JP2010014403A - Measurement device, measurement apparatus, and measurement method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a measuring device with simple construction and capable of speedily and correctly measuring an antigen of objective matter by effectively compensating, per measurement, measurement nonuniformity due to different dissolving conditions of a flocculation accelerator into samples. <P>SOLUTION: The measuring device includes: a space portion, enclosed by an inner wall that supports a sample containing an antigen of inspected matter; a sample feed opening, communicating with the space portion, for feeding the sample into the space portion; an optical measuring portion for optically measuring the sample; and a reagent support portion, provided in the space portion, for supporting an antibody to the antigen and a flocculation accelerator that accelerates the generation of a flocculation complex containing the antigen and the antibody and labeled by a dye. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a measuring device, a measuring apparatus, and a measuring method for analyzing a test substance contained in a sample.

  Conventionally, POCT (Point of Care Testing) equipment has been used as a measuring equipment used in the field of clinical examination. The POCT device is a device used for a clinical test performed in a medical field excluding a hospital laboratory or a test center, and includes a device used for home medical care (see, for example, Patent Document 1 and Patent Document 2). .

  Examples of the POCT device include a blood glucose sensor, a pregnancy diagnostic agent, an ovulation test agent, and an HbA1c / trace albumin measuring device (for example, DCA2000 manufactured by Bayer). Since the POCT instrument can focus on a marker substance specific to a certain disease state and measure the marker substance easily and quickly, it is effectively used for screening and monitoring of a subject. In addition, since the POCT device is small, it is excellent in portability, can be introduced at low cost, and can be used by anyone without requiring special expertise in terms of operability.

At present, there are many measurement items for clinical examinations, but when body fluid such as urine is used as a specimen, the measurement methods are mainly classified into an optical measurement method and an electrochemical measurement method. In conventional large-scale automated equipment and POCT equipment, measurement is performed using either measurement method.
For example, in Patent Document 3, as an example of measurement of an antigen using an antibody, the antigen albumin or CRP (C-reactive protein) is mixed with the antibody or polyethylene glycol as an aggregation promoter to obtain the antigen. -It has been proposed to generate antibody aggregates and to measure turbidimetry using 470 nm light.
JP 07-248310 A Japanese Patent Laid-Open No. 03-046566 JP-T-2002-509247

  However, in the conventional measurement method based on the production of an antigen-antibody aggregate using a reagent such as a dried antibody or an aggregation promoter as described above, the dissolution of the reagent, particularly the aggregation promoter, in the sample is not necessarily performed. Since it is not easy, the dissolution state differs for each measurement, and as a result, the measurement may be adversely affected.

  Accordingly, the present invention has a simple configuration in view of the above-described conventional problems, and efficiently corrects the measurement non-uniformity caused by the difference in the dissolution state of the aggregation promoter in the sample for each measurement, thereby enabling quick and accurate measurement. It is another object of the present invention to provide a measuring device, a measuring apparatus, and a measuring method capable of measuring an antigen as a measurement target substance.

  In order to solve the above-described conventional problems, a measurement device of the present invention is surrounded by an inner wall, communicates with a space part for holding a sample containing an antigen as a test substance, and communicates with the space part. A sample supply port for supplying the sample to the substrate, an optical measurement unit for optically measuring the sample, an agglutination provided in the space, and comprising an antibody against the antigen, and the antigen and the antibody A reagent holding unit that promotes the formation of the complex and holds the aggregation promoter labeled with the dye.

In addition, the measuring apparatus of the present invention includes a measuring device mounting portion for mounting the measuring device, a light source for emitting light that enters the space portion from outside the space portion, and the space portion outside the space portion. A light receiving portion for receiving the light emitted to the light receiving portion, the intensity of the light reflected by the light receiving portion and reflecting the amount of the aggregated complex, and the light receiving portion received by the dye labeled with the dye And an arithmetic unit for quantifying the concentration of the test substance contained in the sample based on the intensity of light reflecting the concentration of the aggregation promoter in the sample.
According to a first preferred aspect of the measuring apparatus of the present invention, the light source includes a first light source and a second light source, and the light receiving unit receives the first emitted light emitted from the first light source. An intensity of the first emitted light received by the first light receiving unit, and a second light receiving unit that receives the second emitted light emitted from the second light source. Reflects the amount of the aggregation complex, the intensity of the second emitted light received by the second light receiving portion reflects the concentration of the aggregation accelerator in the sample, and further the aggregation A plurality of first calibration curves representing a relationship between the concentration of the test substance and the intensity of the first emitted light received by the first light receiving unit when the concentrations of the accelerators are different from each other; and The concentration of the aggregation accelerator and the second emitted light received by the second light receiving unit A storage unit for storing a second calibration curve showing the relationship between the degree.
According to a second preferred embodiment of the measuring apparatus of the present invention, the light source includes a first light source and a second light source, and the light receiving unit receives the first emitted light emitted from the first light source. An intensity of the first emitted light received by the first light receiving unit, and a second light receiving unit that receives the second emitted light emitted from the second light source. Reflects the amount of the aggregated complex, the intensity of the second emitted light received by the second light receiving unit reflects the concentration of the aggregation accelerator in the sample, and A plurality of third calibration curves representing a relationship between the concentration of the aggregation promoter and the intensity of the first emitted light received by the first light receiving unit when the concentrations of the test substances are different from each other; and The concentration of the aggregation accelerator and the second emitted light received by the second light receiving unit A storage unit for storing a second calibration curve showing the relationship between the degree.

  A first measurement method of the present invention is a measurement method for measuring the test substance contained in the sample using the measurement device and the measurement apparatus according to the first preferred embodiment, wherein the measurement device includes: A first step of supplying the sample into the space, a second step of irradiating the space to which the sample is supplied with the first emitted light through the optical measurement unit, and through the optical measurement unit. A third step of measuring the intensity of the first emitted light emitted from the space portion to which the sample is supplied; and the second emitted light is passed through the optical measuring portion to the space portion to which the sample is supplied. Reference is made to the fourth step of irradiating, the fifth step of measuring the intensity of the second emitted light emitted from the space where the sample is supplied through the optical measuring unit, and the second calibration curve. Measured in the fifth step A sixth step of converting the intensity of the emitted second emitted light into the concentration of the aggregation promoter, and the aggregation promoter converted in the sixth step from the plurality of first calibration curves. The seventh step of extracting the first calibration curve corresponding to the concentration, and the first calibration curve extracted in the seventh step, the first calibration curve measured in the third step is referred to And an eighth step of converting the intensity of the emitted light into the concentration of the test substance.

  A second measurement method of the present invention is a measurement method for measuring the test substance contained in the sample using the measurement device and the measurement apparatus according to the second preferred embodiment, wherein the measurement device includes: A first step of supplying the sample into the space, a second step of irradiating the space to which the sample is supplied with the first emitted light through the optical measurement unit, and through the optical measurement unit. A third step of measuring the intensity of the first emitted light emitted from the space portion to which the sample is supplied; and the second emitted light is passed through the optical measuring portion to the space portion to which the sample is supplied. Reference is made to the fourth step of irradiating, the fifth step of measuring the intensity of the second emitted light emitted from the space where the sample is supplied through the optical measuring unit, and the second calibration curve. In the fifth step A sixth step of converting the determined intensity of the second emitted light into the concentration of the aggregation accelerator, and a concentration of the aggregation promoter converted in the sixth step from the third calibration curve In the seventh step, the seventh step of extracting a third calibration curve in which the intensity of the first emitted light corresponding to 1 is equal to the intensity of the first emitted light measured in the third step, and And an eighth step of determining the concentration of the test substance corresponding to the extracted third calibration curve.

  According to the present invention, it has a simple configuration, efficiently corrects the measurement heterogeneity due to the difference in the dissolution state of the aggregation promoter in the sample for each measurement, and quickly and accurately detects the antigen as the measurement target substance. A measurement device, a measurement apparatus, and a measurement method capable of performing measurement can be provided.

The measuring device of the present invention is surrounded by an inner wall and holds a sample containing an antigen as a test substance, and a sample supply that communicates with the space and supplies the sample into the space. Mouth, an optical measurement unit for optically measuring the sample, and provided in the space, promote the production of an antibody against the antigen and an aggregate complex containing the antigen and the antibody, And a reagent holding part for holding the labeled aggregation promoter.
With this configuration, it is possible to realize generation of an aggregate complex by a reaction between an antigen, which is a test substance contained in a sample supplied in the space portion, and an antibody supported on the reagent holding portion. After the light incident on the optical measurement unit of the measurement device is transmitted through the sample supplied in the space, the intensity of scattered light and transmitted light emitted from the optical measurement unit of the measurement device is generated in the sample. Depends on the amount of aggregated complex. For this reason, by measuring the intensity of the emitted light, it is possible to measure the amount of the aggregated complex, that is, the sample test substance.

Furthermore, in the measurement device of the present invention, since the dye is labeled on the aggregation promoter carried on the reagent holding unit, the dye is dissolved with the dissolution of the aggregation promoter in the sample. The intensity of transmitted light (absorbed light) or fluorescence, which is light that is incident on the measurement device and is emitted from the space where the sample is supplied, depends on the amount of the dye dissolved in the sample. For this reason, the quantity of the aggregation promoter dissolved in the sample can be measured by measuring the emitted light.
Therefore, since the amount of aggregation promoter dissolved in the sample can be measured with a simple configuration, the measurement heterogeneity due to the difference in the dissolution state of the aggregation promoter in the sample for each measurement is efficient. The amount of antigen (test substance), which is a substance to be measured, can be measured quickly and accurately with sufficient correction.

The optical measurement unit preferably includes a light incident part for allowing light to enter the space part from outside the space part, and a light emitting part for emitting light from the space part to the outside of the space part. .
The reagent holding unit preferably includes a first reagent holding unit that holds the antibody and a second reagent holding unit that holds the aggregation promoter labeled with the dye. With this configuration, the antibody and the aggregation promoter are held separately, so that the dissolution of the antibody becomes smoother and stable measurement can be performed.
The aggregation accelerator is preferably a polymer compound containing polyethylene glycol as the main chain.

Examples of the dye for labeling the aggregation promoter include phenoxazine and phenoxazine derivatives, phenazine and phenazine derivatives, acridine and acridine derivatives, fluorescein and fluorescein derivatives, and phenothiazine derivatives. The above dyes are chemically stable and well known for their absorption coefficient, fluorescence yield, and spectroscopic properties such as absorption, excitation, and fluorescence wavelength. Therefore, aggregation promoters labeled with these dyes Can be measured stably and accurately.
Examples of phenothiazine derivatives include thionine and thionine derivatives, azure A and azure A derivatives, azure C and azure C derivatives, and toluidine blue and toluidine blue derivatives. The phenothiazine derivatives are chemically stable and well-known for their spectroscopic properties, and have an amino group in the molecule, so that they can be covalently labeled to aggregation promoters. The amount of the aggregation accelerator labeled with these dyes can be measured stably and accurately.

In addition, the measuring apparatus of the present invention includes a measuring device mounting portion for mounting the measuring device, a light source for emitting light that enters the space portion from outside the space portion, and the space portion outside the space portion. A light receiving portion for receiving the light emitted to the light receiving portion, the intensity of the light reflected by the light receiving portion and reflecting the amount of the aggregated complex, and the light receiving portion received by the dye labeled with the dye And an arithmetic unit for quantifying the concentration of the test substance contained in the sample based on the intensity of light reflecting the concentration of the aggregation promoter in the sample.
The light source includes a first light source and a second light source, and the light receiving unit emits a first light receiving unit that receives the first emitted light emitted from the first light source and the second light source. And a second light receiving unit that receives the second emitted light, wherein the intensity of the first emitted light received by the first light receiving unit reflects the amount of the aggregated complex, It is preferable that the intensity of the second emitted light received by the second light receiving unit reflects the concentration of the aggregation accelerator in the sample.

As a first preferred aspect of the measuring apparatus of the present invention, the measuring apparatus further receives the concentration of the test substance and the first light receiving unit when the concentrations of the aggregation accelerators are different from each other. A plurality of first calibration curves representing the relationship with the intensity of the first emitted light, the concentration of the aggregation accelerator, and the intensity of the second emitted light received by the second light receiving unit; A storage unit for storing a second calibration curve representing the relationship.
With this configuration, the second calibration curve stored in the storage unit is referenced based on the signal from the second light receiving unit corresponding to the intensity of the second emitted light emitted from the sample held in the space. Thus, the dissolution concentration of the aggregation promoter labeled with the dye can be determined. A first calibration curve corresponding to the determined concentration of the aggregation promoter can be extracted from the plurality of first calibration curves. By referring to the extracted first calibration curve, the signal from the first light receiving part corresponding to the intensity of the first emitted light emitted from the sample held in the space part is obtained as the concentration of the test substance. Can be converted to Therefore, it is possible to efficiently correct the measurement non-uniformity caused by the difference in the dissolution state of the aggregation promoter in the sample for each measurement, and to quickly and accurately measure the antigen as the measurement target substance.

As a second preferred embodiment of the measuring apparatus of the present invention, the measuring apparatus further receives the concentration of the aggregation promoter and the first light receiving unit when the concentrations of the test substances are different from each other. A plurality of third calibration curves representing the relationship with the intensity of the first emitted light, the concentration of the aggregation accelerator, and the intensity of the second emitted light received by the second light receiving unit. A storage unit for storing a second calibration curve representing the relationship.
With this configuration, the agglutination labeled with the dye with reference to the second calibration curve stored in the storage unit based on the signal output from the second light receiving unit corresponding to the intensity of the second outgoing light The dissolution concentration of the accelerator can be determined. A third output is selected such that the intensity of the first emitted light corresponding to the determined concentration of the aggregation accelerator is equal to the measured intensity of the first emitted light among the plurality of third calibration curves. A calibration curve can be extracted. And the density | concentration of the to-be-tested substance corresponding to the extracted 3rd calibration curve can be calculated | required. Therefore, it is possible to efficiently correct the measurement non-uniformity caused by the difference in the dissolution state of the aggregation promoter in the sample for each measurement, and to quickly and accurately measure the antigen as the measurement target substance.

  Moreover, it is preferable that the measuring apparatus of the present invention further includes a suction unit for sucking the sample into the space of the measuring device. With this configuration, the sample can be sucked from the sample supply port of the measurement device using the suction unit, and the sample can be easily supplied into the sample holding unit of the measurement device.

  The first measurement method of the present invention is a method for measuring the test substance contained in the sample using the measurement device and the measurement apparatus of the first preferred embodiment. That is, a first step of supplying the sample into the space portion of the measurement device, and a second step of irradiating the space portion to which the sample is supplied with the first emitted light through the optical measurement portion; A third step of measuring the intensity of the first emitted light emitted from the space part to which the sample is supplied through the optical measurement part, and the space part to which the sample is supplied through the optical measurement part. A fourth step of irradiating the second outgoing light, a fifth step of measuring the intensity of the second outgoing light emitted from the space where the sample is supplied through the optical measuring unit, and the second A sixth step of converting the intensity of the second emitted light measured in the fifth step into the concentration of the aggregation accelerator, and a plurality of the first calibration curves. From inside, the aggregate converted in the sixth step It was measured in the third step by referring to the seventh calibration step for extracting the first calibration curve corresponding to the concentration of the accelerator and the first calibration curve extracted in the seventh step. And an eighth step of converting the intensity of the first outgoing light into the concentration of the test substance.

  With this configuration, the second calibration curve stored in the storage unit is referenced based on the signal from the second light receiving unit corresponding to the intensity of the second emitted light emitted from the sample held in the space. Thus, the dissolution concentration of the aggregation promoter labeled with the dye can be determined. A first calibration curve corresponding to the determined concentration of the aggregation promoter can be extracted from the plurality of first calibration curves. By referring to the extracted first calibration curve, among the light emitted from the first light source, the first light receiving unit corresponding to the intensity of the first emitted light from the sample held in the space Can be converted into the concentration of the test substance. Therefore, it is possible to efficiently correct the measurement non-uniformity caused by the difference in the dissolution state of the aggregation promoter in the sample for each measurement, and to quickly and accurately measure the antigen as the measurement target substance.

  The second measurement method of the present invention is a method for measuring the test substance contained in the sample using the measurement device and the measurement apparatus of the second preferred embodiment. That is, a first step of supplying the sample into the space portion of the measurement device, and a second step of irradiating the space portion to which the sample is supplied with the first emitted light through the optical measurement portion; A third step of measuring the intensity of the first emitted light emitted from the space part to which the sample is supplied through the optical measurement part, and the space part to which the sample is supplied through the optical measurement part. A fourth step of irradiating the second outgoing light, a fifth step of measuring the intensity of the second outgoing light emitted from the space where the sample is supplied through the optical measuring unit, and the second From the third calibration curve, the sixth step of converting the intensity of the second emitted light measured in the fifth step into the concentration of the aggregation accelerator by referring to the calibration curve of Of the aggregation accelerator converted in the sixth step A seventh step of extracting a third calibration curve in which the intensity of the first emitted light corresponding to the degree is equal to the intensity of the first emitted light measured in the third step; and the seventh step And an eighth step of obtaining the concentration of the test substance corresponding to the third calibration curve extracted in step (b).

  With this configuration, the agglutination labeled with the dye with reference to the second calibration curve stored in the storage unit based on the signal output from the second light receiving unit corresponding to the intensity of the second outgoing light The dissolution concentration of the accelerator can be determined. The third emission light intensity corresponding to the determined concentration of the aggregation accelerator among the plurality of third calibration curves is equal to the first emission light intensity measured in the third step. Can be extracted. And the density | concentration of the to-be-tested substance corresponding to the extracted 3rd calibration curve can be calculated | required. Therefore, it is possible to efficiently correct the measurement non-uniformity caused by the difference in the dissolution state of the aggregation promoter in the sample for each measurement, and to quickly and accurately measure the antigen as the measurement target substance.

Examples of samples in clinical tests include body fluids such as serum, plasma, urine, interstitial fluid, lymph fluid, and liquid samples such as culture supernatant. In particular, urine is a very effective sample for non-invasive daily health management at home. In addition, a reagent that reacts with a specific component in these body fluids, for example, an enzyme, an antibody, or a dye, and a mixture of body fluids may be supplied as a sample to the measurement device.
Examples of the test substance include albumin, hCG, LH, CRP, and IgG.
In urine qualitative tests, renal function tests, pregnancy tests, ovulation tests, etc. performed at the first stage of health care, there are requests for measurement of proteins, trace albumin, hormones such as hCG, LH, etc. Is suitable for optical measurement based on antigen-antibody reaction. Examples of the optical measurement based on the antigen-antibody reaction include those that measure the turbidity generated in the sample based on the antigen-antibody reaction, such as an immunoratio method, an immunoturbidimetric method, and a latex immunoaggregation method.

Moreover, since the antibody contained in the reagent can be produced by a known method, it is advantageous in that the reagent is easy to produce. For example, an antibody against an antigen can be obtained by immunizing a mouse or rabbit using a protein such as albumin or CRP or a hormone such as hCG or LH as an antigen. Examples of the antibody include an antibody against a protein contained in urine such as albumin and CRP, and an antibody against a hormone contained in urine such as hCG and LH.
Hereinafter, embodiments of the present invention will be described with reference to the drawings, using an example in which urine is used as a sample and the test substance is human albumin.

(Embodiment 1)
A measuring device for sucking and holding a sample for measuring a test substance contained in the sample will be described. FIG. 1 is a perspective view of a measurement device according to Embodiment 1 of the present invention, and FIG. 2 is a cross-sectional view taken along line AA in FIG. 1 and 2, the measuring device 1 is composed of a base 2 having a space 2a made of a transparent material, for example, polystyrene. The base 2 includes a hollow quadrangular prism portion 2d and a hollow quadrangular pyramid portion 2e, and the space 2a inside the base 2 functions as a sample holding portion. A suction port 2c is provided at the upper end of the hollow quadrangular column portion 2d, and a sample supply port 2b is provided at the lower end of the hollow quadrangular pyramid portion 2e.

The side surface of the hollow rectangular column portion 2d has four surfaces, that is, a first surface 3, a second surface 4, a third surface 5, and a fourth surface 6. Among these surfaces, the second surface 4 functions as a light incident portion, and the third surface 5 facing the second surface 4 functions as a light emitting portion. The light incident part and the light emitting part correspond to the optical measurement part.
The light incident portion and the light emitting portion are preferably formed of an optically transparent material or a material that does not substantially absorb visible light. For example, in addition to the polystyrene described above, quartz, glass, polymethyl methacrylate, and the like can be given. When the measuring device is made disposable, it is preferable to use a resin material such as polystyrene from the viewpoint of cost. Further, in the measurement device, at least only the light incident part and the light emission part need only be optically transparent, and parts other than the above need not necessarily be optically transparent.

Of the inner wall surface surrounding the space 2a, the first reagent holding part 7 and the second reagent holding part 8 are provided on the inner wall surface of the fourth surface 6 of the hollow quadrangular prism portion 2d.
The first reagent holding unit 7 holds a reagent for reacting with the sample supplied in the space 2a in a dry state. As the reagent, an antibody that reacts with an antigen as a test substance in a sample is used. The first reagent holding unit 7 is preferably arranged at a position where the reagent is easily dissolved in the sample when the sample is supplied into the space 2a.
In addition, the second reagent holding unit 8 carries an agglutination promoter for promoting the agglutination reaction between the antigen and the antibody as a reagent in a dry state. The aggregation promoter is labeled with a dye. As the aggregation promoter labeled with a dye, for example, a compound having a structure represented by the following general formula (1) is used.
General formula (1):

In formula (1), X and Y are each independently hydrogen (—H) or a methyl group (—CH ₃ ). n is preferably 30 to 350.
The general formula (1) shows an example in which polyethylene glycol is used as an aggregation accelerator and a phenothiazine derivative is used as a dye for labeling the aggregation accelerator. Polyethylene glycol and a phenothiazine derivative are bound by a peptide bond at the main chain end of polyethylene glycol, which is an aggregation accelerator. In the general formula (1), the dye is thionine when X = Y = H, Azure C when X = H and Y = CH ₃ , and Azure A when X = Y = CH ₃ .

Examples of the aggregation promoter include compounds containing as a main chain a water-soluble polymer such as the above-described polyethylene glycol and polysaccharides (dextran, cellulose, hyaluronic acid, chondroitin sulfate, etc.). The binding form of the aggregation promoter and the dye may be a C—C bond, a C═C bond, a C═N bond, a S—S bond or the like in addition to the peptide (N—C) bond.
Moreover, the C1-C10 side chain may couple | bond with the main chain of the aggregation promoter. Examples of the side chain include carboxyalkyl, alkyl, alkylamine, alkylamide and the like.

  As the dye for labeling the aggregation promoter, for example, phenoxazine and phenoxazine derivatives, phenazine and phenazine derivatives, acridine and acridine derivatives, and fluorescein and fluorescein derivatives are used in addition to the phenothiazine derivatives described above. Examples of the phenothiazine derivative include thionin and thionine derivatives, Azure A and Azure A derivatives, Azure C and Azure C derivatives, and Toluidine blue and Toluidine blue derivatives.

FIG. 4 shows an absorption spectrum of thionine, which is an example of a dye used for labeling. In FIG. 4, the horizontal axis represents wavelength (nm) and the vertical axis represents absorbance (arbitrary intensity). FIG. 4 shows that the absorbance decreases with decreasing thionin concentration.
Phenothiazine derivatives, phenoxazine and phenoxazine derivatives, phenazine and phenazine derivatives, acridine and acridine derivatives, and fluorescein and fluorescein derivatives are chemically stable, their extinction coefficient, fluorescence yield, and absorption / excitation / fluorescence wavelength. Since the spectroscopic characteristics such as these are well known, the aggregation promoter labeled with these dyes can be measured stably and accurately. Further, thionine and thionine derivatives, azure A and azure A derivatives, azure C and azure C derivatives, and toluidine blue and toluidine blue derivatives are chemically stable and have well-known spectroscopic characteristics. At the same time, these dyes have an amino group in the molecule and can be easily labeled with a covalent bond to an aggregation promoter.

In addition, in the 2nd reagent holding | maintenance part 8, you may hold | maintain not only the aggregation promoter labeled with the pigment | dye but the unlabeled aggregation promoter not labeled with the pigment | dye.
The molecular weight of the aggregation accelerator and the chemical structure of the side chain can be appropriately selected depending on the type of aggregation reaction. As the aggregation accelerator, for example, one having a molecular weight in the range of 1000 to 10,000 is used. Since the aggregation promoter is used to promote the antigen-antibody reaction, the second reagent holding part 8 is preferably arranged in the vicinity of the first reagent holding part 7 containing the antibody. Similarly to the first reagent holding unit 7, the second reagent holding unit 8 is arranged at a position where the aggregation promoter is easily dissolved in the sample when the sample is supplied into the space 2a. It is preferable.
As described above, in the measuring device 1 of the present embodiment, the first reagent holding unit 7 and the second reagent holding unit 8 provided in the space 2a are loaded with the antibody and the aggregation promoter, respectively. The dissolution of the antibody in the sample becomes very smooth and the measurement of the test substance can be performed stably.

When measuring a test substance using the measuring device 1, after immersing a part of the measuring device 1 in, for example, urine collected in a container, the measuring device 1 is used to measure the inside of the space 2a from the suction port 2c. By sucking air, urine is supplied into the space 2a.
In the present embodiment, the surfaces that function as the light incident portion and the light emitting portion are the second surface 4 and the third surface 5, but the present invention is not limited to this. For example, the second surface 4 is The fourth surface 6 may function as a light emitting portion for light that has been made to function as a light incident portion and is scattered in the space portion 2a after entering the space portion 2a from the second surface 4. Good.

Next, a method for manufacturing the measuring device 1 will be described with reference to FIGS.
FIG. 3 is an exploded perspective view of the measuring device according to the first embodiment. As shown in FIG. 3, the first member 31 and the second member 32 constituting the base 2 of the measuring device 1 are each made of a transparent material, such as polystyrene, and have a recess. By combining the first member 31 and the second member 32 with each other, the base body 2 having the hollow quadrangular prism portion 2d and the hollow quadrangular pyramid portion 2e is configured.
The first member 31 and the second member 32 are obtained by molding using a mold. Further, by forming the first member 31 and the second member 32 symmetrically, the first member 31 and the second member 32 can be formed with a single mold, and the cost can be reduced. A known resin molding technique may be used for molding. Although the dimension of the 1st member 31 and the 2nd member 32 should just be set suitably, they are width A10mm, length B84mm, length C6mm, and thickness 1mm, for example.

First, the first reagent holding unit 7 and the second reagent holding unit 8 are provided on the bottom surface of the concave portion of the second member 32 as follows.
For example, a certain amount of an aqueous solution of an antibody against human albumin, which is a reagent for optical measurement, is dropped on the bottom surface of the recess of the second member 32 using a microsyringe or the like, and this is placed in an environment of room temperature to 30 ° C. By allowing to stand and evaporate the water, the antibody against human albumin can be supported in a dry state to form the first reagent holding unit 7. For example, an aqueous solution of an antibody having a concentration of 8 mg / dL may be used and dropped into a region having an area of 5 cm ² with a dropping amount of 0.7 mL.

Similarly, a certain amount of an aqueous solution of the aggregation accelerator is dropped on the bottom surface of the recess of the second member 32 using a microsyringe or the like, and this is left in an environment of about room temperature to 30 ° C. to evaporate water. Thus, the second reagent holding unit 8 can be formed by supporting the aggregation accelerator in a dry state. For example, an aqueous solution of a coagulation accelerator having a concentration of 16% by weight may be dropped into a region having an area of 5 cm ² with a dropping amount of 0.7 mL.
The concentration and amount of the aqueous solution containing the reagent to be applied may be appropriately selected according to the required device characteristics and the spatial restriction of the formation position on the second member 32. Further, the positions and areas of the first reagent holding part 7 and the second reagent holding part 8 in the second member 32 may be appropriately selected in view of the solubility of the reagent in the sample, the position of the optical measurement part, and the like. .

An antibody against human albumin can be obtained by a conventionally known method. For example, an anti-human albumin antibody can be obtained by purifying rabbit antiserum immunized with human albumin by protein A column chromatography and then dialysis using a dialysis tube.
Further, as described above, instead of forming the first reagent holding part 7 and the second reagent holding part 8, for example, a porous carrier made of glass fiber or filter paper is impregnated with a solution of each reagent. After that, the reagent may be supported by drying, and the carrier may be provided in the space portion 2a by sticking the carrier to the bottom surface of the concave portion of the second member 32, respectively. Alternatively, the reagent may be arranged by directly applying a solution of the reagent to a part of the wall surface constituting the space 2a and then drying. Furthermore, a carrier carrying a lyophilized reagent outside the space 2a may be installed in the space 2a.

Next, the second member 32 provided with the first reagent holding unit 7 and the second reagent holding unit 8 obtained above and the first member 31 are positioned at a position indicated by a one-dot chain line in FIG. The measurement device 1 is assembled by joining in relation.
Specifically, after an adhesive such as an epoxy resin is applied to the joint portion between the first member 31 and the second member 32 and the first member 31 and the second member 32 are bonded together, It may be allowed to stand and dry for adhesion. In addition, after combining the first member 31 and the second member 32 without applying an adhesive, a joint portion between the first member 31 and the second member 32 is formed using a commercially available welding machine. It may be welded by heat or ultrasonic waves.

  In the present embodiment, two reagent holding units, that is, a first reagent holding unit 7 and a second reagent holding unit 8 are provided as the reagent holding units so as to carry the antibody and the aggregation promoter as the reagents, respectively. As described above, one reagent holding portion may be formed by supporting a mixture of an antibody and an aggregation promoter in a dry state on the wall surface in the space portion 2a.

  As described above, by using the measurement device 1 according to Embodiment 1 of the present invention, the antigen that is the test substance contained in the sample supplied in the space 2a and the first reagent holding unit 7 are used. It is possible to realize the formation of aggregates by reaction with the antibody supported on the. The intensity of scattered light or transmitted light, which is light emitted from the light emitting portion of the measuring device 1 after being incident from the light incident portion of the measuring device 1 and transmitted through the sample supplied into the space 2a, Depending on the amount of aggregates produced. Therefore, by measuring the emitted light, the amount of the aggregate, that is, the test substance in the sample can be measured.

Furthermore, in the measurement device 1 of the present embodiment, since the dye is labeled on the aggregation accelerator, the dye is dissolved with the dissolution of the aggregation accelerator in the sample. The intensity of transmitted light (absorbed light) or fluorescence, which is light emitted from the light emitting portion of the measuring device after being incident from the light incident portion of the measuring device 1 and transmitted through the sample supplied into the space 2a, is Depends on the amount of dye dissolved in the sample. For this reason, the quantity of the aggregation promoter dissolved in the sample can be measured by measuring the emitted light.
Therefore, the amount of the aggregation promoter dissolved in the sample can be easily measured with the simple configuration of the measuring device 1 in the present embodiment. In addition, measurement heterogeneity due to differences in the state of dissolution of the aggregation promoter in the sample for each measurement is efficiently corrected, and the amount of antigen (test substance) that is the measurement target substance is measured quickly and accurately. be able to.

The shape of the measuring device of the present invention is not limited to the measuring device 1 of the present embodiment as long as it satisfies the constituent requirements of the present invention and can obtain the effects of the present invention. For example, the measurement device 11 shown in FIGS. 5 and 6 may be used. FIG. 5 is a perspective view showing another example of the measuring device according to Embodiment 1 of the present invention, and FIG. 6 is a cross-sectional view taken along the line BB of FIG. In addition, in the measurement device 11 of FIG.5 and FIG.6, the same code | symbol is attached | subjected to the same component as the measurement device 1 of FIG.1 and FIG.2.
5 and 6, the measuring device 11 is composed of a base 2 having a bottomed hollow rectangular parallelepiped shape having a space 2a therein. A sample supply port 2 b is provided below the first surface 3 of the base 2.
The base 2 corresponds to the first surface 3, the second surface 4, and the third surface 5 corresponding to the third surface 5, the first member 31 having the bottom 33, and the fourth surface 6. The second member 32 is configured. A first reagent holding unit 7 and a second reagent holding unit 8 similar to those of the measurement device 1 of FIGS. 1 and 2 are arranged on the inner wall surface of the second member 32.

(Embodiment 2)
In the present embodiment, an example of the measurement apparatus according to the first preferred embodiment of the present invention and the first measurement method of the present invention using the same will be described with reference to the drawings.
First, the configuration of the measuring apparatus will be described. FIG. 7 is a perspective view showing the appearance of the measuring apparatus according to the present embodiment. As shown in FIG. 7, the measurement device 80 includes a measurement device attachment portion 81 for attaching the measurement device 1 in Embodiment 1 to the measurement device 80, a display portion 82 that is a display for displaying measurement results, a sample aspiration A start button 83 and a measurement device removal button 84 are provided. As will be described later, inside the measuring apparatus 80, a device mounting port 94 communicating with the measuring device mounting portion 81 and a piston mechanism 103 connected to the device mounting port 94 via the frame member 92 are provided. The measurement device 1 is preferably attached to the measurement device 80 in a detachable state. Moreover, it is preferable that the measuring device 1 is disposable.

FIG. 8 is a longitudinal sectional view showing an internal configuration when the measuring device is inserted in the measuring apparatus of the present embodiment. As shown in FIG. 8, when the measuring device 1 is inserted into the measuring device 80 from the measuring device mounting portion 81, the suction port 2c of the measuring device 1 is engaged with the convex device mounting port 94 and a slide member. 91 is pushed upward. Then, the measuring device insertion detection switch 95 is pressed by the pressing protrusion 91 a of the slide member 91, and the mounting of the measuring device 1 to the measuring device 80 is completed.
The device mounting port 94 includes a device mounting projection 92a provided in a convex shape on the frame member 92, and a device mounting sealing member 93 fitted into the outer periphery of the device mounting projection 92a. The shape of the outer periphery of the device mounting projection 92 a and the device mounting sealing member 93, that is, the cross-sectional shape of the device mounting projection 92 a and the device mounting sealing member 93 in the direction perpendicular to the center line 96 is the measurement device 1. It has a shape similar to the inner shape of the suction port 2c. The outer diameter dimension of the device mounting sealing member 93 is slightly larger than the diameter dimension of the suction port 2 c of the measuring device 1. For this reason, the adhesiveness of the device mounting sealing member 93 and the suction port 2c is improved, and the leakage of air at those joint portions can be prevented.

The device mounting sealing member 93 is made of an elastic material such as isoprene rubber, fluorine rubber, silicon rubber, Teflon (registered trademark) -coated rubber, or the like. Alternatively, the device mounting port 94 in which the device mounting protrusion 92a and the device mounting sealing member 93 are integrated may be used, and the device mounting port 94 itself may be made of an elastic material such as isoprene rubber.
In the frame member 92, a convex cylinder mounting projection 92 b is provided on the opposite side of the device mounting projection 92 a, and the outer peripheral portion is made of a material having elasticity similar to that of the device mounting sealing member 93. A cylinder mounting sealing member 97 is fitted. A cylinder 100 having a piston 99 to which a piston rod 98 is fixed is mounted on the cylinder mounting protrusion 92 b of the frame member 92 via the cylinder mounting sealing member 97. Thereby, it has airtightness between the cylinder 100 and the frame member 92.

The piston mechanism 103 is a suction part for sucking a sample to the measuring device 1, and is screwed into a cylinder 100 having a piston 99 to which the piston rod 98 is fixed and a female screw part 98 a provided on the piston rod 98. The male screw portion 102 and the motor 101 to which the male screw portion 102 is fixed are configured.
Specifically, a female screw portion 98a is provided at the center of the piston rod 98, and a key groove 98b is formed in the longitudinal direction of the outer peripheral portion thereof. A protrusion 100 a provided on the upper end surface of the cylinder 100 on the side opposite to the joint with the cylinder mounting sealing member 97 is engaged with the key groove 98 b. As a result, the piston rod 98 does not rotate and moves straight without fail.
Further, a male screw portion 102 fixed to the rotating shaft of the motor 101 is screwed into the female screw portion 98a. Therefore, the piston rod 98 can linearly move up and down along the center line 96 by the rotation of the male screw portion 102 accompanying the rotation of the motor 101. In addition, a hole 100b is formed in the upper end surface of the cylinder 100 where the projecting portion 100a is formed so as to remove air in the cylinder 100 that flows in as the piston 99 moves.

The piston mechanism 103 which is a suction portion may be manually operated or automatically operated, but it is preferable to automate it because the burden on the operator can be reduced. As an automatic method, there is a method of operating the piston 99 with the motor 101 as described above. As the motor 101, a step motor, a DC motor or the like is used.
The step motor is a motor that rotates by a specific rotation angle per input pulse signal, and the rotation angle can be determined by the number of pulses, so that an encoder for positioning is not required. That is, the operating distance of the piston 99 can be controlled by the number of input pulses. There is also a linear step motor. This type of motor incorporates a linear mechanism in which a male screw and a female screw are combined in the motor, and is configured such that a rod-like movable portion moves linearly depending on the number of input pulses. For this reason, it is only necessary to directly connect the piston 99 to the rod-like movable portion, and the configuration becomes simple.
On the other hand, in the case of a DC motor, an encoder that detects the rotational position of the motor 101 is required to control the operating distance of the piston 99. Therefore, it is preferable that the piston mechanism 103 in the measuring apparatus according to the present embodiment has a configuration in which a step motor is used as the motor 101 and the piston 99 is operated by the step motor.

Thus, in the present invention, with the measurement device 1 attached so that the suction port 2c of the measurement device 1 is connected to the measurement device 80, the piston mechanism 103 that is a suction portion is used to measure the measurement device 1. A sample can be sucked from the sample supply port 2 b and the sample can be easily supplied into the space 2 a of the measurement device 1.
Further, the measuring apparatus 80 receives the first light source 104 and the second light source 106 for emitting light incident on the optical measurement unit of the measurement device 1 and the light emitted from the optical measurement unit. A first light receiver 105 that is a first light receiver and a second light receiver 107 that is a second light receiver are provided. Each light (first emitted light and second emitted light) emitted from the first light source 104 and the second light source 106 is a light incident portion on the second surface 4 of the measuring device 1, and the measuring device. The sample supplied to the first space portion 2a and the light emitting portion on the third surface 5 of the measuring device 1 are sequentially transmitted and received by the first light receiver 105 and the second light receiver 107, respectively. The first light receiver 105 and the second light receiver 107 each output an electrical signal corresponding to the intensity of the received light to the CPU 121 described later.

In the light path where the light emitted from the first light source 104 is received by the first light receiver 105 and the light path where the light emitted from the second light source 106 is received by the second light receiver 107, The position of the light spot that passes through the light incident portion and the light emitting portion of the measuring device 1 is positioned away from the joint portion between the first member 31 and the second member 32 constituting the measuring device 1. In addition, it is preferable to set the arrangement positions of the first light source 104, the first light receiver 105, the second light source 106, and the second light receiver 107.
As the first light source 104 and the second light source 106, for example, a semiconductor laser that emits light with wavelengths of 780 nm and 600 nm can be used. Alternatively, a light emitting diode (LED) may be used.

In the present embodiment, assuming that measurement by an immunoturbidimetric method is applied, the irradiation and reception wavelength of 780 nm are selected as the wavelengths of the first light source 104 and the first light receiver 105. This wavelength can be appropriately selected according to the measurement method and the measurement target.
The wavelength of the second light source 106 and the second light receiver 107 is 600 nm, but this wavelength is preferably near the absorption peak of the dye that labels the aggregation promoter, and depends on the absorption characteristics of the dye used for the measurement. Thus, an appropriate wavelength may be selected as appropriate.
As the first light receiver 105 and the second light receiver 107, for example, a photodiode can be used. Alternatively, a charge coupled device (CCD), a photomultimeter, or the like may be used.

Here, FIG. 9 is a front view showing a measurement device removal mechanism in the measurement apparatus of the present embodiment, and FIG. 10 is a side view showing the measurement device removal mechanism in the measurement apparatus of the present embodiment. As shown in FIGS. 9 and 10, the measurement device removal mechanism 117 is provided below the frame member 92 in the measurement apparatus 80. The measurement device removal mechanism 117 includes a solenoid 115 having a slide member 91, drive levers 111 and 112, a fulcrum shaft 113, an operation pin 114, and a plunger 116.
Specifically, on the surface of the slide member 91 on the first surface 3 side of the measuring device 1 and the surface on the fourth surface 6 side opposite thereto, pressing pin portions 91b and 91c are provided, respectively. . The pressing pin portion 91 b is fitted so as to be slidable with an elongated hole portion 111 a provided on the end portion side of the drive lever 111. The pressing pin portion 91c is fitted so as to be slidable with an elongated hole portion (not shown) provided on the end portion side of the drive lever 112. Further, on the side of the lower portion of the slide member 91 where the pressing pin portions 91b and 91c are formed, pressing protrusion portions 91d and 91e are provided.

  The drive lever 111 has a substantially Z shape, and the drive lever 112 has a substantially inverted Z shape. The drive levers 111 and 112 are opposed to and in contact with each other in a long hole portion 111b provided in the intermediate portion of the drive lever 111 and a long hole portion (not shown) provided in the intermediate portion of the drive lever 112. The operation pins 114 are overlapped and slidably fitted. The operating pin 114 is also fitted with the plunger 116 of the solenoid 115. Therefore, the drive levers 111 and 112 and the plunger 116 are connected by the operation pin 114. The hole 111c of the drive lever 111 and the hole (not shown) of the drive lever 112 are provided at the ends opposite to the long hole 111a of the drive lever 111 and the long hole of the drive lever 112, respectively. The fulcrum shafts 113 are overlapped so as to face each other and are engaged with each other. Thereby, the drive levers 111 and 112 can rotate about the fulcrum shaft 113 as a fulcrum.

  When the measurement device removal button 84 is pressed after the measurement is completed, the piston mechanism 103 operates, and urine in the space 2a is discharged from the measurement device 1 into a container such as a toilet bowl or a paper cup. Thereafter, the measurement device removal mechanism 117 is operated, current is supplied to the solenoid 115, and the plunger 116 is attracted by the solenoid 115, thereby rotating the drive levers 111 and 112 counterclockwise. By the rotation, the pressing pin portions 91b and 91c are pressed downward, and the slide member 91 is moved downward. Thereby, the pressing protrusions 91d and 91e are in direct contact with the upper end surface of the measuring device 1, and the measuring device 1 is moved downward. In this way, the measuring device 1 can be detached from the device mounting opening 94 and the measuring device 1 can be automatically removed from the measuring device 80.

FIG. 11 is a block diagram showing a configuration of the measuring apparatus according to the present embodiment. As shown in FIG. 11, a CPU 121 is provided inside the measuring device 80. The CPU 121 corresponds to the above calculation unit. The CPU 121 includes a first calculation unit for detecting or quantifying a test substance contained in the sample based on an electrical signal corresponding to the first outgoing light received by the first light receiver 105, And a second arithmetic unit for detecting or quantifying the concentration of the aggregation promoter dissolved in the sample based on the electric signal corresponding to the second outgoing light received by the second light receiver 107.
In addition, a memory 122 is provided inside the measuring device 80. The memory 122 corresponds to the storage unit described above. The memory 122 corrects the influence of the concentration of the aggregation promoter dissolved in the sample on the intensity of the first emitted light received by the first light receiver 105, and the emitted light intensity is the test substance. Data for converting into the concentration of human albumin is stored. More specifically, the memory 122 of the measurement device 80 stores the human albumin concentration and the first light received by the first light receiver 105 at various aggregation promoter concentrations as data for conversion into the human albumin concentration. The first calibration curve group representing the relationship between the intensity of the emitted light and the second calibration representing the relationship between the intensity of the second emitted light received by the second light receiver 107 and the concentration of the aggregation accelerator. Lines are stored.

In the measurement apparatus of the present embodiment, the first light source 104 and the second light source 106, and the first light receiver 105 and the second light receiver 107 are provided, but the measurement apparatus of the present invention is not limited to this. . One light source that emits at least light having an appropriate wavelength according to the measurement method, the measurement target (test substance), and the absorption characteristics of the dye used for measurement may be used. In that case, one light receiver that receives light emitted from one light source, an electric signal corresponding to the wavelength selected for detection of the test substance in the sample from the light receiver, and the dye What is necessary is just to provide CPU which extracts the electrical signal corresponding to the wavelength selected according to the absorption characteristic. In addition, a first light receiver that receives a wavelength selected for detecting a test substance in a sample with respect to one light source, and a second light that receives a wavelength selected according to the absorption characteristic of the dye. A light receiver may be provided.
In the measurement apparatus of the present embodiment, the piston mechanism 103 and the device removal mechanism 117 are provided, but the measurement apparatus of the present invention is not limited to this. Instead of providing such a sample discharge and device removal mechanism in the measurement apparatus 80, the user may manually remove the measurement device 1 from the measurement device mounting portion 81 and discharge the sample.

Next, a measurement method for measuring a test substance in a sample using the measurement device 1 and the measurement apparatus 80 will be described.
First, the suction port 2 c of the measuring device 1 is joined to the device mounting port 94 in the measuring device mounting portion 81 of the measuring device 80, and the measuring device 1 is attached to the measuring device 80. At this time, the measurement device insertion detection switch 95 provided in the measurement device 80 is activated, and the CPU 121 functioning as a control unit detects the insertion of the measurement device 1, and the power supply of the measurement device 80 is turned on.

Next, the measurement device 1 is immersed in at least a position where the sample supply port 2b is immersed in urine collected in a container such as a urine receiving container or a paper cup provided in the toilet bowl. Subsequently, when the sample suction start button 83 is pressed, the piston 99 in the piston mechanism 103 moves, and a predetermined amount (for example, 3 mL) of urine is sucked into the space 2a from the sample supply port 2b of the measurement device 1. (First step).
The urine supplied into the space 2a of the measurement device 1 is an anti-human albumin antibody, which is a dry reagent carried on the first reagent holder 7 provided in the space 2a, and the second reagent. The aggregation promoter, which is a dry reagent carried on the holding unit 8, is dissolved, and the immune reaction between human albumin, which is an antigen in urine, and anti-human albumin antibody proceeds. When the supply of the sample into the space 2a is completed, the CPU 121 causes the timer 123, which is a timer, to start timing.

Thereafter, when the CPU 121 determines that a predetermined time (for example, 2 minutes) has elapsed from the completion of the supply of the sample into the space 2a by the signal from the time measuring unit 123, the CPU 121 determines that the first light source 104 and the second light source Let 106 perform light irradiation.
The first outgoing light emitted from the first light source 104 enters the space 2a through the second surface 4 of the measuring device 1, that is, the light incident part (second step). Thereafter, the first emitted light that has been transmitted and scattered in the urine held in the space portion 2a and emitted from the third surface 5 of the measurement device 1, that is, the light emitting portion, is emitted for a predetermined time (for example, 3 minutes). ), The light is received by the first light receiver 105 provided in the measuring device 80, and the intensity of the light is measured (third step).
Similarly, the second emitted light emitted from the second light source 106 enters the space 2a through the light incident part (fourth step). Thereafter, the second emitted light that has been absorbed by the dye labeled with the aggregation promoter in urine and emitted from the light emitting portion is supplied to the second device provided in the measuring device 80 for a predetermined time (for example, 3 minutes). The light receiver 107 receives the light and measures the intensity of the light (fifth step).

The CPU 121 reads out a second calibration curve representing the relationship between the second emitted light intensity stored in the memory 122 and the dye-labeled aggregation accelerator concentration, and refers to the second calibration curve. The light intensity received by the second light receiver 107 is converted into the dissolution concentration (C _A ) of the dye-labeled aggregation accelerator (sixth step).
Here, an example of the second calibration curve is shown in FIG. FIG. 12 shows the relationship between the aggregation promoter concentration and the emitted light intensity when the human albumin concentration is 20 mg / dL. 12, the horizontal axis represents the polyethylene glycol concentration (wt%) in the sample, and the vertical axis represents the absorbance (arbitrary intensity, hereinafter abbreviated as au). For example, when the second emitted light intensity (absorbance) received by the second light receiver 107 is 1.5, the concentration of polyethylene glycol labeled with the second calibration curve is determined to be 3.0 wt%. .

Next, the CPU 121 stores the relationship between the concentration of human albumin and the intensity of the first emitted light received by the first light receiver 105 at various dye-labeled aggregation promoter concentrations stored in the memory 122. from the first calibration line group representing a represents the dissolution concentration C _a of the dye-labeled cohesion promoter, and the concentration of human albumin, the relationship between the emission intensity of light received by the first photodetector 105 A first calibration curve is selected and read (seventh step). Then, by referring to the first calibration curve in the dissolution concentration C _A of the aggregation accelerator, CPU 121 is the emission intensity of light received by the first photodetector 105, dissolution of the dye-labeled cohesion promoters The influence of the concentration is corrected, and the emitted light intensity is converted into the human albumin concentration (eighth step).
Here, FIG. 13 shows an example of the first calibration curve group. FIG. 13 shows the relationship between the concentration of human albumin and the intensity of the first outgoing light received by the first light receiver 105 at various dye-labeled aggregation promoter concentrations. 13, the horizontal axis represents the human albumin concentration (mg / dL) in the sample, and the vertical axis represents the first emitted light intensity (scattered light intensity) (au) received by the first light receiver 105. Show. In FIG. 13, the first calibration curves A to G constituting the first calibration curve group have dye-labeled polyethylene glycol concentrations (wt%) of 1.0, 1.5, 2.0, and 2, respectively. The calibration curves for .5, 3.0, 3.5, and 4.0 are shown.
For example, as described above, when the polyethylene glycol concentration converted in the sixth step is 3.0 wt%, the CPU 121 selects the first calibration curve E from the first calibration curves A to G, read out. Then, as shown in FIG. 14, by estimating the human albumin concentration on the horizontal axis from the point on the first calibration curve E indicating the intensity of the first emitted light received by the first light receiver 105. The human albumin concentration corresponding to the intensity of the emitted light can be obtained. The horizontal axis in FIG. 14 indicates the human albumin concentration (mg / dL) in the sample, and the vertical axis indicates the scattered light intensity (au) received by the first light receiver 105. For example, when the emitted light intensity (scattered light intensity) received by the first light receiver 105 is 66, the human albumin concentration corresponding to the emitted light intensity is determined to be 14 mg / dL.

  In the present embodiment, the fourth calibration curve representing the relationship between the first emitted light intensity and the human albumin concentration obtained when all of the carried aggregation promoter is dissolved is the first calibration curve. In addition to the group and the second calibration curve, etc., it may be stored in the memory 122. The CPU 121 reads out the fourth calibration curve and refers to the fourth calibration curve, so that it is assumed that the intensity of the emitted light received by the first light receiver 105 has dissolved all of the carried aggregation accelerator. Can be converted to the human albumin concentration. With this configuration, it is possible to know how much the human albumin concentration is different between the case where the dissolution state of the aggregation promoter in the sample is not considered and the case where it is considered.

The human albumin concentration thus obtained is displayed on the display unit 82 as a measurement result, and can be preferably stored in the memory 122 together with the time measured by the time measuring unit 123. Furthermore, the measurement result can be recorded on a removable storage medium such as an SD card by the recording unit 124 provided in the measurement device 80. As a result, the measurement result can be easily taken out from the measuring device 80, so that the storage medium can be brought to an analysis specialist or mailed, and the analysis can be easily requested.
Through the above process, after the measurement of the amount of the test substance is completed, when the user presses the measurement device removal button 84, the piston mechanism 103 is operated, and the urine in the space 2a is converted into the sample supply port 2b of the measurement device 1. From inside the toilet bowl or paper cup. Thereafter, the measurement device removal mechanism 117 is operated to automatically remove the measurement device 1 from the measurement apparatus 80.

Note that the measuring apparatus 80 according to the present embodiment includes a transmission unit 125 that can transmit to an analysis-related department or analysis-related trader in a hospital outside the measurement apparatus 80, an analysis-related department or analysis-related trader in the hospital, and the like. A receiving unit 126 is provided for receiving the result of analysis in FIG. As a result, the measurement result and the analysis result can be directly transmitted / received to / from the analysis-related department or the analysis-related business in the hospital, and the analysis result can be quickly fed back to the user, thereby shortening the time.
In the present embodiment, light irradiation is performed by the first light source 104 and the second light source 106 after a predetermined time from the completion of the supply of the sample (urine) into the space portion 2a. Light irradiation may be started simultaneously with detection of supply completion. In this case, the electrical signals from the first light receiver 105 and the second light receiver 107 that receive the light emitted from the first light source 104 and the second light source 106 are converted into a CPU 121 after a predetermined time from the completion of sample supply. You may set to input to.
As described above, according to the present embodiment, the measurement non-uniformity due to the difference in the dissolution state of the aggregation promoter in the sample for each measurement is efficiently corrected with a simple configuration, and the measurement target substance. Antigen measurement can be performed quickly and accurately.

(Embodiment 3)
In the present embodiment, an example of the measurement apparatus according to the second preferred embodiment of the present invention and the second measurement method of the present invention using the same will be described.
In the measuring apparatus according to the present embodiment, the memory 122 includes the same second calibration curve as in the second embodiment, the concentration of the aggregation promoter labeled with the dye at the various antigen concentrations (test substances), and the first. This is the same as the measurement apparatus of the second embodiment except that the third calibration curve group representing the relationship with the intensity of the first emitted light received by the light receiver 105 is stored.
Hereinafter, a measurement method using the measurement apparatus of the present embodiment will be described. The measurement method of the present embodiment corrects the influence of the concentration of the aggregation accelerator dissolved in the sample on the intensity of the emitted light received by the first light receiver 105, and the intensity of the emitted light is measured. Except for the step of converting to the concentration of human albumin, which is a substance, it is the same as the first step to the fifth step in the measurement method of Embodiment 2, and therefore the description thereof is omitted.

The memory 122 stores, for example, the same second calibration curve shown in FIG. 12 as in the second embodiment and the third calibration curve group shown in FIG. FIG. 15 shows a third calibration curve group representing the relationship between the concentration of the aggregation promoter labeled with a dye and the intensity of the first outgoing light received by the first light receiver 105 at various human albumin concentrations. An example is shown. The horizontal axis in FIG. 15 represents the polyethylene glycol concentration (wt%) in the sample, and the vertical axis represents the scattered light intensity (au) received by the first light receiver 105. In FIG. 15, the third calibration curves AK constituting the third calibration curve group have human albumin concentrations (mg / dL) of 0, 2, 4, 6, 8, 10, 12, 14 respectively. , 16, 18, and 20 are calibration curves.
First, the CPU 121 reads from the memory 122 a second calibration curve representing the relationship between the second emitted light intensity and the concentration of the aggregation promoter labeled with the dye, as shown in FIG. By referring to the calibration curve, the emitted light intensity (absorbance) received by the second light receiver 107 is converted into the concentration of polyethylene glycol labeled with a dye (sixth step).

Next, the CPU 121 reads out a third calibration curve group as shown in FIG. 15 from the memory 122 and refers to the third calibration curve group, so that the CPU 121 receives the output received by the first light receiver 105. The first emission light intensity is converted into the human albumin concentration by correcting the influence of the dissolution intensity of the aggregation promoter labeled with the dye on the emission intensity. That is, from the third calibration curve group, the intensity of the first outgoing light corresponding to the polyethylene glycol concentration obtained in the sixth step is measured in the first output measured in the third step. A third calibration curve that is equal to the intensity of the incident light is extracted (seventh step).
Then, the CPU 121 determines the human albumin concentration by determining how much the extracted third calibration curve is obtained for the human albumin concentration (eighth step).

For example, when the polyethylene glycol concentration converted in the sixth step is 3.0 wt% and the intensity of the first outgoing light (scattered light intensity) measured in the third step is 66, the CPU 121 Is the horizontal axis value indicating the polyethylene glycol concentration (3.0 wt%) determined by the second calibration curve from the third calibration curve groups A to K shown in FIG. A third calibration curve H including an intersection with the vertical axis value indicating the intensity (66) of the second is extracted. Since the third calibration curve H shows a calibration curve when the human albumin concentration is 14 mg / dL, the human albumin concentration is determined to be 14 mg / dL.
As described above, according to the present embodiment, the measurement non-uniformity due to the difference in the dissolution state of the aggregation promoter in the sample for each measurement is efficiently corrected with a simple configuration, and the measurement target substance. Antigen measurement can be performed quickly and accurately.

  The measuring device, the measuring apparatus, and the measuring method according to the present invention measure an accurate amount of a test substance by adding correction according to the influence of the dissolution state of the aggregation accelerator in the sample supplied to the measuring device. Therefore, it is useful in the medical and medical-related examination fields, particularly when measuring the amount of a test substance in a urine sample.

It is a perspective view which shows the measuring device in Embodiment 1 of this invention. It is the sectional view on the AA line of FIG. It is a disassembled perspective view which shows the structure of the measuring device in Embodiment 1 of this invention. It is a figure which shows the absorption spectrum of the pigment | dye which labels the aggregation promoter in Embodiment 1 of this invention. It is a perspective view which shows another example of the measuring device in Embodiment 1 of this invention. FIG. 6 is a sectional view taken along line B-B in FIG. 5. It is a perspective view which shows the measuring apparatus in Embodiment 2 of this invention. It is sectional drawing which shows an internal structure at the time of inserting the measuring device of FIG. 1 into the measuring apparatus in Embodiment 2 of this invention. It is a front view which shows the measurement device removal mechanism of the measuring apparatus in Embodiment 2 of this invention. It is a side view which shows the measurement device removal mechanism of the measuring apparatus in Embodiment 2 of this invention. It is a block diagram which shows the structure of the measuring apparatus in Embodiment 2 of this invention. It is a figure which shows the 2nd calibration curve stored in the memory of the measuring apparatus in Embodiment 2 of this invention. It is a figure which shows the 1st calibration curve group stored in the memory of the measuring apparatus in Embodiment 2 of this invention. It is a figure which shows the 1st calibration curve extracted in Embodiment 2 of this invention. It is a figure which shows the 3rd calibration curve group stored in the memory of the measuring apparatus in Embodiment 3 of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Measuring device 2 Base | substrate 2a Space part 2b Sample supply port 2c Suction port 2d Hollow square column part 2e Hollow square pyramid part 3 1st surface 4 2nd surface 5 3rd surface 6 4th surface 7 1st reagent Holding unit 8 Second reagent holding unit 31 First member 32 Second member 33 Bottom 80 Measuring device 81 Measuring device mounting unit 82 Display unit 83 Sample aspiration start button 84 Measuring device removal button 91 Slide member 91a Pressing protrusion 91b 91c Pressing pin portion 91d, 91e Pressing protrusion 92 Frame member 92a Device mounting protrusion 92b Cylinder mounting protrusion 93 Device mounting sealing member 94 Device mounting port 95 Measuring device insertion detection switch 96 Center line 97 Cylinder mounting sealing Member 98 Piston rod 98a Female thread 98b Keyway 99 Piston 100 Linda 100a Protruding portion 100b, 111c Hole portion 101 Motor 102 Male screw portion 103 Piston mechanism 104 First light source 105 First light receiver 106 Second light source 107 Second light receiver 111, 112 Drive lever 111a, 111b Long hole portion 111c hole 113 fulcrum shaft 114 operating pin 115 solenoid 116 plunger 117 measuring device removal mechanism 121 CPU
122 memory 123 timing unit 124 recording unit 125 transmission unit 126 reception unit

Claims (11)

A space part that is surrounded by an inner wall and holds a sample containing an antigen as a test substance;
A sample supply port for communicating with the space and supplying the sample into the space;
An optical measurement unit for optically measuring the sample;
A measurement device comprising a reagent holding unit that is provided in the space and promotes the production of an antibody against the antigen and an aggregation complex containing the antigen and the antibody, and holds an aggregation promoter labeled with a dye. .   The optical measurement unit includes a light incident unit for allowing light to be incident from outside the space unit into the space unit, and a light emitting unit for emitting light from the inside of the space unit to the outside of the space unit. The measuring device according to 1.   The measuring device according to claim 1, wherein the reagent holding unit includes a first reagent holding unit that holds the antibody and a second reagent holding unit that holds the aggregation promoter.   The measurement device according to claim 3, wherein the aggregation accelerator is a polymer compound containing polyethylene glycol as a main chain. A measuring device mounting portion for mounting the measuring device according to claim 1;
A light source for emitting light that enters the space from outside the space;
A light receiving portion for receiving light emitted from the inside of the space portion to the outside of the space portion;
Light reflected by the light receiving part reflecting the amount of the aggregated complex and light reflected by the light receiving part and reflecting the concentration of the aggregation accelerator labeled with the dye in the sample. And a computing unit for quantifying the concentration of the test substance contained in the sample based on the intensity of the sample. The light source includes a first light source and a second light source,
The light receiving unit receives a first light output from the first light source and receives a second light output from the second light source; and a second light receiving unit receives the second light emitted from the second light source. With
The intensity of the first emitted light received by the first light receiving part reflects the amount of the aggregated complex,
The measurement apparatus according to claim 5, wherein the intensity of the second emitted light received by the second light receiving unit reflects the concentration of the aggregation accelerator in the sample.   Furthermore, when the concentrations of the aggregation accelerators are different from each other, a plurality of first values representing the relationship between the concentration of the test substance and the intensity of the first emitted light received by the first light receiving unit. A storage unit is provided for storing a calibration curve and a second calibration curve representing a relationship between the concentration of the aggregation accelerator and the intensity of the second emitted light received by the second light receiving unit. 6. The measuring device according to 6.   Further, when the concentrations of the test substances are different from each other, a plurality of third relationships representing the relationship between the concentration of the aggregation accelerator and the intensity of the first emitted light received by the first light receiving unit. A storage unit is provided for storing a calibration curve and a second calibration curve representing a relationship between the concentration of the aggregation accelerator and the intensity of the second emitted light received by the second light receiving unit. 6. The measuring device according to 6.   Furthermore, the measuring apparatus in any one of Claims 5-8 provided with the suction part for attracting | sucking the said sample in the said space part of the said measurement device. A measurement method for measuring the test substance contained in the sample using the measurement device according to claim 1 and the measurement apparatus according to claim 7,
A first step of supplying the sample into the space of the measuring device;
A second step of irradiating the first outgoing light through the optical measurement unit to the space part to which the sample is supplied;
A third step of measuring the intensity of the first emitted light emitted from the space to which the sample is supplied through the optical measuring unit;
A fourth step of irradiating the second outgoing light through the optical measurement unit to the space to which the sample is supplied;
A fifth step of measuring the intensity of the second emitted light emitted from the space to which the sample is supplied through the optical measuring unit;
A sixth step of converting the intensity of the second emitted light measured in the fifth step into a concentration of the aggregation accelerator by referring to the second calibration curve;
A seventh step of extracting a first calibration curve corresponding to the concentration of the aggregation promoter converted in the sixth step from the plurality of first calibration curves;
By referring to the first calibration curve extracted in the seventh step, the eighth step of converting the intensity of the first emitted light measured in the third step into the concentration of the test substance And a measuring method. A measurement method for measuring the test substance contained in the sample using the measurement device according to claim 1 and the measurement apparatus according to claim 8,
A first step of supplying the sample into the space of the measuring device;
A second step of irradiating the first outgoing light through the optical measurement unit to the space part to which the sample is supplied;
A third step of measuring the intensity of the first emitted light emitted from the space to which the sample is supplied through the optical measuring unit;
A fourth step of irradiating the second outgoing light through the optical measurement unit to the space to which the sample is supplied;
A fifth step of measuring the intensity of the second emitted light emitted from the space to which the sample is supplied through the optical measuring unit;
A sixth step of converting the intensity of the second emitted light measured in the fifth step by referring to the second calibration curve into a concentration of the aggregation accelerator;
From the third calibration curve, the intensity of the first outgoing light corresponding to the concentration of the aggregation accelerator converted in the sixth step is measured in the third step. A seventh step of extracting a third calibration curve equal to the intensity;
And an eighth step of determining the concentration of the test substance corresponding to the third calibration curve extracted in the seventh step.

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