JPH11142403A - Immunodetection method of alimentary allergen - Google Patents

Abstract

PROBLEM TO BE SOLVED: To easily detect a alimentary allergen by measuring an antibody reaction of an allergen specific antibody in saliva or reactivity with an immunoglobulin(Ig). SOLUTION: The subject immunodetection method is so composed that reactivity with an allergen specific Ig in saliva, which is produced by an antigen/ antibody reaction with an antigen in saliva after adding a alimentary allergen to the saliva gathered from the inside of a mouth of a man to be detected, is measured, or a specific antibody reaction of the specific antibody is measured, and then the measured value is compared with the reactivity with the allergen specific Ig in saliva of a healthy man, which is obtained by the antigen/antibody reaction with the same antigen in the same conditions, or with a measured average value of the specific antibody reaction of the specific antibody, and then the immunodetection is performed base on the significant difference and the carrelation.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for easily detecting a dietary allergen substance by measuring the antibody reaction of an allergen-specific antibody in saliva and the reactivity with immunoglobulin (Ig).

[0002]

2. Description of the Related Art Conventionally, the RAST method has been widely used as a method for detecting allergens in dietary allergies. This measurement method has already been established and is one of the popular methods. However, the detection of allergens by this method requires blood, which is very painful for patients. Other methods for detecting allergens include skin tests, which also have disadvantages that can be very painful for patients. For this reason, these conventional allergen detection methods are performed by special institutions such as medical facilities, so that initial discrimination at the individual level (FAR)
It is not suitable as a means for screen screening. Dietary allergens-It is important for patients to know allergens at the individual level, to prevent allergic outbreaks, to assemble food intake, and to provide early and accurate treatment. There is an urgent need to establish a simple test method for allergen substances.

[0003]

SUMMARY OF THE INVENTION The present inventor studied the method of detecting allergens of dietary allergens more easily and more easily at the individual level of a patient based on the above-mentioned concept. developed.

The inventor of the present invention, Tomasi et al., J. Exp. Med., 121, 101-125 (1961)
5)) is an immunoglobulin (Ig) A in saliva
Since reporting that it is present as secretory IgA bound to retorycomponent, immunoglobulin (Ig) in saliva
Various studies have been conducted, and up to this day, various immunoglobulins such as IgM, IgG, and IgD have been found in saliva.
(Ig) has been clarified (Shigeaki Ishizaka: Journal of the Salivary Gland Society of Japan, 32, 11-12 (1991)). These immunoglobulins (Ig) are considered to react to those taken orally. Since dietary allergens are also taken orally, immunoglobulin (I
I thought that g) could be used.

As an experiment, saliva was collected from pediatric patients diagnosed with dietary allergy among outpatients,
Ovalbumin (0A), ovomucoid (OM), casein, which are considered as major allergens of eggs and milk using this saliva,
Specific antibody reaction against β-lactoglobulin (β-Lg)
The reactivity with Ig in saliva was measured using a sensor chip on which the antigen was immobilized.
An attempt was made to measure with Acore. As a result, the present inventors have obtained the technical knowledge that the following significant difference and a predetermined correlation exist between the allergic patient group and the healthy subject group, and have reached the present invention.

[0006] Specific antibodies in saliva collected from milk and egg allergic patients tend to have significantly higher specific antibody responses to β-lactoglobulin (β-Lg) and ovomucoid (OM) than those of healthy subjects. Is shown. Ovalbumin (OA) and ovomucoid, which are egg-derived antigens
(OM), the measured values of specific IgE and specific IgG responses in saliva of the allergic patient group were significantly higher than those of healthy subjects, and specific sIgA was conversely higher in the allergic patient group. Shows a low trend. Specificity in saliva of allergic patient group against casein and β-lactoglobulin (β-Lg), which are milk-derived antigens
The measured value of IgE is significantly higher than that of the healthy group,
Conversely, specific sIgA tends to be lower in the allergic patient group.

[0007] Considering the results, the idea was obtained that saliva can be used for simple measurement of dietary allergen substances. Therefore, the present inventor has completed a simple immunoassay for a dietary allergen substance using saliva according to the present invention based on the above-mentioned new technical knowledge and idea.

[0008]

According to a first aspect of the invention, a dietary allergen substance is added to saliva collected from the mouth of a test subject to cause an antigen-antibody reaction with the antibody in saliva. Measure the reactivity with allergen specific Ig in the resulting saliva or measure the specific antibody reaction of the specific antibody,
The measured value is compared with the measured average value of the reactivity with allergen-specific Ig in the saliva or the specific antibody reaction of the specific antibody in a healthy subject who has undergone an antigen-antibody reaction under the same conditions for the same antigen, and correlates with the significant difference. An immunoassay for a dietary allergen substance, characterized in that an immunoassay is performed based on the relationship.

[0009] The second invention to be patented is that saliva collected from the mouth of a test subject contains β-lactoglobulin (β-Lg), which is said to be a major allergen of milk, and major allergen of egg white. Ovomucoid is said to be
(OM) was added to allow an antigen-antibody reaction with salivary antibody, and the specific antibody reaction of the salivary allergen-specific antibody, Resolution Unit (RU), was measured.The measured value was the same for the same antigen. In comparison with the measured average value of the specific antibody reaction of the resolution unit (RU), which is an allergen-specific antibody in the saliva of a healthy subject who has undergone an antigen-antibody reaction under the conditions, the immunoassay is performed by the significant difference and correlation. This is an immunoassay for a dietary allergen substance.

[0010] A third invention which is to be patented is that casein or β-lactoglobulin, which is said to be a major allergen of milk, is added to saliva collected from the mouth of a test subject.
(β-Lg) to cause an antigen-antibody reaction with the antibody in saliva to measure the reactivity with allergen-specific IgE and specific IgG in saliva, and to measure the measured value of the antigen / antibody under the same conditions for the same antigen under the same conditions Allergen specificity in saliva of reacted healthy subjects
This is an immunoassay method for a dietary allergen substance, characterized in that an immunoassay is carried out by comparing the measured average value of reactivity with IgE and specific IgG with a significant difference and correlation.

A fourth invention to be patented is that ovalbumin (OA) or ovomucoid (OM), which is said to be a major allergen of egg white, is added to saliva collected from the mouth of a subject to be examined. Measure the reactivity with allergen specific sIgA, specific IgE or specific IgG in saliva generated by the antigen-antibody reaction with the antibody, and measure the measured value with the same antigen under the same conditions for the antigen-antibody reaction. Immunity of a dietary allergen substance, characterized in that the immunoreactivity is compared with the measured average value of the reactivity with allergen-specific sIgA, specific IgE or specific IgG in the saliva of the elderly, and the significant difference and correlation are used. Inspection method.

A fifth invention to be patented is to add a colloidal gold solution to saliva collected from the mouth of a subject to be tested, bind the antibody and the colloidal gold, and preliminarily produce egg white, soybean, or soybean as an antigen on a cellulose membrane. The milk was coated, colloidal gold solution saliva was dropped on one end with a spoid, and after 30 minutes, the colloidal gold deposition was visually judged,
The colloidal gold was determined to be positive (+), non-colloidal gold was determined to be negative (-), and non-colloidal gold was determined to be false positive (±). This is an immunoassay for a dietary allergen substance, characterized in that:

[0013]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail based on embodiments. In order to confirm the effects of the present invention, the inventor prepared a group of 19 patients diagnosed with allergy as test subjects and a group of 10 healthy healthy subjects as controls, and prepared them in advance. The amount of specific Ig antibody in the serum for the test subject's egg white and milk was measured, and on the other hand, saliva was collected from the mouth of those test subjects, and the specific Ig and allergen specificity in saliva in response to various allergens were measured. Antibody to BIA
Core and ELISA measurements were performed, and the two were compared. A clinical experiment was conducted to confirm that a dietary allergen substance could be detected by measurement using saliva.

First, Table 1 shows the clinical background in which the results of measuring the amounts of specific Ig antibodies in the serum of the 29 test subjects were summarized. The breakdown of the test subjects was a total of 19 allergy patients, including 10 males and 9 females up to 2 to 6 years old, and the healthy subjects were 5 males and 5 females up to 2 to 5 years old.
There are 10 people. For these test subjects, the amount of specific IgE antibody in the serum against egg white and milk was measured, and from 0 to 6
Up to seven levels. Incidentally, ND indicates that no measurement was performed.

[0015]

[Table 1]

<Example 1> 1) Subject Subject to be tested 29 shown in Table 1 shown as the clinical background
First name.

2) Saliva Collection Method and Preparation Method Saliva was collected by including a 1 cm square cotton wool in the mouth, and collecting saliva without stimulating secretion. The collected saliva was stored at −20 ° C. until subjected to the experiment. Deionized water was added to the collected cotton wool, and the supernatant was collected after stirring. This operation was performed three times, and the collected supernatant was freeze-dried and used for the following measurement.

3) Measurement of reaction The test subjects (19 allergic patients and 10 healthy subjects)
The saliva preparation supernatant collected from the mouth of the name) contains β-lactoglobulin (β-Lg), which is said to be the major allergen substance of milk, and ovomucoid (OM), which is said to be the major allergen of egg white, as antigens. ), And the specific antibody reaction of the resolution unit (RU), which is an allergen-specific antibody in saliva, produced by causing an antigen-antibody reaction with the antibody in saliva is measured using BIAcore manufactured by Pharmacia. The result of the measurement is the reaction between the antigen in FIG. 1 and the antibody in saliva.

Further, for the purpose of verification, an allergen-specific antibody produced by subjecting the same antigen β-lactoglobulin (β-Lg) and ovomucoid (OM) to an antigen-antibody reaction with an antibody in serum under the same conditions. Measure the resolution unit (RU). The result of the measurement is the reaction between the antigen in FIG. 2 and the antibody in the serum.

4) Results of immunoassay for dietary allergens As can be seen from the measured values in FIG. 1, the measured values of the resolution unit (RU) when saliva was used showed that the patient group had healthy subjects. It shows a higher tendency than the group. The average reading for the specific antigen β-lactoglobulin (β-Lg) in the patient group is about 530 and the mean reading for ovomucoid (OM) is about 480.
The average measured value of β-lactoglobulin (β-Lg) in the healthy group is about 110, and the average measured value of ovomucoid (OM) is about 120. As a result, when saliva was used, a significant difference was recognized between the allergic patient group and the healthy group at a risk factor of 0.01.

Accordingly, the measured values of the resolution unit (RU), which is an allergen-specific antibody in the saliva of an allergic patient group, and the measured average values of the resolution unit (RU), which is an allergen-specific antibody in the saliva of a healthy subject, By comparison, the immunological test such as whether or not the test subject is allergic to milk or whether it is allergic to egg white can be simplified by the significant difference.

5) Verification of the results of the immunoassay according to the present invention Next, in order to verify the reliability of the immunoassay for the dietary allergen substance of the present invention, the reaction with the antibody in serum, which is a conventional method, was carried out in the same manner. Measurement under appropriate conditions and compare with the results. FIG. 2 shows the result. Looking at the measured values in FIG. 2, the measured values of the resolution unit (RU) tend to be higher in the patient group than in the healthy group, as in the case of using saliva. Specific antigen β-lactoglobulin (β-Lg) of patient group when using serum
The mean measured value for ovomucoid (OM) is about 950, the mean measured value for β-lactoglobulin (β-Lg) in a healthy group is about 460, and the mean measured value for ovomucoid
The average measurement of (OM) is about 390. As a result, when saliva was used, a significant difference was recognized between the allergic patient group and the healthy group at a risk factor of 0.01. As a result, it was found that the results of the immunoassay for whether or not the test subject had milk allergy or egg white allergy based on the measured value of the specific antibody in the serum were exactly the same.

As described above, the results of the immunological test using saliva and the immunological test using serum for milk allergy and egg white allergy are the same. We verified that immunoassay for sex allergens was a reliable method.

Example 2 1) Subjects 29 subjects to be tested shown in Table 1 shown as the clinical background
First name.

2) Saliva Collection Method and Preparation Method Saliva was collected by containing a 1 cm square cotton wool in the mouth and collecting saliva without stimulating secretion. The collected saliva was stored at −20 ° C. until subjected to the experiment. Deionized water was added to the collected cotton wool, and the supernatant was collected after stirring. This operation was performed three times, and the collected supernatant was freeze-dried and used for the following measurement.

3) Measurement of the reaction The salivary IgE, IgG and sIgA which react with ovalbumin (OA) and ovomucoid (OM), which are the major allergens of egg white, are analyzed by ELISA.
Measured by A method.

That is, ovalbumin (OA) and ovomucoid (OM), which are said to be the major allergens of egg white, are added to the supernatant of saliva collected from the mouth of the test subject, and the antigen of salivary antibody and The reactivity between allergen-specific IgE and specific IgG in saliva produced by the antibody reaction was measured. The results of the measurement are shown in FIGS. 3 (a) and 3 (b), which show the reaction between the salivary antibody and the egg-derived antigen ovalbumin in allergic patients and healthy subjects, and FIGS. 4 (a) and 4 (b) show the allergy. It is the reaction of the salivary antibody of a patient and a healthy person with the egg-derived antigen ovomucoid.

Next, for the purpose of verification, ovalbumin (OA) and ovomucoid (OM), which are said to be the major allergens of egg white, were added to the serum of the test subject, and the antigen-antibody reaction between the antibody in saliva was performed. Allergen specific in serum produced by aging
The reactivity between IgE and specific IgG was measured by ELISA. The results of the measurement are shown in FIGS. 5 (a) and 5 (b), showing the reaction between the antibody in the serum of an allergic patient and a healthy person and the ovalbumin derived from the egg. FIGS. 6 (a) and (b) show the allergy. It is a reaction between the antibody in the serum of a patient and a healthy person and the egg-derived antigen ovomucoid.

4) Results of immunoassay for dietary allergens. As is clear from the measured values in FIGS. 3 (a) (b) and 4 (a) (b), allergen specificity when saliva reacting with the antigens ovalbumin (OA) and ovomucoid (OM) is used Ig
Measurements of E and IgG antibodies tend to be significantly higher in the patient group than in the healthy group.

As shown in FIG. 3, the average value of the specific IgE in response to the specific antigen ovalbumin (OA) of the patient group is about 0.19, and the specific value of the specific group that responds to ovalbumin (OA) in the healthy subject group. The average measurement of IgE is about 0.075. The value of the patient group is about 2.5 times higher than that of the healthy group. Similarly, the average measurement value of specific IgG that reacts with the antigen ovalbumin (OA) of a healthy person is about 0.3, and the ovalbumin (O
The average measured value of specific IgG responding to A) is about 0.09. The value of the patient group is about three times higher than that of the healthy group. As a result, when saliva was used, a significant difference was recognized between the allergic patient group and the healthy group at a risk factor of 0.01.

As shown in FIG. 4, the average value of the specific IgE that reacts with the specific antigen ovomucoid (OM) of the patient group is about 0.35, and the specific IgE that reacts with the ovomucoid (OM) of a healthy subject is about 0.35. The average measurement is about 0.08. The value of the patient group is about four times higher than that of the healthy group. Similarly, the average measured value of specific IgG that reacts to the antigen ovomucoid (OM) in the patient group is about 0.43, and the average measured value of specific IgG that reacts to ovomucoid (OM) in healthy subjects is about 0.1. Thus, the value of the patient group is about four times higher than that of the healthy group. As a result, when saliva was used, a significant difference of about 4.5 times was observed between the allergic patient group and the healthy group at a risk factor of 0.01.

Therefore, the specificity which reacts with the antigens ovalbumin (OA) and ovomucoid (OM) in the saliva of a group of allergic patients
Comparing the measured value of the reactivity between the IgE antibody and the specific IgG antibody with the average measured value of the reactivity between the specific IgE and the specific IgG that reacts with the antigens ovalbumin (OA) and ovomucoid (OM) in the serum The immunological test such as whether or not the test subject has egg white allergy can be simplified by the significant difference.

5) Verification of immunological test results according to the present invention. Next, in order to verify the reliability of the immunoassay for the dietary allergen substance of the present invention, the reaction with the antibody in serum, which is a conventional method, is measured under similar conditions, and the results are compared with the results. . FIGS. 5A and 6B and FIGS. 6A and 6B show the results. The measured values in FIGS. 5 and 6 show that the specific IgE antibody and the specific
The IgG antibody readings tend to be higher in the patient group than in the healthy group.

As shown in FIGS. 5 (a) and 5 (b), the average measured value of the reactivity of the patient group with the specific IgE antibody reacting with the specific antigen ovalbumin (OA) when serum was used was about 0.28. And the average measured reactivity with the specific IgG antibody is about 0.36. Ovalbumin (OA) in a healthy group to control this
The average measured value of reactivity with specific IgE antibodies that react with
07, and the average measured value of the reactivity with the specific IgG antibody is about 0.
08.

Further, as shown in FIGS. 6 (a) and 6 (b), specific antigen ovomucoid (OM) of a patient group when serum was used
The average measured value of reactivity with specific IgE antibodies that react with
The average value of the reactivity with the specific IgG antibody was about 0.
42. The average measured value of the reactivity with the specific IgG antibody reacting with the antigen ovomucoid (OM) of the healthy group serving as the control is about 0.08, and the average measured value of the reactivity with the specific IgG antibody is about 0.09. .

Considering from the measurement results, when saliva is used, a significant difference is recognized between the allergic patient group and the healthy group at a risk factor of 0.01. Then, it was found that the result of the immunoassay for whether or not the test subject had egg white allergy based on the measured value of the specific antibody in the serum was exactly the same as that using saliva.

As described above, the same results were obtained when the egg white allergy was subjected to immunoassay using saliva and when the immunoassay was performed using serum. It shows that immunoassay for substances is a reliable method.

<Example 3> 1) Subjects 29 subjects to be tested shown in Table 1 shown as the clinical background
First name.

2) Method of collecting and preparing saliva Saliva was collected by including a cotton swab of 1 cm square in the mouth, and collecting saliva without stimulating secretion. The collected saliva was stored at −20 ° C. until subjected to the experiment. Deionized water was added to the collected cotton wool, and the supernatant was collected after stirring. This operation was performed three times, and the collected supernatant was freeze-dried and used for the following measurement.

3) Measurement of Reaction Saliva IgE, IgG and sIgA which react with casein and β-lactoglobulin (β-Lg), which are the major allergens of milk, are converted to E.
It was measured by the LISA method.

That is, casein, which is said to be a major allergen of milk, and β-lactoglobulin (β-Lg) are added to the supernatant of saliva collected from the mouth of the test subject, and the salivary antibody is reacted with the antibody in saliva. The reactivity of allergen-specific IgE, IgG and sIgA in saliva produced by the antigen-antibody reaction was measured. The results of the measurement are shown in FIGS. 7 (a) and 7 (b) showing the reaction between the salivary antibody and the milk-derived antigen casein of the allergic patients and healthy subjects, as shown in FIGS. 8 (a) and (b). It is a reaction between the antibody in saliva of a healthy subject and the antigen derived from milk, β-lactoglobulin.

Next, for the purpose of verification, casein, which is said to be a major allergen of milk, and β-lactoglobulin (β-Lg) were added to the serum of the test subject, and the antigen and the serum antibody were compared with each other. The reactivity between allergen-specific IgE and specific IgG in the serum produced by the antibody reaction was measured by ELISA.
The results of the measurement are shown in FIGS. 9 (a) and 9 (b) showing the reaction between the antibody in serum and the milk case-derived antigen casein of the allergic patients and healthy subjects. It is a reaction between the antibody in serum of a healthy subject and the milk-derived antigen β-lactoglobulin.

4) Results of immunoassay for dietary allergens As can be seen from the measured values in FIGS. 7 and 8, allergens using saliva that reacts with casein and β-lactoglobulin (β-Lg). Measurements of specific IgE and specific IgG antibodies tend to be significantly higher in the patient group than in the healthy group.

First, as shown in FIG. 7, the average measured value of the specific IgE that reacts with the antigen casein in the saliva of the patient group is about 0.4.
The average measured value of specific IgE in response to casein in a healthy group is about 0.07. Similarly, the average measured value of specific IgG in response to the antigen casein in the patient group is about 0.32, and the average measured value of specific IgG in the healthy group is about 0.06. As a result, when saliva was used, there was a significant difference that the allergic patient group was higher than the healthy group at a risk rate of 0.01.

Next, as shown in FIG. 8, β-lactoglobulin (β-Lg), which is a major allergen of milk,
Similarly, the mean specific IgE was about 0.43 in the allergic patient group.
The average value of specific IgG is 0.4 in the group of allergic patients and 0.09 in the group of healthy subjects. As described above, in the case of β-lactoglobulin (β-Lg), a significant difference is higher in the patient group than in the healthy group as in casein.

On the other hand, as shown in FIGS. 7 and 8, the average value of the specific sIgA for casein was about 0.1 in the allergic patient group and about 0.4 in the specific sIgA of the healthy group. Was. Furthermore, the average value of specific sIgA is β-lactoglobulin.
(β-Lg) is about 0.12 in the allergic patient group,
It was about 0.43 in the healthy group. That is, in the case of specific sIgA,
For both the antigen casein and β-lactoglobulin (β-Lg), the healthy group showed a tendency to be significantly higher than the patient group. That is, in the case of specific sIgA, specific IgE or specific IgG
It turns out that there is an inverse correlation with.

Therefore, the measured values of the reactivity of specific IgE, specific IgG and specific sIgA reacting with the antigen casein and β-lactoglobulin (β-Lg) in the saliva of each allergic patient group and the salivary value of the healthy subject group Casein and β-lactoglobulin
By comparing the average reactivity of specific IgE, specific IgG, and specific sIgA in response to (β-Lg), it is possible to determine whether the test subject is allergic to milk based on the significant difference and correlation. Immune tests can be simplified.

5) Verification of the results of the immunoassay according to the present invention Next, in order to verify the reliability of the immunoassay for the dietary allergen substance of the present invention, the reaction with the antibody in serum, which is a conventional method, was performed in the same manner. Measurement under appropriate conditions and compare with the results. 9 and 10 show the results. Looking at the measured values in FIG. 9, the average values of serum specific IgE and specific IgG that react with casein are about 0.4 and 0.32 in the allergic patient group, and about 0.07 and
0.06. FIG. 10 shows that β-lactoglobulin (
The average values of serum specific IgE and specific IgG in response to β-Lg) were about 0.43 and 0.4 in the allergic patient group, and about 0.08 and 0.09 in the healthy group. That is, as in the case of using saliva, the measured values of the specific IgE antibody and the specific IgG antibody are:
The patient group shows a higher tendency than the healthy group.

Considering the results of the measurement, the result of the immunoassay for whether or not the test subject has a milk allergy based on the measured value of the specific antibody in the serum is the same as that using saliva. In this way, the results of the same test for milk allergy using saliva and those using serum for immunoassays are the same, indicating that the method of immunoassay for dietary allergen substances according to the present invention is the same. Indicates a reliable method.

Example 4 1) Subject Subject 29 to be tested shown in Table 1 shown as the clinical background
First name.

2) Saliva Collection Method and Adjustment Method 1 ml of saliva was collected from the mouth using a spoid, and a colloidal gold solution (pH 9.0) was added to bind the antibody and the colloidal gold. Next, egg white, soybean, and milk were previously coated as antigens on a cellulose membrane (1 cm × 10 cm), and colloidal gold solution saliva was dropped on one end with a dropper. The judgment was made visually.

3) Results The results are as shown in Table 2.

[0053]

[Table 2]

Table 2 shows that the salivary sample reacted with each antigen, and the deposition of colloidal gold was recognized as positive (+).
Negative (-) is not observed, and false positive (±) is neither. According to Table 2, about 70% of allergic patients were positive for egg white, and about 90% were positive when combined with false positives. In addition, about 90% of healthy subjects showed negative. For soy, about 50% of allergic patients were positive, including about 65% including false positives. In addition, about 80% of healthy subjects showed negative, and positive and false positive were 10% each. Approximately 50% of the positive and false-positive people responded to milk, but about 80% of healthy subjects were negative. These results indicate that allergen screening can be performed easily and in a short time on an individual level.

[0055]

[Effects] As described above, a dietary allergen substance is added to saliva collected from the mouth of a test subject to cause an antigen-antibody reaction with the antibody in saliva, and the reaction with allergen-specific Ig in saliva produced. By measuring sex or measuring a specific antibody reaction, an immunoassay for a dietary allergen can be easily performed.

Since saliva can be easily collected from a subject without pain, the immunoassay for a dietary allergen substance can be easily performed by the existing measurement method. Moreover, the results are almost the same as those obtained by measuring a specific antibody in serum and performing an immunoassay.
Therefore, performing an immunoassay using a specific antibody in saliva is suitable for primary screening at the patient level.

[Brief description of the drawings]

FIG. 1 shows a reaction between an antigen and an antibody in saliva.

FIG. 2 shows a reaction between an antigen and an antibody in serum.

[FIG. 3] (a) and (b) show the reaction between salivary antibody and egg-derived antigen ovalbumin of allergic patients and healthy subjects.

[FIG. 4] (a) and (b) are reactions of salivary antibodies of allergic patients and healthy subjects with egg-derived antigen ovomucoid.

[FIG. 5] (a) and (b) show the reaction between the antibody in the serum of an allergic patient and a healthy subject and the ovalbumin derived from the egg.

[FIG. 6] (a) and (b) show the reaction between the antibody in serum of an allergic patient and a healthy subject and the egg-derived antigen ovomucoid.

[FIG. 7] (a) and (b) show the reaction between the salivary antibody and the milk-derived antigen casein of allergic patients and healthy subjects.

FIG. 8 (a) and (b) show the reaction between the salivary antibody of an allergic patient and a healthy subject and the milk-derived antigen β-lactoglobulin.

[FIG. 9] (a) and (b) are the reactions of serum antibodies of allergic patients and healthy subjects with milk-derived antigen casein.

[FIG. 10] (a) and (b) are the reactions of serum antibodies of allergic patients and healthy subjects with milk-derived antigen β-lactoglobulin.

Claims (5)

[Claims] Claims: 1. Saliva collected from the mouth of a test subject,
A dietary allergen substance is added and the antigen-antibody reaction with the antibody in saliva is performed to measure the reactivity with allergen-specific Ig in saliva or the specific antibody reaction of the specific antibody is measured. Immunoassay based on the significant difference and correlation with the reactivity with allergen-specific Ig in the saliva or the specific antibody reaction of the specific antibody in the saliva of a healthy subject who has undergone an antigen-antibody reaction under the same conditions for the same antigen An immunoassay for a dietary allergen substance, wherein the immunoassay is carried out. 2. Saliva collected from the mouth of the test subject,
Addition of β-lactoglobulin (β-Lg), which is said to be the major allergen of milk, and ovomucoid (OM), which is said to be the major allergen of egg white, causes an antigen-antibody reaction with antibodies in saliva. The specific antibody reaction of the resolution unit (RU), which is an allergen-specific antibody in saliva, was measured, and the measured values were compared for the same antigen under the same conditions.
Compared to the measured average value of the specific antibody reaction of the resolution unit (RU), which is an allergen-specific antibody in the saliva of a healthy subject who has undergone an antibody reaction, immunoassay is performed based on the significant difference and correlation. Immunoassay for dietary allergens. 3. Saliva collected from the mouth of the test subject,
Casein or β-lactoglobulin (β-Lg), which is said to be the major allergen of milk, is added to cause an antigen-antibody reaction with antibodies in saliva, and the reactivity with allergen-specific IgE and specific IgG in saliva is increased. Measure and compare the measured value with the measured average value of the reactivity with allergen-specific IgE and IgG in saliva of healthy subjects who have undergone an antigen-antibody reaction under the same conditions for the same antigen. An immunoassay for a dietary allergen substance, wherein the immunoassay is performed. 4. Saliva collected from the mouth of a test subject,
Ovalbumin (OA) or ovomucoid (OM), which is said to be the major allergen of egg white, is added to ovalbumin (OM) to cause an antigen-antibody reaction with salivary antibodies.
The reactivity with IgE or specific IgG was measured, and the measured value was compared with the measured average value of the reactivity with allergen-specific IgE or specific IgG in saliva of healthy subjects who had an antigen-antibody reaction with the same antigen under the same conditions. An immunoassay method for a dietary allergen substance, wherein the immunoassay is carried out in comparison with a significant difference and a correlation. 5. A colloidal gold solution is added to saliva collected from the mouth of the test subject, the antibody and the colloidal gold are bound, and egg white, soybean or milk is coated as an antigen on a cellulose membrane in advance, and Colloidal gold solution saliva was dropped with a drop, and after 30 minutes, colloidal gold deposition was visually determined. Positive colloidal gold deposition was observed (positive) and negative colloidal gold deposition was negative (-). ,
An immunoassay for a dietary allergen substance, characterized by visually determining that neither of them is a false positive (±), thereby performing an immunoassay.