JP5603114B2 - Reader and diagnostic system - Google Patents

Description

  The present invention relates to a fluorescence reading apparatus and a diagnostic system for reading the state of fluorescent color development on a flat test piece of fluorescent immunochromatography.

The immunochromatography method is efficient because the test substance moves through the porous support by capillary action, is captured by the labeled particles, and is immobilized locally (for example, in a line) on the porous support. This is an immunoassay method for determining the presence or absence of a test substance by concentrating the test substance on contact with the sample and coloring the line on which the capture substance is immobilized.
The features of the immunochromatography method include at least the following two points.
(1) The time required for determination is 20 minutes or less, and rapid inspection is possible.
(2) The measurement can be performed simply by dropping the sample, and the operation is simple.
Utilizing these characteristics, immunochromatography is used for pregnancy test drugs and influenza test drugs, and is attracting attention as a new POCT (Point Of Care Testing) method.
Here, POCT refers to a test for diagnosis as close as possible to the patient. Conventionally, collected specimens such as blood, urine, and affected tissue are sent to a central laboratory in a hospital or a specialized examination center for examination, so it takes time to confirm the diagnosis. According to POCT, prompt and accurate treatment is possible based on examination information provided instantaneously. Therefore, an emergency inspection at a hospital and an inspection during an operation become possible, and the needs in the medical field are increasing.

In conventional immunochromatography, gold nanoparticles are most often used as a labeling substance. In the case of gold nanoparticles, although a detection device is not required, there is a problem that detection sensitivity is low.
Therefore, the present inventors have invented an immunochromatographic labeling reagent using a fluorescent substance as a labeling substance, an immunochromatographic test strip, and the like (see Patent Document 1). The labeling reagent silica nanoparticles for immunochromatography described in Patent Document 1 can impart various light absorption characteristics and fluorescence characteristics. And by using the immunochromatographic test strip described in Patent Document 1, it is possible to perform a test with high detection sensitivity.

Furthermore, the present inventors have invented a fluorescence reading apparatus 301 shown in FIG. 11 in order to visually confirm an immunochromatographic test strip using a fluorescent substance as a labeling substance.
FIG. 11 is a schematic perspective view of a fluorescence reading apparatus 301 of the prior art. An excitation light source (not shown) is installed inside the housing 311 of the fluorescence reader 301. An input unit 312 (switch) for turning on and off the excitation light source is provided on the front surface of the housing 311. A viewing window 313 for viewing the inside of the housing 311 is formed on the upper surface of the housing 311. An insertion port 314 for inserting a flat test piece (test strip) 303 is formed in one of the side surfaces of the housing 311.
The observer inserts the flat test piece 303 into the insertion port 314, presses the input unit (switch) 312 to emit excitation light from the excitation light source, and looks into the housing 311 from the observation window 313, thereby The state of fluorescent color development on the test piece 303 is visually observed to determine the presence or absence of the test substance.
Since the fluorescence reading apparatus 301 shown in FIG. 11 has a simple configuration, the apparatus is light and excellent in portability, and the manufacturing cost can be kept low.

Japanese Patent No. 4360652

However, the fluorescence reading apparatus 301 shown in FIG. 11 has the following problems.
(1) Multiple persons (for example, a doctor and a patient) cannot confirm at the same time.
(2) Since ambient light enters from the viewing window 313, the accuracy of the inspection result is lowered depending on the inspection environment.
(3) In the case of an infectious disease test, a patient suspected of having an infectious disease touches the fluorescence reading device 301 to possibly spread the infection.
(4) The state of fluorescent color development on the flat test piece 303 cannot be stored electronically, and direct evidence of the inspection result cannot be left.

  The present invention has been made in view of the above-described problems, and an object of the present invention is to provide a fluorescence reading apparatus or the like that allows a plurality of persons to simultaneously confirm the inspection result. It is another object of the present invention to provide a fluorescence reading apparatus and the like that can improve the accuracy of inspection results. Another object of the present invention is to provide a fluorescence reading apparatus and the like that can prevent the spread of infection in the case of infectious disease inspection. It is another object of the present invention to provide a fluorescence reader that can leave direct evidence of the inspection result.

In order to achieve the above-mentioned object, the first invention is a fluorescence reading device for reading the state of color development in a fluorescent immunochromatographic flat test piece, and the membrane of the flat test piece adsorbs a fluorescent labeling reagent. Control area, capture area that captures the target substance is immobilized, and the test area that adsorbs the fluorescent labeling reagent only when the fluorescent labeling reagent is bound to the target substance, and the control area and the test area are excluded A housing having a background region, having an insertion port for inserting a test container in which the flat test piece is accommodated, and having all surfaces formed by members that shield light; A light emitting unit that emits excitation light toward the test container inserted from the insertion port, and an imaging unit that captures an area including at least the test region and the control region of the planar test piece. It is, wherein the outside of the housing, a display unit in which the imaging unit for displaying the fluorescence image captured, is provided, in the interior of the housing, on the basis of an output value outputted from the imaging unit, the following control unit which calculates quantitative evaluation index Rt showing a content and a validity index Rv of the measurement results of the target substance represented by the formula, but is further provided, wherein, each of the image which the image pickup unit has captured In order to determine whether the pixel is the pixel in the control area, the pixel in the test area, or the pixel in the background area, two peak values in the output value for each pixel output from the imaging unit are specified. Then, a pixel related to the first peak value on the upstream side with respect to the flow direction of the liquid sample is set as a pixel of the test area, and is a periphery of the pixel related to the first peak value, and has a predetermined constant α (0 < α <1) × output greater than the first peak value The pixel of the value is also the pixel of the test area, the pixel related to the second peak value that is downstream in the flow direction of the liquid sample is the pixel of the reference area, and the periphery of the pixel related to the second peak value The fluorescent reading apparatus is characterized in that a pixel having an output value larger than the predetermined constant × the second peak value is also used as the pixel in the reference region.
Rt = {[statistic of output value of the test area] − [statistic of output value of the background area]} / {[statistic of output value of the control area] − [statistic of output value of the background area] amount]}
Rv = {[statistics of output value of the control region] − [statistics of output value of the background region]} / [statistics of output value of the background region]
According to the first invention, a plurality of persons can confirm the inspection result at the same time. In addition, the accuracy of the inspection result can be improved. In addition, in the case of an infectious disease test, it is possible to prevent the spread of infection. In addition, the content of the target substance can be quantitatively evaluated.
The display unit may further display an output value output from the imaging unit.

In the first aspect of the present invention, it is preferable that a storage unit that is electrically connected to the imaging unit and stores an image captured by the imaging unit is further provided inside the casing.
This can leave direct evidence of the test results.

According to a second aspect of the present invention, there is provided a diagnostic system in which a fluorescence reader for reading the state of color development in a flat specimen of a fluorescent immunochromatography method and a diagnostic apparatus are connected via a network, the membrane of the planar specimen A control region that adsorbs the fluorescent labeling reagent, a test region that adsorbs the fluorescent labeling reagent only when the capture substance that captures the target substance is immobilized and the fluorescent labeling reagent is bound to the target substance, and the control Having a background region excluding the region and the test region, having an insertion port for inserting a test container in which the flat test piece is accommodated, and having a housing formed by a member that shields light on all surfaces, The housing includes at least a light emitting unit that emits excitation light toward the test container inserted from the insertion port, and the test region and the control region of the planar test piece. An imaging unit that captures the frequency, is provided outside the housing, a display unit in which the imaging unit for displaying the fluorescence image captured, is provided, wherein the diagnostic device is output from the imaging unit A control unit that calculates a quantitative evaluation index Rt indicating the content of the target substance represented by the following formula based on the output value and a validity index Rv of the measurement result is provided . In order to determine whether each pixel in the image is a pixel in the control region, a pixel in the test region, or a pixel in the background region, two output values for each pixel output from the imaging unit A peak value is specified, and a pixel related to the first peak value on the upstream side with respect to the flow direction of the liquid sample is set as a pixel of the test region, and a predetermined constant is around the pixel related to the first peak value. α (0 <α <1) × the first peak A pixel having a larger output value is also a pixel in the test region, and a pixel related to the second peak value on the downstream side in the flow direction of the liquid sample is a pixel in the reference region, and the second peak value is The diagnostic system is characterized in that a pixel having an output value larger than the predetermined constant × the second peak value is also a pixel in the reference region around the pixel .
Rt = {[statistic of output value of the test area] − [statistic of output value of the background area]} / {[statistic of output value of the control area] − [statistic of output value of the background area] amount]}
Rv = {[statistics of output value of the control region] − [statistics of output value of the background region]} / [statistics of output value of the background region]
According to the second invention, a plurality of persons can confirm the inspection result at the same time. In addition, the accuracy of the inspection result can be improved. In addition, in the case of an infectious disease test, it is possible to prevent the spread of infection. In addition, the content of the target substance can be quantitatively evaluated.

The fluorescence reading device according to a second aspect of the invention further includes a communication unit that transmits the fluorescence image to the diagnostic device, and the diagnostic device includes a communication unit that receives the fluorescence image from the fluorescence reading device; It is desirable to include a display unit that displays the fluorescent image and a storage unit that stores the fluorescent image.
This can leave direct evidence of the test results. In addition, the patient and the medical staff are not in contact until the test is completed, and the spread of infection can be completely prevented.

  With the fluorescence reading apparatus of the present invention, a plurality of people can confirm the inspection result at the same time. In addition, the accuracy of the inspection result can be improved. In addition, in the case of an infectious disease test, it is possible to prevent the spread of infection. In addition, direct evidence of test results can be left.

1 is a schematic perspective view of a fluorescence reading apparatus 1 according to a first embodiment. (A) Schematic plan view of the flat test piece 5, (b) Longitudinal sectional view of the schematic plan view of the flat test piece 5 Schematic plan view of test container 3 Block diagram showing the configuration of the fluorescence reading apparatus 1 Schematic sectional view showing the internal arrangement of the light emitting unit 17 and the imaging unit 18 of the fluorescence reader 1 Display example of the display unit 13 of the fluorescence reading device 1 Schematic configuration diagram of a diagnostic system 10 according to the second embodiment The block diagram which shows the structure of the fluorescence reader 1a which concerns on 2nd Embodiment. Block diagram showing the configuration of the diagnostic device 7 (A) An example schematically showing an image displayed on the display unit 13, (b) An example of an output value of the imaging unit 18 Schematic perspective view of prior art fluorescence reader 301

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.
In the present invention, the fluorescence immunochromatography method means that a target substance moves in a porous support (membrane) by capillary action, is captured by a fluorescent labeling reagent, and is locally (for example, in a line) on the porous support. The target substance is concentrated by efficient contact with an immobilized capture substance (for example, an antigen or antibody that specifically binds to the target substance), and the line on which the capture substance is immobilized develops a color. An immunoassay that determines the presence or absence of a substance.
Here, the fluorescent labeling reagent means an arbitrary substance composed of a fluorescent part that emits fluorescence when irradiated with excitation light and a binding part that specifically binds to a target substance.
A region where a substance that directly binds to the binding portion is applied on the membrane of the flat test piece is a control region (control line), and the fluorescent labeling reagent is captured in the control region regardless of the presence or absence of the target substance. That is, the amount of the fluorescent labeling reagent captured in the control region is an index representing the total amount of the fluorescent labeling reagent used in the test.
On the other hand, only when the binding part of the fluorescent labeling reagent is bound to the target substance, the area coated with the substance that binds to the binding part via the target substance (preferably, the substance that specifically binds) is the test area ( Test line), where only the fluorescent labeling reagent that captured the target substance is captured. That is, a capture substance that captures the target substance (for example, an antigen or an antibody that specifically binds to the target substance) is immobilized in the test region. Therefore, when both the control region and the test region are colored, it can be determined that there is a test substance. Further, when only the control region is colored, it can be determined that there is no test substance. If the control area does not develop color, re-inspection is performed. However, there is a certain amount of fluorescent labeling reagent that does not flow with the liquid sample and is left behind in the region other than the control region and the test region (non-specific binding), and the fluorescence emitted from these is the background.

<First Embodiment>
First, an outline of the fluorescence reading apparatus 1 according to the first embodiment will be described. FIG. 1 is a schematic perspective view of a fluorescence reading apparatus 1 according to the first embodiment.
Inside the housing 11 of the fluorescence reading device 1, a light emitting unit that emits excitation light and an imaging unit that captures the fluorescence color development state of the flat test piece 5 are installed. An external part (for example, the front part) of the housing 11 is electrically connected to an input part (switch) 12 for turning on / off the power of the fluorescence reading apparatus 1 and an imaging part, and the imaging part takes an image. A display unit 13 for displaying the state of fluorescent color development is provided. An insertion port 14 for inserting the test container 3 in which the flat test piece is accommodated is formed in one of the side portions of the housing 11. In order to prevent light from entering the inside of the housing 11 with the test container 3 inserted, it is desirable that the housing 11 is formed of a member that shields light except for the insertion port 14. Moreover, it is desirable that the insertion port 14 be formed in accordance with the shape of the test container 3. Specifically, it is desirable that the size of the insertion port 14 is formed in accordance with the shape of the cross section perpendicular to the longitudinal direction of the test container 3.
The observer inserts the test container 3 in which the flat test piece is accommodated into the insertion port 14 and presses the input unit (switch) 12 to turn on the power. When the fluorescence reader 1 is turned on, the light emitting unit inside the housing 11 emits excitation light toward the test container 3, the imaging unit images the fluorescent color development state of the flat test piece, and the display unit 13 shows the fluorescent color development state of the flat test piece imaged.

By using the fluorescence reading device 1, the patient and the medical staff (doctor, nurse, etc.) can simultaneously confirm the fluorescent color development state of the flat test piece displayed on the display unit 13.
In addition, since the fluorescent reader 1 does not allow ambient light to enter the housing 11 with the test container 3 inserted, the accuracy of the inspection result can be maintained in any inspection environment.
Further, even in the case of an infectious disease test, the patient can check the fluorescent color development state of the flat test piece without touching the fluorescence reader 1, and thus the spread of infection can be prevented.

Next, the planar test piece of the fluorescence immunochromatography method applied to the present invention will be described. FIG. 2 is a schematic plan view of the flat test piece 5 (FIG. 2A) and a longitudinal sectional view of the schematic plan view of the flat test piece 5 (FIG. 2B).
The membrane 53 of the flat test piece 5 has a control region (control line) 57 that adsorbs the fluorescent labeling reagent, and a capture substance that captures the target substance is immobilized and the fluorescent labeling reagent is bound to the target substance only. It has a test area (test line) 56 that adsorbs the fluorescent labeling reagent, and a background area excluding the control area 57 and the test area 56. In addition, as shown in FIG.
<1> A test addition member (sample pad) 51 and a reagent-containing member (conjugate pad) 52 containing a fluorescent labeling reagent,
<2> Conjugate pad 52 and membrane 53,
<3> It is more preferable that the membrane 53 and the absorption pad 54 are connected in series so that a capillary phenomenon occurs.
Next, each member described above will be described.
1) Test addition member (sample pad) 51
The sample pad 51 is a constituent member that drops a liquid sample containing a target substance.
2) Test-containing member (conjugate pad) 52
The conjugate pad 52 is a component containing a fluorescent labeling reagent, and the target substance contained in the sample liquid that has moved from the sample pad 51 by capillary action is captured by the fluorescent labeling reagent in a molecular recognition reaction such as an antigen-antibody reaction. The part that is labeled.
3) Membrane 53
The membrane 53 is a structural member in which a target substance labeled with a fluorescent labeling reagent moves by capillary action. For example, a test region in which a sandwich-type immune complex formation reaction comprising a capture antibody-target substance-fluorescent labeling reagent is performed 56.
The shape of each of the control region 57 and the test region 56 in the membrane 53 is not particularly limited as long as the capture antibody or the like is locally immobilized, and examples thereof include a line shape, a circular shape, and a belt shape. A line shape of 0.5 to 1.5 mm is preferable.
By a sandwich-type immunocomplex formation reaction comprising a capture antibody-target substance-fluorescent labeling reagent, the target substance labeled with, for example, silica particles is captured in the test region 56, and the fluorescent labeling reagent in the complex thus formed is captured. The target substance can be quantitatively detected based on the degree of fluorescence derived therefrom.
The amount of antibody immobilized in each of the control region 57 and the test region 56 is not particularly limited. However, when the shape is a line, it is preferably 0.5 μg to 5 μg per unit length (cm).
4) Absorption pad 54
The absorption pad 54 is a constituent member that absorbs the liquid sample and the fluorescent labeling reagent that have moved through the membrane 53 by capillary action and always generates a constant flow.

  As a preferred embodiment of a planar specimen for fluorescent immunochromatography applied to the present invention, for example, a test strip for immunochromatography described in Japanese Patent No. 4360652, which uses labeled silica nanoparticles containing a fluorescent substance as a target substance, Examples thereof include an immunochromatography test kit described in JP-A-2005-031029 and an immunological test kit described in JP-A-2000-0119175. Other labeling particles include latex particles colored with a fluorescent dye.

In the present specification and claims, the liquid sample containing the target substance is not particularly limited, and examples thereof include urine and blood.
In the present specification and claims, the target substance as a control for detection, quantification, examination, diagnosis, and judgment is antigen, antibody, DNA, RNA, sugar, sugar chain, ligand, receptor, peptide, chemical substance, etc. Is mentioned.
As a fluorescent labeling reagent, the surface of a fluorescent dye compound-containing colloidal silica particle described in International Publication No. 2007 / 074722A1 is adsorbed by a substance that recognizes a target substance (for example, a biomolecule such as an antibody, an antigen, DNA, or RNA). Examples thereof include silica particles that have been modified.
The fluorescence wavelength of the fluorescent labeling reagent is not particularly limited as long as it is in a range that can be detected by the fluorescence reader 1, but is preferably in the visible light band of 400 to 750 nm.

Next, a test container that houses the flat test piece 5 will be described. FIG. 3 is a schematic plan view of the test container 3.
The test container 3 has an elongated shape made of, for example, a resin, is composed of a lid and a container, and accommodates the flat test piece 5. The test container 3 has a dropping unit 32 and a fluorescence reading window 33. The dropping unit 32 is a part for dropping a liquid sample containing the target substance. The dripping portion 32 is formed with a circular through hole 34 that exposes a part of the sample pad 51, and is formed in a three-dimensional shape in a mortar shape so that the diameter decreases toward the bottom (back in the drawing). Since the dropping part 32 is three-dimensionally formed in a mortar shape, the liquid sample is guided to the through-hole and reliably dropped onto the sample pad 51 of the flat test piece 5. The fluorescence reading window 33 is a window for the fluorescence reading apparatus 1 to read the state of fluorescence development of the flat test piece. The fluorescent reading window 33 is formed with an elongated through-hole 35 that exposes a part of the membrane 53 including the test region 56 and the control region 57, and the peripheral length decreases as it goes down (backward in the drawing). In addition, each side is three-dimensionally formed in a tapered shape. Since each side is formed in a tapered shape, even if the excitation light source is emitted from an oblique direction with respect to the fluorescence reading window 33, the excitation light source is applied to all the areas of the membrane 53 exposed from the fluorescence reading window 33. Is irradiated.

Next, the configuration of the fluorescence reading apparatus 1 will be described. FIG. 4 is a block diagram showing a configuration of the fluorescence reading apparatus 1.
The fluorescence reading apparatus 1 includes a control unit 15, an input unit 12, a power supply unit 16, a light emitting unit 17, an imaging unit 18, a display unit 13, a storage unit 19, and the like.
The control unit 15 includes a CPU (Central Processing Unit), a ROM (Read Only Memory), a RAM (Random Access Memory), and the like, and is provided inside the housing 11. The CPU calls and executes a program stored in the ROM, the recording unit 19 and the like in the work memory area on the RAM, drives and controls each device connected via the bus, and controls the entire fluorescence reader 1. The ROM is a non-volatile memory and permanently holds programs, data, and the like. The RAM is a volatile memory, and temporarily holds a program, data, and the like, and includes a work area used by the control unit 15 to perform various processes.
In the first embodiment, the control unit 15 converts the analog current intensity (analog value) obtained by converting the received fluorescence intensity by the imaging unit 18 into a digital value and displays it on the display unit 13 as a fluorescence image. However, in the first embodiment, the imaging unit 18 converts the analog current intensity into a value that can be displayed on the display unit 13 and transmits the analog current intensity to the display unit 13, so that the control unit 15 is not necessarily required.

The input unit 12 is, for example, a switch for turning on / off the power source of the fluorescence reading device 1, and is provided on the front portion of the housing 11. The input unit 12 may have other switches as necessary.
The power supply unit 16 supplies electromotive force of the fluorescence reading device 1. The power supply unit 16 is, for example, a battery provided in the housing 11. Further, the power supply unit 16 may convert external power into a form suitable for the fluorescence reading device 1 via an adapter and supply the converted power as an electromotive force of the fluorescence reading device 1.

  The light emitting unit 17 emits excitation light toward the test container 3 inserted from the insertion port 14 and is provided inside the housing 11. Examples of the light emitting unit 17 include a white lamp such as a mercury lamp, a halogen lamp, or a xenon lamp, in addition to an LD (laser diode) and an LED (light emitting diode) matched to the wavelength of the fluorescent labeling reagent to be used. In order to make the fluorescence reading apparatus 1 compact, it is desirable that the light emitting unit 17 is an LD (laser diode) that can be easily downsized as compared with other excitation light sources.

  The imaging unit 18 captures the fluorescence color development state of the flat test piece 5 and reads it two-dimensionally, and is provided inside the housing 11. Examples of the imaging unit 18 include a CCD (Charge Couple Device) image sensor, a CMOS (Complementary Metal Oxide Semiconductor) image sensor, and a PD (Photodiode) array. The imaging unit 18 images an area including at least the test area 57 and the control area 56 of the flat test piece 5.

  The display unit 13 displays the fluorescent color development state of the flat test piece 5 imaged by the imaging unit 18, and is provided on the front part of the housing 11. Examples of the display unit 13 include a liquid crystal display and an organic EL (Electro-Luminescence) display.

The storage unit 19 stores the fluorescent color development state of the flat test piece 5 imaged by the imaging unit 18, and is provided inside the housing 11. Examples of the storage unit 19 include an auxiliary storage device such as a hard disk, a memory card reading device incorporating a flash memory, an ultra-small hard disk, and the like. Examples of the memory card include a USB (Universal Serial Bus) memory and an SD card. The storage unit 19 may be provided outside the housing 11 by providing a port such as USB in the housing 11 so that the storage unit 19 can be detached from the housing 11.
Since the fluorescence reading apparatus 1 includes the storage unit 19, it is possible to electronically store the fluorescent color development state of the flat test piece 5 and leave direct evidence of the inspection result.

Next, the internal arrangement of the light emitting unit 17 and the imaging unit 18 of the fluorescence reading device 1 will be described. FIG. 5 is a schematic cross-sectional view showing the internal arrangement of the light emitting unit 17 and the imaging unit 18 of the fluorescence reading device 1.
As shown in FIG. 5, a light emitting unit 17 and a photographing unit 18 are arranged inside the fluorescence reading apparatus 1. Further, when performing the inspection, the test container 3 is inserted from the insertion port 14.
The imaging unit 18 includes a condenser lens 21 (for example, a biconvex lens) that collects fluorescence from the flat test piece 5, and a bandpass filter that removes excitation light from the light emitting unit 17 and transmits only fluorescence from the flat test piece 5. 22, a high-sensitivity image sensor 23 that receives the fluorescence transmitted through the band-pass filter 22 and captures a fluorescence image is provided.
Since the image becomes clearer as the intensity of the excitation light applied to the flat test piece 5 becomes stronger, it is desirable to make the distance between the light emitting unit 17 and the tester 3 as close as possible. There is no restriction | limiting in particular about the irradiation angle of the excitation light by the light emission part 17. FIG.
The test container 3 is fixed at, for example, an intersection 26 of a center line 24 (illustrated by a dotted line) indicating the center of excitation light from the light emitting unit 17 and a center line 25 indicating the center of the condenser lens 21. From the right end of the region (test line) 56 (end on the downstream side with respect to the flowing direction of the liquid sample) and the left end of the control region (control line) 57 (end on the upstream side with respect to the flowing direction of the liquid sample), etc. It is determined to be a distance position.

FIG. 6 is a display example of the display unit 13 of the fluorescence reading apparatus 1. FIG. 6 shows a display example in which the test result is positive, that is, both the control region 57 and the test region 56 are colored and it is determined that there is a test substance. FIG. 6 shows a color image converted into a gray scale image.
Since the fluorescence reading apparatus 1 according to the first embodiment displays the fluorescent color development state of the flat test piece 5 on the display unit 13 as in the display example shown in FIG. can do. In addition, since the image displayed on the display unit 13 of the fluorescence reading device 1 is captured by the imaging unit 18 installed inside the housing 11 where light does not enter the interior, in any inspection environment. In addition, the accuracy of the inspection result can be maintained. Further, even in the case of an infectious disease test, it is possible to confirm the state of fluorescence development of the flat test piece without the patient touching the fluorescence reading device 1.
Furthermore, the fluorescence reading apparatus 1 can store the state of fluorescence development of the flat test piece 5 electronically by storing the image shown in FIG. 6 in the storage unit 19.

<Second Embodiment>
Next, the diagnostic system 10 according to the second embodiment will be described. The same elements as those in the first embodiment are denoted by the same reference numerals, and redundant description is omitted.
FIG. 7 is a schematic configuration diagram of a diagnostic system 10 according to the second embodiment. As shown in FIG. 7, the diagnostic system 10 includes a fluorescence reader 1 a installed in the examination room 101 and a diagnostic apparatus 7 installed in the diagnostic room 103. The fluorescence reading device 1 a and the diagnostic device 7 are connected via a network 9.
A patient 105 is present in the examination room 101, and a medical worker (doctor, nurse, etc.) 107 is present in the diagnosis room 103. Further, the examination room 101 and the diagnosis room 103 are spatially separated, and the patient 105 and the medical worker 107 do not come into contact until the examination is completed.
The network 9 may be wired or wireless. The network 9 may be an in-hospital LAN (Local Area Network) or the Internet.

FIG. 8 is a block diagram showing the configuration of the fluorescence reading apparatus 1a according to the second embodiment.
As shown in FIG. 8, the fluorescence reading apparatus 1a includes a control unit 15, an input unit 12, a power supply unit 16, a light emitting unit 17, an imaging unit 18, a display unit 13, a communication unit 20, and the like.
The communication unit 20 controls communication with the diagnostic device 7 via the network 9 and includes a communication control device, a communication port, and the like. The communication unit 20 may be an external device such as a NIC (network interface card), for example, and a NIC slot may be provided in the housing 11 so as to be removable from the fluorescence reader 1a.

FIG. 9 is a block diagram showing the configuration of the diagnostic device 7. Note that the configuration in FIG. 9 is an example, and various configurations can be adopted depending on the application and purpose.
As shown in FIG. 9, in the diagnostic device 7, a control unit 71, a storage unit 72, a display unit 73, an input unit 74, a communication unit 75, and the like are connected via a bus.
The controller 71 includes a CPU (Central Processing Unit), a ROM (Read Only Memory), a RAM (Random Access Memory), and the like.
The CPU calls a program stored in the storage unit 72, ROM, or the like to a work memory area on the RAM and executes it, drives and controls each device connected via the bus, and realizes processing performed by the diagnostic device 7. .
The ROM is a non-volatile memory, and permanently stores programs such as a boot program and BIOS for the diagnostic device 7, data, and the like.
The RAM is a volatile memory, and temporarily stores a program, data, and the like loaded from the storage unit 72, ROM, recording medium, and the like, and includes a work area used by the control unit 71 for performing various processes.
The storage unit 72 is an HDD (hard disk drive), and stores a program executed by the control unit 71, data necessary for program execution, an OS (operating system), and the like. As for the program, a control program corresponding to an OS (operating system) and an application program are stored. Each of these program codes is read by the control unit 71 as necessary, transferred to the RAM, read by the CPU, and executed as various means.
The display unit 73 includes a display device such as a liquid crystal panel, and a logic circuit or the like (video adapter or the like) for realizing the video function of the diagnostic device 7 in cooperation with the display device.
The input unit 74 inputs data and has an input device such as a pointing device such as a keyboard and a mouse. An operation instruction, an operation instruction, data input, and the like can be performed on the diagnostic device 7 via the input unit 74.
The communication unit 75 includes a communication control device, a communication port, and the like, and is a communication interface that mediates communication between the diagnosis device 7 and the network 9, and performs communication control with the fluorescence reading device 1 a via the network 9.

In the diagnostic system 10 according to the second embodiment, a patient 105 occupying the examination room 101 inserts the test container 3 in which the flat test piece 5 is accommodated into the insertion port 14 of the fluorescence reader 1a, and inputs the input unit. Press 12 to turn on the power. In the fluorescence reading device 1a, when the power is turned on, the light emitting unit 17 inside the housing 11 emits excitation light toward the test container 3, and the imaging unit 18 images the fluorescent color development state of the flat test piece 5. The display unit 13 displays a fluorescence image indicating the state of fluorescent color development of the flat test piece 5 on which the image is taken. In addition, the communication unit 20 of the fluorescence reading device 1 a transmits a fluorescence image to the diagnostic device 7 via the network 9. When the communication unit 75 of the diagnostic device 7 receives the fluorescent image, the control unit 71 displays the received fluorescent image on the display unit 73. The medical worker 107 confirms the fluorescent image displayed on the display unit 73 and determines whether it is positive or negative.
Thus, in the diagnostic system 10 according to the second embodiment, the patient 105 and the medical worker 107 do not contact each other until the examination is completed, and the spread of infection can be completely prevented. The place where the fluorescence reading apparatus 1a is installed is not particularly limited as long as it can be connected to the network 9, and may be the home of the patient 105 or the like.

<Third Embodiment>
Next, quantitative evaluation according to the third embodiment will be described. In addition, the same code | symbol is attached | subjected to the same element as 1st Embodiment and 2nd Embodiment, and suppose that the duplicate description is abbreviate | omitted.
In the fluorescence immunochromatography method, the fluorescent color development on the flat test piece can be evaluated with high sensitivity and quantitative. In order to evaluate quantitatively, the test area (test line) 56, the control area (control line) 57 in the membrane 53, and the background area which is the other area are read two-dimensionally, and the obtained data group is appropriately selected. It is necessary to uniquely derive the target substance content by processing. Therefore, in the quantitative evaluation according to the third embodiment, the output value output from the imaging unit 18 of the fluorescence reader 1 according to the first embodiment or the fluorescence reader 1a according to the second embodiment is used. Based on the quantitative evaluation.
Here, the output value output from the imaging unit 18 is not only the analog current intensity (analog value) obtained by converting the received fluorescence intensity by the imaging unit 18, but also the analog current intensity is digitized by an A / D converter or the like. It is also assumed that the digital value obtained and the value obtained by converting the digital value based on a predetermined calculation formula are included.

FIG. 10A is an example schematically showing an image displayed on the display unit 13, and FIG. 10B is an example of an output value of the imaging unit 18.
The image shown in FIG. 10A includes an examination area 56 and a control area 57, and shows an example in which the test result is positive.
In the graph shown in FIG. 10B, an average value of pixel values is calculated for pixels having the same x coordinate and displayed as an output value. The output value is not limited to this, and may be a pixel value related to a pixel at a specific y coordinate, for example. The specific y coordinate is, for example, the center coordinate of the through hole 34 of the dropping unit 32 formed in the test container 3, the center line 25 indicating the center of the excitation light from the light emitting unit 17 of the fluorescence reading device 1 (1a), and the collection point. For example, the coordinates of the intersection 26 (see FIG. 5) with the center line 25 indicating the center of the optical lens 21 may be used. The coordinates of the center position of the through-hole 34 are considered to have high fluorescence intensity in the test region 56 and the control region 57 because the distance from the position where the liquid sample is dropped is the shortest among the pixels of the same x coordinate.
The display unit 13 of the fluorescence reading device 1 (1a) may display the graph shown in FIG. 10B by superimposing the image captured by the imaging unit.

Below, the output value of the imaging part 18 is considered divided into three types. That is, the output value of the test area pixel 205 which is a pixel corresponding to the test area 56, the output value of the control area pixel 207 which is a pixel corresponding to the control area 57, and the background area pixels 209 and 211 which are pixels corresponding to the background area. 213 is an output value.
As a reference for determining whether each pixel corresponds to the test region pixel 205, the control region pixel 207, or the background region pixels 209, 211, and 213, for example, the peak values 201 and 203 in the graph shown in FIG. Can be used. First, the pixel having the peak value 201 on the upstream side with respect to the flowing direction of the liquid sample is a test area pixel 205. A pixel having an output value larger than α × peak value 201 (0 <α <1) around the pixel having the peak value 201 is also set as the test region pixel 205. Similarly, the pixel having the peak value 203 on the downstream side with respect to the flowing direction of the liquid sample is set as a control region pixel 207. A pixel having an output value larger than α × peak value 203 (0 <α <1) around the pixel having the peak value 203 is also set as the control region pixel 207. The pixels that are not the test region pixel 205 or the control region pixel 207 are background region pixels 209, 211, and 213, which are pixels corresponding to the background region, according to the x coordinate. It is preferable that the value of α can be finely adjusted based on a calibration sample having a clear boundary between the test region 56, the control region 57, and the background region.
If the test result is negative, the peak value 201 does not exist. However, if the test result is negative, there is no need to perform quantitative evaluation. There is no.

The control unit 15 of the fluorescence reading device 1 (1a) or the control unit 71 of the diagnostic device 7 performs quantitative evaluation indicating the content of the target substance represented by the following formula based on the output value output from the imaging unit 18. The index R _t and the validity index R _v of the measurement result are calculated.
R _t = {[statistic of output value of test area] − [statistic of output value of background area]} / {[statistic of output value of control area] − [statistic of output value of background area]}
R _v = {[statistic of output value of control area] − [statistic of output value of background area]} / [statistic of output value of background area]}
The calculated quantitative evaluation index R _t and the validity index R _v of the measurement result are displayed on the display unit 13 of the fluorescence reading device 1 (1a) or the display unit 73 of the diagnostic device 7.
The quantitative evaluation index _Rt is not a value that directly represents the target substance, but in order to obtain the content of the target substance, it is necessary to multiply this by a conversion coefficient. Since the conversion coefficient is a value determined by the combination of the target substance and the fluorescent labeling reagent used, it is desirable to calculate the conversion coefficient corresponding to the combination of the target substance and the fluorescent labeling reagent in advance.
Validity index R _v is some amount of fluorescent labeling reagent to the liquid sample, in which a fixed position of the Flat Specimen 5 indicates whether appropriate.
Due to the restriction due to the size of the fluorescence reader 1 (1a), the irradiation angle of the excitation light from the light emitting unit 17 is oblique as shown in FIG. 5, and if the intensity of the excitation light irradiation is uneven, the validity index error occurs in the setting determination by the R _v. Therefore, providing a calibration sample that shows the same fluorescence intensity in both area of the test region 56 and control region 57, it is preferable to finely adjust the validity index R _v based thereon.

The statistic of the output value of the test area is, for example, the average value, median value, maximum value, minimum value, etc. of the test area pixel 205. Further, the statistic of the output value of the test area may be an output value related to a specific x coordinate, for example. Specific x coordinates include the peak value 201, the center of the pixel determined as the test area pixel 205, the left end, the right end, and the like.
The statistic of the output value of the control area is also the average value, median value, maximum value, minimum value, etc. of the control area pixel 207, for example. Further, the statistic of the output value of the control area may be an output value related to a specific x coordinate, for example. Specific x coordinates include the peak value 203, the center of the pixel determined to be the control region pixel 207, the left end, the right end, and the like.
The statistic of the output value of the background area is, for example, an average value, median value, maximum value, minimum value, or the like of any one of the background area pixels 209, 211, and 213. Further, the statistic of the output value of the background area may be an average value, median value, maximum value, minimum value, or the like of any two of the background area pixels 209, 211, and 213. Further, the statistic of the output value of the background area may be the average value, median value, maximum value, minimum value, etc. of all the background area pixels 209, 211, and 213. Further, the statistic of the output value of the background area may be an output value related to a specific x coordinate, for example. Specific x-coordinates include, for example, the midpoint between the peak value 201 and the peak value 203, the center of the pixel determined as the background region pixel 209 (211 and 213), the left end, and the right end.

In the conventional immunochromatographic test kit, as shown in FIG. 10 (a), the test region 56 and the control region 57 are arranged in series with respect to the flowing direction of the liquid sample, and the test region 56 is located upstream. The control region 57 is in the positional relationship of the downstream side. In this case, if an excessive amount of target substance is contained in the liquid sample, most of the fluorescent target reagent bound to them will be adsorbed specifically to the test region 56, and they will reach the control region 57. I can't. Therefore, the control region 57 does not represent the total amount of the fluorescent labeling reagent mixed in the liquid sample.
Therefore, when the target substance is quantitatively evaluated, the planar test piece 5 is arranged such that the control region 57 and the test region 56 are arranged at an equal distance from the reagent-containing member, in other words, the test region 56 and The control region 57 is preferably arranged in parallel with the liquid sample flow.
Note that the arrangement of two lines arranged in parallel is not very suitable for visually recognizing them. On the other hand, the fluorescence reading device 1 (1a) captures the fluorescent color development state of the flat test piece 5 in a state where ambient light does not enter the inside of the housing 11, and displays the captured image on the display unit 13. Even two lines arranged in parallel are sufficiently distinguishable.

  The preferred embodiments of the fluorescence reading apparatus and the like according to the present invention have been described above with reference to the accompanying drawings, but the present invention is not limited to such examples. It will be apparent to those skilled in the art that various changes or modifications can be conceived within the scope of the technical idea disclosed in the present application, and these naturally belong to the technical scope of the present invention. Understood.

DESCRIPTION OF SYMBOLS 1 ......... Fluorescence reader 3 ......... Test container 5 ......... Plane test piece 7 ......... Diagnosis device 9 ......... Network 10 ......... Diagnostic system 11 ......... Case 12 ......... Input part 13 ......... Display unit 14 ......... Insertion port 15 ......... Control unit 16 ......... Power supply unit 17 ......... Light emitting unit 18 ......... Imaging unit 19 ......... Storage unit 20 ......... Communication unit 56 ... ... test area 57 ..... control area 205 ..... test area pixel 207 ..... control area pixel 209, 211, 213 ..... background area pixel

Claims (5)

A fluorescence reader for reading the state of fluorescent color development in a flat specimen of a fluorescent immunochromatography method,
The membrane of the flat test piece adsorbs the fluorescent labeling reagent only when the control region for adsorbing the fluorescent labeling reagent and the capture substance for capturing the target substance are immobilized and the fluorescent labeling reagent is bound to the target substance. A test area, and a background area excluding the control area and the test area;
It has an insertion port for inserting a test container in which the flat test piece is housed, and includes a housing formed by a member that shields light on all surfaces,
Inside the housing,
A light emitting unit that emits excitation light toward the test container inserted from the insertion port;
An imaging unit for imaging an area including at least the test area and the control area of the planar test piece;
Is provided,
Outside the housing,
A display unit for displaying a fluorescent image captured by the imaging unit;
Is provided ,
Based on the output value output from the imaging unit, the quantitative evaluation index Rt indicating the content of the target substance represented by the following formula and the validity index Rv of the measurement result are calculated inside the casing. A control unit is further provided ,
The control unit outputs from the imaging unit to determine whether each pixel of the image captured by the imaging unit is a pixel in the control region, a pixel in the test region, or a pixel in the background region. Two peak values in the output value for each pixel to be identified are specified, a pixel related to the first peak value on the upstream side with respect to the flow direction of the liquid sample is set as a pixel in the test region, and the pixel related to the first peak value A pixel having an output value larger than a predetermined constant α (0 <α <1) × the first peak value is also a pixel in the test region and is downstream of the liquid sample flow direction. A pixel related to the second peak value on the side is a pixel in the control region, and a pixel having an output value larger than the predetermined constant × the second peak value around the pixel related to the second peak value The pixel in the reference area Fluorescence reader to symptoms.
Rt = {[statistic of output value of the test area] − [statistic of output value of the background area]} / {[statistic of output value of the control area] − [statistic of output value of the background area] amount]}
Rv = {[statistics of output value of the control region] − [statistics of output value of the background region]} / [statistics of output value of the background region]   The fluorescence reading apparatus according to claim 1, wherein the display unit further displays an output value output from the imaging unit. 3. The storage unit according to claim 1, further comprising a storage unit that is electrically connected to the imaging unit and stores an image captured by the imaging unit. Fluorescence reader. A diagnostic system in which a fluorescence reading device for reading the state of fluorescent color development in a flat specimen of a fluorescent immunochromatography method and a diagnostic device are connected via a network,
The membrane of the flat test piece adsorbs the fluorescent labeling reagent only when the control region for adsorbing the fluorescent labeling reagent and the capture substance for capturing the target substance are immobilized and the fluorescent labeling reagent is bound to the target substance. A test area, and a background area excluding the control area and the test area;
It has an insertion port for inserting a test container in which the flat test piece is housed, and includes a housing formed by a member that shields light on all surfaces,
Inside the housing,
A light emitting unit that emits excitation light toward the test container inserted from the insertion port;
An imaging unit for imaging an area including at least the test area and the control area of the planar test piece;
Is provided,
Outside the housing,
A display unit for displaying a fluorescent image captured by the imaging unit;
Is provided ,
The diagnostic device includes a control unit that calculates a quantitative evaluation index Rt indicating the content of the target substance represented by the following formula and a validity index Rv of the measurement result based on an output value output from the imaging unit. Prepared ,
The control unit outputs from the imaging unit to determine whether each pixel of the image captured by the imaging unit is a pixel in the control region, a pixel in the test region, or a pixel in the background region. Two peak values in the output value for each pixel to be identified are specified, a pixel related to the first peak value on the upstream side with respect to the flow direction of the liquid sample is set as a pixel in the test region, and the pixel related to the first peak value A pixel having an output value larger than a predetermined constant α (0 <α <1) × the first peak value is also a pixel in the test region and is downstream of the liquid sample flow direction. A pixel related to the second peak value on the side is a pixel in the control region, and a pixel having an output value larger than the predetermined constant × the second peak value around the pixel related to the second peak value to a pixel of said control region Diagnostic system and butterflies.
Rt = {[statistic of output value of the test area] − [statistic of output value of the background area]} / {[statistic of output value of the control area] − [statistic of output value of the background area] amount]}
Rv = {[statistics of output value of the control region] − [statistics of output value of the background region]} / [statistics of output value of the background region] The fluorescence reading device further includes a communication unit that transmits the fluorescence image to the diagnostic device,
The diagnostic device includes a communication unit that receives the fluorescence image from the fluorescence reading device, wherein a display unit for displaying the fluorescence image, claim 4, characterized in that it comprises a storage unit that stores the fluorescence image The diagnostic system described in 1.