JPH10239312A - Method for measuring sensitivity and simple kit for measuring sensitivity - Google Patents

Abstract

PROBLEM TO BE SOLVED: To measure objectively and quantitatively a state of sensitivity by measuring a concentration of immunoglobulin in saliva a predetermined time later after a start of testing of subject receiving an external stimulus. SOLUTION: A concentration of immunoglobulin in saliva is measured a predetermined time later after a start of testing of a subject receiving an optional external stimulus influencing sensitivity. A state of sensitivity which is correlative to immunoglobulin can thus be quantitatively measured. A liquid-storing cylinder 13 is set standing at one end of a slim rectangular sheet-like supporting body 12 of a measuring element 11 of the simple sensitivity-measuring kit, and an antibody-fixing part 14 is arranged at an inner bottom part of the cylinder where an anti immunoglobulin antibody is fixed. A sample saliva and distilled water are added to a liquid reservoir on the fixing part 14, and the liquid is disposed of after left stationary. Then, an anti immunoglobulin secondary antibody labelled with an enzyme is added before a coloring enzyme substrate solution is added, so that a degree of comfort is detected from an intensity of coloration.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sensitivity measurement method and a simple sensitivity measurement kit, and more particularly, to a sensitivity measurement method for measuring any known or unknown human sensitivity having a correlation with the amount of immunoglobulin secreted. Measurement method, and
The present invention relates to a simple sensibility measurement kit suitable for easily performing these measurement methods. The present invention can be preferably used for various medical diagnoses, or for designing emotional engineering of a living space such as a house or various vehicles, and for providing basic data for other purposes.

[0002]

2. Description of the Related Art Sensitivity of human beings, particularly comfort and relaxation based on audiovisual stimuli, has a strong subjective character, and a psychological evaluation in the form of a questionnaire has been widely used for evaluation. However, in order to apply the results of kansei evaluation to various purposes with sufficient reliability, the changes in kansei are detected as objective changes in the living body to external stimuli, and the significance of the changes is physiologically explained. Something that can be done.

For this reason, it is conceivable to detect physiological changes in addition to psychological evaluation. That is, one is an electrophysiological measurement method such as an electroencephalogram, and the other is a biochemical measurement method for measuring a biological component. As an application example of the former, an attempt has been made to measure the ratio of α-waves appearing in brain waves and so-called 1 / f fluctuation. However, as the application example of the latter, a report on a feeling of comfort and a feeling of relaxation based on audiovisual stimulation has not yet been made.

[0004] The preference of a scent, which is a kind of sensation different from audiovisual, ie, the sensitivity evaluation based on the olfactory stimulus, is described in Japanese Patent Application Laid-Open No. 7-20115, "Method of Measuring Preference of Scent". Some have tried biochemical measurement methods. According to this measurement method, the concentration of immunoglobulins, particularly secretory immunoglobulins, in the saliva of a person who smells the odor is measured, and the preference of the scent can be measured from the increase rate of the secretory amount.

[0005]

However, the psychological evaluation based on the above-mentioned questionnaire has a strong subjectivity, and it is necessary to prepare a subject skilled in psychological measurement in order to perform a quantitative and excellent reproducibility measurement. Yes, it is not easy.

[0006] Next, the data obtained by the above-mentioned electroencephalogram measurement is questionable in its quantitativeness, and equipment such as a shield room is required for accurate measurement.

Further, the technique disclosed in the above-mentioned Japanese Patent Application Laid-Open No. 7-20115 has the following insufficient points. That is, although "the amount of secretory immunoglobulin secretion in saliva is compared before and after smelling," the secretion curve over time has not been elucidated,
Moreover, it is not known how many minutes after the smell was taken, the saliva was collected.

[0008] Therefore, the measurement results in the prior art can be obtained only when the undisclosed assumption is made that saliva was collected at the same time after the test for all the subjects, and furthermore, the relative results between the subjects were different. Only in an evaluative sense is the objectivity maintained. This is because, at the time of saliva collection, the secretory amount of secretory immunoglobulin of the subject may have already entered the falling period of the secretory amount curve, or may have not yet reached the peak of secretion.

Further, in the above prior art, the relationship between the olfactory stimulus and the secretory immunoglobulin secretion amount is only clarified, so that it can be used only in the technical field related to the olfactory stimulus, for example, in the development of fragrance and the like. .

Furthermore, the above-mentioned prior art only discloses a pure laboratory method for measuring the degree of preference, but does not disclose any practical device for simply and quickly performing the method.

Therefore, the present invention firstly provides a biochemical measurement method for objectivity and quantification of general sensibility, and a biochemical measurement method for comfort and relaxation based on audiovisual stimulation. Second, it is an object to solve the problem to provide a practical device for performing measurement easily and quickly.

[0012]

The present inventor has experimentally found that the amount of secreted immunoglobulin has a relationship with the feeling of comfort and relaxation based on the audiovisual stimulus. The present invention was completed by recognizing that the amount corresponds to various sensitivities, and elucidating that the amount of immunoglobulin secretion follows a pattern of a secretion amount curve that peaks after a predetermined time has elapsed since the start of the test. .

[0013]

[Means for Solving the Problems]

(Structure of the First Invention) A first invention of the present application for solving the above problems.
The configuration of the invention (the invention according to claim 1) is to measure the concentration of immunoglobulin in saliva of a subject who has received any external stimulus that affects sensitivity at a point in time after a predetermined period of time from the start of the subject. Is a sensitivity measuring method for quantitatively measuring the state of the sensitivity having a correlation with the secretion amount of the immunoglobulin.

(Structure of the Second Invention) The structure of the second invention (the invention according to claim 2) for solving the above-mentioned problem is as follows.
In a subject who has received any audiovisual stimulus that affects the feeling of comfort or relaxation, the comfort or relaxation is measured by measuring the concentration of immunoglobulin in saliva at a point in time after the start of the test. This is a measurement method of the sensitivity to measure.

(Structure of Third Invention) The structure of the third invention (the invention described in claim 3) for solving the above-mentioned problem is as follows.
A measurement piece in which an antibody fixing portion with a liquid storage structure is provided on a support and an anti-globulin antibody is fixed to the antibody fixing portion, and a substance for label expression prepared so as to be administrable to the antibody fixing portion, This is a simple kansei measurement kit that is configured and used for kansei measurement.

[0016]

【The invention's effect】

(Effect of the First Invention) In the first invention, the concentration of immunoglobulin in saliva at a point in time after a predetermined time has elapsed after the start of the test is measured in consideration of the curve of the amount of immunoglobulin secreted by an external stimulus. The state of sensitivity can be objectively and quantitatively measured.

(Effect of the Second Invention) In the second invention, a feeling of comfort or a sense of relaxation is measured when the external stimulus in the first invention is an audiovisual stimulus. Measurement can be performed quantitatively or qualitatively.

(Effect of Third Invention) According to the third invention, a practical device for easily and quickly performing the measurement according to the first invention or the second invention is provided.

[0019]

Next, embodiments of the first to third inventions will be described.

(Sensitivity, feeling of comfort, feeling of relaxation) Sensitivity means various senses of human beings in general. In particular, in the present invention, it is preferable that it is found that there is a correlation with the amount of immunoglobulin secreted at the present time, or that it is a valid sense that can be reasonably estimated.

The sensitivity in this sense can be objectively and quantitatively measured for the first time by the first invention, and the result can be used as reliable data in all application fields.

The sense of comfort or relaxation based on the audiovisual stimulus is at least a kind of human sensibility, and is at least a sensibility that is clearly distinguished from the sense of taste of the scent based on the olfactory stimulus. It is difficult to precisely define the content of comfort or relaxation, but it is difficult, for example, in a state without mental stress, a stable or positive mental state, or a mental state in which the environment is comfortable. It refers to a state that can be illustratively described.

The feeling of comfort or relaxation in such a sense can be measured for the first time by the second invention. Moreover, depending on the accuracy of the required data, the timing of saliva collection after the start of the test is strictly defined or loosely performed, depending on whether it is easy to obtain qualitative measurement data, objective and quantitative Since data can be obtained freely, the results should be widely used as a very effective basic data for various medical diagnoses, emotional engineering design of living spaces such as houses and various vehicles, and other purposes. Becomes possible.

(External Stimulus) The external stimulus referred to in the first invention is:
As long as it affects the broad sense described above, touch, touch, and a tangible stimulus such as taste, smell, music, environmental sound, landscape, regardless of being an intangible stimulus such as a display, The type is not limited.

The audio-visual stimulus according to the second invention is at least one that appeals to the sight, such as a beautiful natural scenery, a masterpiece appreciation, an excellent image or display, or a preferable environmental sight.
Further, it includes those that appeal to hearing, such as α music, famous tunes, and favorable environmental sounds that have a high effect of increasing the so-called α wave.

(Sampling Time of Saliva from Subject) In the first invention, the timing of collecting saliva from a subject subjected to an external stimulus is not arbitrary, and each subject in the experimental example and the comparative example in the same measurement test has the same timing. Saliva is collected at The timing is determined based on a secretion curve of immunoglobulin to the external stimulus, which is known by a preliminary test or the like of the measurement test.

In general, the peak time of the immunoglobulin secretion amount curve in the case of the sensitivity measurement may be adopted, but as long as the immunoglobulin secretion amount has reached an amount that can be effectively evaluated, the peak time point may be used. Other points in time may be adopted.

In the second invention, when it is necessary to obtain objective and quantitative data on the feeling of comfort or relaxation, the timing of saliva collection is the same as in the first invention, and the secretion curve of immunoglobulin is obtained. It is necessary to specify precisely based on. However, when the purpose is to easily obtain qualitative measurement data, loose measurement that does not specifically define the timing of saliva collection, such as the technique disclosed in Japanese Patent Application Laid-Open No. 7-20115, is used. But it is enough.

(Immunoglobulin) The immunoglobulin is intended to be present or secreted in saliva, and immunoglobulin A is particularly preferable. However, it is not denied that immunoglobulins other than immunoglobulin A and their analogous substances or decomposed substances can be targeted. However, in the case of a measurement method using an anti-immunoglobulin antibody, the range is limited to a substance that specifically binds to the antibody.

(Measurement of Immunoglobulin Concentration) As a method for measuring the concentration of immunoglobulin or immunoglobulin A in saliva, various known methods may be arbitrarily adopted.

One example is the antibody method. This method can measure an immunoglobulin concentration by using an anti-immunoglobulin antibody or an immunoglobulin A concentration by using an anti-immunoglobulin A antibody, respectively. , An enzyme immunoassay using a radioactive substance, a fluorescent antibody method using a fluorescent dye, and the like.

In addition, a direct method using an antibody of only one step (for example, a predetermined amount of a labeled competitive immunoglobulin is separately prepared and added to a saliva sample to express a label. And an indirect method using an immunoglobulin concentration in a saliva sample indirectly) and an immunoglobulin captured by an antibody using a secondary antibody further labeled.

(Measurement of state of sensitivity) The state of sensitivity based on the measurement results of the immunoglobulin concentration can be measured by comparing the experimental example of the present invention with an appropriate blank test, or without performing the blank test. The determination may be made based on the immunoglobulin secretion amount curve data that has already been obtained. Also,
The psychological evaluation for the subject may be performed in parallel. For further accuracy, these methods may be used in combination.

(Simplified Kansei Measurement Kit) The support of the measuring piece in the simple Kansei measurement kit is not particularly limited in terms of its shape, dimensions, material and the like. Is preferred. The liquid storage structure of the antibody fixing part provided on the support,
Any structure may be used as long as saliva or a diluent administered to the antibody immobilizing part is not washed away. As an example, a cylindrical structure provided so as to surround the antibody immobilization part can be considered.

The antibody immobilization part may be an anti-immunoglobulin antibody in the direct method or an anti-immunoglobulin primary antibody in the indirect method, as long as it is immobilized by appropriate means. The structure and surface shape are not particularly limited. . For example, a flat film body provided on the support and surrounded by the liquid storage structure can be used.

A substance for expressing a label, which is prepared so as to be administrable to the antibody immobilization part together with the measurement piece, constitutes a kit.
Here, "prepared to be administrable" means that it is in a state where it can be distributed on the market as an integrated set with the measuring piece, taking into account the storage stability such as enclosing the container.

The “substance for expressing a label” includes an enzyme substrate, a fluorescent luminescent agent, etc., according to the classification of the antibody method described above.
The direct method includes, for example, a labeled competitive immunoglobulin, and the indirect method includes a labeled anti-immunoglobulin secondary antibody.

(High-Speed Measurement Method) FIG. 1 shows a high-speed measurement method using flow injection analysis, which enables accurate quantification of immunoglobulin in about 10 to 20 minutes per sample.

In this method, first, saliva as a measurement sample is injected into the sample loop 4, the valve 5 is switched, and the measurement sample is supplied to the reaction section 9 by the pump 8 using the carrier buffer 2.
Send to Since the primary anti-immunoglobulin antibody is fixed in the reaction section 9, immunoglobulins in the sample are captured.

Next, an enzyme-labeled anti-immunoglobulin antibody (secondary antibody) is injected into the sample loop 4 and sent to the reaction section 9 in the same manner. After the unreacted sample and enzyme-labeled anti-immunoglobulin antibody are washed away in the reaction section 9, the valve 6 is switched to send the enzyme substrate solution 1.

In this manner, measurement is performed by detecting the substrate reaction product of the labeling enzyme captured in the reaction section 9 by the detector 10. If the regeneration liquid 3 is sent by switching the valve 7 to remove the immunoglobulin or the enzyme-labeled anti-immunoglobulin antibody bound to the reaction section 9, the reaction section 9 can be regenerated for the next use.

[0042]

【Example】

(Method Used in Examples) In Examples 1 to 3 below, the concentration of immunoglobulin A in saliva was measured, and the measurement was performed using a biosensor (Pharmacia BIAcore-registered trademark) using surface plasmon resonance. )It was used.
In this method, an antibody against immunoglobulin A is immobilized on the sensor surface, and a sample is added thereto to detect an increase in weight due to immunoglobulin A binding on the sensor surface.

The amount of the saliva sample used, the pretreatment before use, and the method of converting the measurement result into the immunoglobulin A concentration (μg / ml) are as follows.

With respect to collection and storage of saliva, saliva was collected using a cotton ball salivetti for saliva sampling (Salstedt, Germany). Immediately after collecting the saliva, store it on ice, and then centrifuge at 2,800 rpm for 10 minutes to collect saliva and keep it at -20 ° C until use for measurement.
And stored frozen.

Regarding the method for measuring the immunoglobulin A concentration, prior to the measurement, an anti-human immunoglobulin antibody was bound to a sensor chip CM-5 (Pharmacia) according to the instruction manual of the biosensor of Pharmacia.
And the above-mentioned cryopreserved saliva is naturally thawed at room temperature,
The supernatant obtained by centrifugation at 15,000 rpm for 10 minutes was used for HB for the same device.
The solution was diluted 100-fold with S buffer (Pharmacia), and 100 μl of the solution was used for measurement.

The elevation value (ΔRU) from the baseline of the sensorgram obtained by the antigen-antibody reaction for 5 minutes was read. The concentration of immunoglobulin A in saliva is determined using a human immunoglobulin A solution of known concentration (0, 0.5, 1, 2, 3 μm).
g / ml), and was calculated based on a calibration curve prepared based on the ΔRU observed at the time of measurement using (g / ml).

In each example, a questionnaire based on psychological evaluation was also given to the subject in parallel. Pleasant and relaxed scores in psychological evaluation are +3 to
It is a 7-point scale of -3, and the most comfortable
Or +3 in a relaxed state, on the contrary, most anxious,
Alternatively, the score is -3 when the nervous state is set.

Example 1 In this example, an increase in immunoglobulin A concentration due to presentation of a comfortable image having a high relaxing effect was measured.

As a comfortable image, a Canadian Rocky scenery image ("Kawabe" Apollon APLJ-4008) was presented to five subjects using a video projector.
As a result, as shown in FIG. 2, the presentation of the video showed an increase in the reported score of comfort and relaxation and a corresponding increase in the immunoglobulin A concentration for all the subjects.

According to this data, after starting the video presentation,
It is considered that the time when 0 to 30 minutes has elapsed is a timing appropriate for evaluating the feeling of comfort and the feeling of relaxation. This change in immunoglobulin A concentration was statistically significant at a 5% risk factor.

Example 2 In this example, an increase in immunoglobulin A concentration due to listening to so-called α music was measured.

After subjecting the three subjects to a 30-minute calculation problem as a stress load, the subjects were placed in a relaxation chair and listened to α music (“daytime music” Apollon APCE5369) for 20 minutes. As a result, as shown in FIG. 3, there was a slight difference in behavior between the graph of the psychological evaluation score during the stress load and the graph of the immunoglobulin A concentration. The graphs started to rise almost synchronously, and the same behavior on the graphs was seen.

According to this data, after starting the music listening,
It is considered that the time when about 0 minutes has passed is the timing appropriate for evaluating the feeling of comfort and the feeling of relaxation, and the result was almost similar to that of Example 1. This change in immunoglobulin A concentration was statistically significant at a 5% risk factor.

From the above Examples 1 and 2, when measuring the general sensitivity not only by the feeling of comfort or relaxation but also by the change in the concentration of immunoglobulin A, it is necessary to measure
Minutes, 20 minutes or 30 minutes, or 10-3
It is effective to objectively and quantitatively evaluate the sensitivity by collecting a subject's saliva at an appropriate fraction of 0 minutes and measuring the change in the immunoglobulin A concentration in the saliva. It is suggested to say.

Example 3 In this example, audiovisual stimuli having different tastes (presence or absence of comfort and how to receive the degree of comfort) differed depending on the subject, and the increase in the immunoglobulin A concentration due to the stimulation was measured.

That is, an action movie (“Raiders Lost Arc”) laser disc S
F078-0052) was presented using a video projector for 40 minutes, and the comfort before and after the presentation was measured.

FIG. 4 shows changes in the salivary immunoglobulin A concentration before and after the presentation and the report score for psychological evaluation by the subject. The numerical value in parentheses in each graph in the figure indicates the transition of the report score of the psychological evaluation. For example, (0 →
1) indicates that the declaration score before the video presentation was 0 and the declaration score after the video presentation was 1.

According to FIG. 4, in subjects who reported that the feeling of comfort increased with the presentation of images in the psychological evaluation (indicated by a circle in the figure), immunoglobulin A in saliva was
The concentration also increased correspondingly, and subjects who showed a negative increase in comfort with video presentation in the psychological evaluation (in the figure,
In (plotted with ●), the concentration of immunoglobulin A in saliva either leveled off or decreased correspondingly.

Therefore, the sensitivity measuring method according to the present invention is as follows.
Even if the specific external stimulus that affects the sensitivity is a type of stimulus that is received differently depending on the personality of the subject,
It can be seen that the measurement can accurately reflect the way of receiving.

(Embodiment 4) In this embodiment, an example of a measuring piece according to a simple feeling measuring kit is shown. FIG. 5 (a) and FIG.
In (b), a measuring piece 11 is provided with an elongate rectangular plate-shaped support body 12 made of an arbitrary rigid material, and a cylinder 13 constituting a liquid storage structure is erected on one end side thereof. It has a structure in which the fixing portion 14 is provided.

The antibody fixing section 14 is a membrane, on which an anti-immunoglobulin antibody is fixed. As evident in the figure,
A fixed volume reservoir is formed by the cylinder 13 above the antibody fixing portion 14. Note that, as described above, the measurement piece 11 constitutes a simple sensitivity measurement kit in combination with a label expression substance (not shown).

The substance for label expression may be prepared as a simple ampule package made of, for example, glass or plastic, and the ampule package may be formed integrally with the measuring piece 11. Even if it is formed on the body, it is sufficient if it is enclosed in the same packaging box or the like as the measuring piece 11.

The contents of the labeling substance are also as described above. According to the measurement method, such as an enzyme substrate or a fluorescent agent, the direct method includes, for example, a labeled immunoglobulin for competition, and the indirect method optionally includes a labeled anti-immunoglobulin secondary antibody. It may contain distilled water for saliva dilution or washing.

The method of using the simple sensitivity measuring kit according to the present embodiment is as follows, for example. That is, first, about 0.5 ml of the sample saliva and an equal amount of distilled water are added to the liquid reservoir on the antibody immobilizing section 14, lightly immersed, left standing for about 5 minutes, and then the liquid is discarded. . Then, about 0.5 ml of a solution of the enzyme-labeled anti-immunoglobulin secondary antibody is added, and the mixture is allowed to stand for about 5 minutes. Then, 3 ml of 1 ml of distilled water
After washing twice, a color-developing enzyme substrate solution is added and allowed to stand for about 5 minutes, and comfort is determined from the intensity of the color development.

As can be understood from the above operation, the use of the simple kansei measurement kit makes it possible to perform kansei measurement extremely easily without using any special machinery and equipment. With the structure, there is no need to worry about spilling saliva, etc., causing measurement failure or soiling the floor.

[Brief description of the drawings]

FIG. 1 is a diagram showing a flowchart of a high-speed measurement method for sensitivity evaluation.

FIG. 2 is a graph showing changes in immunoglobulin A, a feeling of comfort and a feeling of relaxation accompanying presentation of a comfortable image.

FIG. 3 is a graph showing changes in immunoglobulin A, a feeling of comfort and a feeling of relaxation by listening to α music.

FIG. 4 is a graph showing changes in immunoglobulin A and a feeling of comfort when an action movie is presented.

5A and 5B are views showing a simple feeling measurement kit according to the present invention, wherein FIG. 5A is a plan view thereof, and FIG.
FIG. 3A is a sectional view taken along line AA of FIG.

[Explanation of symbols]

 4 Sample loop 9 Reaction part 11 Measurement piece 12 Support 13 Tube 14 Antibody fixing part

Claims (3)

[Claims] 1. A method for measuring the secretion amount of said immunoglobulin by measuring the concentration of immunoglobulin in saliva in a subject who has received any external stimulus that affects sensitivity, after a lapse of a predetermined time from the start of the test. A sensitivity measurement method characterized by quantitatively measuring the state of the sensitivity having a correlation. 2. In a subject who has received any audio-visual stimulus that affects the feeling of comfort or relaxation, the concentration of immunoglobulin in saliva is measured at an arbitrary time after the start of the test, whereby the comfort is measured. A method for measuring sensibility, characterized by measuring sensation or relaxation. 3. A measuring piece having an antibody fixing portion with a liquid storage structure provided on a support and an anti-globulin antibody fixed to the antibody fixing portion, and a label expression preparation prepared so as to be administrable to the antibody fixing portion. A simple sensibility measurement kit, comprising: a substance;