JP2016191567A - Fluorescence detection system, immunochromatographic device, immunochromatographic method, and fluorescence detection method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a fluorescence detection system which enables high-S/N detection of a target fluorescent label without being affected by autofluorescence of a porous support body with an immunochromatographic device or the like using an inexpensive and simple configuration.SOLUTION: A fluorescence detection system 5 comprises a fluorescence excitation light source 2, a support body 1 on which a fluorescent label is spread, and fluorescence detection means 3 for detecting fluorescence of the fluorescent label. The fluorescence detection means 3 is a color sensor, which allows for detecting the fluorescent label spread on the support body 1 that generates autofluorescence, without using time-resolved fluorescence and wavelength range cut filters.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a fluorescence detection system, an immunochromatography device, an immunochromatography method, and a fluorescence detection method. In particular, in an immunochromatography apparatus or the like, a target fluorescent label is not affected by the autofluorescence of a porous support. The present invention relates to a fluorescence detection system, an immunochromatography device, and the like that can detect at a high S / N ratio.

  In medical settings such as hospitals, clinics, nursing homes, and home medical care, there is a growing need for tests for point-of-care testing (POCT) to diagnose patients as close to the patient as possible without the need for clinical laboratory specialists. ing. This is because POCT enables rapid and accurate treatment based on immediately obtained examination information. As an inspection apparatus for POCT, an immunochromatography apparatus based on an immunochromatography method has been conventionally used.

  In immunochromatography, when analytes (specimens) such as collected blood, urine, and affected tissue penetrate the porous support body by capillary action, they are captured by labeled particles (antibodies) such as gold colloid, color latex, and fluorescent substances. In addition, an immune complex is formed by efficient contact with a capture substance (immobilized antibody) locally immobilized on the porous support, and the specimen is detected as a visible or visible accumulation. This is the principle. The time required for the determination is short (20 minutes or less), rapid examination is possible, and measurement can be performed only by a simple operation of dropping the specimen onto the apparatus.

  Conventionally, many technical proposals have been made on immunochromatography apparatuses. For example, in Patent Document 1 listed below, as an immunochromatographic sensor capable of easily confirming that a specimen (sample solution) has entered the development layer through the visualization substance-binding antibody layer, a sample solution absorption part, a visualization substance-binding antibody layer, And a test strip having a development layer large enough for an antigen-antibody reaction, and a housing containing the test strip, wherein the development layer has a determination unit provided with a fixed antigen, and the housing has a determination unit. An immunochromatographic sensor has been disclosed in which a window portion that allows visual observation from the outside and a window portion that allows at least part of the visualization substance-bound antibody layer to be visible from the outside are provided.

  Patent Document 2 discloses an immunochromatography kit containing organic silver salt particles, a reducing agent for silver ions, and a metal colloid label or a metal sulfide label as a simple, quick and highly sensitive immunochromatography kit. Yes. Patent Document 3 discloses a configuration of a reader using a cartridge provided with a plurality of depressions having a shape corresponding to the outer shape of the casing of the immunochromatographic test device as a reader for an immunochromatographic test piece that can be applied to casings having different shapes. Has been.

JP 2010-156642 A "Immunochromatographic sensor" JP 2008-139297 A "Immunochromatographic kit" Japanese Patent Application Laid-Open No. 2005-114677 “Immunochromatographic Test Piece Reader”

  In an immunochromatography apparatus, there are several types of sensors for qualitative / quantitative detection of immobilized capture substances, but photodiodes and phototransistors are often used when the system is configured at low cost. For example, when measuring fluorescent labels using these sensors, when the excitation light in the ultraviolet region is irradiated, the porous support also generates autofluorescence at the same time as the fluorescent label, which is a factor that the S / N ratio cannot be increased. Yes.

  One of the measures for improving the S / N ratio is “time-resolved fluorescence measurement method”. To do this, a substance that emits “phosphorescence” is used for the label. Phosphorescence persists for a certain period of time even when the excitation light is turned off. The autofluorescence of the porous support is not “phosphorescence”, but extinguishes at the same time as the excitation light is extinguished. Therefore, it is possible to measure only the luminescence amount of the fluorescent label without increasing the S / N ratio.

  In addition to “time-resolved fluorescence”, a self-fluorescence of the porous support can be removed by placing a filter that can cut light in a limited wavelength region, such as a bandpass filter, in front of the sensor. However, in any case, in order to perform these methods, it is inevitable to add functions in terms of software and hardware, and the measuring instrument becomes expensive.

  Therefore, the problem to be solved by the present invention is to eliminate such problems of the prior art, and in an immunochromatography apparatus and the like, by a cheaper and simple configuration, without being affected by autofluorescence of the porous support, It is intended to provide a fluorescence detection system, an immunochromatography device, an immunochromatography method, and a fluorescence detection method that can detect a fluorescent label to be detected at a high S / N ratio.

  As a result of studying the above problems, the inventor of the present application has found that it can be solved by using a color sensor capable of individually recognizing R, G, and B color signals instead of the conventional photodiodes and phototransistors. The present invention has been completed. That is, the invention claimed in the present application, or at least the disclosed invention, as means for solving the above-described problems is as follows.

[1] A fluorescence detection system comprising a fluorescence excitation light source, a support on which a fluorescence label is developed, and fluorescence detection means for detecting fluorescence of the fluorescence label, the fluorescence detection means being a color sensor, A fluorescence detection system capable of detecting a fluorescent label developed on the support that generates autofluorescence without using time-resolved fluorescence or a wavelength region cut filter.
[2] The fluorescence detection system according to [1], wherein a fluorescent label that emits fluorescence having a color region different from the autofluorescence color region of the support is used.
[3] The fluorescence detection system according to [1] or [2], wherein the support is a porous support, and europium is used for the fluorescent label.

[4] The fluorescence detection system according to any one of [1] to [3], which is used for immunochromatography.
[5] An immunochromatography device using the fluorescence detection system according to any one of [1] to [3].
[6] An immunochromatography method using the fluorescence detection system according to any one of [1] to [3].
[7] The fluorescent label developed on the support that generates autofluorescence by detecting the fluorescent label developed on the support and fluorescently excited using a color sensor that is a fluorescence detection means is time-resolved. A fluorescence detection method characterized by being able to detect without using fluorescence or a wavelength region cut filter.

  Since the fluorescence detection system, the fluorescence detection method, and the like of the present invention are configured as described above, according to these, a cheaper and simpler configuration can be achieved in a technique / apparatus / system that requires fluorescence detection such as immunochromatography. Thus, the target fluorescent label can be detected with a high S / N ratio without being affected by the autofluorescence of the porous support. Moreover, the detection accuracy can be improved more inexpensively and easily than before by an immunochromatography device, an immunochromatography reader, or an immunochromatography method using such a fluorescence detection system.

It is a conceptual diagram which shows the basic composition of the fluorescence detection system of this invention. It is a perspective view which shows the Example of this invention immunochromatography device. It is a perspective view which shows the Example of the immunochromatography reader for the immunochromatography device of FIG. It is a graph which shows the example of fluorescent color measurement by a color sensor.

Hereinafter, the present invention will be described in detail with reference to the drawings.
FIG. 1 is a conceptual diagram showing the basic configuration of the fluorescence detection system of the present invention. As shown in the figure, the present fluorescence detection system 5 comprises a light source 2 for excitation of fluorescence, a support 1 on which a fluorescent label is developed, and a fluorescence detection means 3 for detecting the fluorescence of the fluorescence label. The main configuration is that it is a sensor and can detect a fluorescent label developed on the support 1 that generates autofluorescence without using time-resolved fluorescence or a wavelength region cut filter.

  That is, in the present invention, a color sensor capable of individually recognizing R, G, B color signals is used as the fluorescence detection means 3 instead of the conventional photodiode or phototransistor. In the present fluorescence detection system 5, a fluorescent label that emits fluorescence in a color region different from the autofluorescence color region of the support 1 is used. Since a porous support is usually used as the support 1, examples of fluorescent labels that emit fluorescence in different color regions include a europium complex. In addition, this invention is not limited to this, If it is a fluorescent label which emits the fluorescence of a color area different from the autofluorescence of a support body, it can use suitably.

  With this configuration, the fluorescence detection method can be performed by the fluorescence detection system 5. That is, the support 1 on which the fluorescent label is developed is irradiated with excitation light L from the fluorescence excitation light source 2 to emit fluorescence F, which is detected by the color sensor as the fluorescence detection means 3. At this time, autofluorescence from the support 1 is also generated and detected by the color sensor. However, the fluorescence F by the fluorescent label is a fluorescence having a color region different from the autofluorescence color region of the support 1. Both are detected separately. Therefore, the fluorescent label developed on the support 1 can be immediately detected without using time-resolved fluorescence detection. Of course, it is not necessary to use a wavelength region cut filter.

FIG. 2 is a perspective view showing an embodiment of the immunochromatography device of the present invention. Also,
FIG. 3 is a perspective view showing an embodiment of an immunochromatography reader for the immunochromatography device of FIG. In these, the above-described fluorescence detection system and fluorescence detection method of the present invention can be used.

  FIG. 4 is a graph showing an example of fluorescent color measurement by a color sensor. In the figure, (I) “Pure” is autofluorescence of the porous support (membrane) itself, and (II) “Phosphorescent paint (white light emission)” is a porous support coated with white light emitting paint. (III) “Fluorescence (2 μl)”, (IV) “Fluorescence (5 μl)”, respectively, in accordance with the method of the present invention, the fluorescence label developed on the support is converted into the autofluorescence of the porous support. The color detection result of fluorescence when a europium complex (excitation light 340 nm, detection wavelength 615 nm) emitting fluorescence R (red) in a color region different from the color region B (blue) is used at different concentrations is shown.

  As shown in the figure, in (II), the fluorescence by the phosphorescent paint (white light emission) is in the B (blue) region, which is not different from the fluorescence color region of the porous support. Therefore, since the two cannot be clearly distinguished and cannot be taken out as separate signals, the signal detection of the fluorescent label will be influenced by the fluorescence of the porous support, and as a result, the S / N ratio. The improvement effect cannot be obtained.

  On the other hand, in (III) and (IV), the fluorescence of the europium complex, which is a fluorescent label, is detected separately in the R (red) region, and the fluorescence of the porous support is detected in the B (blue) region. Thus, since both can be taken out as separate signals, the signal of the fluorescent label can be detected without being influenced by the fluorescence of the porous support, and as a result, the S / N ratio is improved.

  According to the fluorescence detection system, the fluorescence detection method, etc. of the present invention, the influence of the autofluorescence of the porous support can be achieved with a cheaper and simpler configuration in a technique / apparatus / system that requires fluorescence detection such as immunochromatography. The target fluorescent label can be detected with a high S / N ratio. Therefore, the invention is highly industrially applicable in this field and all related fields.

DESCRIPTION OF SYMBOLS 1 ... Support body 2, 22 ... Light source 3 for fluorescence excitation, 23 ... Fluorescence detection means (color sensor)
5 ... Fluorescence detection system 24 ... Immunochromatography device 25 ... Immunochromatography reader F ... Fluorescence L ... Excitation light

Claims (7)

A fluorescence detection system comprising a light source for fluorescence excitation, a support on which a fluorescence label is developed, and a fluorescence detection means for detecting fluorescence of the fluorescence label, wherein the fluorescence detection means is a color sensor. A fluorescence detection system capable of detecting a fluorescent label developed on the support generating fluorescence without using time-resolved fluorescence or a wavelength region cut filter. The fluorescence detection system according to claim 1, wherein a fluorescent label that emits fluorescence of a color region different from the color region of autofluorescence of the support is used. The fluorescence detection system according to claim 1 or 2, wherein the support is a porous support, and europium is used for the fluorescent label. The fluorescence detection system according to any one of claims 1 to 3, wherein the fluorescence detection system is used for immunochromatography. An immunochromatography device using the fluorescence detection system according to any one of claims 1 to 3. An immunochromatography method using the fluorescence detection system according to any one of claims 1 to 3. The fluorescent label developed on the support that generates self-fluorescence is detected by detecting the fluorescence label developed on the support and fluorescently excited using a color sensor as a fluorescence detection means. A fluorescence detection method characterized by being able to detect without using a region cut filter.