JP3240183B2 - Method and apparatus for measuring antigen or antibody in sample - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for measuring an antigen or an antibody in a sample by immunological agglutination, and a measuring apparatus used in the method.

[0002]

2. Description of the Related Art Immunological measurement methods have high specificity and sensitivity and have become extremely useful clinically in the medical field in recent years. In particular, a method utilizing immunological agglutination is used for various diagnostic uses such as detection of fecal occult blood and rheumatoid factor, pregnancy diagnosis, etc., because the test can be easily performed.

Conventionally, to carry out these inspections,
A suspension (reagent) of latex particles sensitized with an antibody or antigen corresponding to the antigen or antibody that is the test substance, and a test solution are placed in a reservoir (a cavity provided in the test plate) of the test plate. Alternatively, mix the solution in a test plate surrounded by a hydrophobic ring, etc.), shake the reaction solution by hand or with a rocking device for several minutes, and observe the presence or absence of aggregation with the naked eye. ing. In general, those in which aggregation is observed are regarded as positive, and those in which it is not observed are regarded as negative.

[0004] In these methods, the final judgment is made by observing the presence or absence of aggregation with the naked eye. In particular, in the vicinity of a concentration where a fine positivity / negative determination is required (so-called determination suspension area), it is necessary to determine whether or not a slight aggregation has occurred. The judgment may be erroneously made depending on the reaction time, the degree of swing, and the like.

[0005] As a method of solving the problem of the variance and the erroneous determination by the naked eye determination, various methods of performing measurement using an apparatus including optical means and the like have been proposed. For example,
Japanese Patent Publication No. 58-11575 proposes a method in which an agglutination reaction is performed in a cell for measuring absorbance, and the increase in absorbance of the reaction solution accompanying the agglutination reaction is measured.

However, in this method, since the absorbance measurement is performed simultaneously with the progress of the reaction, it is necessary to homogenize the reaction solution by performing the reaction under stirring or shaking, and is susceptible to the influence of insoluble substances derived from the specimen. In the measurement of a sample containing a high concentration of an antigen or an antibody, the absorbance may not correctly correspond to the amount of the antibody or the antigen, and the measurable concentration range is limited. For this reason, when measuring a high-concentration sample, it is necessary to pre-dilute the sample, which has the drawback that the operation is complicated and errors due to dilution are likely to occur.

[0007]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel measuring method capable of quantitatively measuring an antigen or an antibody in a sample without visual judgment. It is a further object of the present invention to provide a measuring device suitably used in the above measuring method.

[0008]

Means for Solving the Problems The present inventors have proposed that the agglutination reaction be carried out in the liquid reservoir of the test plate and that the reaction liquid in the liquid reservoir be swirled horizontally, and that the liquid reservoir be substantially omitted. Focusing on the fact that the light transmittance of the central reaction solution temporarily decreases and then increases over time as shown in FIG. 6, it is possible to measure the amount of antigen or antibody in the sample by using this phenomenon. It has been found that the present invention is possible, and the present invention has been completed.

The method for measuring an antigen or antibody in a sample according to the present invention comprises the steps of: agglutinating a sample containing an antigen or antibody with carrier particles sensitized with an antibody or antigen corresponding to the antigen or antibody; A method for measuring an antigen or an antibody, wherein the agglutination reaction is performed in a liquid reservoir of a test plate, and the reaction liquid in the liquid reservoir is swirled horizontally while light transmission of the reaction liquid substantially at the center of the liquid reservoir is performed. rate was continuously measured after the predetermined time T ₁ elapses, by determining the characteristic value of the time change of the light transmittance, and measuring the amount of antigen or antibody.

Further, the apparatus for measuring an antigen or an antibody in a sample of the present invention, which is preferably used in the above method, includes a light source, a sample mounting portion, and a light receiving portion. A measuring unit that irradiates light from the light source to a substantially central portion of the sample accommodated in the liquid reservoir, detects the transmitted light by the light receiving unit, and measures the light transmittance of the sample, and a driving unit provided with a driving source And a driving force transmitting unit that connects the measuring unit and the driving unit, transmits the driving force of the driving source, and rotates the measuring unit in a horizontal direction, and preferably the measuring unit is A lid provided with a light source, an inspection plate mounting pedestal arranged so that light emitted from the light source passes through substantially the center of the liquid reservoir, and a substantially center of the liquid reservoir provided at a lower portion of the pedestal. A slit for passing light transmitted through the portion downward,
A measuring unit main body including a light receiving unit that is disposed immediately below the slit and senses light passing through the slit, and more desirably, the light receiving unit is connected to an arithmetic unit and measured by the measuring unit. It is configured to calculate the amount of antigen or antibody in the sample from the transmittance.

Hereinafter, the present invention will be described in more detail. FIG. 6 shows the change in light transmittance due to the aggregation reaction. Suspension (reagent) of carrier particles obtained by sensitizing a sample containing an antigen or an antibody with an antibody or an antigen corresponding to the antigen or the antibody
Was added drop wise, at first reagent is light transmittance is small because of the vicinity of the liquid reservoir center, by pivoting horizontally, the light transmittance is increased until the time T ₁ to dispersed throughout the liquid reservoir. T ₁ after the particles with the progress of the agglutination reaction are aggregated,
Since the apparent particle diameter increases and the scattered light increases, the light transmittance decreases. On the other hand, the agglomerated particles gradually move to the periphery of the liquid reservoir due to centrifugal force by turning horizontally, so that the central portion becomes transparent and the light transmittance increases. Since the latter phenomenon occurs later than the former phenomenon, the light transmittance decreases and then increases.

As shown in FIG. 7, when the amount of the antigen or antibody in the reaction solution is large, the curve of the light transmittance approximated to a quadratic curve after the lapse of T ₁ becomes sharp, and conversely, the amount of the antigen or antibody increases. When the number is small, the curve of the light transmittance becomes gentle. Therefore the measurement method of the present invention, (1) the reaction solution the light transmittance continuously measured after the predetermined time T ₁ elapses, quadratic the light transmittance _{y = a 0 + a 1 x} + a
₂ x ² [where x represents time and y represents light transmittance], and the amount of antigen or antibody is measured by the value of the quadratic coefficient a ₂ .

When the amount of the antigen or antibody in the reaction solution is large as shown in FIG. 7, the time until the light transmittance becomes minimum in the light transmittance curve after the lapse of T ₁ is short, and the antigen or antibody is short. When the amount of the antibody is small, the time to the minimum becomes longer. Therefore, another measurement method of the present invention comprises: (2) measuring the light transmittance of the reaction solution continuously after a lapse of a predetermined time T ₁ , and measuring the amount of the antigen or antibody by the time when the light transmittance is minimized. Is what you do. The time at which the light transmittance is minimized may be determined based on the actually measured value, or the light transmittance continuously measured in the same manner as in the above (1) is calculated by a quadratic expression y =
a ₀ + a ₁ x + a ₂ x ² [where x represents time and y represents light transmittance], and x obtained by differentiating this quadratic expression
= −a ₁ / 2a ₂ may be the time when y becomes the minimum.

Further, the measuring method of the present invention provides the above-described light transmission
Focusing on the characteristics of the transmittance curve, the light transmittance is highest in FIG.
Predetermined time T from time when it becomes small_TwoOf light transmittance increase
(L _Two-L₁) And FIG. 6, the light transmittance is minimized.
Time T after the time_ThreeOf light transmittance at
(L_Three)) To determine the amount of antigen or antibody. Sand
According to another measurement method of the present invention, (3) the light transmittance of the reaction₁Continuous after elapse
For a predetermined time from the time when the light transmittance is minimized.
T_TwoBy measuring the increase in light transmittance of the antigen or
It measures the amount of antibody. Further, another measuring method of the present invention comprises: (4) measuring the light transmittance of the reaction₁After the elapse
Measured continuously, and after the time when the light transmittance becomes the minimum,
Time T_ThreeOf the antigen or antibody depending on the value of the light transmittance at
It measures the amount. In these measurement methods (3) and (4), light is transmitted.
The time at which the rate is at a minimum may be determined based on actual measurements.
X = −a obtained by differentiating the above quadratic equation₁/
2a_TwoMay be a time when y is minimized.

Here, the light transmittance is a ratio of intensity of transmitted light to incident light, and the light transmittance T (%) is expressed by the following equation. T = (I / I ₀ ) × 100 (where I represents the intensity of transmitted light and I ₀ represents the intensity of incident light.) In the present specification, “continuous measurement” means continuous measurement. This means that the measurement is performed not only while monitoring the temperature, but also at regular time intervals.

In each of these methods, a calibration curve is prepared by measuring the light transmittance using a standard solution dilution series with a known concentration, and the light transmittance of an unknown sample is measured under the same conditions as in the measurement of the standard dilution series. Is measured, and the amount (or concentration) of the antigen or antibody in the unknown sample can be quantified by comparing with the calibration curve.

Note that all the measuring methods of the present invention are based on the light transmittance, but since the transmittance (T) and the absorbance (A) have a relationship of A = -log (1 / T). ,
By measuring the absorbance, the amount of the antigen or antibody in the sample can be similarly measured using the methods (1) to (4) of the present invention. Therefore, the case where the absorbance is used instead of the light transmittance is also included in the claims of the present invention.

According to the measuring method of the present invention, an antigen or an antibody in a sample can be quantitatively measured without relying on the naked eye. In addition, since the change in the light transmittance at the approximate center of the liquid reservoir is measured, even when an insoluble material is mixed into the sample, the insoluble material moves to the peripheral portion of the liquid reservoir by centrifugal force while rotating, and the light is emitted. The measurement of the transmittance has the advantage that it is not disturbed.

The antibody or antigen to which the method of the present invention can be applied is not particularly limited, and the present invention relates to the measurement of serum proteins such as fecal occult blood reaction and tumor markers, the measurement of hormones such as pregnancy diagnosis, and the detection of viruses and autoantibodies. Used for measurement and the like.

The carrier particles used in the present invention are not particularly limited, and any carrier particles that are biologically inert, insoluble in water, and function as a carrier for an antigen or an antibody can be used. However, the average particle size is 0.05 to 10 μm, preferably 0.1 to 1.0 for reasons such as low light scattering of the carrier particles themselves in the light transmittance measurement.
Latex particles of 0 μm are preferably used. There is no particular limitation on the type of latex, but styrene, olefin, vinyl, acrylate, methacrylate, diene, and other latexes, and a modifying monomer such as acrylic acid or acrylonitrile are reacted with these. Latex and the like are exemplified, and among them, polystyrene latex and the like are preferably used.

As the aqueous solvent in which the carrier particles are dispersed, a pH buffer (eg, a phosphate buffer, a borate buffer, etc.) or the like is preferably used, and sodium chloride, a preservative, bovine serum albumin are appropriately used. Such a protein may be added.

The inspection plate used in the present invention is made of a material that does not affect the measurement of light transmittance. For example, a transparent test plate provided with a liquid reservoir having a hydrophobic ring on the peripheral portion, a glass or plastic plate provided with a shallow U-shaped liquid reservoir, and the like, among which a shallow U-shape A transparent plastic inspection plate provided with a mold liquid reservoir is preferably used because it has little effect on the measurement of light transmittance.

The shape of the liquid reservoir of the inspection plate is as follows.
Preferably, the horizontal cross-sectional shape is a circle or an ellipse. It is preferable that the side wall of the liquid reservoir has a smooth curved surface. This is to make the swirling smooth when the reaction liquid in the liquid reservoir is swirled in the method of the present invention. In addition, since the reacting liquid being swirled swirls along the side wall of the reservoir, the shape of the transparent or translucent portion generated near the center of the reservoir due to the swirl is determined by the horizontal cross-sectional shape of the reservoir. Shows a shape similar to. For the measurement of the light transmittance at substantially the center of the liquid reservoir, it is desirable that the shape of the transparent or translucent portion is a perfect circle or a shape close to a perfect circle. Therefore, in the present invention, an inspection plate in which the horizontal cross-sectional shape of the liquid reservoir is a perfect circle or a circle close thereto is most preferably used.

In the present invention, the measurement of the light transmittance is carried out using an aqueous solvent used for the agglutination reaction and a light ray having a wavelength that is little absorbed by the carrier particles.
nm, preferably using light having a wavelength in the vicinity of 600 to 1300 nm. In the region where the light transmittance is measured, it is desirable to measure about 1 to 25%, preferably about 1 to 10%, of the reaction liquid near the center of the liquid reservoir with the liquid surface area being 100%. When a plate having a circular cross section with a diameter of 20 mm is used, it is preferable to measure the light transmittance of a concentric region having a diameter of about 2 to 7 mm.

The number of revolutions and radius of rotation of the reaction solution are as follows:
Depending on the shape of the liquid reservoir, the amount of the reaction solution, the specific gravity of the latex particles, and the like, the optimum rotation speed and rotation radius change, so that the rotation speed is appropriately set.
pm, preferably 80 to 140 rpm, and the turning radius is 5 to 5.
It may be selected from a range of 50 mm, preferably 10 to 30 mm.

As a light source for irradiating the above-mentioned light beam, a tungsten lamp, a halogen lamp, a silicon carbide rod, a nichrome wire, etc. combined with a filter for setting the above-mentioned wavelength, or a light emission having a specific wavelength characteristic is used. A diode (LED) or the like is used, and an LED is particularly preferable because it can emit sharp linear light without scattered light.

In the present invention, a sample containing an antigen or an antibody is mixed with carrier particles sensitized with the antibody or the antigen corresponding to the antigen or the antibody in the liquid reservoir of the test plate to cause an agglutination reaction. At the same time, since the reaction solution in the liquid reservoir is swirled on a horizontal plane, the inspection plate needs to be swirled in a horizontal direction at a constant cycle. For this purpose, an oscillating device for giving a rotational movement to the reaction liquid in the liquid reservoir is required. However, if a tilting motion or a translational motion is added to the rotational motion, the formation of a transparent or translucent portion of the reaction liquid generated substantially at the center of the liquid reservoir is not preferable. As such a swinging device, for example, a shaft is fixed perpendicularly to the axial direction of the rotating power take-off shaft, and at an arbitrary position on the shaft different from the power take-out axis, parallel to the axial direction of the power take-out shaft. A device can be used in which the shaft is fixed and the shaft is supported on the bearing of the rotary table for mounting the inspection plate, and the table is held horizontally and turned in a horizontal manner.

FIG. 1 is a plan view showing the movement of an inspection plate which is mounted on a rotary table (not shown) of the swinging device and is horizontally turned. In the drawing, reference numeral 1 denotes an inspection plate having ten liquid reservoirs 2. A liquid sample for measurement (not shown) is contained in the liquid reservoir 2. For example, by setting the position indicated by I in the drawing as a start position and rotating the inspection plate horizontally in the order of I → II → III → IV → I, the sample liquid is horizontally set in the liquid reservoir 2. Can be turned.

FIG. 2 shows a part of the cross section taken along the line AA ′ of I of the inspection plate 1 being rotated as described above. Sample liquid 3 swirled horizontally in liquid reservoir 2
At the position I is as shown in FIG. By continuing the swirling of the sample liquid in this way, the agglomerated particles generated by the agglutination reaction move to the periphery of the liquid reservoir 2 under centrifugal force. It is thought that a transparent or translucent portion is generated depending on the degree.

An apparatus that can be suitably used in the above-mentioned method includes an oscillating section that can rotate the inspection plate in a horizontal direction at a constant cycle and that applies a rotating motion to the sample liquid in the liquid reservoir, And a measuring unit for irradiating the sample liquid in the reservoir with light and measuring its light transmittance. The present invention provides an apparatus for measuring an antigen or an antibody in a sample having the above two functions. Hereinafter, this measuring device will be described in detail with reference to the drawings showing one embodiment.

In FIG. 3, (a) is a perspective view of a measuring device showing one embodiment of the present invention, and (b) is a partially cutaway front view thereof. In FIG. 3A, reference numeral S denotes a light transmittance measuring device, which includes a lid 11 having a light source 10, a test plate mounting portion 12, and a light receiving portion 13, and measures a light transmittance of a sample liquid. A measuring unit 20 composed of a main body 14, a driving unit 21 having a driving source 15 disposed thereon, and connecting the measuring unit 20 and the driving unit 21 to each other to transmit a driving force of the driving source 15 to the measuring unit Driving force transmitting means 2 for rotating 20 in the horizontal direction
And 2.

As shown in FIG. 4, the measuring section 20 includes a lid 11 for blocking light from the outside,
A test plate mounting pedestal 16 arranged so that irradiation light from the light source 10 passes through substantially the center of the liquid reservoir 2 of the test plate 1, and a lower portion of the pedestal 16 And a slit 16a for guiding light transmitted through substantially the center of the liquid reservoir 2 downward.
a measuring section main body 14 which is disposed immediately below the light receiving section 13a and senses transmitted light passing through the slit 16a.

As the light source 1, the light source exemplified above is used, and particularly, an LED is suitably used. As shown by phantom lines in FIG.
If the shielding plate 17 formed with a is installed and the light emitted from the light source 1 is guided, the liquid reservoir 2 of the inspection plate 1 can be provided.
It is preferable because light can be accurately irradiated to substantially the center of the light emitting device.

The drive source 15 includes a power take-out shaft 18.
Any means can be used as long as it can apply a rotational motion to the and a force for rotating the measuring unit 20, and an electric motor or the like is generally used.

As the driving force transmitting means 22, a shaft 19 is fixed to a power take-out shaft 18 directly connected to the drive source 15 perpendicularly to the axial direction thereof. One end of a connecting shaft 19a is fixed to an eccentric arbitrary position in parallel with the axial direction of the power take-out shaft 18, and the other end is a bearing 19 provided at the bottom of the measuring unit main body.
b to support the measurement unit 20.

Next, the operation of this apparatus will be described.
, The power take-out shaft 18 is rotated by the operation of the drive source 15 such as a motor.
Rotates. This shaft 19 has an eccentric connecting shaft 1
Since the shaft 9a is fixed, the connecting shaft 19a makes a circular motion about the axis of the power take-out shaft 18. This connecting shaft 19
Since a is pivotally supported by the bottom of the measuring section main body 14, the measuring section main body 14 is rotated in the horizontal direction by this circular motion.
Therefore, the sample liquid contained in the liquid reservoir 2 of the inspection plate 1 attached to the pedestal 16 of the measuring unit main body 14 shown in FIG. Aggregates in the center move to the periphery.

On the other hand, light is emitted downward from the light source 10 at the same time as the turning operation of the measuring section 20 or at a certain time interval. The irradiated light passes through the substantially central portion of the sample liquid 3 attached to the base 16 and rotated along the side wall of the liquid reservoir 2, and the transmitted light is sensed by the light receiving unit 13 disposed thereunder. Then, the light transmittance of the sample liquid 3 is measured.

In this device, the light transmittance is calculated as described above from the irradiation amount radiated from the light source 10 and the transmitted light amount sensed by the light receiving unit 13. The antigen or antibody can be measured by determining the characteristic value of the change.

According to the above-described measuring device, the inspection plate can be rotated horizontally at a constant cycle, and the oscillating device for giving a rotational movement to the reaction liquid in the liquid reservoir and the light in the reaction liquid in the liquid reservoir can be used. And the light transmittance of the reaction solution can be accurately measured.

As shown in the system diagram of FIG. 5, when the light receiving unit 13 is connected to the arithmetic unit 32, the amount of antigen or antibody in the sample is automatically determined based on the value of the light transmittance. This is particularly preferable because the calculation can be performed more accurately, so that the antibody or antigen can be measured more accurately. At this time, it is preferable to use the amplifier 30 and the A / D converter 31 together in the connection circuit.

[0041]

EXAMPLES Examples of the present invention will be shown below, and will be described more specifically. Example 1 Measurement of Human Hemoglobin (Material) 1 mg / ml of 1-ethyl-3- (3-diaminopropyl) carbodiimide 1 was added to 10 ml of 5% carboxylated polystyrene latex (average particle size: 0.30 μm).
After adding 0 ml and reacting for 20 minutes, 0.01 mol /
It was centrifugally washed twice with 1 liter of borate buffer (pH 8.0). To this latex, 7 ml of an anti-human hemoglobin antibody (rabbit IgG.5 mg / ml) was added, reacted for 5 hours, and further dissolved in 0.01 mol of 0.1% bovine serum albumin.
The mixture was centrifugally washed three times with 1 / liter borate buffer (pH 8.0) to obtain an anti-human hemoglobin antibody-sensitized latex reagent having a latex concentration of 1%. The test plate has a diameter of 2
A transparent polystyrene plate provided with a U-shaped liquid reservoir having a 0 mm round opening was used.

(Apparatus) As the measuring apparatus, a light transmittance measuring apparatus shown in FIG. 3 was used, and L having a wavelength of 940 nm was used.
An ED was used as a light source, and a silicon photodiode was used as a light receiving unit. The optical axis of the light source was set perpendicular to the inspection plate, and (4.8) concentric with the center of the liquid reservoir.
(mmφ) region was set to irradiate light.

(Measurement) Human hemoglobin was added to 0.1% bovine serum albumin, 0.9% sodium chloride, 0.1%
It was dissolved in a 0.1 mol / liter borate buffer (pH 8.0) containing sodium azide to obtain a standard solution. The standard solution was appropriately diluted with the buffer solution to prepare a standard solution dilution series. Standard solution diluent 1 injected into the test plate liquid reservoir
After adding 25 μl of the latex reagent prepared above to 00 μl, this was attached to the pedestal of the measuring device, the measuring device was started, and the measuring section was rotated horizontally at a rotation radius of 25 mm and a rotation speed of 110 rpm, and light was emitted from the LED. The light transmitted through the sample that was irradiated and injected into the liquid reservoir was allowed to be sensed by a silicon photodiode, and the light transmittance was measured at intervals of 5 to 6 seconds over 3 minutes. FIG. 7 shows the change over time in the light transmittance at each antigen concentration.

(1) The light transmittance after the lapse of 33 seconds is minimized.
The quadratic equation y = a₀+ A₁x + a _Twox^Two[Where x
Represents time and y represents light transmittance], and a quadratic coefficient a_Two
Find the value of Table 1 shows the results.

[0045]

[Table 1]

FIG. 8 shows a calibration curve obtained by plotting the above data on a graph with the antigen concentration of the standard solution on the horizontal axis and the value of the quadratic coefficient a _{2 on} the vertical axis. The value of the antigen concentration and the secondary coefficient a ₂ is shows a good correspondence.

(2) The light transmittance after 33 seconds has passed is minimized.
The quadratic equation y = a₀+ A₁x + a _Twox^Two[Where x
Represents time and y represents light transmittance].
To obtain a time x at which the light transmittance y becomes minimum (that is, a time x).
X = -a₁/ 2a_TwoSeek). Table 2 shows the results.

[0048]

[Table 2]

FIG. 9 shows a calibration curve obtained by plotting the above data on a graph with the antigen concentration of the standard solution as the horizontal axis and the light transmittance minimum time x as the vertical axis. The antigen concentration and the time x show a good correspondence.

(3) The time when the light transmittance becomes minimum for each density is determined based on the actually measured value, and the increase in the light transmittance for 22 seconds is determined from the time. Table 3 shows the results.

[0051]

[Table 3]

FIG. 10 shows a calibration curve obtained by plotting the above data on a graph with the antigen concentration of the standard solution on the horizontal axis and the increase (%) in light transmittance on the vertical axis. The antigen concentration and the increase in light transmittance show a good correspondence.

(4) The time at which the light transmittance becomes minimum for each density is determined based on the actually measured value, and after that time 1
Table 4 shows the values of the light transmittance at 25 seconds.

[0054]

[Table 4]

FIG. 11 shows a calibration curve obtained by plotting the above data on a graph with the antigen concentration of the standard solution on the horizontal axis and the light transmittance (%) at 125 seconds on the vertical axis. The antigen concentration and the value of the light transmittance show a good correspondence.

[0056]

According to the present invention, it is possible to quantitatively measure an antigen or an antibody in a sample without relying on the naked eye judgment, and the variation in the judgment result by the examiner as in the conventional method using the naked eye judgment. And the problem of erroneous determination in the determination suspension area is eliminated, and accurate measurement with high reproducibility becomes possible. Unlike the conventional method, the method of the present invention measures a change in light transmittance at a substantially central portion of the liquid reservoir while rotating the reaction solution. Since the insoluble matter derived from the specimen moves to the periphery of the liquid reservoir due to the centrifugal force during the rotation, there is an advantage that the measurement is not hindered by the insoluble matter. It is suitably used for measurement of a sample in which insoluble matter derived from a specimen such as a fecal occult blood reaction may be mixed.

Another advantage is that the test plates and reagents used in the conventional visual inspection method can be used as they are. In the conventional method of measuring the increase in absorbance while stirring, a sample containing a high concentration of antigen or antibody may be measured at a lower concentration than the actual concentration. According to the measurement method of the present invention, the accuracy is slightly lowered even with a high-concentration sample, but it is easy to determine whether or not an antigen or antibody at a certain concentration or more is contained (determination of negative or positive). The measurement can be performed without pre-dilution of the sample, and the operation is simple.

According to the measuring apparatus of the present invention, the sample can be irradiated with light while rotating the inspection plate in the horizontal direction at a constant period, so that the light transmittance of the sample solution can be continuously and precisely measured. it can. In addition, when the light transmittance measurement device is connected to a calculation device, calculation processing of the measurement result is automatically performed, so that quick measurement is possible. This eliminates the need for conventional human operation or visual determination, shortening the time, and enabling precise and accurate determination.

[Brief description of the drawings]

FIG. 1 is a plan view showing the movement of a horizontally rotating inspection plate.

FIG. 2 is a diagram showing A-
It shows a part of an A ′ cross section.

FIG. 3A is a perspective view of a measuring apparatus showing one embodiment of the present invention, and FIG. 3B is a partially cutaway front view thereof.

FIG. 4 is a partial cross-sectional view showing a measurement unit of the device.

FIG. 5 is a system diagram of a measuring apparatus showing another embodiment of the present invention.

FIG. 6 shows the state when the reaction solution in the liquid reservoir is swirled horizontally.
It is a graph which shows the time change of the light transmittance of the liquid storage part substantially center.

FIG. 7 is a graph showing a change in light transmittance with time for each antigen concentration in Example 1.

FIG. 8 is a graph showing a calibration curve obtained in Example 1 (1).

FIG. 9 is a graph showing a calibration curve obtained in Example 1 (2).

FIG. 10 is a graph showing a calibration curve obtained in Example 1 (3).

FIG. 11 is a graph showing a calibration curve obtained in Example 1 (4).

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 10 Light source 12 Inspection plate mounting part 13 Light receiving part 15 Drive source 20 Measuring part 21 Drive part 22 Driving force transmission device S Light transmittance measuring device

Continuation of the front page (58) Field surveyed (Int.Cl. ⁷ , DB name) G01N 33/543 G01N 21/59

Claims (5)

(57) [Claims] 1. A method for measuring an antigen or antibody in a sample by subjecting a sample containing an antigen or antibody to an agglutination reaction with an antibody or a carrier particle sensitized with the antigen corresponding to the antigen or antibody, together causing agglutination in liquid pooling portion of the test plate, continuously the light transmittance of the reaction solution reservoir portion substantially center while turning horizontally reaction liquid in the liquid reservoir after a predetermined time T ₁ elapses The light transmittance is measured by a quadratic expression y
= A ₀ + a ₁ x + a ₂ x ² [where x represents time and y represents light transmittance], and the amount of antigen or antibody is measured by the value of quadratic coefficient a _2. A method for measuring an antigen or an antibody in a sample. 2. A method for measuring an antigen or antibody in a sample by subjecting a sample containing an antigen or antibody to an agglutination reaction with an antibody or a carrier particle sensitized with the antigen corresponding to the antigen or antibody, together causing agglutination in liquid pooling portion of the test plate, continuously the light transmittance of the reaction solution reservoir portion substantially center while turning horizontally reaction liquid in the liquid reservoir after a predetermined time T ₁ elapses A method for measuring an antigen or an antibody in a sample, comprising measuring the amount of the antigen or the antibody based on the time at which the light transmittance is minimized. 3. A method for measuring an antigen or antibody in a sample by subjecting a sample containing an antigen or antibody to an agglutination reaction with an antibody or a carrier particle sensitized with the antigen corresponding to the antigen or antibody, together causing agglutination in liquid pooling portion of the test plate, continuously the light transmittance of the reaction solution reservoir portion substantially center while turning horizontally reaction liquid in the liquid reservoir after a predetermined time T ₁ elapses measured, the measurement method of an antigen or antibody in a sample, which comprises measuring the amount of antigen or antibody by light transmittance measuring the increase from smallest time of the light transmittance of the predetermined time T ₂ . 4. A method for measuring an antigen or an antibody in a sample by subjecting a sample containing an antigen or an antibody to an agglutination reaction with an antibody or a carrier particle sensitized with the antigen corresponding to the antigen or the antibody, together causing agglutination in liquid pooling portion of the test plate, continuously the light transmittance of the reaction solution reservoir portion substantially center while turning horizontally reaction liquid in the liquid reservoir after a predetermined time T ₁ elapses measured, the antigen or the measuring method of an antibody in a sample, wherein the light transmittance is to measure the amount of antigen or antibody by the value of light transmittance in smallest time after a predetermined time T _3. 5. The sample according to claim 1, wherein
Measuring apparatus for performing the method for measuring antigen or antibody
A light source, a sample mounting portion, and a light receiving portion, wherein the light source irradiates light to the substantially central portion of the sample housed in the liquid reservoir of the inspection plate mounted on the sample mounting portion, and transmits the light. A measuring unit for measuring the light transmittance of the sample by detecting the light receiving unit, a driving unit provided with a driving source, and connecting the measuring unit and the driving unit to transmit a driving force from the driving source. An apparatus for measuring an antigen or an antibody in a sample, comprising: a driving force transmitting means for rotating the measuring section in a horizontal direction.