JP2015152491A - Solid phase target substance collection member, solid phase target substance detection system, solid phase target substance collection method and solid phase target substance detection method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new solid phase target substance collection member capable of collecting a solid phase target substance in a measurement object sample gas into a liquid, a solid phase target substance detection system, a solid phase target substance collection method, and a solid phase target substance detection method.SOLUTION: A solid phase target substance collection member 10 comprises a liquid cell 17 having a storage part 171, a water-repellent porous member 16 which is arranged so as to contact a liquid 175 supplied to the storage part 171 and arranged so as to cover the storage part 171 in order to prevent leakage of the liquid 175 from the storage part 171, and a hydrous member 15 which is arranged so as to contact the water-repellent porous member 16. A solid phase target substance detection system 30 comprises the solid phase target substance collection member 10, supply means 20 which can supply a measurement object sample gas to the hydrous member 15, and detection means which can detect the solid phase target substance in the liquid 175 from the storage part 171.

Description

  The present invention relates to a solid phase target substance collection member, a solid phase detection system, a solid phase target substance collection method, and a solid phase detection method.

  Conventionally, methods for collecting and detecting solid phase target substances such as suspended matters (for example, allergens and microorganisms) have been developed.

  Patent Document 1 discloses a method for collecting a solid phase target substance floating in the air with an adhesive sheet.

  Patent Document 2 discloses a method of collecting a solid phase target substance in an aerosol floating in the air with a filter.

  Patent Document 3 discloses a system and method for detecting a solid phase target substance by using a scattered light of particles in the air while directing a convergent beam while the solid phase target substance is present in an air sample. Has been.

JP 2007-135476 A JP 2005-291987 A JP 2006-511822 A

  Unlike the prior art, the present invention provides a novel solid phase target substance collection member, a solid phase target substance detection system, a solid phase, which can recover a solid phase in a sample gas to be measured in a liquid. It is an object of the present invention to provide a method for collecting a target substance and a method for detecting a solid phase target substance.

  The present inventors supply the solid phase target substance to the water repellent porous member and pass through the water repellent porous member in order to recover the solid phase target substance in the liquid via the water repellent porous member. As a result, it was difficult for the solid phase target substance to pass through the water-repellent porous member and it was difficult to collect the solid phase target substance in the liquid, but the liquid-containing member was in contact with the water-repellent porous member. The solid phase target substance is found to move to the liquid through the water repellent porous member by supplying the solid phase target substance from the side of the water-containing member, and completing the present invention. It came to. More specifically, the present invention provides the following.

(1) A collecting member for a solid phase target substance,
A liquid cell having a reservoir;
A water repellent porous member disposed so as to be in contact with the liquid supplied to the reservoir, and disposed so as to cover the reservoir so as to prevent leakage of liquid from the reservoir;
And a water-containing member disposed so as to be in contact with the water-repellent porous member.

  (2) The solid-phase target substance collection according to (1), further comprising a mesh-like member disposed on the opposite side of the water-repellent porous member with respect to the water-containing member and having higher rigidity than the water-containing member. Element.

  (3) The solid phase target substance collecting member according to (2), wherein the mesh member has a porosity of 40 to 70%.

  (4) The water-containing member is further provided with a water leakage inhibiting member that is disposed on the opposite side of the water-repellent porous member and that surrounds the exposed portion of the water-containing member. ) The solid phase target substance collecting member according to any one of the above.

(5) A solid phase target substance detection system,
(1) to (4) the solid phase target substance collecting member, supply means capable of supplying the sample gas to be measured to the hydrous member, and the solid phase target in the liquid from the reservoir A solid phase target substance detection system comprising: a detection means capable of detecting a substance.

(6) A method for collecting a solid phase target substance in a sample gas to be measured,
The water-containing member is disposed so as to contact the water-repellent porous member,
The liquid cell has a reservoir,
The water-repellent porous member is disposed so as to be in contact with the liquid supplied to the reservoir, and is disposed so as to cover the reservoir so as to prevent leakage of liquid from the reservoir.
The said collection method is a collection method of a solid-phase target substance which has the process of supplying the said measuring object sample gas toward the said hydrous member, and collect | recovering the said solid-phase target substance in the said liquid.

  (7) The collection method according to (6), wherein the mesh member is disposed on the opposite side of the water-repellent porous member with respect to the water-containing member and has rigidity.

  (8) The collection method according to (7), wherein the mesh member has a porosity of 40 to 70%.

  (9) A water leakage preventing member is disposed on the opposite side of the water-repellent porous member with respect to the water-containing member, and is disposed so as to surround an exposed portion of the water-containing member. Or the collection method described.

(10) A method for detecting a solid phase target substance in a sample gas to be measured,
Collecting the solid phase target substance in the liquid by the method according to any one of (6) to (9);
Detecting the solid phase target substance in the liquid after collection, and detecting the solid phase target substance in the measurement target sample gas.

  According to the present invention, a novel solid phase target substance collection member, a solid phase target substance detection system, and a solid phase target substance that can recover a solid phase target substance in a sample gas to be measured in a liquid. A collection method and a detection method for a solid phase target substance can be provided.

It is a disassembled perspective view of the collection member for solid phase target substances concerning one example of the present invention. It is a figure which shows the solid-phase target substance detection system which concerns on one Example of this invention. It is a figure which shows the aspect which enables repeated detection in the solid-phase target substance detection system which concerns on one Example of this invention. It is a figure which shows the relative value of the light absorbency at the time of detecting the collect | recovered solid-phase target substance by the solid-phase target substance detection system of FIG. It is a figure which shows the result of having detected Der f1 for every mesh-like member using the mesh-like member of different porosity by the solid-phase target substance detection system of FIG. It is a figure which shows the relationship between the density | concentration of Der f1 in the measurement object sample gas supplied to the hydrous member, and the result of having detected collect | recovered Der f1 by fluorescence using the solid-phase target substance detection system of FIG. The Der f1 in house dust was collected and detected using the solid phase target substance detection system of FIG. 2, and the detection was performed by immunochemical fluorescence, and the ELISA method using a solution prepared directly from house dust It is a figure which shows the graph which compared with the result of having detected Der f1 by. The Der f1 in house dust was collected and detected using the solid phase target substance detection system of FIG. 2, and the detection was performed by immunochemical fluorescence, and the ELISA method using a solution prepared directly from house dust It is a figure which shows the correlation with the result of having detected Der f1 by. It is the figure which showed the selectivity with respect to Der f1 of the detection means using an immunochemical fluorescence method.

  Hereinafter, although one embodiment of the present invention is described with reference to drawings, the present invention is not limited to this.

<Collecting member for solid phase target substance>
As shown in FIG. 1, a solid phase target substance collecting member 10 according to an embodiment of the present invention includes a gas phase cell 11, an exhaust member 12, a water leakage suppression member 13, a mesh-like member 14, and a water content. The permeable member 15, the water repellent porous member 16, and the liquid phase cell 17 are provided. The solid phase target substance collecting member 10 is a member for recovering the solid phase target substance into a liquid 175 (see FIG. 2) supplied to the storage unit 171 of the liquid phase cell 17. As shown in FIG. 1, each component includes a gas phase cell 11, an exhaust member 12, a water leakage suppression member 13, a mesh member 14, and a water-containing member 15 from the suction / discharge member 21 (see FIG. 2) side. The water repellent porous member 16 and the liquid phase cell 17 are arranged in this order. Each component will be described in detail below.

  The liquid phase cell 17 includes a storage part 171, a fixing part 172, a liquid supply part 173 (see FIG. 2), and a liquid outlet part 174 (see FIG. 2). A liquid 175 is supplied to the reservoir 171 of the liquid phase cell 17 (see FIG. 2). The fixing part 172 of the liquid phase cell 17 is a part for connecting the liquid phase cell 17 and the exhaust member 12 via a fixing part 1212 of the main body part 121 of the exhaust member 12 described later. In addition, the fixing portion 172 includes the fixing portion 112 of the gas phase cell 11 described later and the fixing portion 1212 of the main body portion 121 of the exhaust member 12, and each component located between the gas phase cell 11 and the liquid phase cell 17. It is used for fixing with a pinch. More specifically, the gas phase cell 11, the exhaust member 12, and the liquid phase cell 17, which will be described later, are a fixing portion 112 of the gas phase cell 11, a fixing portion 1212 of the main body 121 of the exhaust member 12, A rod-shaped member is disposed so as to pass through the fixing portion 172 of the liquid phase cell 17, and the respective components located between the gas phase cell 11 and the liquid phase cell 17 are sandwiched via the rod-shaped member. Is done. Thereby, each component arranged between the gas phase cell 11 and the liquid phase cell 17 can be fixed. The liquid supply part 173 is a part for supplying the liquid 175 to the storage part 171, and the liquid outlet part 174 is a part for taking the liquid 175 supplied to the storage part 171 out of the storage part 171.

  The water repellent porous member 16 prevents the liquid 175 from leaking from the reservoir 171. Specifically, it is disposed so as to cover the reservoir 171 so as to prevent leakage of the liquid 175 from the reservoir 171, and is in contact with the liquid 175 supplied to the reservoir 171. The water-containing member 15 is disposed in contact with the water-repellent porous member 16. In the present specification, the “water-containing member” refers to a member that can contain water, and includes both a state containing water and a state not containing water.

  As described above, the water repellent porous member 16 is disposed so as to be in contact with the liquid 175 supplied to the storage portion 171, and the water-containing member 15 is disposed so as to be in contact with the water repellent porous member 16. Then, the water-containing member 15 is kept in a state containing water. Here, when the solid phase target substance is supplied to the water-containing member 15 by the supply means 20 (details will be described later) capable of supplying the sample gas to be measured to the water-containing member 15, the water-repellent porous member 16 is interposed. Thus, the solid phase target substance is recovered in the liquid 175 in the reservoir 171. The reason why the solid phase target substance is recovered in the liquid 175 even though the water repellent porous member 16 is sandwiched is that the solid phase target substance is contained in the water contained in the water-containing member 15, so that the liquid 175 It is estimated that this is because a solid phase target substance migrates into the liquid 175 having a lower concentration of the solid phase target substance due to a concentration gradient.

  Although the material which comprises the liquid phase cell 17 is not specifically limited, For example, acrylic resin (polymethylmethacrylate etc.), a polycarbonate, a polyethylene terephthalate, a polystyrene-type resin, polyamide, etc., fluororesins (perfluoroalkoxyalkane (PFA), polytetra) Examples thereof include resins such as fluoroethylene (PTFE) and ethylene-tetrafluoroethylene copolymer (ETFE), and various metals and alloys such as stainless steel, nickel steel, chromium steel, aluminum alloy, and titanium alloy.

  Examples of the material of the water-repellent porous film 16 include fluorine resins such as perfluoroalkoxyalkane (PFA), polytetrafluoroethylene (PTFE), and ethylene-tetrafluoroethylene copolymer (ETFE). The pore size of the water repellent porous membrane 16 is not particularly limited, but is preferably 30 μm or more in that the portion for moving the solid phase target substance from the water in the water-containing member 17 to the liquid 175 becomes large. 50 μm or more is more preferable, and 70 μm or more is most preferable. Further, from the viewpoint of preventing the liquid 175 from leaking, the pore diameter of the water repellent porous membrane 16 is preferably 200 μm or less, preferably 150 μm or less, preferably 120 μm or less, and most preferably 100 μm or less.

  Although the raw material of the water-containing member 15 will not be specifically limited if it is a raw material which can contain water, For example, a cellulose, cotton, etc. are mentioned.

  The mesh member 14 is disposed on the opposite side of the water-repellent porous member 16 with respect to the water-containing member 15. Thus, since the shape is a mesh shape, it is supplied through a ventilation portion 111 of the gas phase cell 11 described later, a ventilation portion 1211 of the main body portion 121 of the exhaust member 12, and a ventilation portion 131 of the water leakage suppression member 13. The measured sample gas can be passed from the mesh gap to the water-containing member 15.

  If the water-containing member 15 contains water, it may be bent. When the water-containing member 15 is bent, the contact area of the water-containing member 15 with respect to the water-repellent porous membrane 16 is reduced, and thus the amount of the solid phase target substance recovered in the liquid 175 may be reduced. Therefore, it is preferable to use a material having a rigidity that can suppress the bending of the water-containing member 15 as the mesh member 14, and specifically, a material having a higher rigidity than the water-containing member 15 is preferable.

  The material of the mesh member 14 is not particularly limited, but it is preferable to use a material having higher rigidity than the water-containing member 15 as described above. Examples of the material of the mesh member 14 include fluororesin (ethylene-tetrafluoroethylene copolymer (ETFE), perfluoroalkoxyalkane (PFA), polytetrafluoroethylene (PTFE), etc.), acrylic resin (polymethyl methacrylate, etc.). Examples thereof include resins such as polycarbonate, polyethylene terephthalate, polystyrene resin, polyamide, and various metals and alloys such as stainless steel, nickel steel, chromium steel, aluminum alloy, and titanium alloy.

  Further, the porosity of the mesh member 14 is not particularly limited. However, as the porosity decreases, it becomes difficult to pass the supplied measurement target sample gas from the mesh gap to the water-containing member 15, and the solid phase target substance to be recovered is collected. The amount may be reduced. From the viewpoint of facilitating passage from the mesh gap to the water-containing member 15, the mesh member 14 has a porosity of preferably 15% or more, more preferably 40% or more, and most preferably 50% or more. From another point of view, when the porosity of the mesh-like member 14 is increased, the portion of the water-repellent porous membrane 16 that can be prevented from being bent is reduced, and the amount of the recovered solid phase may be reduced. The porosity of the mesh member 14 is preferably 80% or less, more preferably 70% or less, and still more preferably 60% or less in that the deflection of the water repellent porous film 16 can be further suppressed. The “porosity” in the present invention refers to the ratio of the area of the hole to the entire area (including the hole) of the mesh member on the surface in contact with the water-containing member.

  The gas phase cell 11 is used to supply the measurement target sample gas to the water-containing member 15. The material of the gas phase cell 11 is not particularly limited. For example, acrylic resin (polymethyl methacrylate, etc.), polycarbonate, polyethylene terephthalate, polystyrene resin, polyamide, etc., fluororesin (perfluoroalkoxyalkane (PFA), polytetrafluoro). Examples thereof include resins (ethylene (PTFE), ethylene-tetrafluoroethylene copolymer (ETFE) and the like), various metals and alloys such as stainless steel, nickel steel, chromium steel, aluminum alloy, and titanium alloy.

  The gas phase cell 11 has a ventilation part 111 and a fixing part 112. The ventilation part 111 forms a space through which a gas sample gas supplied from a suction / discharge member 21 described later passes through the ventilation part 1211 of the main body 121 of the exhaust member 12 arranged on the opposite side of the suction / discharge member 21. The connecting part 113 is a part where the gas phase cell 11 and the discharge part 243 of the suction / discharge part agent 24 are connected. The fixing part 112 is a part for connecting the gas phase cell 11 and the exhaust member 12 via the fixing part 1212 of the main body part 121 of the exhaust member 12.

  The exhaust member 12 forms a space for exhausting the measurement target sample gas supplied from the vent 111 of the gas phase cell 11. Further, the exhaust member 12 is disposed between the gas phase cell 11 and the water leakage member 13 as shown in FIG. The material constituting the exhaust member 12 is not particularly limited. For example, acrylic resin (polymethyl methacrylate, etc.), polycarbonate, polyethylene terephthalate, polystyrene resin, polyamide, etc., fluororesin (perfluoroalkoxyalkane (PFA), polytetra Examples thereof include resins such as fluoroethylene (PTFE) and ethylene-tetrafluoroethylene copolymer (ETFE), and various metals and alloys such as stainless steel, nickel steel, chromium steel, aluminum alloy, and titanium alloy.

  The exhaust member 12 includes a main body part 121 and a space forming part 122. The main body 121 includes a ventilation part 1211 and a fixing part 1212. The ventilation unit 1211 forms a space through which the measurement target sample gas supplied from the ventilation unit 111 of the gas phase cell 11 passes through the ventilation unit 131 of the water leakage suppression member 13. The fixing part 1212 connects the exhaust member 12 to the gas phase cell 11 at one end via the fixing part 112 of the gas phase cell 11 and connects to the fixing part 172 via the fixing part 172 of the liquid phase cell 17 at the other end. It is a part to do.

  The space forming part 122 is a member for forming a space between the gas cell 11 and the main body part 121 by being arranged between the gas cell 11 and the main body part 121, and the formed space is This is the exhaust part 123. The gas sample gas supplied from the vent 111 of the gas phase cell 11 is exhausted from the exhaust unit 123. The space forming part 122 is not particularly limited as long as the exhaust part 123 can be formed. For example, the space forming part 122 can be formed using a washer-like member. When a washer-shaped member is used, the washer-shaped member is passed through the rod-shaped member in a state where it is disposed between the gas-phase cell 11 and the exhaust member 12, and one end of the rod-shaped member is fixed to the gas-phase cell 11. The gas phase cell 11 and the main body 121 are fixed so that the washer-shaped member is sandwiched between the gas phase cell 11 and the exhaust member 12 by passing the other end through the fixing portion 1212 of the main body portion 121. Thus, the exhaust part 123 is formed. The exhaust part 123 is a part for escaping the supplied measurement target sample gas to the outside of the solid phase target substance collecting member 10, whereby the measurement target sample gas can be supplied to the water-containing member 15. .

  The water leakage suppression member 13 is disposed on the opposite side of the water repellent porous member 16 with respect to the water-containing member 15 and is disposed so as to surround the exposed portion of the water-containing member 15. Thereby, the water leakage suppression member 13 prevents water from leaking from between the water-containing member 15 and the main body 121 of the exhaust member 12. The exposed part of the water-containing member 13 refers to a part of the water-containing member 13 facing the ventilation part of the gas phase cell 11 and the ventilation part 1211 of the main body part 121 of the exhaust member 12.

  The water leakage suppression member 13 has a ventilation part 131. The ventilation part 131 forms a space through which the measurement target sample gas supplied from the ventilation part 1211 of the main body part 121 of the exhaust member 12 passes through the ventilation part 131 of the water leakage suppression member 13. The space formed by the ventilation portion 131 is a space formed by surrounding the exposed portion of the water-containing member 15.

  Although the material which comprises the water leak inhibiting member 13 is not specifically limited, For example, packing materials, such as silicone rubber, fluorine rubber, ethylene-propylene rubber, acrylic rubber, nitrile rubber, chloroprene rubber, butyl rubber, styrene-butadiene rubber, urethane rubber Is mentioned. Moreover, the shape of the water leak suppression member 13 will not be specifically limited if it has the ventilation part 131, A sheet form may be sufficient and the shape of an O-ring may be sufficient. The shape of the water leakage suppression member 13 is preferably a sheet shape having an area that can cover the upper surface 1711 of the storage portion 171 of the liquid phase cell 17.

<Solid phase target substance detection system>
As shown in FIG. 2, the solid phase target substance detection system 30 includes the above-described solid phase target substance collection member 10, supply means 20, and detection means (not shown). Each component will be described in detail below. In addition, description of the solid-phase target substance collection member 10 will be omitted with respect to portions overlapping with the above description.

(Supply means)
The supply unit 20 is a unit that can supply the measurement target sample gas to the water-containing member 15, and includes a suction / discharge member 21.

  The suction / discharge member 21 includes an air supply unit 211, a suction unit 212, and a discharge unit 213. The suction unit 212 is a part that sucks the measurement target sample gas. The air supply unit 211 is a part that supplies air into the suction / discharge member. The measurement target sample gas sucked by the suction unit 212 is supplied from the discharge unit 213 to the solid phase target substance collecting member 10 by the gas fed from the gas feeding unit 211. The discharge part 213 is connected to the connection part 113 of the gas cell 11.

  As long as the suction / discharge member 21 is capable of sucking the measurement target sample gas and supplying the measurement target sample gas to the solid phase target substance collecting member 10, any of the conventional known ones may be used. For example, you may use granular material conveyance apparatuses, such as blur slider FK-6 (made by Breath).

  The flow rate at which the aspirated measurement target sample gas is supplied to the solid phase target substance collecting member 10 is not particularly limited, and may be appropriately set according to the supply time or the like. For example, in the case of supplying for 2 minutes, in order to improve the amount of the solid phase target substance to be recovered, the flow rate of supplying the measurement target sample gas is preferably 0.5 L / min or more, and 1.0 L / min or more. More preferably, 1.5 L / min or more is still more preferable.

(Detection means)
The detection means refers to means capable of detecting the solid phase target substance in the liquid 175 from the reservoir 171.

  The detection means is not particularly limited as long as it can be appropriately selected according to the solid phase target substance to be detected and the purpose. As the detection means, for example, a means for detecting using an antibody that captures a solid phase target substance (a means for detecting using an ELISA method, a means for detecting a fluorescent substance produced by reacting a substrate with an enzyme-labeled detection antibody ( Hereinafter, in this specification, it is referred to as immunochemical fluorescence method)) and the like. For detection by immunochemical fluorescence, for example, streptavidin peroxidase (HRP) may be used as a target enzyme, and 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) may be used as a substrate. It is possible to detect fluorescence of resorufin which is a sex product. In the immunochemical fluorescence method, for example, an optical fiber can be used as the sensitive part. The optical fiber may be made by subjecting the end face of a polystyrene optical fiber to mirror treatment (dry polishing, wet polishing, etc.), or a commercially available glass fiber such as F1000-900 PROBE (manufactured by Ocean Optics). Good. Detection means using immunochemical fluorescence is preferable in that detection can be performed with high sensitivity even when the solid phase target substance is at a low concentration. Further, when the solid phase target substance is a nucleic acid probe, the nucleic acid probe may be detected using hybridization utilizing complementarity to the nucleic acid.

  If the detection means is a means that can detect the solid phase target substance collected in the liquid 175 of the reservoir 171 while being held in the liquid 175, as in the case of immunochemical fluorescence described above, the detection means is directly connected to the reservoir 171. You may prepare. Alternatively, a detection unit may be provided separately from the storage unit 171 and the recovered solid phase target substance in the liquid 175 may be taken out and detected by the detection unit.

  The conventional method of collecting the solid phase target substance in the sample gas to be measured by the air bubbling (impinger method, cyclone method) in the liquid is the bubbling, so that the solid phase target substance is detected in the bubbling state. Difficult to connect with measurement technology is difficult, and it is difficult to perform continuous detection in real time. In contrast, according to the present invention, since the solid phase target substance in the measurement target sample gas can be recovered without air bubbling, the solid phase target substance can be detected while recovering the solid phase target substance in the liquid. Is possible. That is, as long as the solid phase target substance can be detected while the solid phase target substance in the liquid 175 is contained in the liquid 175, such as a detection means based on immunochemical fluorescence, the detection means is placed in the reservoir 171. By directly providing, the solid phase target substance can be easily detected continuously in real time.

  Further, in order to replace the supplementary antibody that has already captured the solid phase target substance, the detection means and the liquid phase cell 17 may be configured to be able to replace the antibody. Thereby, it is possible to detect a solid-phase target substance repeatedly. Specifically, for example, as shown in FIG. 3, a sensing unit 1714 serving as a detection unit is provided on the bottom surface 1712 of the storage unit 171, a magnetic control unit 1713 is provided so as to sandwich the sensing unit 1714 from both sides, and a liquid 175 is further provided. By solidifying the capture antibody B therein, the solid phase target substance A can be repeatedly detected. That is, by turning on the magnetism of the magnetic control unit 1713, the capturing antibody B can be fixed to the sensitive unit 1714. The capturing antibody B captures the solid phase target A and detects the solid phase target A. By subsequently turning off the magnetism of the magnetic control unit 1713, the capture antibody B can be separated from the sensitive unit 1714 and removed from the liquid outlet unit 174. Further, by subsequently turning on the magnetism of the magnetic control unit 1713 again, a new capture antibody supplied from the liquid supply unit 173 can be fixed to the sensitive unit 1714. As described above, the solid phase target substance can be repeatedly detected by exchanging the supplementary antibody already supplemented with the solid phase target substance.

  The solid phase target substance refers to both the target solid phase to be collected by the collection member of the present invention and the target solid phase to be detected by the detection system of the present invention. The solid phase target substance is not particularly limited, and examples thereof include allergens such as Der f1, nucleic acid probes, microorganisms, and viruses.

<Effect>
According to this embodiment, the following effects can be obtained.

  The solid-phase target substance collecting member 10 is disposed so as to be in contact with the liquid cell 17 having the reservoir 171 and the liquid 175 supplied to the reservoir 171 and prevents leakage of the liquid 175 from the reservoir 17. Thus, the water-repellent porous member 16 disposed so as to cover the storage portion 171 and the water-containing member 15 disposed so as to be in contact with the water-repellent porous member 16 are configured. Thereby, when the solid phase target substance is supplied to the water-containing member 15, the solid phase target substance can be recovered in the liquid 175 in the reservoir 171 through the water repellent porous member 16. In addition, since the solid phase target substance in the measurement target sample gas can be collected in the liquid 175 without bubbling air, the solid phase target substance can be detected while collecting the solid phase target substance in the liquid 175. It is. Therefore, it can be used as a collecting member for simply and continuously detecting a solid phase target substance in real time.

  In the solid phase target substance collecting member 10, the mesh member 14 is disposed on the opposite side of the water repellent porous member 16 with respect to the water-containing member 15. Thereby, the measurement object sample gas supplied through the ventilation part 131 of the water leakage suppression member 13 can be passed from the mesh gap to the water-containing member 15. Further, the mesh member 14 is configured to have higher rigidity than the water-containing member 15. Thereby, bending of the water-containing member 15 containing water is suppressed, and the amount of the solid phase target substance recovered in the liquid 175 is improved.

  In the solid phase target substance collecting member 10, the water leakage suppressing member 13 is disposed on the opposite side of the water repellent porous member 16 with respect to the water-containing member 15, and is disposed so as to surround the exposed portion of the water-containing member 15. . Thereby, the water leakage suppression member 13 can prevent water from leaking from between the water-containing member 15 and the main body 121 of the exhaust member 12.

  The solid-phase target substance detection system 30 includes a solid-phase target substance collection member 10, a supply unit 20 that can supply a measurement target sample gas to the water-containing member 15, and a solid-phase target in the liquid 175 from the reservoir 171. And a detecting means capable of detecting the substance. Thus, the solid phase target substance contained in the measurement target sample gas can be recovered in the liquid 175 of the solid phase target substance collection member 10, and the recovered solid phase target substance can be detected.

  The preferred embodiment of the solid-phase target substance collection member 10 and the solid-phase target substance detection system 30 according to the present invention has been described above. However, the present invention is not limited to the above-described embodiment, and may be modified as appropriate. Is possible.

  For example, although the solid-phase target substance collection member 10 includes the exhaust member 12, the present invention is not limited thereto. That is, it is sufficient that a portion for allowing the supplied measurement target sample gas to escape to the outside of the solid phase target substance collection member 10 is provided in any of the solid phase target substance collection members 10. The gas phase cell 11 may be provided with an opening that can be evacuated, and the measurement target sample gas may be released from the opening to the outside of the solid phase target substance collecting member 10. In the case where the exhaust member 12 is not included, the water leakage suppression member 13 may be disposed between the gas phase cell 11 and the water leakage suppression member 13. 11 is prevented from leaking between the surface of the water leakage suppressing member 13 and the water-containing member 15.

  Further, in the solid phase target substance collecting member 10, the liquid supply part 173 and the liquid outlet part 174 are provided in the storage part 171, but the invention is not limited thereto. That is, even when the liquid supply part 173 and the liquid outlet part 174 are not provided, the liquid cell 17 is supplied after supplying the liquid 175 to the storage part 171 in advance when using the solid phase target substance collection member 10. You may assemble with another member.

  Although the liquid 175 supplied to the storage part 171 of the liquid phase cell 17 is not specifically limited, For example, buffer liquids, such as a potassium phosphate buffer, may be sufficient, and the case where a detection means is directly provided in the storage part 171. May contain components necessary for detecting the solid phase target substance.

  The supply unit 20 is configured by providing the suction / discharge member 21, but is not limited thereto. That is, any configuration may be used as long as the solid phase target substance can be supplied into the gas phase cell. For example, the solid phase target substance can be manually inserted into the gas phase cell without using the suction / discharge member 21. You may comprise so that supply is possible.

  Although the supply unit 20 includes the suction / discharge member 21, the supply unit 20 may include other members depending on the purpose. For example, in order to confirm the collection capability and detection sensitivity of the solid phase target substance detection system 30, a spray member 22, a gas chamber 23, and a gas supply path member 24 are further provided as shown in FIG. May be. The spray member 22, the gas chamber 23, and the gas supply path member 24 will be described below.

  The spray member 22 has a spray part 221 and sprays the solid phase target substance from the spray part 221 toward the outlet part 223. Specifically, as shown in FIG. 2, the spray liquid 222 is accumulated in the spray member 22, and the spray liquid 222 is sprayed from the spray section 221 toward the outlet section 223. The spray liquid 222 contains a solid phase target substance.

  The spray member 22 may be any conventionally known one as long as it can spray the spray liquid 222 containing the solid phase target substance. For example, NE-C26 (manufactured by OMRON Healthcare), etc. The nebulizer may be used.

  As shown in FIG. 2, the gas chamber 23 has a space for uniformly diffusing the solid phase target substance supplied from the outlet portion 223 of the spray member 21 into the gas, and includes a gas chamber inlet portion 231 and a gas chamber outlet. Part 232. The gas chamber inlet portion 231 is a portion opened so that gas can be sucked by the suction and discharge member 21. The solid phase target substance supplied from the spray unit 231 is uniformly diffused into the gas in the gas chamber 23, and a gas (measurement target sample gas) containing the solid phase target substance is supplied from the gas chamber outlet 232 as a gas. It is supplied to the road member 24.

  The material of the gas chamber 23 is not particularly limited. For example, acrylic resin (polymethyl methacrylate, etc.), polycarbonate, polyethylene terephthalate, polystyrene resin, polyamide, etc., fluorine resin (perfluoroalkoxyalkane (PFA), polytetrafluoroethylene, etc.) (PTFE), ethylene-tetrafluoroethylene copolymer (ETFE) and the like), various metals and alloys such as stainless steel, nickel steel, chromium steel, aluminum alloy, and titanium alloy.

  The gas supply path member 24 has one end connected to the gas chamber outlet 232 and the other end connected to the suction part 212 of the suction / discharge member 21. Thus, the measurement target sample gas supplied from the gas chamber outlet 232 to the gas supply path member 24 is supplied to the suction part 212 of the suction / discharge member 21 through the inside of the gas supply path member 24.

  The material of the gas supply path member is not particularly limited, and examples thereof include flexible materials such as polyethylene, polypropylene, silicone resin, vinyl chloride, and synthetic rubber.

<Example 1>
The solid phase target substance was collected and detected using the solid phase target substance detection system. The solid phase target substance detection system used in the above-described embodiment is configured without providing the suction / discharge member, the spray member, the gas chamber, and the gas supply path member. Further, as the solid phase target substance collecting member, the one configured without providing the exhaust member in the above-described embodiment was used. In the above-described embodiment, the gas phase cell is configured such that a portion corresponding to the connecting portion is closed without being provided with a connecting portion with the discharge portion of the suction / discharge member and is not opened. As the water repellent porous member, PFA net (F-3220-01, pore diameter 89 μm, manufactured by Freon Industries Co., Ltd.) is used, and as the water containing member, a hydrous cellulose membrane (M-3II, manufactured by Asahi Kasei Fibers Co., Ltd.) is used. As the mesh member, ETFE mesh (F-3006-9A-1800, manufactured by Freon Industries, Ltd., porosity 44%) was used. The liquid phase cell uses the lower side of a commercially available dialysis cell (capacity: 1.5 ml, FA-1, manufactured by Sampleratech Co., Ltd.), and an O-ring (size: inner diameter 25 mm, material: silicone rubber) as a water leakage inhibiting member Was used.

  The solid phase target substance was detected as Der f1 in house dust. First, house dust was sealed on the gas phase cell side, the liquid in the liquid phase cell (phosphate buffer solution) was stirred for 30 minutes, and then the liquid in the liquid phase cell was recovered to detect Der f1. . Detection was performed by ELISA. Specifically, using a supplemental anti-Der f1 antibody, horseradish peroxidase (HRP) as a target enzyme, and TMB chromogenic substrate (3,3 ′, 5,5′-tetramethylbenzidine) as a substrate The wavelength of 3,3 ′, 5,5′-tetramethylbenzidinediimine, which is a product, was detected. Further, a microplate reader (SH-1000Lab) manufactured by Corona Electric Co., Ltd. was used as a detector.

<Comparative Example 1>
As Comparative Example 1, Der f1 was collected and detected under the same conditions as in Example 1 except that the hydrated cellulose membrane and the ETFE mesh were not used.

  Those without Der f1 were detected as negative controls. FIG. 4 shows the detection results of Example 1, Comparative Example 1, and negative control. As shown in FIG. 4, in Comparative Example 1, the amount of absorption wavelength was smaller than that of negative control, and in Comparative Example 1, it was confirmed that Der f1 could not be absorbed. On the other hand, in Example 1, compared with Comparative Example 1 and negative control, a large amount of Der f1 was confirmed, that is, Der f1 in the house dust supplied to the water-containing member was water repellent. It was confirmed that the liquid in the reservoir was recovered through the porous member. The reason why the solid phase target substance was recovered in the liquid in spite of sandwiching the water-repellent porous member is that the solid phase target substance is contained in the water contained in the water-containing member. It is presumed that a concentration gradient occurs and the solid phase target substance moves into a liquid having a lower concentration.

<Example 2>
In the above-described embodiment, the solid phase target substance detection system is configured by providing a suction / discharge member, a spray member, a gas chamber, and a gas supply path member. As the solid-phase target substance collecting member, the solid-phase target substance collecting member of Example 1 described above is provided with an exhaust member, and the water leakage suppressing member is not an O-ring but a silicone sheet (thickness). : 0.5 mm, having an area that can cover the upper surface of the storage part of the liquid phase cell), the point that the porosity of the ETFE mesh was changed to 29%, and the gas phase cell, The same thing as the collection member for the solid phase target substance of Example 1 is used except that the upper side of the same thing as the cell is machined and a connecting part that can be connected to the discharging part of the suction and discharging member is provided. The solid phase target substance was collected and detected. As the exhaust member, an acrylic plate (thickness 3 mm) was used as the main body, and two stainless steel washers (each thickness: 1.6 mm) were used as the space forming portion. NE-C26 (made by OMRON Healthcare) was used as the spray member. As the gas chamber, a conductive resin container (1000 ml, 25007, manufactured by Sampleratech Co., Ltd.) was used. A vinyl tube was used as the gas supply path member. As a suction and discharge member, a blur slider FK-6 (manufactured by Breath) was used. Supply is performed by using a horseradish peroxidase (HRP) solution of 0.6 μg / ml (enzyme activity 0.06 units / ml) as a spray, spraying the spray rate of the HRP solution at 400 μl / min for 2 minutes, and a suction discharge member The measurement sample gas was supplied to the gas phase cell at a flow rate of 2 L / min, and the HRP supplied to the gas phase was 0.48 μg (0.048 units). The liquid in the liquid phase cell was the same as in Example 1. In the detection, the HRP taken in the liquid is collected, and the wavelength of the product 3,3 ′, 5,5′-tetramethylbenzidinediimine is obtained using a TMB chromogenic substrate in the same manner as in Example 1. The detection was performed using a microplate reader (SH-1000Lab) manufactured by Corona Electric Co., Ltd.

<Example 3>
Der f1 was collected and detected under the same conditions except that the porosity of the ETFE mesh of Example 2 was changed from 29% to 44%.

<Example 4>
Der f1 was collected and detected under the same conditions except that the porosity of the ETFE mesh of Example 2 was changed from 29% to 59%.

<Example 5>
Der f1 was collected and detected under the same conditions except that the porosity of the ETFE mesh of Example 2 was changed from 29% to 61%.

  The detection results of Examples 2 to 5 are shown in FIG. When Examples 2 to 4 were compared, it was confirmed that Example 4 (59%), which has a high porosity of the ETFE mesh, has the largest recovery amount of Der f1. This is because the porosity of Example 4 is higher than that of Examples 2 and 3, so it is easy to pass the supplied measurement target sample gas from the mesh gap to the water-containing member, and the amount of Der f1 to be recovered is high. This is thought to be because of On the other hand, when Example 4 (59%) was compared with 5 (61%), it was confirmed that Example 4 having a low porosity of the ETFE mesh has a large amount of Der f1 recovered. This is because the porosity of Example 5 is higher than that of Example 4 and it is easy to pass from the mesh gap to the water-containing member, but the portion that can suppress the deflection of the water-repellent porous membrane is reduced, so that the recovered Der This is probably because the amount of f1 has decreased. That is, Example 5 had the largest amount of Der f1 recovered because of the ease of passing the supplied measurement target sample gas from the mesh gap to the water-containing member and the suppression of the bending of the water-repellent porous membrane. This is presumably because of its excellent balance.

<Example 6>
Der f1 was collected and detected using the same solid phase target substance collecting member as in Example 2 except that the porosity of the mesh member was 44%. In addition, the Der f1 solution was used as the spray liquid from the spray member, and the concentration of the Der f1 solution (spray rate: 400 μl / min) was set in the same manner as in Example 2 except that it was set as shown in Table 1 below. went. The right column of Table 1 shows the Der f1 concentration in the measurement target sample gas.

  The point that the solid phase to be detected was changed to Der f1 was the same as in Example 2. First, the solid phase target substance recovered in the liquid in the reservoir of the liquid phase cell was stirred for 8 minutes in the liquid, and then a sample was taken to detect Der f1. Detection was performed by immunochemical fluorescence.

  Specifically, first, a capture anti-Der f1 antibody was immobilized on the inner wall of a polyethylene measurement cell. Next, after the blocking treatment, each recovered Der f1 solution (0.49 to 250 ng / ml, room temperature 2 h), biotinylated anti-Der f1 antibody (room temperature 1 h), streptavidin peroxidase (room temperature 0.5 h) The reaction was performed in the order of to form an immune complex. Thereafter, 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) is added as a substrate, and immediately after the reaction is stopped, the end face of the optical fiber, which is the sensitive part, is immersed in a measurement cell, and resorufin (excitation light: excitation light: (570 nm, fluorescence: 585 nm) was detected. As a sensitive part, dry polishing (silica film, particle diameter 30 μm) and wet polishing (silica film, particle diameter 30 μm) on both surfaces of a polystyrene optical fiber (Φ1.0 mm, 12.0 cm, Polystyrene plastic optical fiber, Shenzhen Corporate Photoelectric) (Diameter: 9 μm alumina film, particle diameter: 0.3 μm) was used. In this example, excitation light from an LED (OSPG 5111P, manufactured by OptSupply) is passed through a bandpass filter (BPF: 570 ± 10 nm (MX0570), manufactured by Asahi Spectroscopic Co., Ltd.) from the output side of the bifurcated optical fiber to the sample solution. Irradiation and the fluorescence of resorufin generated in the sample are passed through a fluorescence bandpass filter (600 ± 10 nm (MC600), manufactured by Asahi Spectroscopic Co., Ltd.) to a photomultiplier tube (PMT; H7421, manufactured by Hamamatsu Photonics). The amount of Der f1 was measured. The result is shown in FIG.

As shown in FIG. 6, it was confirmed that in the case where the Der f1 solution having each concentration was used as the spray solution, the respective fluorescence outputs increased with the increase of the Derf concentration (0.125-2.0 mg). / M ³ ). Thus, it was confirmed that Der f1 could be detected in a concentration range of 0.49 to 250 ng / ml by using an immunochemical fluorescence method using an optical fiber as a detection means. Further, it was confirmed that the average recovery rate of Der f1 was 0.23% with respect to the concentration of Der f1 in the measurement target sample gas.

(Test Example 1)
Using the solid phase target substance detection system used in Example 6, four types of dust were collected from carpets, mattresses, blankets, and pillows using a vacuum cleaner, and other house dust (product number D9, manufactured by Greer, USA). ) Was collected and collected for each 5 mg of dust. As a comparison, 5 mg of each dust was added to 100 μl of 0.1% Tween20-PB solution, soaked at room temperature for 4 h, centrifuged at 2000 g for 10 minutes, and the supernatant was collected. Using this, the ELISA method was used. Detection was performed. The result is shown in FIG. FIG. 8 shows the correlation between the detection result of Der f1 by the immunochemical fluorescence method using the solid phase target substance detection system of Example 6 and the detection result by the ELISA method. “House dust” in FIG. 7 indicates the other house dust (product number D9, manufactured by Greer, USA). In addition, "fiber-optic chemistry immunoassay" in FIG. 7 shows the result detected by the immunochemical fluorescence method using the solid-phase target substance detection system of Example 6.

  As shown in FIG. 7, when the detection result (immunochemical fluorescence method) using the solid phase target substance detection system of Example 6 is compared with the detection result by ELISA method, detection by the immunochemical fluorescence method of Example 6 is performed. Was confirmed to have a quantitative property equivalent to that detected by the ELISA method, and to have better detection sensitivity on the low concentration side. Further, as shown in FIG. 8, the correlation between the detection results of both was confirmed, and the detection by the immunochemical fluorescence method of Example 6 was shown to be consistent with the detection result by the ELISA method.

(Test Example 2)
The selectivity for detection by Der f1 of the immunochemical fluorescence method used in Example 6 was tested. Seven single antigen solutions and two mixed solutions were used for the test. Specifically, a single antigen solution of each of Der f1, Der f2, Der p1, Cry j1, Amb a1, Alt a1, and Can f1, a mixed solution of Der f1 and Der f2, and Der f1 and Der p1 Using the mixed solution, the allergen concentration at the time of detection of each solution was adjusted to 10 ng / ml. The detection result is shown in FIG.

  As a result, a significant difference (p <0.05) was observed in the output between the Der f1 single solution and the other single antigen solutions. Further, it was confirmed that the fluorescence output of the two mixed solutions containing Der f1 was comparable to that of the Der f1 single antigen solution. From the above results, it was confirmed that detection by immunochemical fluorescence has high selectivity for Der f1.

DESCRIPTION OF SYMBOLS 10 Collecting member for solid phase target substance 11 Gas phase cell 111 Vent part of gas phase cell 112 Gas phase cell fixing part 113 Connection part with suction / discharge member 12 Exhaust member 121 Main body part 1211 Exhaust member vent part 1212 Exhaust member Fixed portion 122 space forming portion 123 exhaust portion 13 water leakage inhibiting member 131 ventilation portion of water leakage inhibiting member 14 mesh-like member 15 water-containing member 16 water repellent porous member 17 liquid phase cell 171 reservoir portion 1711 upper surface of reservoir portion 1712 reservoir portion 1713 Magnetic control unit 1714 Sensitive unit 172 Liquid phase cell fixing unit 173 Liquid supply unit 174 Liquid outlet unit 175 Liquid 20 Sample gas supply means 21 for measurement 21 Suction discharge member 211 Air supply unit 212 Suction unit 213 Discharge unit 22 Spray member 221 Spraying part 222 Spraying liquid 223 Outlet part of spraying member 23 Gas chamber 231 Gas inlet portion 232 Gas outlet portion 24 Gas supply path member 30 Solid phase target substance detection system A Solid phase target substance B Supplementary antibody

Claims (10)

A collecting member for a solid phase target substance,
A liquid cell having a reservoir;
A water repellent porous member disposed so as to be in contact with the liquid supplied to the reservoir, and disposed so as to cover the reservoir so as to prevent leakage of liquid from the reservoir;
And a water-containing member disposed so as to be in contact with the water-repellent porous member.   The solid-state target substance collecting member according to claim 1, further comprising a mesh-like member disposed on the opposite side of the water-repellent porous member with respect to the water-containing member and having higher rigidity than the water-containing member.   The solid phase target substance collecting member according to claim 2, wherein the mesh member has a porosity of 40 to 70%.   4. The water leakage member according to claim 1, further comprising a water leakage suppressing member that is disposed on the opposite side of the water repellent porous member with respect to the water containing member and that surrounds an exposed portion of the water containing member. Collection member for solid phase target substance. A solid phase target substance detection system comprising:
A solid-state target substance collecting member according to any one of claims 1 to 4, supply means capable of supplying a measurement target sample gas to the hydrous member, and the solid-phase target substance in a liquid from the reservoir A solid phase target substance detection system comprising: a detectable detection means. A method for collecting a solid phase target substance in a sample gas to be measured,
The water-containing member is disposed so as to contact the water-repellent porous member,
The liquid cell has a reservoir,
The water-repellent porous member is disposed so as to be in contact with the liquid supplied to the reservoir, and is disposed so as to cover the reservoir so as to prevent leakage of liquid from the reservoir.
The said collection method is a collection method of a solid-phase target substance which has the process of supplying the said measuring object sample gas toward the said hydrous member, and collect | recovering the said solid-phase target substance in the said liquid.   The collection method according to claim 6, wherein the mesh-shaped member is disposed on the opposite side of the water-repellent porous member with respect to the water-containing member and has higher rigidity than the water-containing member.   The collection method according to claim 7, wherein the mesh member has a porosity of 40 to 70%.   9. The collection according to claim 6, wherein a water leakage preventing member is disposed on the opposite side of the water-repellent porous member with respect to the water-containing member and surrounds an exposed portion of the water-containing member. Method. A method for detecting a solid phase target substance in a sample gas to be measured,
Collecting the solid phase target substance in the liquid by the method according to claim 6;
Detecting the solid phase target substance in the liquid after collection, and detecting the solid phase target substance in the measurement target sample gas.

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