state of the inner surface 66 of the first electrode 22 that has deteriorated due to the sampling operation of the microorganisms 1.

[0034] The first outer collar portion 58 protrudes radially outward from one end of the main body 56 in the axial direction of the main body 56 and is formed integrally with the main body 56. The first outer collar portion 58 is annular around the axis B of the main body 56. In other words, the first outer collar portion 58 is annular when viewed from the axial direction of the main body 56. The first outer collar portion 58 is disposed inside the first bearing seal 14.

[0035] The second outer collar portion 60 protrudes radially outward from the other end of the main body 56 in the axial direction, and is formed integrally with the main body 56. The second outer collar portion 60 is annular around the axial center B of the main body 56. In other words, the second outer collar portion 60 is annular when viewed from the axial direction of the main body 56. The second outer collar portion 60 is disposed inside the second bearing seal 16.

[0036] The first inner flange 62 protrudes radially inward from one end of the main body 56 in the axial direction of the main body 56 and is formed integrally with the main body 56. The first inner flange 62 is annular around the axial center B of the main body 56. In other words, the first inner flange 62 is annular when viewed from the axial direction of the main body 56.

[0037] The second inner flange 64 protrudes radially inward from the other end of the main body 56 in the axial direction, and is formed integrally with the main body 56. The second inner flange 64 is annular around the axial center B of the main body 56. In other words, the second inner flange 64 is annular when viewed from the axial direction of the main body 56.

[0038] The first electrode 22 is installed in an orientation in which the axis B of the main body 56 is parallel to the horizontal direction. The first electrode 22 is supported so as to be rotatable around the axis B of the main body 56 (see arrow C in FIG. 4). In other words, the first electrode 22 is supported so as to be rotatable on its own axis.

[0039] The first electrode 22 stores a hydrophilic treatment liquid 67 or a collection liquid 68 on the inner surface 66 of the main body 56. Specifically, the first electrode 22 stores the hydrophilic treatment liquid 67 or the collection liquid 68 in a part of the inner surface 66 in the direction around the axis B of the main body 56 (see arrow D in FIG. 4). The stored hydrophilic treatment liquid 67 or the collection liquid 68 is located below the axis B of the main body 56. The first electrode 22 stores the hydrophilic treatment liquid 67 or the collecting liquid 68 in that portion of the inner surface 66 along the axial direction of the main body 56. The first inner flange 62 holds the hydrophilic treatment liquid 67 or the collecting liquid 68 so that the hydrophilic treatment liquid 67 or the collecting liquid 68 stored in that portion of the inner surface 66 of the main body 56 does not spill out from one end of the main body 56 in the axial direction. The second inner flange 64 holds the hydrophilic treatment liquid 67 or the collecting liquid 68 so that the hydrophilic treatment liquid 67 or the collecting liquid 68 stored in that portion of the inner surface 66 of the main body 56 does not spill out from the other end of the main body 56 in the axial direction. In this way, the first electrode 22 stores the hydrophilic treatment liquid 67 or the collecting liquid 68 in that portion of the inner surface 66 of the main body 56 so that the hydrophilic treatment liquid 67 or the collecting liquid 68 does not leak out of the main body 56. Within the main body 56, above the stored hydrophilic treatment liquid 67 or the collected liquid 68, an internal space 69 is formed that passes through the main body 56 in the axial direction.

[0040] The second electrode 24 is linear and extends in the axial direction of the body 56 of the first electrode 22. The second electrode 24 passes through the radially inner portion of the body 56 of the first electrode 22 and is positioned inside the body 56. That is, the second electrode 24 protrudes outward from one end of the body 56 in the axial direction of the body 56 and protrudes outward from the other end of the body 56. The second electrode 24 is spaced apart from the inner surface 66 of the body 56 of the first electrode 22 and is located near the center of the first electrode 22. The second electrode 24 is located within the internal space 69. In this embodiment, the second electrode 24 is installed in an orientation in which the axis of the second electrode 24 coincides with the axis B of the body 56 of the first electrode 22. For example, the second electrode 24 is formed of tungsten or the like.

[0041] The voltage application unit 26 applies a voltage between the first electrode 22 and the second electrode 24. The voltage application unit 26 has a first support 70, a second support 72, a first electric wire 74, and a second electric wire 76.

[0042] The first support 70 is fixed to the first flange member 18 and is located inside the first flange member 18. The first support 70 is connected to one axial end of the second electrode 24 and supports the second electrode 24. The second support 72 is fixed to the second flange member 20 and is located inside the second flange member 20. The second support 72 is connected to the other axial end of the second electrode 24 and supports the second electrode 24. The first support 70 and the second support 72 are conductive and electrically connected to the second electrode 24. The first electric wire 74 is electrically connected to the second electrode 24 via the second support 72. The second electric wire 76 is electrically connected to the first electrode 22 via a gear 86 or the like.

[0043] The voltage application unit 26 can apply electricity of any magnitude and waveform to the first electrode 22 and the second electrode 24, which is installed near the center of the first electrode 22, via the first electric wire 74 and the second electric wire 76. This allows the microorganism sampling device 10 to perform electrostatic collection of microorganisms. The second electrode 24 does not have to be linear; it can be plate-shaped or needle-shaped, for example. There are no limitations on its structure or installation position, as long as it is capable of generating an uneven electric field. For example, the voltage application unit 26 can be implemented by a power supply circuit including a converter. For example, the voltage application unit 26 applies a DC voltage of 6 kV.

[0044] For example, the voltage application unit 26 applies a voltage between the first electrode 22 and the second electrode 24 so that the second electrode 24 side has a higher potential than the first electrode 22 side. As a result, an electric field is generated in the internal space 69 from the second electrode 24 toward the first electrode 22 (see arrow E in FIG. 3 and arrow E in FIG. 4).

[0045] The supply unit 28 supplies the hydrophilic treatment liquid 67 or the collection liquid 68 into the first electrode 22, and causes the hydrophilic treatment liquid 67 or the collection liquid 68 to accumulate in a part of the inner surface 66 of the first electrode 22 in the direction around the axis B. In other words, the supply unit 28 supplies the hydrophilic treatment liquid 67 or the collection liquid 68 to the first electrode 22 in the direction around the axis B. The hydrophilic treatment liquid 67 or the collecting liquid 68 is supplied into the first electrode 22 so as to accumulate in a portion of the inner surface 66 of the main body 56 of the first electrode 22. In this way, the hydrophilic treatment liquid 67 or the collecting liquid 68 supplied by the supply unit 28 is accumulated on the inner surface 66 of the main body 56 of the first electrode 22. The supply unit 28 has a tank 78, a tank 79, and an injection unit 80. The supply of the liquid by the supply unit 28 may be from the opening side of the first electrode 22 (one end side in the axial direction of the main body 42) as shown in FIG. 3, or a hole may be formed in a portion of the side surface of the first electrode 22 and the liquid may be supplied through the opening.

[0046] The tank 79 holds the hydrophilic treatment liquid 67 to be supplied into the first electrode 22. The hydrophilic treatment liquid 67 held in the tank 79 is discharged from an injection part 80 by a pump (not shown) or the like, and supplied into the main body 56 of the first electrode 22. In this way, the tank 79 is provided to store in advance the hydrophilic treatment liquid 67 for the hydrophilic treatment of the inner surface 66 of the first electrode 22, and the hydrophilic treatment liquid 67 is supplied into the inside of the first electrode 22 through the injection part 80. Note that although shown in an isolated state in FIG. 3 and other figures, the tank 79 is provided with a structure similar to that of the tank 78.

[0047] The tank 78 holds the collection liquid 68 to be supplied into the first electrode 22. The collection liquid 68 held in the tank 78 is discharged from the injection part 80 by a pump (not shown) or the like, and supplied into the main body 56 of the first electrode 22. In this way, the tank 78 is provided to store the collection liquid 68 for capturing microorganisms in advance, and the collection liquid 68 is supplied into the first electrode 22 through the injection part 80. Furthermore, the supply of the collection liquid 68 is not limited to once per sampling, but may be supplied multiple times depending on the amount of liquid in the first electrode 22, or in conjunction with collection before starting sampling.

[0048] In this embodiment, the supply unit 28 supplies a liquid for hydrophilic treatment of the inner surface 66 of the first electrode 22 as a hydrophilic treatment liquid 67, collects fine particles adhering to the inner surface 66 of the first electrode 22, and supplies a liquid for analyzing the collected fine particles as a collection liquid 68.

[0049] The hydrophilic treatment liquid 67 is a liquid containing an amphipathic substance, such as an amphipathic polymer. An amphipathic polymer is a polymer with both hydrophilic and hydrophobic regions within its polymer. For example, PEG (Polyethylene Glycol) or dextran is used as the hydrophilic region, and PLA (Polylactic Acid) or PLGA (Polylactic Co Glycolic Acid) is used as the hydrophobic region. In this case, the amount of substance contained in the liquid, such as pure water, is preferably at least 0.1 weight percent (wt%). If the amount of substance is less than 0.1 wt%, there is a concern that the hydrophilicity required for efficient capture by the collection liquid 68 may not be maintained. By supplying the hydrophilic treatment liquid 67 containing the amphipathic polymer and rotating the first electrode 22, the hydrophobic region of the amphipathic polymer is adsorbed onto the inner surface 66 of the first electrode 22, and the hydrophilic region is oriented so as to be exposed to the collection liquid 68, which will be supplied later. This increases the affinity at the interface between the inner surface 66 of the first electrode 22 and the collection liquid 68, making it possible to hydrophilize the surface of the inner surface 66 of the first electrode 22. In other words, the hydrophilic state of the inner surface 66 of the first electrode 22 can be maintained by the hydrophilization treatment liquid 67.

[0050] The trapping liquid 68 is a liquid for trapping the microorganisms 1 in the target space R. The trapping liquid 68 is, for example, a liquid that maintains the activity of the microorganisms 1 or a liquid used for analysis, such as a buffer solution. More specifically, the buffer solution may contain sodium chloride, potassium chloride, sodium hydrogen phosphate, and potassium dihydrogen phosphate.

[0051] In this embodiment, the collection liquid 68 further contains a labeling substance capable of labeling the microorganisms in the liquid for collecting the microorganisms 1. Here, the "labeling substance capable of labeling the microorganisms" is a fluorescent reagent. The labeling substance is one or more selected from labeling substances including drugs, staining reagents, luminescent particles, and color-producing particles. The labeling substance will be described in detail below.

[0052] By including a labeling substance capable of labeling the microorganisms 1 in the collection liquid 68, it is possible to label the microorganisms 1 while they are being collected. In other words, since the microorganisms 1 can be labeled without a separate process for labeling the collected microorganisms 1 after collection, the time required to visualize the collected microorganisms 1 can be reduced. Furthermore, by labeling the microorganisms 1 while they are being collected, it is possible to label the microorganisms 1 based on their state at the time of collection. Conventionally, when the microorganisms 1 were labeled after collection, even if they were active immediately after collection, the activity could be lost before labeling, making it difficult to accurately evaluate the state of the microorganisms 1 in the target space R. In contrast, in the present embodiment, by including a labeling substance capable of labeling the microorganisms 1 in the collection liquid 68 and labeling the microorganisms 1 while they are being collected, it is possible to more accurately evaluate the state of the microorganisms 1 in the target space R compared to conventional labeling of the microorganisms 1 after collection.

[0053] The labeling substance used to label the microorganism 1 may be one or a combination of multiple labeling substances selected from multiple labeling substances depending on the type of microorganism 1 to be evaluated. Specifically, the labeling substance used to label the microorganism 1 may be selected as follows:

[0054] When the microorganism 1 to be evaluated is a bacterium, archaea, or fungus, the microorganism 1 can be labeled using a labeling substance capable of labeling live bacteria or a labeling substance capable of labeling dead bacteria.

[0055] A fluorescent reagent or a staining reagent may be used as a labeling substance capable of labeling live bacteria. Specifically, for example, a fluorescent indicator of intracellular esterase activity, a fluorescent indicator of nutrient uptake activity, or a staining reagent capable of staining live cells may be used.

[0056] Specific examples of fluorescent indicators for intracellular esterase activity include 5-carboxyfluorescein diacetate acetoxymethyl ester, 5-(6-)carboxyfluorescein diacetate, 2',7'-bis-(2-carboxyethyl)-5-(6-)carboxyfluorescein acetoxymethyl ester, 5-(6-)sulfofluorescein diacetate, fluorescein diacetate, 5-chloromethylfluorescein diacetate, 5-(6-)carboxyfluorescein diacetate succinimidyl ester, and fluorescein-5-carbonylazide diacetate.

[0057] Specific examples of fluorescent indicators for nutrient uptake activity include 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose and 6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose.

[0058] Alternatively, a fluorescent indicator or a staining reagent for a cell membrane permeable indicator may be used as a labeling substance capable of labeling dead bacteria. Specific examples of fluorescent indicators for cell membrane permeable indicators include propidium iodide, ethidium bromide, and 2',6-diamidino-2-phenylindole. Specific examples of staining reagents for cell membrane permeable indicators include trypan blue.

[0059] When the microorganism 1 to be evaluated is a virus, the labeling substance may be, for example, a fluorescent reagent, a light-emitting particle, or a color-producing particle that can label the activity of a surface protein of the virus.

[0060] Specific examples of luminescent particles capable of labeling the activity of viral surface proteins include fluorescent particles labeled with fluorescent dyes such as FITC (Fluorescein isothiocyanate) or RITC (Tetramethylrhodamine isothiocyanate). These are antibodies that exhibit specificity through an antigen-antibody reaction using viral surface proteins as antigens. Another specific example of a luminescent particle that can label the activity of viral surface proteins is fluorescent beads containing a high concentration of fluorescent dye, and the surface of these fluorescent beads is modified with antibodies that exhibit specificity through an antigen-antibody reaction using viral surface proteins as antigens. In other words, fluorescent beads are, for example, antibody-modified fluorescent polystyrene particles.

[0061] A specific example of a color-forming particle capable of labeling the activity of a viral surface protein is an antibody that is labeled with an enzyme such as horseradish peroxidase or alkaline phosphatase that forms a pigment, and that exhibits specificity through an antigen-antibody reaction using the viral surface protein as an antigen.

[0062] In addition to the labeling substance, the collection liquid 68 may contain a substance that assists in labeling the microorganisms 1. For example, the collection liquid 68 may contain ethylenediaminetetraacetic acid, a chelating agent, as a component that increases the cell membrane permeability of the fluorescent indicator.

[0063] Details will be described later, but in this embodiment, when sampling microorganisms 1, a hydrophilic treatment liquid 67 is supplied for each sampling to hydrophilize the inner surface 66 of the first electrode 22, and then a collection liquid 68 is supplied to sample the microorganisms 1.

[0064] By initializing the hydrophilic state of the inner surface 66 of the first electrode 22 with the hydrophilization treatment liquid 67, the subsequent collection liquid 68 spreads uniformly within the first electrode 22, making it possible to maintain a constant and highly reproducible distance between the central second electrode 24 and the collection liquid 68. This suppresses dielectric breakdown between the collection liquid 68 and the second electrode 24, allowing for normal discharge of the second electrode 24 (described below) and the formation of a desired electric field between the second electrode 24 and the first electrode 22, enabling normal sampling.

[0065] The recovery unit 30 recovers the hydrophilic treatment liquid 67 or the collection liquid 68 stored in a part of the inner surface 66 in the direction around the axis B of the first electrode 22. The recovery unit 30 has a tank 82, a tank 83, and an extraction unit 84. The hydrophilic treatment liquid 67 or the collection liquid 68 stored in a part of the inner surface 66 in the direction around the axis B of the first electrode 22 is sucked from the extraction unit 84 by a pump (not shown) or the like, and is held in and recovered in the tank 82 or the tank 83.

[0066] The tank 82 is a container for storing the hydrophilic treatment liquid 67 recovered as waste liquid, and is connected to the extraction unit 84. After the hydrophilic treatment liquid 67 supplied for hydrophilization has performed a hydrophilic treatment operation on the inner surface 66 of the first electrode 22, it is sucked through the extraction unit 84 and held in the tank 82. In addition, the collection liquid 68 supplied for the purpose of cleaning, such as co-washing, on the inner surface 66 of the first electrode 22 is also sucked through the extraction unit 84 and held in the tank 82 after performing a cleaning operation.

[0067] The tank 83 is a container for storing the collection liquid 68 collected as a specimen sample containing the target microorganisms 1, and is connected to the extraction unit 84 in a manner that switches it with the tank 82. Although the tank 83 is shown in an isolated state in FIG. 3 etc., it is installed with the same structure as the tank 82. The collection liquid 68 as a specimen sample containing the microorganisms 1 is sucked through the extraction unit 84 and held in the tank 83 after a processing operation for capturing the microorganisms 1 in the collection liquid 68 is performed.

[0068] 3, each liquid may be collected by the collection unit 30 from the opening side of the first electrode 22 (the other end side in the axial direction of the main body 42), or the side surface of the first electrode 22 may be partially opened and the liquid may be collected from the opening. Note that the collection of the capture liquid 68 does not necessarily have to be carried out after one supply, and collection may be carried out after multiple supplies.

[0069] By doing so, the amount of trapped liquid 68 stored is kept constant, thereby suppressing variations in concentration between samplings due to the amount of trapped liquid 68 stored, thereby improving sampling accuracy. Similarly, by keeping the amount of trapped liquid 68 stored constant, it is possible to maintain a constant distance between the trapped liquid 68 and the second electrode 24, thereby suppressing variations between and within samplings in the electric field generated between the trapped liquid 68 and the second electrode 24 when a voltage is applied. This improves sampling accuracy and simplifies estimation of sampling performance. Furthermore, when the environmental humidity is higher or lower than normal, the amount of evaporation of the trapped liquid 68 varies depending on the environmental humidity, which can lead to discharge failure at the second electrode 24 due to an excess of trapped liquid 68, and to loss or inactivation of sampled microorganisms due to depletion of the trapped liquid 68. However, by supplying the trapped liquid 68 at a fixed time interval that can be set arbitrarily, these occurrences can be suppressed.

[0070] The driver 32 rotates the first electrode 22 around a rotation axis that extends in the axial direction of the body 56 of the first electrode 22 and passes through the first electrode 22. In this embodiment, the rotation axis coincides with the axis B of the body 56. That is, in this embodiment, the driver 32 rotates the first electrode 22 around the axis B of the body 56 of the first electrode 22. The driver 32 includes a gear 86 and a motor 88 for rotating the gear 86. The gear 86 has external teeth (not shown) that mesh with external teeth (not shown) of the first electrode 22. As the motor 88 rotates the gear 86 (see arrow F in FIG. 4), the first electrode 22 rotates around the axis B of the body 56 (see arrow C in FIG. 4). In this manner, the first electrode 22 rotates due to the gear 86 driven by the motor 88.

[0071] In addition, gas outside the microbial sampling device 10 may pass through the inside of the second flange member 20, pass through the inside of the main body 56 of the first electrode 22, and be released from the first flange member 18 to the outside of the microbial sampling device 10.

[0072] (Microbial Treatment Device) The microorganism treatment device 90 will be described below with reference to Fig. 5. Fig. 5 is a schematic diagram showing the microorganism treatment device 90. In the first sampling step (step S101) and second sampling step (step S103) described below, a pretreatment is performed in preparation for detecting the target microorganisms 1 contained in the collected specimen sample (collecting liquid 68). The pretreatment is performed using the microorganism treatment device 90 shown in Fig. 5. The microorganism treatment device 90 is a device that separates microorganisms 1 contained in a specimen sample (collection liquid 68) from the collection liquid 68. Specifically, the microorganism treatment device 90 includes a filter 91, a filter support 92, and a pressurizing unit 93. In the microorganism treatment device 90, the specimen sample (collection liquid 68) containing the microorganisms 1 is filtered by the filter 91. As a result, the microorganisms 1 (microorganisms 1 labeled with a labeling substance) are separated onto the filter 91. Here, the filter 91 is a member that separates the microorganisms 1 contained in the specimen sample (collection liquid 68) from the collection liquid 68. The filter 91 has a mesh size that allows the collection liquid 68 and a cleaning liquid 94 (described later) to pass through but not the target microorganisms 1. The filter support 92 is a member that supports and fixes the filter 91. The filter support 92 clamps and fixes the filter 91 from above and below. The pressurizing unit 93 applies pressure to the liquid while or after supplying the collection liquid 68 and a cleaning liquid 94 (described later) to the filter 91, thereby performing filtration. The pressurizing unit 93 is, for example, a syringe, a cylindrical tool used to inject liquid. Note that filtration of the microorganisms 1 onto the filter 91 may be performed manually using a syringe, or automatically using a device (not shown) equipped with a pump or the like. As a result of filtration, the target microorganisms 1 remain on the filter 91, and the collection liquid 68 passes through the filter 91 and is collected as waste liquid. The microorganisms 1 remaining on the filter 91 are washed away with a cleaning liquid 94 .

[0073] The cleaning liquid 94 used to clean the microorganisms 1 remaining on the filter 91 is, for example, a buffer solution, such as pure water, physiological saline, a liquid that maintains the activity of the microorganisms 1, or a liquid used for analysis. More specifically, the buffer solution may contain sodium chloride, potassium chloride, sodium hydrogen phosphate, and potassium dihydrogen phosphate.

[0074] The washing solution 94 may also contain a substance that assists in labeling the microorganisms 1. For example, the washing solution 94 may contain ethylenediaminetetraacetic acid, which is a chelating agent, as a component that increases the cell membrane permeability of the fluorescent indicator.

[0075] (Microbial detection device) The microorganism detection device will now be described. In the first sampling step (step S101) and second sampling step (step S103) described below, detection of target microorganisms 1 is carried out using a microorganism detection device (not shown). The microorganism detection device is, for example, a device that optically, electrically, or chemically detects a labeling substance that labels target microorganisms 1, and detects the number of labeled microorganisms 1. More specifically, the microorganism detection device captures an image of the fluorescent labeling substance that labels the microorganisms 1 using an imaging element such as a CMOS (Complementary Metal Oxide Semiconductor) image sensor or a CCD (Charge Coupled Device) image sensor, and detects the number of labeled microorganisms 1. Specifically, when the microorganism 1 to be evaluated is a bacterium, an archaea, or a fungus, the microorganism detection device detects the number of live bacteria or the number of dead bacteria. When the microorganism 1 to be evaluated is a virus, the microorganism detection device detects the number of viruses whose surface proteins are in an active state.

[0076] (Space Purification Device) The space purification device will be described. The space purification step (step S102) according to this embodiment is performed using a space purification device. The space purification device is, for example, a hypochlorous acid release device that purifies the target space R by releasing hypochlorous acid gas. The hypochlorous acid release device purifies the target space R, including removing, sterilizing, and inhibiting the activity of microorganisms 1, by releasing hypochlorous acid gas. Since a general device can be used as the space purification device, a detailed description will be omitted. Note that the target space R may be purified using something other than hypochlorous acid gas, such as ozone gas. The target space R may also be purified by spraying disinfecting water (for example, hypochlorous acid water or ozone water).

[0077] (Microbial evaluation method) The microbial evaluation method according to this embodiment will be described below with reference to Figures 6, 7, 8, and 9. Figure 6 is a flowchart showing the entire microbial evaluation method according to this embodiment. Figure 7 is a flowchart showing detailed steps in the first sampling step (step S101) and the second sampling step (step S103). Figure 8 is a cross-sectional view taken along line III-III in Figure 1, and is an explanatory diagram for explaining an example of the operation of the microbial sampling device 10 of Figure 1, showing the movement of viruses within the microbial sampling device 10 until the viruses are actually collected. Figure 9 is a schematic diagram showing a detailed flow in the microbial treatment step (step S9).

[0078] As shown in FIG. 6, the microbial evaluation method according to this embodiment includes a first sampling step (step S101), a space purification step (step S102), a second sampling step (step S103), and an evaluation step (step S104).

[0079] The first sampling step (step S101) is a step of collecting and detecting microorganisms 1 from the target space R before space purification. This is a step in which the target space R is purified using an air purification device, including removing, sterilizing, or inhibiting the activity of microorganisms 1. The second sampling step (step S103) is a step in which microorganisms 1 are captured and detected from the target space R after the space purification. The evaluation step (step S104) is a step in which the states of the microorganisms 1 captured and detected in the first sampling step (step S101) and the second sampling step (step S103) are compared, and the change in the state of the microorganisms due to the space purification is evaluated.

[0080] The first sampling step (step S101) and the second sampling step (step S103) each include multiple steps (see FIG. 7), so below we will explain the multiple steps included in the first sampling step (step S101) as a representative.

[0081] (First sampling step: step S101) As shown in FIG. 7, the first sampling step (step S101) includes a hydrophilic treatment liquid supply step (step S1), a hydrophilic treatment step (step S2), a hydrophilic treatment liquid recovery step (step S3), a collection liquid supply step (step S4), an air flow start step (step S5), a collection step (step S6), an air flow stop step (step S7), a collection liquid recovery step (step S8), a microorganism treatment step (step S9), and a microorganism detection step (step S10).

[0082] First, the supply unit 28 supplies the hydrophilic treatment liquid 67 into the first electrode 22, and causes the hydrophilic treatment liquid 67 to accumulate in a part of the inner surface 66 in the direction around the axis B of the first electrode 22 (step S1: hydrophilic treatment liquid supply step).

[0083] When step S1 is performed, as shown in Figure 8, a hydrophilic treatment liquid 67 is supplied into the main body 56 of the first electrode 22, and the hydrophilic treatment liquid 67 is stored in a portion of the inner surface 66 in the direction around the axis B of the main body 56 of the first electrode 22.

[0084] The driving unit 32 rotates the first electrode 22 around the axis B (step S2: hydrophilic treatment step).

[0085] 8, the first electrode 22 rotates around the axis B in a state in which the hydrophilic treatment liquid 67 is stored in a portion of the inner surface 66 of the first electrode 22 in the direction around the axis B of the main body 56. In other words, the first electrode 22 rotates around the axis B in a state in which the hydrophilic treatment liquid 67 is stored below the axis B so that the hydrophilic treatment liquid 67 does not flow out of the main body 56. As a result, the inner surface 66 of the first electrode 22 sequentially comes into contact with the stored hydrophilic treatment liquid 67. As a result, the amphiphilic polymer contained in the hydrophilic treatment liquid 67 comes into contact with and adheres to the surface of the inner surface 66 of the first electrode 22, and the inner surface 66 of the first electrode 22 is subjected to a hydrophilic treatment. In other words, the hydrophilic state of the inner surface 66 of the first electrode 22 is initialized and maintained. This prevents the specimen sample containing microorganisms 1 from adhering, adsorbing, or adhering to the inner surface 66 of the first electrode 22 in the capture step (step S6), reducing the loss of microorganisms 1 captured in the capture liquid 68 supplied after the hydrophilization treatment. Furthermore, the capture liquid 68 supplied after the hydrophilization treatment spreads uniformly within the first electrode 22, allowing the distance between the central second electrode 24 and the capture liquid 68 to be maintained constant with good reproducibility. This prevents dielectric breakdown between the capture liquid 68 and the second electrode 24, allowing for normal discharge of the second electrode 24 (described below) and the formation of a desired electric field between the second electrode 24 and the first electrode 22, enabling normal sampling.

[0086] Furthermore, by using the hydrophilic treatment liquid 67 containing an amphipathic polymer, it becomes unnecessary to treat the electrodes each time a hydrophilic treatment such as plasma treatment is performed.

[0087] Next, in step S2, the drive unit 32 is stopped, and the rotation of the first electrode 22 is stopped. Thereafter, the stored hydrophilic treatment liquid 67 is recovered (step S3: hydrophilic treatment liquid recovery step). For example, at any timing after a certain period of operation in step S2, the hydrophilic treatment liquid 67 is recovered into the recovery unit 30 (tank 82) through the extraction unit 84.

[0088] When the hydrophilic treatment liquid supply step (step S1), the hydrophilic treatment step (step S2), and the hydrophilic treatment liquid recovery step (step S3) are performed, sampling performance is improved by initializing the hydrophilic state of the surface of the inner surface 66 of the first electrode 22. Note that in the first sampling step (step S101) and the second sampling step (step S103), steps S1 to S3 relating to the hydrophilic treatment described above may be omitted, and the process may start from step S4 described below.

[0089] Next, the supply unit 28 supplies the collection liquid 68 into the first electrode 22, and causes the collection liquid 68 to accumulate in a part of the inner surface 66 in the direction around the axis B of the first electrode 22 (step S4: collection liquid supply step).

[0090] When step S4 is performed, a collection liquid 68 is supplied into the body 56 of the first electrode 22, and the collection liquid 68 is stored on a part of the inner surface 66 in the direction around the axis B of the body 56 of the first electrode 22. Prior to step S4, a separate cleaning process may be performed to prevent residual liquid of the hydrophilization treatment liquid 67 containing the amphipathic polymer from being contained in the collection liquid 68, thereby suppressing a decrease in the infectivity of the microorganisms 1 due to the amphipathic polymer when analyzing the collection liquid 68. The cleaning process here refers to, for example, supplying the collection liquid 68 to wash the inside of the first electrode 22, rotating the first electrode 22 for a certain period of time after supplying the collection liquid 68, and then recovering the collection liquid 68.

[0091] Next, a gas such as air is circulated from the target space R into the interior of the microorganism sampling device 10 in a direction that passes through the microorganism sampling device 10 (the direction shown by arrow A in FIG. 8) (step S5: air circulation start step). For example, air is drawn directly into the microorganism sampling device 10 using a pump (not shown) or the like, and the gas is circulated within the microorganism sampling device 10.

[0092] The driving unit 32 rotates the first electrode 22 around the axis B, while the voltage application unit 26 applies a voltage between the first electrode 22 and the second electrode 24 (step S6: collection step).

[0093] When step S6 is performed, the first electrode 22 rotates around the axis B in a state in which the collecting liquid 68 is stored on a part of the inner surface 66 of the first electrode 22 in the direction around the axis B of the main body 56. In other words, the first electrode 22 rotates around the axis B in a state in which the collecting liquid 68 is stored below the axis B so as to prevent the collecting liquid 68 from flowing out of the main body 56. As a result, the inner surface 66 of the first electrode 22 sequentially comes into contact with the stored collecting liquid 68.

[0094] Microorganisms 1 in gas introduced into the microorganism sampling device 10 by a given air flow (see arrow A in Figure 8) are first charged either positively or negatively by ions 2 released by discharge from the second electrode 24 to which a high voltage is applied. Here, we will explain the case where the microorganisms 1 are positively charged.

[0095] The microorganisms 1, now in an electrically charged state, move along trajectory 3 due to the electric field (see arrow E in FIG. 8) formed between the second electrode 24 and the first electrode 22, and are collected on the inner surface 66 of the first electrode 22. In this way, the microorganisms 1 adhere to the inner surface 66 of the first electrode 22 and are captured on the inner surface 66.

[0096] The microorganisms 1 collected on the inner surface 66 are collected at any timing with the collection liquid 68 stored in the main body 56 of the first electrode 22. Specifically, the microorganisms 1 attached to the inner surface 66 of the main body 56 of the first electrode 22 come into contact with the stored collection liquid 68, and are detached from the inner surface 66 and collected in the collection liquid 68. The movement (rotation) of the first electrode 22, which is rotated by the motor 88 and gear 86, makes it possible to wash away the entire surface of the inner surface 66 of the first electrode 22 with the accumulated (stored) collection liquid 68.

[0097] In addition, when the inner surface 66 of the first electrode 22 has been hydrophilized, a voltage may be applied between the first electrode 22 and the second electrode 24 before rotating the first electrode 22 around the axis B, and the first electrode 22 may be rotated around the axis B with the voltage applied between the first electrode 22 and the second electrode 24.

[0098] By including a labeling substance capable of labeling microorganisms 1 in the collection liquid 68, it becomes possible to label the microorganisms 1 while collecting them, and a first specimen sample containing microorganisms 1 labeled with the labeling substance can be obtained.

[0099] Furthermore, by including a labeling substance capable of labeling the microorganisms 1 in the collection liquid 68, it becomes possible to label the microorganisms 1 while they are being collected, thereby reducing the time required to visualize the collected microorganisms 1. Furthermore, by labeling the microorganisms 1 while they are being collected, it becomes possible to label them based on the state of the microorganisms 1 at the time of collection, and the state of the microorganisms 1 can be evaluated more accurately compared to the conventional method of recovering the microorganisms 1 in a collection liquid and then labeling the microorganisms 1 contained in the collection liquid.

[0100] The driving unit 32 is stopped, the rotation of the first electrode 22 is stopped, and the flow of gas such as air into the inside of the microorganism sampling device 10 is stopped (step S7: air flow stopping step).

[0101] The recovery unit 30 recovers the trapping liquid 68 stored in the first electrode 22 (step S8: trapping liquid recovery step). For example, when the voltage application in step S6 reaches a certain time period, the trapping liquid 68 is recovered into the recovery unit 30 (tank 83) through the extraction unit 84. This makes it possible to obtain a first specimen sample (trapping liquid 68) containing the target microorganisms 1 separated from the gas.

[0102] Next, pretreatment, which is preparation for detecting target microorganisms 1 contained in the collected specimen sample (collection liquid 68), is carried out using a microorganism treatment device 90 (step S9: microorganism treatment step). As shown in Fig. 9, the microorganism treatment step (step S9) includes a specimen sample supply step (step S9a), a specimen sample filtering step (step S9b), a cleaning liquid supply step (step S9c), and a cleaning liquid filtering step (step S9d).

[0103] More specifically, in the microorganism treatment step (step S9), a specimen sample is supplied onto the filter 91 (step S9a), and pressure is applied to the specimen sample by the pressurizing unit 93 to filter the specimen sample (step S9b). If the pressurizing unit 93 is a syringe, the specimen sample supply step (step S9a) and the specimen sample filtering step (step S9b) can be performed simultaneously. The filter 91 has a mesh size that allows the collection liquid 68 to pass through but not the target microorganisms 1. As a result, when the specimen sample filtering step (step S9b) is performed, the target microorganisms 1 remain on the filter 91, and the collection liquid 68 passes through the filter 91 and is collected as waste liquid.

[0104] Subsequently, in the microorganism treatment step (step S9), a cleaning solution 94 is supplied onto the filter 91 on which the microorganisms 1 remain (step S9c), and pressure is applied to the cleaning solution 94 by the pressurizing unit 93, thereby filtering the cleaning solution 94 (step S9d). In this case, the cleaning liquid supply step (step S9c) and the cleaning liquid filtration step (step S9d) can be performed simultaneously. The filter 91 has a mesh size that allows the cleaning liquid 94 to pass through but not the target microorganisms 1. As a result, when the cleaning liquid filtration step (step S9d) is performed, the cleaning liquid 94 passes through the filter 91 while washing away the microorganisms 1 remaining on the filter 91, and is collected as waste liquid.

[0105] By carrying out the microbial treatment step (step S9) in the above manner, the target microorganisms 1 are left on the filter 91, while other substances than the target, such as dust or pollen, and excess marker substance, which may become noise in the subsequent detection of the microorganisms 1, can be reduced, thereby improving the accuracy of the microbial evaluation. Note that in the first sampling step (step S101) and the second sampling step (step S103), depending on the properties of the marker substance that labels the target microorganisms 1, the above-mentioned microbial treatment step (step S9) may be omitted, and the microbial detection step (step S10), which will be described below, may be started after the collection liquid recovery step (step S7).

[0106] Finally, the target microorganisms 1 are detected using a microorganism detection device (step S10: microorganism detection step). The microorganism detection device is, for example, a device that optically, electrically, or chemically detects the labeled substance that has been labeled on the target microorganisms 1, and detects the number of microorganisms. Specifically, the microorganism detection device captures an image of the fluorescent labeled substance that has been labeled on the microorganisms 1 using an imaging element such as a CMOS image sensor or a CCD image sensor.

[0107] This allows the amount of labeled substance contained in the first specimen sample in the captured image to be measured, and based on this, the amount of target microorganisms 1 contained in the first specimen sample can be calculated. The measurement result of the target microorganisms 1 based on the first specimen sample (for example, the number of live bacteria, dead bacteria, or viruses of microorganisms 1 per unit volume) is defined as the first labeled result.

[0108] The microbial processing step (step S9) and the microbial detection step (step S10) may be performed at different locations after the specimen sample is collected, or may be performed in-line by connecting the specimen sample to the microbial sampling device 10.

[0109] (Space purification step: Step S102) As shown in Fig. 6, between the first sampling step (step S101) and the second sampling step (step S103), space purification of the target space R is performed using a space purification device (step S102: space purification step). The space purification device is, for example, a hypochlorous acid release device that purifies the target space R by releasing hypochlorous acid gas. The hypochlorous acid release device purifies the target space R, including removing, sterilizing, or inhibiting the activity of microorganisms 1, by releasing hypochlorous acid gas.

[0110] When the space purification step (step S102) is performed, the hypochlorous acid gas released from the hypochlorous acid release device purifies the target space R, including removing, sterilizing, or inhibiting the activity of microorganisms 1. This reduces the number of live bacteria or viruses with active surface proteins contained in the target space R.

[0111] (Second sampling step: step S103) After the space purification step (step S102), a second sampling step (step S103) is performed. The first sampling step (step S101) and the second sampling step (step S103) include similar steps, so a description of the second sampling step (step S103) will be omitted. In the second sampling step (step S103), the collected specimen sample (collected liquid 68) is used as the second specimen sample, and the measurement result of the target microorganisms 1 based on the second specimen sample is converted into a second labeled result (for example, the number of microorganisms 1 per unit volume). The number of live bacteria, dead bacteria or viruses)

[0112] The first sampling step (step S101) and the second sampling step (step S103) are each performed under the same conditions, such as the composition of the hydrophilic treatment liquid, the time for hydrophilic treatment, the composition of the collection liquid, the time for collection, the rotation speed of the first electrode 22, and the strength of the applied voltage.

[0113] (Evaluation step: Step S104) Finally, the first labeling result based on the first specimen sample in the first sampling step (step S101) is compared with the second labeling result based on the second specimen sample in the second sampling step (step S103), and the change in the state of the microorganisms 1 in the target space R due to the space purification by the space purification device is evaluated (step S104: evaluation step).

[0114] This makes it possible to compare the states of microorganisms in the target space R in the first sampling step (step S101) and the second sampling step (step S103), and to evaluate changes in the cleanliness of the target space R based on this. In other words, the space purification step (step S102) is performed using a hypochlorous acid release device, and the space purification performance of the hypochlorous acid release device can be evaluated by evaluating changes in the state of the microorganisms 1 before and after the space purification step. For example, by comparing the amount of active microorganisms 1 in the target space R before and after the space purification step (step S102), the purification effect on the microorganisms 1 (the effect of suppressing the activity of the microorganisms 1) due to the release of hypochlorous acid gas can be evaluated.

[0115] More specifically, if the microorganisms 1 to be evaluated are bacteria, archaea, or fungi, a labeling substance capable of labeling live bacteria is used in the collection step (step S6) of the first sampling step (step S101) and the second sampling step (step S103), thereby comparing the numbers of live bacteria contained in the first specimen sample and the second specimen sample, and evaluating the change in the state of the bacteria, archaea, or fungi in the target space R. For example, if the number of live bacteria contained in the second specimen sample is reduced compared to the first specimen sample, the sterilization or disinfection effect of the microorganisms 1 by the space purification step (step S102) can be evaluated based on the degree of reduction.

[0116] Furthermore, when the microorganisms 1 to be evaluated are bacteria, archaea, or fungi, by using a labeling substance capable of labeling dead bacteria in the collection step (step S6) of the first sampling step (step S101) and the second sampling step (step S103), the number of dead bacteria contained in the first specimen sample and the second specimen sample can be compared to evaluate the change in the state of the bacteria, archaea, or fungi in the target space R. For example, if the number of dead bacteria contained in the second specimen sample increases compared to the first specimen sample, the sterilization or disinfection effect of the microorganisms 1 by the space purification step (step S102) can be evaluated based on the degree of increase.

[0117] Furthermore, if the microorganism 1 to be evaluated is a virus, by using a labeling substance capable of labeling the activity of the viral surface protein in the collection step (step S6) of the first sampling step (step S101) and the second sampling step (step S103), the activity of the viral surface protein contained in the first specimen sample and the second specimen sample can be compared to evaluate the change in the state of the virus in the target space R. For example, if the number of active viruses contained in the second specimen sample is reduced compared to the first specimen sample, the effect of the space purification step (step S102) in suppressing the activity of the microorganism 1 can be evaluated based on the degree of reduction.

[0118] As described above, the microbial evaluation method according to this embodiment uses the first electrode 22, which is cylindrical and has both ends open in the axial direction, and the second electrode 24, which extends in the axial direction of the first electrode 22 and is disposed within the first electrode 22 at an interval from the inner surface 66 of the first electrode 22, to measure the concentration of the target air. The microbial evaluation method captures and evaluates microorganisms 1 contained in air taken in through the gap R and flowing inside the device, and includes a supply step (step S4: trapping liquid supply step) of supplying a trapping liquid 68 into the first electrode 22 and storing the trapping liquid 68 on a part of the inner surface 66 in a direction around the axis B of the first electrode 22; a capture step (step S6) of applying a voltage between the first electrode 22 and the second electrode 24 while rotating the first electrode 22 around a rotation axis that extends in the direction of the axis B of the first electrode 22 and passes through the first electrode 22, thereby capturing the microorganisms 1 contained in the air flowing inside the device in the trapping liquid 68; The method includes a first sampling step (step S101) of performing a supplying step (step S4), a collecting step (step S6), and a collecting step (step S8) to collect a first specimen sample containing microorganisms 1, a second sampling step (step S103) of performing a supplying step (step S4), a collecting step (step S6), and a collecting step (step S8) to collect a second specimen sample containing microorganisms 1, and an evaluation step (step S104) of evaluating the state of the microorganisms 1 collected in the first sampling step (step S101) and the second sampling step (step S102). In the collecting step (step S6), a collecting liquid containing a labeling substance capable of labeling the microorganisms 1 to be collected is used as the collecting liquid 68, and in the evaluation step (step S104), a first labeling result based on the first specimen sample and a second labeling result based on the second specimen sample are compared to evaluate a change in the state of the microorganisms 1 in the target space R.

[0119] This makes it possible to compare the states of the microorganisms 1 in the target space R in the first sampling step (step S101) and the second sampling step (step S103), and based on this, to evaluate changes in the cleanliness of the target space R. In this case, since it is possible to label the microorganisms 1 while capturing them, it is possible to reduce the time required to evaluate the captured microorganisms 1.

[0120] Specifically, by labeling the microorganisms 1 while capturing them in the target space R, it is possible to label the microorganisms 1 based on their state at the time of capture. Therefore, compared to conventional methods of recovering a capture liquid containing the microorganisms 1 and then labeling the microorganisms 1 contained in the capture liquid, the state of the microorganisms 1 can be evaluated in a shorter time than the time required to label the microorganisms 1 contained in the capture liquid.

[0121] In addition, the microbial evaluation method of this embodiment further includes a space purification step (step S102) for purifying the space of the target space R, and the space purification step (step S102) is performed between the first sampling step (step S101) and the second sampling step (step S103).

[0122] This allows the first labeling result based on the first specimen sample to be compared with the second labeling result based on the second specimen sample, thereby enabling the change in the state of microorganism 1 in the target space R due to the space purification step (step S102) (the effect of sterilization, disinfection, or activity inhibition of microorganism 1) to be evaluated in a short period of time.

[0123] In the microorganism evaluation method according to this embodiment, the space purification in the space purification step (step S102) may be performed by a space purification device.

[0124] This allows the space purification performance of the space purification device (ability to remove, sterilize, or inhibit the activity of microorganisms 1) to be evaluated. In other words, the space purification performance of the space purification device can be visualized at the evaluation site.

[0125] Furthermore, in the microorganism evaluation method according to the present embodiment, when the microorganism 1 to be evaluated is a bacterium, an archaea, or a fungus, the marker substance includes a marker substance capable of labeling live bacteria or a marker substance capable of labeling dead bacteria.

[0126] According to this, the number of live bacteria or the number of dead bacteria contained in the first specimen sample and the second specimen sample are compared. This makes it possible to evaluate changes in the state of bacteria, archaea, or fungi in the target space R.

[0127] Furthermore, in the microorganism evaluation method according to the present embodiment, when the microorganism 1 to be evaluated is a virus, the labeling substance includes a labeling substance capable of labeling the activity of a surface protein of the virus.

[0128] This makes it possible to compare the activity of the surface protein of the virus contained in the first specimen and the second specimen, and evaluate the change in the state of the virus in the target space R.

[0129] Furthermore, in the microorganism evaluation method according to the present embodiment, the labeling substance is one or more labeling substances selected from a staining reagent, a light-emitting particle, and a color-producing particle.

[0130] This allows the specific microorganism 1 that one wishes to visualize and evaluate to be reliably labeled.

[0131] Furthermore, in the microbial evaluation method according to this embodiment, in the first sampling step (step S101) and the second sampling step (step S103), the supply step (step S4), the capture step (step S6), and the recovery step (step S8) are each performed under the same conditions.

[0132] This allows the state change of the microorganism 1 in the target space R to be evaluated by comparing the first labeling result based on the first specimen sample with the second labeling result based on the second specimen sample.

[0133] The present invention has been described above based on the embodiments, but the present invention is not limited to the above embodiments, and it can be easily inferred that various improvements and modifications are possible within the scope of the invention without departing from the spirit of the invention.

[0134] In the microbial evaluation method according to this embodiment, the collection liquid 68 may contain an amphipathic substance, such as an amphipathic polymer. An amphipathic polymer is a polymer that has a hydrophilic and a hydrophobic portion within its polymer. For example, PEG (Polyethylene Glycol) or dextran is used as the hydrophilic portion, and PLA (Polylactic Acid) or PLGA (Polylactic Co Glycolic Acid) is used as the hydrophobic portion. The amount of the substance contained in the collection liquid is approximately 0.1 to 1.0 weight percent (wt%). A substance amount less than 0.1 wt% may not maintain the hydrophilicity required for efficient collection. A substance amount greater than 1.0 wt% may weaken biological properties, such as the infectivity titer or antigenicity, of the microorganisms 1 to be collected. By supplying the capturing liquid 68 containing this amphipathic polymer and rotating the first electrode 22, the hydrophobic portion of the amphipathic polymer is adsorbed onto the surface of the inner surface 66 of the first electrode 22, and the hydrophilic portion is oriented so as to be exposed to the capturing liquid 68. This increases the affinity at the interface between the inner surface 66 of the first electrode 22 and the capturing liquid 68, making the surface of the inner surface 66 of the first electrode 22 hydrophilic. In other words, the hydrophilic state of the inner surface 66 of the first electrode 22 can be maintained. This makes it possible to suppress the adsorption of microorganisms 1 onto the inner surface 66 of the first electrode 22, thereby reducing the loss of microorganisms 1 in the capturing liquid 68. Note that when the capturing liquid 68 described above is used, the steps performed using the hydrophilization treatment liquid 67 (steps S1 to S3) may be omitted.

[0135] Furthermore, by including an amphiphilic polymer in the collecting liquid 68, the collecting liquid 68 spreads evenly within the first electrode 22, and the distance between the central second electrode 24 and the collecting liquid 68 can be maintained constant. This suppresses dielectric breakdown between the collecting liquid 68 and the second electrode 24, allowing for normal discharge of the second electrode 24 (described below) and the formation of a desired electric field between the second electrode 24 and the first electrode 22, enabling normal sampling.

[0136] In the microorganism evaluation method according to this embodiment, the capturing step (step S6) may be performed while controlling the temperature of the capturing liquid 68. For example, the capturing liquid 68 may be heated in order to increase the labeling efficiency of the microorganism 1 with the labeling substance. More specifically, when the microorganism 1 to be evaluated is a bacterium, archaea, or fungus and a labeling substance capable of labeling live bacteria is used, the capturing liquid 68 may be heated to the optimal temperature for the microorganism 1 to be evaluated or its intracellular enzymes.

[0137] Furthermore, in the microorganism evaluation method according to the present embodiment, the space purification step (step S102) is performed using a space purification device, but this is not limiting. For example, the space purification step (step S102) may be performed using an air purifier, a ventilation device, or natural ventilation, and changes in the state of the microorganisms 1 (changes in the number of live cells of the microorganisms 1 or changes in the number of viruses with active surface proteins) before and after the air purifier, ventilation device, or natural ventilation may be evaluated. This also provides similar benefits.

[0138] Furthermore, in the microbial evaluation method according to the present embodiment, the first sampling step (step S101) is performed immediately before the space purification step (step S102), but this is not limited to this. The first sampling step may be any step in which microorganisms 1 are collected and detected from the target space R before space purification. For example, one of the sampling steps (steps S1 to S10) performed multiple times (for example, five times) may be considered to be the first sampling step. The same is true for the second sampling step (step S103). The second sampling step may be any step in which microorganisms 1 are collected and detected from the target space R after space purification. For example, one of the sampling steps (steps S1 to S10) performed multiple times (for example, five times) may be considered to be the second sampling step. Even in this case, the above-mentioned effects can be obtained.

[0139] Furthermore, in the microbial evaluation method according to this embodiment, when a space purification step is performed between the first sampling step (step S101) and the second sampling step (step S103), a predetermined amount of microorganisms 1 (e.g., bacteria that are not harmful to the human body) may be released and diffused into the target space before the first sampling step (step S101) is performed, ensuring that the microorganisms 1 are present in the target space. This allows for more reliable evaluation of the space purification performance of the space purification device (the ability to remove, sterilize, or inhibit the activity of microorganisms 1), and also makes it easier to compare and evaluate the space purification performance in the space purification step using different types of space purification devices.

[0140] Furthermore, in the microbial evaluation method according to the present embodiment, the space purification step (step S102) is performed after the microbial detection step (step S10) in the first sampling step is completed, but this is not limiting. For example, the space purification step (step S102) may be performed after the collected liquid recovery step (step S8) in the first sampling step is completed. In this way, the time required for evaluating the collected microorganisms 1 can be further reduced by the time required for the microbial treatment step (step S9) and the microbial detection step (step S10). [Industrial Applicability]

[0141] The microbial evaluation method according to this embodiment can be widely used to evaluate microorganisms when verifying and checking spatial purification. [Explanation of symbols]

[0142] 1 Microorganisms 2. Ions 3 locus 10 Microbial sampling equipment 12 Duct 14 No. 1 bearing seal 16 No. 2 bearing seal 18 First flange member 20 Second flange member 22 1st electrode 24 2nd electrode 26 Voltage application section 28 Supply section 30 Collection Department 32 Drive unit 42 Main Unit 44 1st support part 46 Second support part 48 Main Unit 50 flange 52 Main Unit 54 flange 56 Main Unit 58 First outer flange 60 Second outer flange 62 First inner flange 64 Second inner flange 66 Inner 67 Hydrophilic treatment liquid 68 Collection liquid 69 Interior Space 70 First support 72 Second support 74 First Electric Wire 76 Second Electric Wire 78 Tank 79 Tank 80 Injection part 82 Tank 83 Tank 84 Extraction part 86 gears 88 Motor 90 Microbial Treatment Equipment 91 filters 92 Filter support 93 Pressure section 94 Cleaning Solution A. Air flow B axis center C Rotation direction of the first electrode D Storage direction of hydrophilic treatment liquid or collection liquid E electrolysis F gear rotation direction R object space

Claims

1. A microbial evaluation method for capturing and evaluating microorganisms contained in air taken in from a target space and circulating inside the target space, using a first electrode that is cylindrical and has both ends in an axial direction that are open, and a second electrode that extends in the axial direction of the first electrode and is disposed within the first electrode at a distance from an inner surface of the first electrode, a supplying step of supplying a collection liquid into the first electrode and storing the collection liquid in a part of the inner surface in a direction around the axis of the first electrode; a capturing step of applying a voltage between the first electrode and the second electrode while rotating the first electrode around a rotation axis that extends in the axial direction of the first electrode and passes through the first electrode, thereby capturing microorganisms contained in air flowing inside the first electrode in the capturing liquid; a first sampling step of collecting the collection liquid in which the microorganisms have been collected in the collection step as a first specimen sample containing the microorganisms; a second sampling step of performing the supplying step, the capturing step, and the recovering step to recover a second specimen sample containing the microorganism; an evaluation step of evaluating the state of the microorganisms collected in the first sampling step and the second sampling step; Equipped with In the capturing step, a capturing liquid containing a labeling substance capable of labeling the microorganisms to be captured is used as the capturing liquid, In the evaluation step, a first labeling result based on the first specimen sample and a second labeling result based on the second specimen sample are compared to evaluate a change in state of the microorganisms in the target space. Microbial evaluation methods.

2. Further comprising a space purification step of purifying the target space, performing the spatial purification step between the first sampling step and the second sampling step; The method for evaluating microorganisms according to claim 1 .

3. The space purification is performed by a space purification device, In the evaluation step, the space purification performance of the space purification device is evaluated by evaluating the state change of the microorganisms. The method for evaluating microorganisms according to claim 2 .

4. the microorganism is a bacterium, an archaea, or a fungus, The labeling substance includes a labeling substance capable of labeling live bacteria or a labeling substance capable of labeling dead bacteria. The method for evaluating microorganisms according to claim 1 .

5. the microorganism is a virus, the labeling substance includes a labeling substance capable of labeling the activity of the surface protein of the virus; The method for evaluating microorganisms according to claim 1 .

6. The labeling substance is one or more labeling substances selected from a staining reagent, a light-emitting particle, and a color-producing particle. The method for evaluating microorganisms according to claim 4 or 5.

7. In the first sampling step and the second sampling step, the supplying step, the collecting step, and the recovering step are each performed under the same conditions. The method for evaluating microorganisms according to claim 1 .

Images