KR101725837B1 - Module type Decision Condition Control Window Diagnostic Kit with Own Light Source for Detecting Cancer Existence - Google Patents

Module type Decision Condition Control Window Diagnostic Kit with Own Light Source for Detecting Cancer Existence Download PDF

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Publication number
KR101725837B1
KR101725837B1 KR1020160075738A KR20160075738A KR101725837B1 KR 101725837 B1 KR101725837 B1 KR 101725837B1 KR 1020160075738 A KR1020160075738 A KR 1020160075738A KR 20160075738 A KR20160075738 A KR 20160075738A KR 101725837 B1 KR101725837 B1 KR 101725837B1
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South Korea
Prior art keywords
diagnostic
window
color
light
leu
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KR1020160075738A
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Korean (ko)
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신동진
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(주)큐브바이오
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/533—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving isomerase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
    • C12Y114/18—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with another compound as one donor, and incorporation of one atom of oxygen (1.14.18)
    • C12Y114/18001—Tyrosinase (1.14.18.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y503/00—Intramolecular oxidoreductases (5.3)
    • C12Y503/02—Intramolecular oxidoreductases (5.3) interconverting keto- and enol-groups (5.3.2)
    • C12Y503/02001—Phenylpyruvate tautomerase (5.3.2.1)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57434—Specifically defined cancers of prostate
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6806—Determination of free amino acids
    • G01N33/6812—Assays for specific amino acids
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/0025—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00 for optical correction, e.g. distorsion, aberration
    • G02B27/005—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00 for optical correction, e.g. distorsion, aberration for correction of secondary colour or higher-order chromatic aberrations
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B5/00—Optical elements other than lenses
    • G02B5/20—Filters
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B6/00—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings
    • G02B6/0001—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings specially adapted for lighting devices or systems
    • G02B6/0011—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings specially adapted for lighting devices or systems the light guides being planar or of plate-like form
    • G02B6/0013—Means for improving the coupling-in of light from the light source into the light guide
    • G02B6/0015—Means for improving the coupling-in of light from the light source into the light guide provided on the surface of the light guide or in the bulk of it
    • G02B6/0016—Grooves, prisms, gratings, scattering particles or rough surfaces

Abstract

The present invention relates to a separable cancer diagnostic kit having a decision condition control window using a self-luminous light source of diagnosing existence of cancer and metabolic diseases. The separable cancer diagnostic kit comprises: a diagnostic module structure having an open window of measuring a color change, and a reaction module inlet of making a reaction module structure inserted and installed; the reaction module structure inserted into the diagnostic module structure and having a diagnostic window to allow the color change by a reaction with an enzyme composition for detecting tyrosine when a sample is inserted to be observed with naked eyes; and a light source and light guiding unit mounted on the diagnostic module structure, irradiating light and selectively changing a lighting color in a reaction area.

Description

[0001] The present invention relates to a detachable cancer diagnostic kit having a judgment environment control window using a self-

The present invention relates to a kit for diagnosing the presence or absence of cancer, and more particularly, to a kit for diagnosing the presence or absence of cancer, which comprises a light source and a light guide plate unit The present invention relates to a detachable cancer diagnostic kit having a judgment environment control window using a light source.

Cancer markers are substances that occur in the malignant tumor cells themselves or are caused by reactions of normal tissues against cancer and exhibit abnormally high concentrations in the blood, urine or tissues, and the cancer marker concentration in the blood, urine or tissue You can diagnose, screen, and track progress and degeneration.

In general, the measurement or detection of the concentration of the cancer marker is carried out through the blood, but the blood has a problem that it is difficult for the general person to collect it.

There has been developed a method for diagnosing cancer by detecting the concentration of cancer biomarker present in the urine by using urine which is not easy to obtain and which is not a biomaterial such as blood or tissue.

On the other hand, it is known that the tyrosine can be used as the cancer biomarker because the concentration of tyrosine present in the urine is high in cancer patients as compared with the general public.

Korean Patent No. 10-0033547 relates to a mixed reagent for diagnosing cancer by detecting aromatic amine (Tyrosine) commonly present in urine of a cancer patient, wherein the mixed reagent is composed of mercury, nickel, nitric acid and distilled water .

However, the above-mentioned mixed reagent contains mercury and is currently prohibited because it has a great influence on the environment and human body.

Therefore, there is an urgent need to develop a composition for diagnosing cancer by eco-friendly detection of the concentration of cancer biomarker tyrosine (Tyrosine) present in the urine by using urine which is not easy to obtain but is not easy to obtain.

Korean Patent No. 10-1323371 discloses a prostate-free cancer diagnostic kit capable of semi-quantitatively measuring the amount of whole prostate-specific antigen in a urine using an immunochromatography method in a kit. Prostate specific antigen (PSA), a serine protease that is expressed at high levels in the prostate epidermis, is produced by the antigen and antibody reaction, To be able to do.

However, the prostate-detached type cancer diagnosis kit of the above-mentioned patent has a problem that only the prostate cancer can be limitedly diagnosed and it is not possible to diagnose whether or not cancer other than prostate cancer is present.

In particular, the diagnostic kits of the prior art have the convenience of being visually diagnosed through the color development by the antigen and the antibody reaction, but the degree of color development is judged by individual subjectivity.

Therefore, when the color displayed by the external influence is distorted, it is difficult to make an accurate judgment.

In other words, it is difficult to determine the degree of color development due to problems such as scattering, refraction, and diffuse reflection due to external light when judging through the window of the diagnostic kit. Particularly, light enters the side of the transparent window constituting the window It is difficult to judge the exact color when scattered in the center of transparent window.

Accordingly, there is a need to develop a new technology that solves the problem of solely relying on external light when judging the degree of color development of the diagnostic kit and improves the visibility.

In the case of using a self-light source to solve the problem of solely relying on external light, it is disadvantageous in terms of manufacturing cost, and thus it is difficult to apply mass production.

In addition, there is a problem that accurate determination is difficult due to a difference in color recognition characteristics of individual users, and development of a new window structure that provides an appropriate judgment environment for each user by controlling the color recognition environment of the reaction region is required.

Korean Patent No. 10-0033547 Korean Patent No. 10-1323371

The present invention solves the problem of such prior art diagnostic kits and includes the enzymes that are characterized in converting L-Tyrosine into Eumelanin, so that the concentration of tyrosine present in the urine The present invention provides a separate type cancer diagnostic kit having an environmental control window using a self-light source using an enzyme composition for detecting cancer biomarkers, which can detect the presence of cancer and metabolic diseases.

The present invention relates to a method and apparatus for diagnosing cancer and metabolic diseases in a home without the inconvenience of users visiting a specialized institution such as a hospital and having to undergo cancer diagnosis by using urine which can be easily collected, It is an object of the present invention to provide a detachable cancer diagnostic kit having an environmental control window.

The present invention has its own light source in a diagnostic kit, and can accurately determine the degree of color change due to a reaction by using the light of its own light source irradiated from the backside of the reaction area without depending on external natural light or external light. The present invention provides a separate type cancer diagnosis kit having a judgment environment control window using a light source.

The present invention relates to a diagnostic module comprising a diagnostic module having a self light source irradiated from the rear side of a reaction area and an enzyme composition for cancer diagnosis detection and having a structure in which a reaction module causing color change due to reaction upon urine injection is separated, And a judgment environment control window using a self light source which is advantageous in terms of utilization.

It is an object of the present invention to provide a separate type cancer diagnosis kit having a judgment environment control window using a self-light source that provides a judgment environment suitable for each user by controlling the color recognition environment of the reaction region of the cancer diagnosis kit.

The present invention has a configuration in which a side of a transparent window member constituting a window and a part of an inner surface in contact with a side face are wrapped to prevent light from being introduced into the side of the transparent window constituting the window of the reaction module and scattering to the center of the transparent window, An object of the present invention is to provide a detachable cancer diagnostic kit having a judgment environment control window using a self light source having a black matrix layer formed on the entire outer periphery of a transparent window member.

The present invention relates to a detachable cancer diagnostic kit having a judgment environment control window using an own light source having an optical film layer for preventing a contrast ratio from being reflected by external light and projecting an external shadow onto a window surface of a reaction module The purpose is to provide.

The objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned can be clearly understood by those skilled in the art from the following description.

To achieve these and other advantages and in accordance with the purpose of the present invention, as embodied and broadly described herein, there is provided a detachable cancer diagnostic kit having a decision environment control window using a self light source, comprising an open window for measuring a color change and a reaction module inlet A reaction module structure having a diagnostic module structure, a diagnostic window that is introduced into the diagnostic module structure and enables visual observation of a color change due to a reaction with the enzyme composition for tyrosine detection when a sample is input, And a light source and a light guide plate unit for irradiating light and selectively changing an illumination color of the reaction region.

Herein, the enzyme composition comprises tyrosine (L-Tyrosine), tyrosinase (DCT) and tyrosinase-related protein 1 (Tyrp1) Is converted into melanin.

The specimen is any one of body fluids derived from a human body, and the body fluids include blood, urine, serum, plasma, lymph fluid, tissue fluid, and secretion.

And a contrast color table for comparing colors observed through a diagnostic window is provided on one surface of the transparent window member constituting the diagnostic window of the reaction module structure. The reference color table is made of a transparent material, The wavelength of the background illumination color is added, and the color is changed in accordance with the added wavelength.

The color change observation is performed such that the reaction module structure is drawn into the interior of the diagnostic module structure and the open window of the diagnostic module structure and the diagnostic window of the reaction module structure are aligned, And then proceeding while irradiating light through the light source.

The light guide plate unit includes a light guide plate having an incident surface on which light emitted from the light source is incident and an exit surface on which the light propagated through the incident surface is deflected, A diffusing sheet for adjusting the diffusion and propagation direction of light passing through the light guide plate; a prism sheet for refracting and diffusing light emitted from the diffusion sheet to increase the brightness; And a reflective sheet for reflecting light is stacked.

The transparent window member has a shape that surrounds a side surface of the transparent window member and a portion of the inner surface contacting the side surface. The transparent window member is formed around the entire outer surface of the transparent window member to allow light to flow into the side surface of the transparent window constituting the diagnostic window And a black matrix layer for suppressing the black matrix layer.

The enzyme composition for detecting tyrosine was prepared by adding 0.5 to 2 parts by weight of a dopa chromium mutant enzyme (DCT) to 1 part by weight of the tyrosinase and 30 parts by weight of tyrosinase-related protein 1 (Tyrp1 The present invention is characterized in that the color of urine is changed from brown to black as the concentration of tyrosine is increased by converting L-tyrosine into Eumelanin.

The enzyme composition for detecting tyrosine may be a phosphate buffer or a Tris-HCl buffer solution having a final concentration of 50 to 100 mM, a Tween 20 having a final concentration of 0.1 to 0.5%, and distilled water And is contained in a liquid having a final reaction pH of 7.0 to 8.5.

And an optical film formed on the surface of the diagnostic window to prevent color distortion due to external light.

The separate type cancer diagnosis kit having the judgment environment control window using the self light source according to the present invention has the following effects.

First, a separate type cancer diagnostic kit using an enzyme composition for detecting cancer biomarkers that can detect the presence or absence of cancer and metabolic diseases by environmentally detecting the concentration of tyrosine present in the urine can be provided.

Secondly, the user can easily diagnose the presence of cancer and metabolic diseases in the home without the inconvenience of visiting the specialist institution such as the hospital and receiving the cancer diagnosis by using the urine which can be collected easily.

Third, a self-light source is provided in the diagnostic kit, and the degree of color change due to the reaction can be accurately determined by using the light of the self-light source irradiated from the backside of the reaction area, without depending on external natural light or illumination.

Fourth, a diagnostic module having a self-light source irradiated from the rear of the reaction area and a reaction module including an enzyme composition for detecting cancer diagnosis and causing color change due to reaction during urination are separated, It is advantageous in terms of utilization.

Fifth, the accuracy of determination can be improved by controlling the color recognition environment of the reaction region of the cancer diagnosis kit to provide an appropriate judgment environment for each user.

Sixth, a black matrix layer is formed on the entire outer periphery of the transparent window member to surround a side of the transparent window member constituting the window and a part of the inner surface in contact with the side surface, Can be prevented from being scattered to the center of the transparent window.

Seventh, when determining the degree of color development of the diagnostic kit, the influence of external light is suppressed as much as possible and the visibility is improved, thereby minimizing the error caused by subjective judgment of the individual and making accurate judgment possible.

1 is a perspective view illustrating a detachable type cancer diagnostic kit having a judgment environment control window using a self light source according to an embodiment of the present invention;
2 is a cross-sectional view illustrating the use of a detachable cancer diagnostic kit having a decision environment control window using the self light source of FIG.
3A is a cross-sectional view showing a window structure of a reaction module according to the present invention;
3B is a cross-sectional view showing a structure of a light guide plate unit of a diagnostic module according to the present invention
FIG. 3C is a plan view of the window area in a state in which the reaction module and the diagnostic module are coupled;
FIG. 3D is a diagram showing an example of R, G and B LEDs applied to a high-definition window using a self light source according to the present invention
FIG. 4 shows a comparison color chart for measuring the color of the produced eumelanin in a white light environment
Figure 5 is a photograph comparing the color of the produced melanin in a white light environment
6 is a color wheel configuration diagram showing a decision color area according to the color recognition environment control of the reaction area according to the present invention
FIG. 7 is a table showing the relationship between the contrast color table for measuring the color of the produced eumelanin in the high-brightness blue (B)
8 is a photograph comparing the color of melanin produced in a high-brightness blue (B) light environment
9 is a view showing an example of a light guide plate unit applied to a high-definition window using a self light source according to the present invention
10A to 10D are diagrams showing an example of an optical film for constructing a high-definition window using a self light source according to the present invention

Hereinafter, a preferred embodiment of a separate type cancer diagnostic kit having a judgment environment control window using its own light source according to the present invention will be described in detail.

Features and advantages of a detachable cancer diagnostic kit having a decision environment control window using its own light source according to the present invention will be apparent from the following detailed description of each embodiment.

FIG. 3A is a sectional view showing a window structure of a reaction module according to the present invention, and FIG. 3B is a sectional view showing the structure of a light guide plate unit of a diagnostic module according to the present invention.

And FIG. 3C is a plan view of a window region in a coupled state of the reaction module and the diagnostic module. FIG. 3D is a schematic view illustrating an example of R, G, and B LEDs applied to a high-definition window using a self- .

A separate type cancer diagnosis kit having a judgment environment control window using a self light source according to the present invention includes the following features in order to accurately determine the degree of color change due to a reaction.

First, a self-light source is provided inside the diagnostic kit so that the degree of color change due to the reaction can be accurately determined by using the light of the self-light source irradiated from the rear side of the reaction area without depending on external natural light or illumination.

Secondly, to solve the problem caused by differences in color recognition characteristics of individual users, the system includes a configuration for selectively controlling the R, G, and B LEDs to change the background color of the reaction region.

Third, it has a structure in which a diagnostic module having a self-light source irradiated from a rear surface of a reaction area, and an enzyme module for cancer diagnosis detection and a reaction module for causing color change due to reaction during urine injection are separated.

Fourth, an optical film layer formed on the surface of the window region of the reaction module is provided to prevent the contrast ratio from being reduced by the reflection of external light and to project an external shadow onto the window surface.

Fifth, in order to prevent light from being introduced into the side of the transparent window constituting the window area of the reaction module and scattering to the center of the transparent window, a form of wrapping a part of the inner side in contact with the side face and the side face of the transparent window member constituting the window And a black matrix layer formed on the entire outer periphery of the transparent window member.

The present invention provides a self-contained light source inside the diagnostic kit in order to minimize the influence of external light and improve the visibility in determining the degree of color development of the diagnostic kit.

In particular, the present invention relates to a diagnostic module having a self light source and a structure in which a reaction module which is introduced into the interior of a diagnostic module at the time of diagnosis and contains an enzyme composition for detecting cancer diagnosis therein will be.

By separating the diagnostic module having its own light source and the reaction module causing the color change by the actual reaction, when the diagnosis module is purchased once, it is necessary to purchase only the reaction module when a family or repeated diagnosis is required Thereby enabling efficient diagnosis.

Also, the R, G, and B LEDs of the self light source provided in the diagnostic kit are selectively controlled to change the background illumination color and brightness of the reaction area to provide a judgment environment suitable for the user's color recognition characteristics.

This makes it possible to further improve the judgment accuracy by comparing the color change in a white light environment and the color change in a different color light environment.

In the following description, the enzyme composition used as the reactant is exemplified by the enzyme composition for tyrosine detection, and the urine is exemplified as the sample to be used. However, it is natural that the kind of the enzyme composition and the kind of the sample can be different.

For example, the specimen applicable to the present invention is any one of body fluids derived from a human body, and the body fluids may include blood, urine, serum, plasma, lymph fluid, tissue fluid, secretion and other body fluids, Lt; / RTI > group.

1 and 2 show an example of a detachable type cancer diagnostic kit to which a high-definition window using the self light source of the present invention can be applied. The diagnosis module structure 10 and the reaction module structure 20 are separated, And the module structure 20 is drawn into and coupled to the inside of the diagnostic module structure 10.

The diagnostic module structure 10 has a light source and a light guide plate unit therein and has an open window 12 for measuring a color change on one side thereof and allows the reaction module structure 20 to be mounted on an upper surface thereof, And a reaction module inlet 14 serving to pull out the reaction module structure 20 after diagnosis.

The diagnostic module structure 10 includes a switch 11 for controlling the power supply to irradiate the light using the self light source in the direction of the open window 12 by using a light source and a light guide plate unit provided inside the one side surface.

The position and shape of the switch 11 are not limited as shown in the drawings, but may vary depending on the shape of the diagnostic kit.

According to the number of times the switch 11 is pressed, all the LEDs of R, G and B are turned on and provided to the white illumination environment in a single cycle. In the case of two cycles, only the red LED is turned on, In the case of three times, only the green (G) LED is turned on so that the background illumination color is provided in the green illumination environment, and in the four times, only the blue (B) LED is turned on so that the background illumination color is provided in the blue illumination environment .

The LEDs may be turned on in the order of red (R) + green (G), red (R) + blue (B), green (G) + blue (B)

Also, the output brightness of the LED may be set differently in steps.

The reaction module structure 20 is provided with an inlet 16 into which urine is injected and is connected to a diagnostic window 10 that is inserted into the diagnostic module structure 10 and aligned with the open window 12 of the diagnostic module structure 10 15), and an enzyme composition (30) for detecting tyrosine which is contained in the urine and converts tyrosine (L-Tyrosine) present in the urine into melanin.

On the upper surface of the reaction module structure 20, an inlet 16 for urine is formed to penetrate in the vertical direction. The inlet 16 is preferably formed in such a manner that the outer periphery thereof is inclined downward so that the urine is smoothly guided to the inside of the inlet 16 and does not flow to the outside.

In order to prevent the enzyme composition for detecting tyrosine 30 from flowing out, a check valve 18 is provided at the lower end of the charging port 16 to allow fluid flow from the outside to the inside of the charging port 16, The inlet 16 may be provided with a stopper 17 for opening and closing the inlet 16.

In this embodiment, the check valve 18 is integrally formed at one end with the charging port 16 for accommodating the enzyme composition 30 for detecting tyrosine, and when the urine is injected through the charging port 16, When the urine is not supplied, the lower end of the charging port 16 is closed by the elastic force of the charging port 16. The rim of the charging port 16 is supported by the lower end of the charging port 16 and is directed toward the charging port 16 Thereby preventing the enzyme composition 30 for tyrosine detection from escaping.

The stopper (17) is inserted into the discharge port (16) to open and close the discharge port (16). It goes without saying that, unlike the embodiment, it is of course possible to open and close the charging port 16 by using a tape or a known sealing mechanism without using the cap 17.

The reaction module structure 20 has a tubular shape and functions as a container for containing the enzyme composition 30 for detecting tyrosine in the liquid, and the enzyme composition 30 for detecting tyrosine reacts with tyrosine in the urine, Is made of a transparent material so that the user can visually observe it from the outside.

The reaction module structure 20 may be made entirely of a transparent material and is made of an opaque material that blocks external light to suppress deformation of the enzyme composition 30 for detecting tyrosine therein, A transparent window may be formed only in the diagnosis window 15 area.

The verification color table 13 can be displayed in the diagnostic window 15 area of the reaction module structure 20 so that the user can easily determine whether the cancer has occurred through the color observed through the open window 12 .

Here, it is preferable that the reference color table 13 is made of a transparent film material.

Since the tyrosine detection enzyme composition 30 is color-changed from brown to black as the tyrosine concentration in the urine increases, colors of the reference color table 13 are basically displayed between brown and black.

The contrast color table 13 displays colors between brown and black in a white illumination environment using a self light source, but selectively controls the R, G, and B LEDs of the self light source provided in the diagnostic module structure 10, When the background illumination color of the area is changed, the wavelength of the background illumination color is added, and the color is changed and displayed accordingly.

The diagnostic module structure 10 further includes a light guide plate 20 for uniformly illuminating light through its own light source to increase the measurement accuracy of the color change through the diagnostic window 15 of the reaction module structure 20 aligned with the open window 12. [ Unit.

In the case of using the separate type cancer diagnostic kit according to the present invention having the structure for separating the diagnostic module having its own light source according to the present invention and the reaction module causing color change due to the actual reaction, The diagnostic module structure 10 may be inserted into the reaction module structure 20 and the urine may be injected into the reaction module structure 20 while the reaction module structure 20 and the diagnostic module structure 10 are coupled.

For correct color determination, urine may be injected into the reaction module structure 20 while the reaction module structure 20 and the diagnostic module structure 10 are combined.

The structure for separating the diagnostic module having its own light source according to the present invention and the reaction module causing the color change due to the actual reaction is applied to the diagnostic kit located on the upper surface in addition to the diagnostic kit in which the window for diagnosis is located on the front side It is natural to be able to.

That is, this type of detachable cancer diagnostic kit is an example of applying a high-definition window using the self light source according to the present invention, and the form and structure of the detachable cancer diagnostic kit are not limited thereto.

The judgment environment control window structure using the self light source according to the present invention will be described in detail as follows.

3A is a cross-sectional view illustrating a window structure of a reaction module according to the present invention.

3A is an area in which an actual user can see the color change visually by being aligned with the open window 12 when coupled with the diagnostic module structure 10 and the reaction module structure 20 in FIG. The diagnosis window structure is formed to illustrate an example of the separate type cancer diagnosis kit according to the present invention and is not limited to this type and structure.

On the outer surface of the diagnostic window member 21 formed on the reaction module structure 20, the contrast ratio is reduced by reflection of external light, and external shadows are prevented from projecting onto the window surface, thereby preventing color distortion appearing in the window An optical film layer 22 is provided on the surface of the window to prevent color distortion due to external light.

The diagnostic window member 21 has a shape that surrounds a part of the inner surface contacting the side surface and the side surface of the diagnosis window member 21 in order to prevent light from being introduced into the side surface of the diagnostic window member 21 and scattering to the central portion. And a black matrix layer 23 formed around the entire outer periphery of the black matrix layer 23.

In order to suppress deformation of the enzyme composition contained in the diagnostic kit, the black matrix layer 23 may be formed using a carbon-based organic material or a photo acryl based material without using a metal such as chromium Resin is preferably used, and the materials used are not limited thereto.

For example, in the case of forming using a pigment dispersion method, a polymer compound that enables the support to function and a certain thickness to be maintained, that is, a binder resin and two components of a photopolymerizable monomer that reacts with light upon exposure to form a photoresist image And may be formed from a photosensitive resin composition containing a pigment, a polymerization initiator, an epoxy resin, a solvent, and other additives in addition to the above components.

And has a shape that surrounds a part of the inner surface contacting the side surface and the side surface of the diagnosis window member 21 about the (a) actual viewing area of the diagnosis window member 21, A black matrix layer 23 is formed on the periphery of the black matrix layer 23 and an optical film layer (not shown) is formed on the surface of the diagnosis window member 21 to prevent color distortion due to external light, 22 are formed.

3B is a cross-sectional view showing the structure of a light guide plate unit of a diagnostic module according to the present invention.

The detachable cancer diagnostic kit according to the present invention can accurately determine the degree of color change due to the reaction without using the external natural light or illumination and by using the light of the self light source irradiated from the backside of the reaction region facing the (A) region A light guiding plate unit 40 having a self light source 40a inside the diagnostic module structure 10 is provided.

The light guide plate unit 40 can be formed on the inner wall surface of the diagnostic module structure 10 having a wider area than the formation area of the optical film layer 22 and opposed to the area of the optical module layer 22, And may be formed on the bottom surface.

A light source 40a is provided on one side of the light guide plate unit 40 and the light source 40a is connected to a battery 11b for power supply and a switch 11 for power supply control.

3C is a plan view of the window region in the coupled state of the reaction module and the diagnostic module.

And FIG. 3D is a view illustrating an LED for providing a judgment environment suitable for the user's color recognition characteristics by selectively controlling the R, G, and B LEDs of the self light source provided in the diagnostic module structure 10, Fig.

The LED circuit configuration shown in FIG. 3D is an example of application to the judgment environment control window using the self light source according to the present invention, but it is not limited thereto, and it is natural that the LED circuit configuration can be another structure.

Further, it may further comprise a control means for selectively controlling the ON and the output combination or the output luminance of the LED.

A separate type cancer diagnostic kit having a judgment environment control window using a self light source according to the present invention selectively controls the R, G and B LEDs of the self light source provided therein to change the background color of the reaction area, The characteristics in the white light environment and the blue light environment will be described as follows.

FIG. 4 is a control color table for measuring the color of the produced eumelanin in the white light environment, and FIG. 5 is a photograph of the color of the produced melanin in the white light environment.

The color change in a white light environment basically changes from a brown area to a black area.

In this state, when the user selectively changes the background illumination color, the color of the background color of the reference color table 13 formed in the diagnostic window 15 of the reaction module structure 20 is added, and the color of the background color is changed.

6 is a color wheel configuration diagram showing a decision color area according to the color recognition environment control of the reaction area according to the present invention.

In FIG. 6, (A) is a region where a color change occurs in a white light environment, and when the background color of the reaction region is changed by selectively controlling the R, G, and B LEDs of the self light source, C) (D).

FIG. 7 is a contrast color chart for measuring the hue of the produced eumelanin in a high-brightness blue (B) light environment, and FIG. 8 is a photograph comparing the colors of melanin produced in a high-brightness blue (B) light environment.

The light guide plate unit 40 applied to the present invention may be configured as follows.

FIG. 9 is a view showing an example of a light guide plate unit applied to a high-definition window using a self light source according to the present invention.

A light guide plate 41 having an incident surface on which light irradiated from the light source is incident and an exit surface on which the light propagated through the incident surface is deflected, A prism sheet 43 for increasing the brightness by refracting and diffusing the light emitted from the diffusion sheet 42 and a light guide plate 43 for guiding the light emitted from the lower side of the light guide plate 41 The light guide plate unit 40 may be configured to have a structure in which the light source or the reflection sheet 44 for reflecting the light directly incident on the light guide plate 41 is laminated and the structure of the light guide plate unit 40 is not limited thereto Do not.

The light guide plate is preferably made of a transparent material such as plastic or resin such as polycarbonate (PC) or polymethylmethacrylate (PMMA), and can be manufactured by injection molding.

A separate type cancer diagnosis kit having a window for judging environmental control using the self light source according to the present invention having such a window structure is characterized in that the color of the reaction after the urine injection does not depend on external natural light or illumination, The degree of color change caused by the reaction can be accurately determined by using the light of the self light source irradiated from the bottom surface.

Therefore, it is possible to improve the accuracy in the negative and positive determination as in the comparison color table of FIG. 4 for measuring the color of the produced eumelanin.

The structure and characteristics of the optical film applied to the detachable cancer diagnostic kit having the judgment environment control window using the self light source according to the present invention will be described in detail as follows.

10A to 10D are block diagrams of optical films formed on the diagnostic window surface of a detachable cancer diagnostic kit having a decision environment control window using its own light source according to the present invention.

The optical film applied to the detachable cancer diagnostic kit having the judgment environment control window using the self light source according to the present invention may be an anti-reflection film, an anti-glare film or a polarizing film And may be a multilayer structure in which these layers are laminated.

The structure of the optical film described below is an example of an optical film applied to a detachable cancer diagnostic kit having a judgment environment control window using its own light source according to the present invention, and is not limited thereto.

In addition, the materials used for the production of the optical film and the kind of the pressure-sensitive adhesive on the window surface of the optical film are not limited to those which can be applied to the present invention.

10A shows an example of an anti-reflection film for minimizing reflection on a window surface.

The antireflection film (51a)

A multilayer thin film 51c made of a material having a different refractive index on the base film 51b such as polyethylene terephthalate (PET) is formed on the surface of the film by a vapor deposition method or a coating method so as to have an antireflection effect May be used.

For example, silicon oxide (SiO 2 ) and titanium oxide (TiO 2 ) can be alternately laminated.

10B shows an example of an anti-glare film for preventing a shadow of an external object from being projected onto a window surface.

The anti-glare film (52a)

And includes an irregular surface 52c on the surface of a base film 52b such as polyethylene terephthalate (PET), so that external light is irregularly reflected.

Such an irregular surface 52c can be formed by spraying silicon particles.

FIG. 10C shows an example of a linear polarizer for preventing reflection of external light.

The linear polarizing film 53a is supported by a support 53b such as triacetyl-cellulose (TAC) around a polymer polarizing medium 53c such as polyvinyl alcohol (PVA) for polarizing incident light. (53d) may be located on both sides of the polymeric polarization medium (53c).

FIG. 10D shows an example of a circular polarizer for enhancing the function of a linear polarizer for preventing reflection of external light.

The circular polarizing film 54a is formed on the retardation film 54b for compensating the retardation with a polymer polarizing medium 54d such as polyvinyl alcohol (PVA) A support 54c (54e) such as tri-acetyl-cellulose (TAC) may be positioned on both sides of the polymeric polarization medium 54d.

The optical film applied to the present invention is adhered to the surface of the diagnosis window 15, and the diagnosis kit is not judged by the naked eye, and may be desorbed if necessary by using a separate analyzer.

In order to adhere and detach the optical film, it is important to control the peeling force of the adhesive.

In the present invention, the pressure-sensitive adhesive used for controlling the peeling force may be a material comprising an acrylic resin, a polyester resin or a silicone resin composition.

Such a resin composition may include an oligomer, a monomer, a photoinitiator, an adhesion promoter, a leveling agent, and other additives.

The diagnosis mechanism using the separate type cancer diagnosis kit having the judgment environment control window using the self light source according to the present invention having the above structure will be described in detail as follows.

The present invention relates to a pharmaceutical composition comprising tyrosinase, Dopachrome tautomerase (DCT) and tyrosinase-related protein 1 (Tyrp1) for detecting the concentration of the cancer biomarker tyrosine present in the urine An enzyme composition (30) for detecting cancer biomarker tyrosine is used as a diagnostic substance.

More specifically, the tyrosine detecting enzyme composition (30) comprises an enzyme composition in which a tyrosinase is supplemented with a Dopachrome tautomerase (DCT) and a tyrosinase-related protein 1 (Tyrp1) enzyme To convert cancer biomarker tyrosine (L-Tyrosine) present in the urine into melanin, and the cancer can be diagnosed by measuring the concentration through the color of the melanin.

In particular, in the present invention, the enzyme composition for detecting tyrosine 30 is used to convert L-tyrosine to melanin, wherein the melanin is eumelanin.

Since the enzyme composition 30 for detecting tyrosine according to the present invention is for diagnosing cancer using urine, it is preferable to use a mixture of melanin (Pheomelanin) or brown melanin (brown melanin) When the concentration is measured through the urine, it may be difficult to measure the concentration because it is similar to the color of the urine. Therefore, it is preferable that the enzyme composition for tyrosine detection (30) according to the present invention is measured for its concentration through black colored eumelanin.

The enzyme composition for detecting tyrosine (30), by adding DCT and Tyrp1 enzyme to tyrosinase, not only accelerates the pathway of eumelanin biosynthesis, but also induces the color of the produced uromelanin to a more black path have.

Since the concentration of tyrosine present in the urine can be measured through the color of the eumelanin produced by the reaction with the tyrosine detecting enzyme composition 30, when the color of the urine produced by the produced eumelanin is close to black, It can be seen that the amount of tyrosine is high.

The tyrosine detection reaction by the tyrosine detection enzyme composition 30 can be carried out in a phosphate buffer or Tris-HCl buffer solution having a final concentration of 50 to 100 mM and a final concentration of 0.1 to 0.5 % Tween 20, and the final reaction pH is 7.0 to 8.5.

For example, when the phosphoric acid buffer solution contained in the tyrosine detecting enzyme composition 30 is 200 mM and the total volume of 500 μl of the enzyme composition and 500 μl of the urine sample is 1000 μl, the tyrosine detection reaction is performed at a final concentration of 50 mM .

In one embodiment of the present invention, the tyrosine detecting enzyme composition (30) comprises tyrosinase, DCT and Tyrp1, and is diluted with a phosphate buffer or a Tris-HCl buffer solution, Tween 20, And adding distilled water.

Preferably, the enzyme composition for detecting tyrosine 30 comprises 40 units of tyrosinase in a weight ratio of 0.5 to 2 parts by weight of DCT and 0.5 to 2 parts by weight of Tyrp1 to 1 part by weight of the tyrosinase.

If the above content is not satisfied, it may be difficult to measure the concentration of tyrosine because the color of the produced melanin may appear to be a yellowish brown color and be mixed with the urine color.

As described above, the tyrosine detecting enzyme composition (30) accelerates the pathway of eumelanin biosynthesis by tyrosinase to DCT and Tyrp1 enzyme, and induces the color of the resulting eumelanin to a more black coloring pathway .

Referring to Reaction Scheme 1 below, L-tyrosine is oxidized by tyrosinase to form 3,4-dihydroxyphenylalanine (DOPA) and then oxidized by tyrosinase to form DOPA quinone , And Leucodopachrome to form Dopachrome.

The dopachrome is a melanin precursor and is tautomerized by DCT to form 5,6-dihydroxyindole-2-carboxylic acid (DHICA).

The 5,6-dihydroxyindole-2-carboxylic acid is oxidized by Tyrp 1 to form indole-5,6-quinonecarboxylic acid and the indole-5,6-quinonecarboxylic acid is polymerized to form uamelanin .

[Reaction Scheme 1]

Figure 112016058509390-pat00001

The tyrosinase is a tyrosinase hydroxylase and a copper-containing enzyme having catalytic activity of dopa oxidase, which can be found in microorganisms and plant and organism tissues. In particular, the tyrosinase promotes the production of melanin and other pigments by oxidation of phenols such as tyrosine.

The Dopachrome tautomerase (DCT), also known as tyrosinase-related protein 2 (Tyrp 2), is a protein that is characterized by the binding of tyrosinase and Tyrp1 to convert L-tyrosine to melanin in melanocytes It is a melanin cell specific enzyme.

The tyrosinase-related protein 1 (Tyrp1) plays a role in stabilizing tyrosinase.

The tyrosine detection enzyme composition (30) according to the present invention is characterized in that the dopachrome is converted to indole-5,6-quinone (5-dihydroxyindole) by means of tyrosinase via 5,6- , 6-quinone) to form melanin near the brown color (see the following reaction formula 2), but is tautomerized by DCT and oxidized by Tyrp 1 to form melanin close to black, By separating them from color, it is possible to perform density measurement effectively.

[Reaction Scheme 2]

Figure 112016058509390-pat00002

In one embodiment of the present invention, when measuring the concentration of tyrosine present in the urine through the hue of the produced eumelanin, the color of the produced eumelanin is compared with that of the control color table (13) to confirm the concentration of tyrosine in the urine . This makes it possible to easily diagnose the occurrence of cancer and metabolic diseases.

Hereinafter, the tyrosine detecting enzyme composition (30) of the present invention will be described in more detail by way of examples. It is to be understood by those skilled in the art that these embodiments are for illustrative purpose only and that the scope of the present invention is not limited to these embodiments.

Manufacturing example  1: Preparation of each component of the enzyme composition

The tyrosinase enzyme was purchased from Sigma.

DCT and Tyrp1 were obtained from mouse blood by reference to the following steps and purified after protein expression in E. coli.

(i) RNA extraction from mouse blood

RiboEx RNA extraction solution kit (Jinol Biotechnology, Korea) was used in this embodiment. The process is as follows.

a. 300 μl of mouse blood and 600 μl of RiboEx RNA extraction solution (Jeolol Biotechnology, Korea) were mixed from the mouse blood, mixed strongly and left at room temperature for 5 ~ 10 minutes to dissolve the cell membrane.

b. 200 μl of chloroform was added and mixed vigorously. The mixture was centrifuged at 13,000 rpm at 4 ° C for 10 minutes using a centrifuge to separate the upper and lower layers.

c. After collecting only the upper layer, the same amount of the column-binding buffer was added and mixed well. Then, the solution was added to the column provided in the kit and spin-down to bind only RNA to the column.

d. The wash buffer provided in the kit was added to the column, and then the material other than RNA was removed by spin-down. The remaining wash buffer was removed by centrifugation for an additional 1 minute.

e. The elution buffer was added to recover the RNA bound to the column, and the RNA was recovered by spinning 5 minutes later.

(ii) preparing cDNA from the obtained RNA

 1 μg RNA, 5 units of reverse transcriptase (reverse transcriptase), oligo dT-50 pmol, 0.1 M DTT- 2 μl, 10x reaction buffer - 2 μl, water - about 20 μl; The mixture was prepared in the above composition and reacted at 45 ° C for 10 minutes and at 38 ° C for 3 hours or longer in a PCR instrument.

(iii) securing the gene (DCT, Tyrp1) from cDNA

a. In order to amplify the gene from the cDNA generated in the above step, the mixture was prepared with the following composition, and the gene amplification reaction (PCR) was performed under the following conditions.

Mixture composition: 2 μl of cDNA solution, 5 units of pfu DNA amplification enzyme, 2 μl of 10 mM dNTP, 4 μl of 10 × reaction buffer, 2 pmol of each primer pair, and water - about 40 μl.

Amplification reaction conditions: 95 ° C 4 min - 1 time; 95 ° C for 20 seconds, 60 ° C for 20 seconds, 72 ° C for 1 minute and 50 seconds - 35 times; 72 占 폚 10 min - 1 time; Continue at 4 ℃.

The information of the primer pair used in the above reaction is shown in Table 1 below.

division Base sequence Primer for DCT amplification Forward primer Atgggccttgtgggatggg (SEQ ID NO: 1) Reverse primer Ctaggcttcctccgtgtatctcttgc (SEQ ID NO: 2) Tyrp1 primer for amplification Forward primer Atgaaatcttacaacgtcctccccct (SEQ ID NO: 3) Reverse primer Tcagaccatggagtggttaggattcg (SEQ ID NO: 4)

The DCT and Tyrp1 enzymes were prepared by inserting the genes obtained by the above method into the pET28a vector and constructing a fusion protein in which 6x His was attached to the N-terminus and transformed into BL21 (DE3) Escherichia coli. The process is as follows.

(i) the step of culturing the transformed E. coli

a. NdeI of the pET28a vector, and DCT or Tyrp1 gene into the XhoI restriction enzyme site, thereby preparing a recombinant DNA which produces 6x His ~ protein.

b. E. coli transformed with BL21 (DE3) was prepared by heat shock transformation.

c. The E. coli strain was cultured in 3 L of Luria broth at 37 DEG C and 200 rpm at OD600 ~ 0.7, followed by treatment with 0.1M IPTG to induce protein production.

d. After further culturing at 30 DEG C for 4 hours, E. coli was precipitated by centrifugation.

   (ii) Escherichia coli disruption step

a. The precipitated E. coli was resuspended in 200 ml of 1x PBS and washed by centrifugation.

b. The precipitated E. coli was suspended in 40 ml of 0.5 M NaCl, 5 mM imidazole, 20 mM Tris-HCl, pH 7.9; Lt; / RTI >

c. Escherichia coli was disrupted in an ultrasonic cell disruptor under the condition of Energy 38% max, total cell breaking time 4 min (2 second sonic treatment / 4 sec pause). The bottle containing the E. coli sample remained ice-filled during the cell disruption process.

d. The shredded E. coli was centrifuged at 13,000 rpm, 30 minutes, and 4 ° C.

e. Only the supernatant was collected in a clean conical tube.

   (iii) Purification of his tag-fusion protein using Ni-NTA agarose resin

a. The water-soluble protein solution obtained in the above step was mixed with Ni-NTA agarose resin and shaken for 30 minutes at 4 ° C to induce binding of the his tag-fusion protein to the Ni-NTA agarose resin.

b. The protein solution and the resin mixture were put into the column, and the column cock was opened to extract the liquid.

c. 400 ml of washing buffer (0.5 M NaCl, 60 mM imidazole, 20 mM Tris-HCl, pH 7.9) was passed through to remove proteins other than impurities and fusion proteins.

d. 3 ml of elution buffer (0.25 M NaCl, 500 mM imidazole, 20 mM Tris-HCl, pH 7.9) was added and allowed to stand for 10 minutes.

e. d process was repeated twice.

(iii) Protein purification using FPLC-size exclusion method

a. The recovered proteins were loaded onto FPLC equipped with a superdex S200 (GE healthcare) column and the proteins were separated by size.

b. Among the obtained fraction samples, fractions corresponding to his tag fusion proteins were identified by polyacrylamide gel electrophoresis and finally the desired his tag fusion protein was obtained.

c. Abs 280 values were measured, and protein concentrations were converted from the obtained values, and they are shown in Table 2 below.

division Protein concentration (OD 280 nm) 1 mg DCT / ml 1.65 1 mg Tyrp1 / ml 1.42

Example 1

40 [micro] l of DCT having an activity of 1000 units / ml prepared in Preparation Example 1 and 40 [micro] l of Tyrp1 having an activity of 1000 units / ml were added to 40 [micro] l of tyrosinase enzyme having activity of 1000 unit / ml, 250 μl of 200 mM Tris buffer pH 8, 25 μl of 10% Tween 20, and 105 μl of distilled water were added to prepare an enzyme composition (stock solution).

Stock solution Volumn (500 μl) 200 mM Tris buffer pH8 250 μl 10% Tween 20 25 μl Tyrosinase (> = 1000 unit / ml) 40 μl DCT (> = 1000 unit / ml) 40 μl TYRP1 (> = 1000 unit / ml) 40 μl D.W 105 μl

Next, urine samples were collected from normal and cancer patients. The method of collecting the urine samples was done by taking 30 ml of sample from the urine without taking the first urine from the urethra in the first urine collection after the morning wake.

In order to measure the concentration of tyrosine present in a certain amount, the same amount of 500 μl of each of the urine collected from the normal and cancer patients was reacted with the enzyme composition for tyrosine detection of Example 1 (1000 μl in total).

Then, the color of the urine produced by each produced eumelanin is compared with the control color table of FIG. 4 and is shown in Table 4 below.

Color of urine by eumelanin Urine of a normal person Yellow to tan Urine of cancer patient black

As shown in Table 4, the urine color produced by the reaction of the normal human urine with the enzyme composition of the present invention was yellow to yellowish brown, while the urine of the cancer patient was reacted with the enzyme composition, The color of the urine was confirmed to be black.

This shows that tyrosine is present in large amounts in urine of cancer patients.

Example 2

When the enzyme composition (30) of the present invention, which additionally contains DCT and Tyrp1, is used as a tyrosinase compared to the conventional enzyme composition containing only tyrosinase, the color of the urine produced by the produced uromelain appears more black The following experiment was carried out.

A mixture (A) in which 500 μl of normal human urine collected in the same manner as in Example 1 was mixed with 40 parts of tyrosinase, 20 units of DCT and 20 units of Tyrp1, A mixture (B) in which 500 μl of the composition was mixed with an existing enzyme composition containing only 40 units of tyrosinase, and 500 μl of urine of a cancer patient were mixed with 40 units of tyrosinase, 20 units of DCT and 20 units of Tyrp1 The mixture (C) mixed with the enzyme composition was reacted at 37 DEG C for 30 minutes, respectively.

4, the mixture (A) in which the enzyme composition according to the present invention is reacted with urine of a normal person exhibits yellow color, while the mixture (B) and (C) in which the urine of the cancer patient is reacted shows a darker tan or black Respectively.

Especially, as a result of urine detection of a cancer patient, it was confirmed that the color of the mixture (C) containing DCT and Tyrp 1 is more black than the mixture (B) containing only tyrosinase.

Therefore, by adding the DCT and Tyrp1 enzyme to the tyrosinase, the enzyme composition according to the present invention not only accelerates the pathway of eumelanin biosynthesis, but also can induce the color of the resulting eumelanin to a more black-colored pathway Respectively.

The separate type cancer diagnosis kit having the judgment environment control window using the self light source according to the present invention can be applied to the reaction using the light of the self light source irradiated from the rear surface or the bottom surface of the reaction area without depending on external natural light or illumination So that it is possible to accurately determine the degree of color change caused by the color change.

In particular, a diagnostic module having a self-light source irradiated from a rear surface or a bottom surface of a reaction area, and a reaction module including an enzyme composition for cancer diagnosis detection and causing a color change due to reaction during urination are separated will be.

In order to prevent scattering of light into the center of the transparent window due to the flow of light into the side of the transparent window constituting the optical film layer and the diagnostic window for preventing color distortion due to external light, A black matrix layer is provided so that the influence of external light can be suppressed as much as possible and the visibility can be improved.

In addition, the present invention includes a configuration for selectively controlling the R, G, and B LEDs to change the background illumination color of the reaction area to solve problems caused by differences in color recognition characteristics of individual users.

As described above, it will be understood that the present invention is implemented in a modified form without departing from the essential characteristics of the present invention.

It is therefore to be understood that the specified embodiments are to be considered in an illustrative rather than a restrictive sense and that the scope of the invention is indicated by the appended claims rather than by the foregoing description and that all such differences falling within the scope of equivalents thereof are intended to be embraced therein It should be interpreted.

10. Diagnostic Module Structure 20. Reaction Module Structure
30. Enzyme composition for tyrosine detection 40. Light guide plate unit

<110> CubeBio <120> Enzyme Compositions for Detecting Cancer Biomarker Tyrosine <130> DP18030 <160> 7 <170> KoPatentin 3.0 <210> 1 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> primer <400> 1 atgggccttg tgggatggg 19 <210> 2 <211> 26 <212> RNA <213> Artificial Sequence <220> <223> primer <400> 2 ctaggcttcc tccgtgtatc tcttgc 26 <210> 3 <211> 26 <212> RNA <213> Artificial Sequence <220> <223> primer <400> 3 atgaaatctt acaacgtcct ccccct 26 <210> 4 <211> 26 <212> RNA <213> Artificial Sequence <220> <223> primer <400> 4 tcagaccatg gagtggttag gattcg 26 <210> 5 <211> 556 <212> PRT <213> Agaricus bisporus <400> 5 Met Ser Leu Ile Ala Thr Val Gly Pro Thr Gly Gly Val Lys Asn Arg   1 5 10 15 Leu Asn Ile Val Asp Phe Val Lys Asn Glu Lys Phe Phe Thr Leu Tyr              20 25 30 Val Arg Ser Leu Glu Leu Leu Gln Ala Lys Glu Gln His Asp Tyr Ser          35 40 45 Ser Phe Phe Gln Leu Ala Gly Ile His Gly Leu Pro Phe Thr Glu Trp      50 55 60 Ala Lys Glu Arg Pro Ser Met Asn Leu Tyr Lys Ala Gly Tyr Cys Thr  65 70 75 80 His Gly Gln Val Leu Phe Pro Thr Trp His Arg Thr Tyr Leu Ser Val                  85 90 95 Phe Glu Gln Ile Leu Gln Gly Ala Ala Ile Glu Val Ala Asn Lys Phe             100 105 110 Thr Ser Asn Gln Thr Asp Trp Ile Gln Ala Ala Gln Asp Leu Arg Gln         115 120 125 Pro Tyr Trp Asp Trp Gly Phe Glu Leu Met Pro Pro Asp Glu Val Ile     130 135 140 Lys Asn Glu Glu Val Asn Ile Thr Asn Tyr Asp Gly Lys Lys Ile Ser 145 150 155 160 Val Lys Asn Pro Ile Leu Arg Tyr His Phe His Pro Ile Asp Pro Ser                 165 170 175 Phe Lys Pro Tyr Gly Asp Phe Ala Thr Trp Arg Thr Thr Val Arg Asn             180 185 190 Pro Asp Arg Asn Arg Arg Glu Asp Ile Pro Gly Leu Ile Lys Lys Met         195 200 205 Arg Leu Glu Glu Gly Gln Ile Arg Glu Lys Thr Tyr Asn Met Leu Lys     210 215 220 Phe Asn Asp Ala Trp Glu Arg Phe Ser Asn His Gly Ile Ser Asp Asp 225 230 235 240 Gln His Ala Asn Ser Leu Glu Ser Val His Asp Asp Ile His Val Met                 245 250 255 Val Gly Tyr Gly Lys Ile Glu Gly His Met Asp His Pro Phe Phe Ala             260 265 270 Ala Phe Asp Pro Ile Phe Trp Leu His His Thr Asn Val Asp Arg Leu         275 280 285 Leu Ser Leu Trp Lys Ala Ile Asn Pro Asp Val Trp Val Thr Ser Gly     290 295 300 Arg Asn Arg Asp Gly Thr Met Gly Ile Ala Pro Asn Ala Gln Ile Asn 305 310 315 320 Asp Glu Thr Pro Leu Glu Pro Phe Tyr Gln Ser Glu Asp Lys Val Trp                 325 330 335 Thr Ser Ala Ser Leu Ala Asp Thr Ala Arg Leu Gly Tyr Ser Tyr Pro             340 345 350 Asp Phe Asp Lys Leu Val Gly Gly Thr Lys Glu Leu Ile Arg Asp Ala         355 360 365 Ile Asp Asp Leu Ile Asp Glu Arg Tyr Gly Ser Lys Pro Ser Ser Gly     370 375 380 Ala Arg Asn Thr Ala Phe Asp Leu Leu Ala Asp Phe Lys Gly Ile Thr 385 390 395 400 Lys Glu His Lys Glu Asp Leu Lys Met Tyr Asp Trp Thr Ile His Val                 405 410 415 Ala Phe Lys Lys Phe Glu Leu Lys Glu Ser Phe Ser Leu Leu Phe Tyr             420 425 430 Phe Ala Ser Asp Gly Gly Asp Tyr Asp Gln Glu Asn Cys Phe Val Gly         435 440 445 Ser Ile Asn Ala Phe Arg Gly Thr Thr Pro Glu Thr Cys Ala Asn Cys     450 455 460 Gln Asp Asn Glu Asn Leu Ile Gln Glu Gly Phe Ile His Leu Asn His 465 470 475 480 Tyr Leu Ala Arg Asp Leu Glu Ser Phe Glu Pro Gln Asp Val His Lys                 485 490 495 Phe Leu Lys Glu Lys Gly Leu Ser Tyr Lys Leu Tyr Ser Arg Glu Asp             500 505 510 Lys Ser Leu Thr Ser Leu Ser Val Lys Ile Glu Gly Arg Pro Leu His         515 520 525 Leu Pro Pro Gly Glu His Arg Pro Lys Tyr Asp His Thr Gln Asp Arg     530 535 540 Val Val Phe Asp Asp Val Ala Val His Val Ile Asn 545 550 555 <210> 6 <211> 517 <212> PRT <213> Mus musculus <400> 6 Met Gly Leu Val Gly Trp Gly Leu Leu Leu Gly Cys Leu Gly Cys Gly   1 5 10 15 Ile Leu Leu Arg Ala Arg Ala Gln Phe Pro Arg Val Cys Met Thr Leu              20 25 30 Asp Gly Val Leu Asn Lys Glu Cys Cys Pro Pro Leu Gly Pro Glu Ala          35 40 45 Thr Asn Ile Cys Gly Phe Leu Glu Gly Arg Gly Gln Cys Ala Glu Val      50 55 60 Gln Thr Asp Thr Arg Pro Trp Ser Gly Pro Tyr Ile Leu Arg Asn Gln  65 70 75 80 Asp Asp Arg Glu Gln Trp Pro Arg Lys Phe Phe Asn Arg Thr Cys Lys                  85 90 95 Cys Thr Gly Asn Phe Ala Gly Tyr Asn Cys Gly Gly Cys Lys Phe Gly             100 105 110 Trp Thr Gly Pro Asp Cys Asn Arg Lys Lys Pro Ala Ile Leu Arg Arg         115 120 125 Asn Ile His Ser Leu Thr Ala Gln Glu Arg Glu Gln Phe Leu Gly Ala     130 135 140 Leu Asp Leu Ala Lys Lys Ser Ile His Pro Asp Tyr Val Ile Thr Thr 145 150 155 160 Gln His Trp Leu Gly Leu Leu Gly Pro Asn Gly Thr Gln Pro Gln Ile                 165 170 175 Ala Asn Cys Ser Val Tyr Asp Phe Phe Val Trp Leu His Tyr Tyr Ser             180 185 190 Val Arg Asp Thr Leu Leu Gly Pro Gly Arg Pro Tyr Lys Ala Ile Asp         195 200 205 Phe Ser His Gln Gly Pro Ala Phe Val Thr Trp His Arg Tyr His Leu     210 215 220 Leu Trp Leu Glu Arg Glu Leu Gln Arg Leu Thr Gly Asn Glu Ser Phe 225 230 235 240 Ala Leu Pro Tyr Trp Asn Phe Ala Thr Gly Lys Asn Glu Cys Asp Val                 245 250 255 Cys Thr Asp Glu Leu Leu Gly Ala Ala Arg Gln Asp Asp Pro Thr Leu             260 265 270 Ile Ser Arg Asn Ser Arg Phe Ser Thr Trp Glu Ile Val Cys Asp Ser         275 280 285 Leu Asp Asp Tyr Asn Arg Arg Val Thr Leu Cys Asn Gly Thr Tyr Glu     290 295 300 Gly Leu Leu Arg Arg Asn Lys Val Gly Arg Asn Asn Glu Lys Leu Pro 305 310 315 320 Thr Leu Lys Asn Val Gln Asp Cys Leu Ser Leu Gln Lys Phe Asp Ser                 325 330 335 Pro Pro Phe Phe Gln Asn Ser Thr Phe Ser Phe Arg Asn Ala Leu Glu             340 345 350 Gly Phe Asp Lys Ala Asp Gly Thr Leu Asp Ser Gln Val Met Asn Leu         355 360 365 His Asn Leu Ala His Ser Phe Leu Asn Gly Thr Asn Ala Leu Pro His     370 375 380 Ser Ala Asn Asp Pro Val Phe Val Val Leu His Ser Phe Thr Asp 385 390 395 400 Ala Ile Phe Asp Glu Trp Leu Lys Arg Asn Asn Pro Ser Thr Asp Ala                 405 410 415 Trp Pro Gln Glu Leu Ala Pro Ile Gly His Asn Arg Met Tyr Asn Met             420 425 430 Val Pro Phe Pro Pro Val Thr Asn Glu Glu Leu Phe Leu Thr Ala         435 440 445 Glu Gln Leu Gly Tyr Asn Tyr Ala Val Asp Leu Ser Glu Glu Glu Ala     450 455 460 Pro Val Trp Ser Thr Thr Leu Ser Val Val Ile Gly Ile Leu Gly Ala 465 470 475 480 Phe Val Leu Leu Leu Gly Leu Leu Ala Phe Leu Gln Tyr Arg Arg Leu                 485 490 495 Arg Lys Gly Tyr Ala Pro Leu Met Glu Thr Gly Leu Ser Ser Lys Arg             500 505 510 Tyr Thr Glu Glu Ala         515 <210> 7 <211> 537 <212> PRT <213> Mus musculus <400> 7 Met Lys Ser Tyr Asn Val Leu Pro Leu Ala Tyr Ile Ser Leu Phe Leu   1 5 10 15 Met Leu Phe Tyr Gln Val Trp Ala Gln Phe Pro Arg Glu Cys Ala Asn              20 25 30 Ile Glu Ala Leu Arg Arg Gly Val Cys Cys Pro Asp Leu Leu Pro Ser          35 40 45 Ser Gly Pro Gly Thr Asp Pro Cys Gly Ser Ser Ser Gly Arg Gly Arg      50 55 60 Cys Val Ala Val Ile Ala Asp Ser Arg Pro His Ser Arg His Tyr Pro  65 70 75 80 His Asp Gly Lys Asp Asp Arg Glu Ala Trp Pro Leu Arg Phe Phe Asn                  85 90 95 Arg Thr Cys Gln Cys Asn Asp Asn Phe Ser Gly His Asn Cys Gly Thr             100 105 110 Cys Arg Pro Gly Trp Arg Gly Ala Ala Cys Asn Gln Lys Ile Leu Thr         115 120 125 Val Arg Arg Asn Leu Leu Asp Leu Ser Pro Glu Glu Lys Ser His Phe     130 135 140 Val Arg Ala Leu Asp Met Ala Lys Arg Thr Thr His Pro Gln Phe Val 145 150 155 160 Ile Ala Thr Arg Arg Leu Glu Asp Ile Leu Gly Pro Asp Gly Asn Thr                 165 170 175 Pro Gln Phe Glu Asn Ile Ser Val Tyr Asn Tyr Phe Val Trp Thr His             180 185 190 Tyr Tyr Ser Val Lys Lys Thr Phe Leu Gly Thr Gly Gln Glu Ser Phe         195 200 205 Gly Asp Val Asp Phe Ser His Glu Gly Pro Ala Phe Leu Thr Trp His     210 215 220 Arg Tyr His Leu Leu Gln Leu Glu Arg Asp Met Gln Glu Met Leu Gln 225 230 235 240 Glu Pro Ser Phe Ser Leu Pro Tyr Trp Asn Phe Ala Thr Gly Lys Asn                 245 250 255 Val Cys Asp Val Cys Thr Asp Asp Leu Met Gly Ser Arg Ser Asn Phe             260 265 270 Asp Ser Thr Leu Ile Ser Pro Asn Ser Val Phe Ser Gln Trp Arg Val         275 280 285 Val Cys Glu Ser Leu Glu Glu Tyr Asp Thr Leu Gly Thr Leu Cys Asn     290 295 300 Ser Thr Glu Gly Gly Pro Ile Arg Arg Asn Pro Ala Gly Asn Val Gly 305 310 315 320 Arg Pro Ala Val Gln Arg Leu Pro Glu Pro Gln Asp Val Thr Gln Cys                 325 330 335 Leu Glu Val Arg Phe Asp Thr Pro Pro Phe Tyr Ser Asn Ser Thr             340 345 350 Asp Ser Phe Arg Asn Thr Val Glu Gly Tyr Ser Ala Pro Thr Gly Lys         355 360 365 Tyr Asp Pro Ala Val Arg Ser Leu His Asn Leu Ala His Leu Phe Leu     370 375 380 Asn Gly Thr Gly Gly Gln Thr His Leu Ser Pro Asn Asp Pro Ile Phe 385 390 395 400 Val Leu Leu His Thr Phe Thr Asp Ala Val Phe Asp Glu Trp Leu Arg                 405 410 415 Arg Tyr Asn Ala Asp Ile Ser Thr Phe Pro Leu Glu Asn Ala Pro Ile             420 425 430 Gly His Asn Arg Gln Tyr Asn Met Val Pro Phe Trp Pro Pro Val Thr         435 440 445 Asn Thr Glu Met Phe Val Thr Ala Pro Asp Asn Leu Gly Tyr Ala Tyr     450 455 460 Glu Val Gln Trp Pro Gly Gln Glu Phe Thr Val Ser Glu Ile Ile Thr 465 470 475 480 Ile Ala Val Ala Ala Leu Leu Ala Val Ala Ile Phe Gly Val                 485 490 495 Ala Ser Cys Leu Ile Arg Ser Ser Ser Thr Lys Asn Glu Ala Asn Gln             500 505 510 Pro Leu Leu Thr Asp His Tyr Gln Arg Tyr Ala Glu Asp Tyr Glu Glu         515 520 525 Leu Pro Asn Pro Asn His Ser Met Val     530 535

Claims (10)

A diagnostic module structure having an open window for measuring color change and a reaction module inlet for allowing the reaction module structure to be inserted and mounted;
A reaction module structure having a diagnostic window that is introduced into the diagnostic module structure and enables visual observation of color change due to reaction with the enzyme composition for tyrosine detection when a sample is added;
And a light source and a light guide plate unit provided in the diagnostic module structure for irradiating light and selectively changing an illumination color of the reaction area,
The enzyme composition comprises tyrosine (L-Tyrosine) present in urine, including tyrosinase, Dopachrome tautomerase (DCT) and tyrosinase-related protein 1 (Tyrp1) Wherein the enzyme control enzyme is an enzyme composition for tyrosine detection. delete The sample according to claim 1,
Wherein the body fluid is blood, urine, serum, plasma, lymph, tissue fluid, secretion, and the like. 2. The method according to claim 1, further comprising a control color table for comparing colors observed through a diagnostic window in one surface area of a transparent window member constituting a diagnostic window of the reaction module structure,
Wherein the reference color table is made of a transparent material, and when the background illumination color of the reaction area is changed, the wavelength of the background illumination color is added so that the color is changed and displayed. . The method according to claim 1,
With the reaction module structure being drawn into the interior of the diagnostic module structure and the open window of the diagnostic module structure and the diagnostic window of the reaction module structure being aligned,
Wherein the diagnostic light is irradiated with light through a light source and a light guide plate unit provided in the diagnostic module structure. The light guide plate unit according to claim 1,
A light guide plate having an incident surface through which light irradiated from a light source is incident and an exit surface through which the light propagated through the incident surface is deflected in the direction of propagation by reflection,
A diffusion sheet for adjusting the diffusion and propagation direction of the light transmitted through the light guide plate,
A prism sheet for refracting and diffusing light emitted from the diffusion sheet to increase brightness,
And a reflective sheet for reflecting the light beam directly below the light source or the light guide plate are laminated on the lower side of the light guide plate. The diagnostic window according to any one of claims 1 to 4, further comprising: a side surface of the transparent window member constituting the diagnostic window and a side surface of the transparent window formed on the entire outer periphery of the transparent window member, Further comprising a black matrix layer for inhibiting light from entering the discriminating environment control window. 2. The enzyme composition according to claim 1,
(DCT) and tyrosinase-related protein 1 (Tyrp1) in a ratio of 0.5 to 2 parts by weight based on 1 part by weight of the tyrosinase, and 40 parts by weight of tyrosinase, , And L-tyrosine into Eumelanin, and the color of the urine is changed from brown to black as the concentration of tyrosine is increased. Diagnostic Kit. 9. The enzyme composition according to any one of claims 1 to 8,
Further comprising a phosphate buffer or Tris-HCl buffer solution having a final concentration of 50 to 100 mM, a Tween 20 having a final concentration of 0.1 to 0.5%, and distilled water, And the liquid is contained in the liquid as the liquid. The detachable cancer diagnostic kit according to claim 1, further comprising an optical film formed on a surface of the diagnostic window to prevent color distortion due to external light.
KR1020160075738A 2016-06-17 2016-06-17 Module type Decision Condition Control Window Diagnostic Kit with Own Light Source for Detecting Cancer Existence KR101725837B1 (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0332444B1 (en) * 1988-03-11 1992-07-01 SAM I1 PHARMACEUTICAL MANUFACTURING CO., Ltd Reagent for diagnosis of cancer
KR101323371B1 (en) 2011-05-03 2013-10-29 김수동 Apparatus and manufacturing method of a diagnostic kit with the sample of urine to diagnose the prostate cancer

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0332444B1 (en) * 1988-03-11 1992-07-01 SAM I1 PHARMACEUTICAL MANUFACTURING CO., Ltd Reagent for diagnosis of cancer
KR101323371B1 (en) 2011-05-03 2013-10-29 김수동 Apparatus and manufacturing method of a diagnostic kit with the sample of urine to diagnose the prostate cancer

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Title
J. Biol. Chem., Vol.85:261-273 (1929) *
J. Clin. Chem. Clin. Biochem.(1986), Vol.24:167-173 *
Journal of Investigative Dermatology Vol.100(2):126-131 *

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