JP2015102421A5 - Protein detection apparatus and protein detection method - Google Patents

Description

The present invention relates to a protein detection apparatus and a protein detection method .

If the vibration apparatus is installed in the inspection apparatus, the inspection apparatus may be increased in size.
An object of the present invention is state, and are able to provide a high protein detection device of small and inspection efficiency and to provide a high protein detection methods of inspection efficiency can contribute to downsizing of the apparatus.

To achieve the above object, a first aspect of the invention, pressurization of vibrating the reaction vessel by transmitting ultrasonic waves to the reaction vessel an annular shape surrounding the permeable Kahikariro of translucent possible reaction vessel (61) and oscillating mechanism (5), wherein the providing the color change detecting part for detecting the degree of antigen-antibody reaction in the reaction vessel as the degree of color change by chromogenic substrate (50), a protein detection device (1) comprising a.
The apparatus main body (3) including a holding part (2) for detachably holding the cartridge (60) having the reaction container as in claim 2, and the reaction container of the cartridge held by the holding part. An optical mechanism (4) 0 including a light emitting unit (14, 18) that emits irradiation light along the transmitted light path, and a light receiving unit (15) that is provided at an end of the transmitted light path and receives the irradiation light; , May be provided.
A control unit (40) for controlling operations of the optical mechanism and the excitation mechanism as in claim 3, wherein the color change detection unit is provided in the control unit and is based on a detection result of the light receiving unit. Thus, the degree of the antigen-antibody reaction may be detected as the degree of color change by the chromogenic substrate.

In addition, although the alphanumeric character in a parenthesis represents the corresponding component etc. in embodiment mentioned later, this does not mean that this invention should be limited to those embodiment as a matter of course. The same applies hereinafter.
In addition, as in claim 4, the liquid supplied to the reaction vessel is a primary antibody-containing liquid in which a large number of magnetic beads (83) having a primary antibody (81) immobilized thereon are dispersed in the liquid ( 82), and the reaction vessel includes a first portion (76; 761) through which the transmission optical path passes, and a second portion (77; 762) surrounding the first portion, and the magnetic beads are magnetized. A magnetic attraction mechanism ( 6) for retracting the magnetic beads from the first part and holding the magnetic beads in the second part by suction may be provided.

According to a fifth aspect of the present invention, the magnetic attraction mechanism may include an electromagnetic coil (6) that is on / off controlled by the control unit.
Further, as in claim 6 , the magnetic attraction mechanism includes a permanent magnet ( 6A) capable of changing a distance to the second part, and the control unit to make the permanent magnet closer to the second part. And a drive member ( 94 ) that is driven and controlled by.

Further, as in claim 7 , the reaction vessel has a bottom wall (76; 76A) and a conical tapered peripheral side wall (77) surrounding the bottom wall, and the excitation mechanism includes An ultrasonic oscillator (23) as an annular excitation source that surrounds the transmitted optical path, and an ultrasonic horn (24) that transmits an ultrasonic wave from the ultrasonic oscillator that forms an annular shape that surrounds the transmitted optical path. In addition, the ultrasonic horn may include a conical tapered transmission surface (24b) along the peripheral side wall.

Moreover, you may provide the elastic member (25) supported by the said apparatus main body and supporting the said excitation mechanism like Claim 8 .
According to a ninth aspect of the present invention, the optical mechanism includes a first light emitting element (14) as the light emitting unit that emits first irradiation light (B) whose absorbance changes due to the color change, and an absorbance due to the color change. A second light-emitting element (18) as the light-emitting section that emits second irradiation light (R) that does not change, and a single detection light-receiving element (15) as the light-receiving section provided at the end of the transmitted light path, The control unit may cause the first light emitting element and the second light emitting element to emit light alternately.

According to a tenth aspect of the present invention, the color change detection unit may calculate the degree of color change based on a comparison between a value of (Bn / B0) and a value of (Rn / R0).
However, Bn is the detected light reception amount of the first irradiation light by the detection light receiving element after the color change, and B0 corresponds to the detection light reception amount of the first irradiation light by the detection light receiving element before the color change. A reference value, Rn is a detected light reception amount of the second irradiation light by the detection light receiving element after the color change, and R0 is a detection light reception amount of the first irradiation light by the detection light receiving element before the color change. Is a reference value corresponding to.

According to the present invention, since the reaction vessel to promote the promotion and washing of antigen antibody reaction can be vibrated at ultrasonic, inspection efficiency is significantly improved. In addition, since the excitation mechanism is arranged in an annular shape that surrounds the transmitted light path, it is possible to achieve downsizing.
According to the second aspect of the present invention, the reaction container can be vibrated to promote the antigen-antibody reaction and the washing while the cartridge is held in the holding portion of the apparatus main body. improves.
In the invention of claim 3, the color change detection unit provided in the control unit detects the color change based on the detection result of the light receiving unit.
According to the invention of claim 4 , in the step of detecting the color change, the magnetically attracted magnetic beads are retracted from the first part through which the transmission optical path passes and are retained in the second part, whereby the light emitting unit The color change can be detected with high accuracy by receiving the irradiation light from the light receiving portion without being obstructed by the magnetic beads. Further, when washing the reaction vessel with the washing liquid, the magnetic beads can be magnetically attracted and held in the second portion.

According to the invention of claim 5 , the magnetic beads can be easily retracted from the first portion or the retracting can be released by turning on and off the electromagnetic coil.
According to the invention of claim 6 , a strong magnetic field by a permanent magnet is applied to the second portion, so that the magnetic beads can be reliably retracted from the first portion and held in the second portion.
Further, according to the invention of claim 7 , the conical taper-shaped transmission surface of the ultrasonic horn is placed along the conical taper-shaped peripheral side wall of the reaction vessel, so that a small but large-area transmission surface can be obtained and strong. Excitation can be performed.

In addition, according to the invention of claim 8 , regardless of variation in dimensional accuracy, the transmission surface of the ultrasonic horn is surely brought into surface contact with the peripheral side wall of the reaction vessel, and sufficient vibration is performed to ensure reaction efficiency and The cleaning efficiency can be improved and the inspection efficiency can be improved.
According to the ninth aspect of the present invention, the first light emitting element and the second light emitting element are caused to emit light alternately, and the first irradiation light and the second irradiation light are received by the common light receiving element for detection. Can be simplified and the apparatus can be miniaturized.

Further, according to the invention of claim 10 , based on a comparison between a change in the amount of received light of the first irradiation light whose absorbance changes due to a color change and a change in the amount of received light of the second irradiation light whose absorbance does not change due to a color change. To detect a color change. Therefore, the influence of disturbance can be suppressed, color change can be detected with high accuracy, and accurate antigen detection can be performed.
According to the method of the invention of claim 11, in order to promote the antigen-antibody reaction and to promote washing, the reaction container is vibrated ultrasonically by using a vibration mechanism arranged in an annular shape surrounding the transmitted light path. Thus, the inspection efficiency can be improved in a small space.

Embodiments of the present invention will be described with reference to the accompanying drawings.
FIG. 1 is a cross-sectional view of a protein detection apparatus 1 according to an embodiment of the present invention, and FIG. 2 is a plan view of a cartridge 60. First, referring to FIG. 2, the cartridge 60 includes a pair of translucent reaction vessels 61 for causing an antigen-antibody reaction, and a primary antibody-containing liquid containing a primary antibody (capture antibody) in each reaction vessel 61. A pair of primary antibody-containing liquid supply paths 62 to be supplied, a pair of first cleaning liquid supply paths 63 to supply the first cleaning liquid to each reaction container 61, and a secondary antibody (enzyme-labeled antibody) to each reaction container 61. secondary antibody-containing liquid containing and a secondary antibody-containing liquid supply path 64 for supplying respectively.

The cartridge 60 also has a second cleaning liquid supply path 65 for supplying a second cleaning liquid containing the second cleaning liquid to each reaction container 61, and a coloring substrate that is an enzyme substrate of the enzyme-labeled antibody labeling enzyme for each reaction container 61. A pair of chromogenic substrate-containing liquid supply paths 66 that respectively supply the chromogenic substrate-containing liquid that is included, and a sample supply path 67 that supplies a sample to each reaction vessel 61 are provided.
In this embodiment, to each reaction vessel 61, while introducing the other sample liquid will comprise an antigen to be detected protein, with reference to the example of setting the cartridge 60 to a protein detection device 1 described To do. Therefore, in this embodiment, the sample supply path 67 is not used.

In addition, a sample solution containing an antigen that is a protein to be detected is placed in one reaction vessel 61 in advance, and a sample solution containing an antigen that is a protein to be detected is not put in the other reaction vessel 61 (that is, the antigen is The cartridge 60 may be set in the protein detection apparatus 1 in a state where a blank liquid not included is charged. Further, the cartridge 60 after setting the protein detection device 1, through the sample supply path 67, may be supplied to each separate sample.

The cartridge 60 has a common waste reservoir 6 9 for storing a waste liquid discharged through the respective discharge passage 68 from each reaction vessel 61. An exhaust port 69 a communicating with an exhaust valve (not shown) is provided at the bottom of the waste liquid storage unit 69.
As shown in FIGS. 2 and 3, liquid insertion holes 62a, 63a, 64a, 65a, 66a, and 67a for inserting corresponding liquids are provided at one ends of the supply paths 62, 63, 64, 65, 66, and 67, respectively. Is provided.

As shown in FIG. 3, the convex portion 70 is fitted in a concave portion 73 provided on the upper surface 12 a of the cartridge receiving plate 12. A space between the lower surface of the cartridge 60 and the upper surface 12a of the cartridge receiving plate 12 is sealed by a sealing member 74 such as an O-ring fitted around the base end of the convex portion 70. Thereby, without leakage to the corresponding liquids insertion hole 62a~67a from liquid paths 71, and to be able to insert the corresponding liquid.

As shown in FIGS. 7A and 7B, the primary antibody (capture antibody) 81 is immobilized on the surface of countless magnetic beads 83 dispersed in the primary antibody-containing liquid 82 supplied into the reaction vessel 61. It is phased. In the magnetic bead 83, the primary antibody 81 is solid-phased on the surface 83a of the magnetic bead 83 formed of a synthetic resin material containing magnetic powder. The diameter of a magnetic bead is 0.1-10 micrometers in diameter, for example, More preferably, it is 0.1-3 micrometers. Referring to FIG. 2, the primary antibody-containing liquid 82 is supplied from the first tank 301 to the reaction vessel 61 through the tube 7 2 , the primary antibody-containing liquid supply path 62 and the common supply path 75.

Referring to FIG. 1, a protein detection device 1 includes a device main body 3 having a holding portion 2 that is, for example, a recess, on which a cartridge 60 is detachably mounted, an optical mechanism 4 supported by the device main body 3, and a device main body. 3, an excitation mechanism 5 supported by the apparatus body 3, an electromagnetic coil 6 as a magnetic attraction mechanism supported by the apparatus main body 3, and an ECU as a control unit that controls operations of the optical mechanism 4, the excitation mechanism 5, and the electromagnetic coil 6. 40 (Electronic Control Unit).

The apparatus main body 3 includes a base 7, front and rear side frames 8 fixed to the base 7, a first support plate 9 supported by the side frame 8, and a first support plate 9 disposed below the first support plate 9. A second support plate 10 facing the support plate 9, a cartridge receiving plate 12 fixed on the first support plate 9 and having a holding portion 2 formed of a recess for receiving and holding the cartridge 60 on the upper surface 12 a, and the holding portion 2 The cartridge holding plate 13 is detachably fixed to the upper surface 12a of the cartridge receiving plate 12 so as to hold the cartridge 60 held by the cartridge.

The blue LED 14 as the first light emitting element is fixed to the lower surface 17 a of the optical path forming block 17. Further, the first side surface 17c of the optical path forming block 17 is a second light emitting element that emits red light R (second irradiation light) whose absorbance does not change due to a color change of a chromogenic substrate (reaction product) by an enzyme reaction described later. Red LED 18 is fixed.
The optical path forming block 17 is disposed on the transmission optical path TK, transmits blue light B (first irradiation light) from the blue LED 14 (first light emitting element), and transmits light from the red LED 18 (second light emitting element). die click Roikkumira 19 for guiding the red light R (the second irradiation light) on the downstream side of the transmission optical path TK incident from a direction perpendicular to the optical path TK is fixed.

A holding recess 22 b for holding the ultrasonic oscillator 23 is formed on the upper surface 22 a of the holder 22. A compression coil spring 25 as an elastic member for elastically supporting the holder 22 is interposed between the lower surface 22c of the holder 22 and the upper surface 10b of the second support plate 10 opposed thereto. The compression coil spring 25 elastically supports the excitation mechanism 5 via the holder 22.
The fixed end 25 a of the compression coil spring 25 is fixed to a locking portion made of, for example, a convex portion of the upper surface 10 b of the second support plate 10. The movable end 25 b of the compression coil spring 25 is locked to a locking portion 27 made of, for example, a recess on the lower surface 22 c of the holder 22.

It is preferable that a plurality (three in this embodiment) of the compression coil springs 25 are provided at equal intervals on the circumference centered on the transmission optical path TK. Due to expansion / contraction deformation and collapse deformation of the compression coil spring 25, variations in dimensional errors among the components are absorbed, and the transmission surface 24b of the vibration exciting mechanism 5 reliably comes into surface contact with the peripheral side wall 77 of the reaction container 61 of the cartridge 60. .
The center hole of the holder 22 is fitted to the upper end of the optical path forming cylinder 31 that forms the transmitted optical path TK so as to be slidable in the longitudinal direction of the optical path forming cylinder 31.

The electromagnetic coil 6 as a magnetic attraction mechanism is arranged in an annular shape so as to surround the ultrasonic horn 24 of the vibration mechanism 5 and is controlled to be turned on / off by the ECU 40 . Electromagnetic coil 6 is turned on, by dispersed magnetic beads magnetic attraction in the antibody-containing liquid contained in the reaction vessel 6 in 1, the peripheral side wall and retracts the magnetic beads from the bottom wall 76 (first part) 77 (second part). As a result, the magnetic beads are retracted from the transmitted light path TK, and the detection accuracy by the optical mechanism 4 is improved. Further, the magnetic beads are held on the peripheral side wall 77 (second portion) at the stage of cleaning the reaction vessel 61 with the cleaning liquid supplied from the cleaning liquid supply path 64.

The ECU 40 also includes an ultrasonic oscillator 23 as a vibration source, an electromagnetic coil 6 as a magnetic attraction mechanism, and a display unit 44 such as a liquid crystal display unit that displays a message display to the operator and a result of antigen detection. If, blue light Studies mechanism 4 LED 14 (the first light emitting element) and red LED18 (second light-emitting element), and the pump motor 201 to 207 are connected.
ECU40 includes a driving unit 45 for driving the ultrasonic oscillator 23, a drive unit 46 for driving the electromagnetic coil 6, a driving unit 47 for driving the display unit 44, a drive unit 48 for driving the blue LED1 4, red The drive part 49 which drives LED18, and the drive parts 401-407 which each drive each pump motor 201-207 are provided.

ECU40 outputs the signal which drives the pump motor 201-207 corresponding to each process to the corresponding drive parts 401-407. In addition, the ECU 40 includes a color change detection unit 50 that receives a signal from the detection light receiving element 15 and the correction light receiving element 20 via the corresponding AD converters 41 and 42 and detects a color change.
The ECU 40 includes a process control unit 51 that inputs a signal from the operation unit 43 and controls the inspection process. Process control unit 51, via the drive unit 45 to 47 corresponding in accordance with a signal from the operation unit 43 ultrasonic oscillator 23 controls the electromagnetic coil 6 and the display unit 4 4. In addition, the ECU 40 operates the color change detection unit 50 in accordance with a signal from the process control unit 51. The color change detection unit 50 outputs drive signals to the corresponding drive units 48 and 49 so as to drive the blue LED 14 and the red LED 18 in the color change detection stage.

In step S3, the optical mechanism 4 is driven. Specifically, the blue LED 14 (first light emitting element) and the red LED 18 (second light emitting element) are driven alternately. In this case, at the time of driving the blue LED 14, the received light amount of the blue light B from the blue LED1 4 by detecting light-receiving element 15 is implemented in the feedback control such that the constant. Further, when the red LED 18 is driven, feedback control is performed so that the amount of red light R received from the red LED 18 by the correction light receiving element 20 is constant.

When the blue LED 14 is driven, the detected received light amount of the blue light (first irradiation light) B from the blue LED 14 by the detection light receiving element 15 is stored as the reference value B0. When the red LED 18 is driven, the detected light reception amount of the red light (second irradiation light) R from the red LED 18 by the correction light receiving element 20 is stored as the reference value R0.
In step S4, the first pump 101 is driven, and the primary antibody-containing liquid 82 stored in the first tank 301 is supplied to the reaction vessel 61 as shown in FIG. 7A (primary antibody supply step). Equivalent). The primary antibody 81 is preliminarily immobilized on the surface of a large number of magnetic beads 83 contained in the primary antibody-containing solution 82.

Then, in step S6, while holding the magnetic beads 83 by turning the electromagnetic coil 6 is magnetically attracted to the peripheral side wall 77 as shown in FIG. 8, in step S7, the second pump 1 02 (second pump motor 2 02) and drives the ultrasonic oscillator 23 for vibrating mechanism 5, with feed Kyusuru the reaction vessel 61 of the first cleaning liquid, to vibrate the reaction vessel 61. That is, the ultrasonic oscillator of the vibration mechanism 5 is turned on in a state where the electromagnetic coil 6 as the magnetic attraction mechanism is turned on and the magnetic beads are held on the peripheral side wall 77 (second portion) of the reaction vessel 61 by the magnetic attraction force. 23 is propagated to the reaction vessel 61 through the ultrasonic horn 24, and the reaction vessel 61 is vibrated to facilitate the cleaning of the reaction vessel 61 (corresponding to the first cleaning step).

Next, in step S10, with the electromagnetic coil 6 turned on, the fourth pump 1 0 4 ( fourth pump motor 2 0 4 ) and the ultrasonic oscillator 23 of the vibration mechanism 5 are driven to react the second cleaning liquid. While feeding to the container 61, the reaction container 61 is vibrated. That is, with the electromagnetic coil 6 as the magnetic attraction mechanism turned on and the magnetic beads are held on the peripheral side wall 77 (second portion) of the reaction vessel 61 as shown in FIG. 5 is propagated to the reaction vessel 61 through the ultrasonic horn 24 to vibrate the reaction vessel 61 and promote cleaning of the reaction vessel 61 (in the second washing step). Equivalent).

Next, in step S12, for example, the ultrasonic oscillator 23 of the vibration mechanism 5 is driven for a predetermined time while the electromagnetic coil 6 is turned off (or can be turned on), and the reaction vessel 61 is vibrated. Using the chromogenic substrate, to perform the enzymatic reaction in the reaction vessel 6 within 1, to facilitate the enzymatic reaction by excitation. The reaction product from the enzyme reaction is colored yellow, for example.
Next, in step S13, the optical mechanism 4 is driven to detect the amount of received light B and L after color development. Specifically, on the electromagnetic coil 6 as a magnetic attraction mechanism, by the magnetic attraction force, as shown in FIG. 8, and the magnetic beads 83 is retracted from the reaction vessel 61 of the bottom wall 76 (first part) In other words, the optical mechanism 4 is driven in a state where it is held on the peripheral side wall 77 (second portion) (withdrawn from the transmitted light path TK).

The blue LED 14 (first light emitting element) and the red LED 1 8 (second light emitting element) are driven alternately and stored each time the amount of received light B, L is measured. Then, the detected light reception amount Bn of the blue light (first irradiation light) B from the blue LED 14 by the detection light receiving element 15 in the measurement n (n is predetermined) is stored. Further, the detected light reception amount Rn of the red light (second irradiation light) R from the red LED 18 by the correction light receiving element 20 at the nth measurement is stored.

When the blue LED 14 is driven, feedback control is performed so that the amount of blue light B received from the blue LED 14 by the detection light receiving element 15 is constant. Further, when the red LED 18 is driven, feedback control is performed so that the amount of red light R received from the red LED 18 by the correction light receiving element 20 is constant.
Next, in step S14, the degree of color change An (corresponding to change in absorbance) at the n-th measurement is calculated based on the following equation (1).

An = (Bn / B0) / (Rn / R0) (1)
Next, in step S15, the detected color change degree An is compared with a threshold value F1. In step S15, if the degree An of the detected color change is determined to be equal to or greater than the threshold value F1 (An ≧ F1) (YES at step S15), the process proceeds to step S16, the display unit 4 4 For example, after displaying that the antigen has been detected, such as “An antigen has been detected”, the processing is terminated.

In step S15, if the degree An of the detected color change is determined to be less than the threshold F1 (An <F1) (NO in step S15), the process proceeds to step S17, the display unit 4 4, For example, after displaying that the antigen is not detected, such as “No antigen was detected”, the processing is terminated.
According to the present embodiment, the reaction container 61 can be vibrated to promote the antigen-antibody reaction and the cleaning while the cartridge 60 is held in the holding unit 2 of the apparatus main body 3, so that the inspection efficiency is improved. Greatly improved. In addition, since the excitation mechanism 5 is arranged in an annular shape surrounding the transmission optical path TK, it is possible to achieve downsizing.

In addition, using the formula (1), the change in the amount of received light of the first irradiation light (blue light B) whose absorbance changes due to color change and the reception of the second irradiation light (red light R) whose absorbance does not change due to color change. A color change is detected based on a comparison with the change in quantity. Therefore, the influence of disturbance can be suppressed, color change can be detected with high accuracy, and accurate antigen detection can be performed.
9 and 10 show another embodiment of the present invention. This embodiment shown in FIGS. 9 and 10 is mainly different from the first embodiment of FIG. 4 in that, in the embodiment of FIG. 4, an electromagnetic coil 6 that is on / off controlled is used as the magnetic attraction mechanism. , in the second embodiment, as shown in FIGS. 9 and 10, that it uses the magnetic attraction Organization comprising a solenoid 94 as a drive member for displacing the permanent magnet 6 a and the permanent magnet 6 a cyclic It is in.

Specifically, the center portion 761 of the bottom wall 76A constitutes a first portion through which the transmitted light path TK passes, and the annular portion 762 surrounding the center portion 761 in the bottom wall 76A constitutes a second portion. The permanent magnet 6A can be inserted through the ultrasonic horn 24, the ultrasonic oscillator 23, and the holder 22 and can change the distance to the annular portion 762 (second portion) of the bottom wall 76A.
Further, the permanent magnet 6A is moved to a position close to the second portion (annular portion 762) as shown in FIG. 12 and a position separated from the second portion (annular portion 762) as shown in FIG. The ECU 40 controls the drive of the solenoid 94 so that the control is performed.

The solenoid 94 includes a control rod 95 that can be expanded and contracted, and the control rod 95 connects the permanent magnet 6 </ b> A through the first connection arm 96 and the second connection arm 97 so as to be integrally movable. The first connecting arm 96 is inserted through a longitudinal groove 98 through which the optical path forming cylinder 31 is inserted, and the second connecting arm 97 is disposed in the optical path forming cylinder 31 and the holder 22.
In the components of the embodiment of FIGS. 9 and 10, the same reference numerals as those of the embodiment of FIG. 4 are attached to the same components as those of the embodiment of FIG.

According to this embodiment, by applying a strong magnetic field by the permanent magnet 6 A relative to the second portion (annular portion 672 of the bottom wall 76A), the first portion (bottom wall 76A reliably magnetic beads 83 It can be withdrawn from the central portion 761) and held in the second portion (the annular portion 672 of the bottom wall 76A). Although not shown as a modification of the embodiment in FIGS. 9 and 10, an electric motor that is driven and controlled by the ECU may be used as the drive member instead of the solenoid. In this case, the output (linear motion) of a motion conversion mechanism (for example, a ball screw mechanism) that converts rotation of the electric motor into linear motion is transmitted to the permanent magnet. For example, by rotating and driving a screw shaft whose axial movement is restricted by an electric motor, the ball nut whose rotation is restricted and whose axial movement is allowed is moved in the axial direction, and is moved integrally with the ball nut in the axial direction. Displace possible permanent magnets.

Claims (11)

A vibration mechanism for vibrating the reaction vessel by transmitting ultrasonic waves to the reaction vessel an annular shape surrounding the permeable Kahikariro of translucent possible reaction vessel,
Protein detection device and a color change detecting part which detects as the degree of color change by chromogenic substrate the extent of antigen-antibody reaction in the reaction vessel. The apparatus body according to claim 1, further comprising a holding unit that removably holds the cartridge having the reaction container;
An optical system comprising: a light emitting unit that emits irradiation light along the transmission light path of the reaction container of the cartridge held by the holding unit; and a light receiving unit that is provided at the end of the transmission light path and receives the irradiation light. A protein detection device comprising: a mechanism; In Claim 2, It has a control part which controls operation of the optical mechanism and the vibration mechanism, The protein change apparatus, wherein the color change detection unit is provided in the control unit and detects the degree of the antigen-antibody reaction as the degree of color change by a chromogenic substrate based on a detection result of the light receiving unit. In any one of Claims 1-3, the liquid supplied to the said reaction container contains the primary antibody containing liquid which disperse | distributed in the liquid many magnetic beads by which the primary antibody was solid-phased on the surface,
The reaction vessel includes a first portion through which the transmitted light path passes, and a second portion surrounding the first portion,
A protein detection apparatus comprising a magnetic attraction mechanism that retracts the magnetic beads from the first portion and holds the magnetic beads in the second portion by magnetically attracting the magnetic beads. 5. The protein detection apparatus according to claim 4 , wherein the magnetic attraction mechanism includes an electromagnetic coil that is on / off controlled by the control unit. 5. The magnetic attraction mechanism according to claim 4 , wherein the magnetic attraction mechanism includes a permanent magnet capable of changing a distance to the second portion, and a drive member that is driven and controlled by the control unit so as to move the permanent magnet closer to the second portion. A protein detection apparatus comprising: In any one of Claim 1 to 6 , the said reaction container has a bottom wall and the conical taper-shaped surrounding side wall surrounding the said bottom wall,
The excitation mechanism includes an ultrasonic oscillator as an annular excitation source that surrounds the transmitted optical path, an ultrasonic horn that transmits an ultrasonic wave from the ultrasonic oscillator that has an annular shape that surrounds the transmitted optical path, and Including
The ultrasonic horn is a protein detection device including a conical tapered transmission surface along the peripheral side wall. The protein detection apparatus according to claim 7 , further comprising an elastic member that is supported by the apparatus main body and supports the excitation mechanism. In any one of claims 1-8, wherein the optical mechanism includes a first light emitting element as the light emitting unit for emitting a first illumination light absorbance changes by the color change, it does not change the absorbance by the color change A second light-emitting element as the light-emitting unit that emits second irradiation light, and a single light-receiving element for detection as the light-receiving unit provided at the end of the transmitted light path,
Wherein, the protein detection devices emit light alternately to the first light emitting element and the second light emitting element. 10. The protein detection apparatus according to claim 9 , wherein the color change detection unit calculates a degree of color change based on a comparison between a value of (Bn / B0) and a value of (Rn / R0).
However, Bn is the detected light reception amount of the first irradiation light by the detection light receiving element after the color change, and B0 corresponds to the detection light reception amount of the first irradiation light by the detection light receiving element before the color change. A reference value, Rn is a detected light reception amount of the second irradiation light by the detection light receiving element after the color change, and R0 is a detection light reception amount of the first irradiation light by the detection light receiving element before the color change. Is a reference value corresponding to. A vibration step of vibrating the reaction container by transmitting ultrasonic waves to the reaction container by an annular vibration mechanism surrounding a transmission light path of the translucent reaction container;
And a color change detecting step of detecting the degree of the antigen-antibody reaction in the reaction vessel as the degree of color change by the chromogenic substrate after the vibration of the vibration step.