JP4391856B2 - Caries risk inspection tool - Google Patents

Description

  The present invention relates to a caries risk testing tool for comprehensively examining caries risk from the two aspects of factors attributable to oral bacteria and factors attributable to a host using an immunochromatography method.

  In recent years, dentistry surgeons have begun to try to grasp the caries risk of patients in order to consider measures to suppress the progression of caries and to make caries treatment plans. It is known that caries is a multifactorial disease caused by oral conditions and lifestyle habits and develops when all three factors of "oral bacteria", "host" and "food" are combined ( For example, refer nonpatent literature 1). Having the patient understand the caries risk is also important to motivate the maintenance of oral care.

  In particular, it is known that there is a close relationship between oral bacteria and the development of dental caries, and the higher the number of mutans streptococcus, a kind of oral bacteria, the higher the probability of developing dental caries. Since then, many attempts have been made to easily determine the number of mutans streptococci and understand the caries risk. A simple method for quantifying the number of Streptococcus mutans in human saliva is to apply a monoclonal antibody that reacts specifically with Streptococcus mutans, a type of Streptococcus mutans. (For example, refer to Patent Documents 1 and 2) and those for visually quantifying the cells grown in a simple culture kit (for example, refer to Patent Document 3).

  In addition, as a method for grasping caries risk from the viewpoint of “host”, for example, saliva secretion amount and saliva buffering capacity have been generally measured, but in recent years, the surface layer of Streptococcus mutans bacteria A specific protein that is a part of the substance was confirmed to be related to the initial adhesion of Streptococcus mutans to the tooth surface. It was elucidated that secretory immunoglobulin antibody A (hereinafter referred to as SIgA) secreted into saliva has a function of protecting the adhesion of Streptococcus mutans to the tooth surface. That is, it has been proposed as a caries risk test method that has been found that Streptococcus mutans bacteria hardly adhere to the tooth surface when the antibody titer of SIgA in human saliva is high, and that the caries risk is low (patent) Reference 4).

  However, since these caries risk tests were performed for each element such as “bacteria in the mouth” and “host”, the risks associated with various factors despite the fact that caries are inherently a multifactorial disease. There was a problem that it was difficult to judge the overall risk from. In general, when examining caries risk using an immunochromatography method, the determination result is indicated by a red line, whether the caries risk is high or low. Often, patients who have been tested often have difficulty understanding at a glance whether their caries risk is high or low.

KeyesPH: Recent advances in dental caries research, bacteriology, bacteriological findings, and biological implications, Int Dent J 1962; 12: 443 Japanese Unexamined Patent Publication No. 2-177898 Japanese Patent Laid-Open No. 10-36400 US Patent 5374538 Japanese Patent Application No. 2002-273125

  The present invention is capable of comprehensively judging the caries risk from the two aspects of oral bacteria and host when grasping the human caries risk, and the patient who has undergone the examination has its own caries risk. It is an object of the present invention to provide a caries risk test tool using an immunochromatography method capable of understanding at a glance.

That is, the present invention is a caries risk test in which two types of porous membrane pieces (X) having a length and a porous membrane piece (Y) are fixed to a base material (Z) made of a non-absorbable material. The porous membrane piece (X) includes an antibody (A ′) that specifically binds to Streptococcus mutans on one end of the porous membrane piece (X) attached with red colored particles (Pr). The specimen dropping section (X ₁ ) is arranged, and the detection section (X ₂ ) containing an antibody (A) different from the antibody (A ′) that specifically binds to Streptococcus mutans bacteria in the middle. Is disposed, and a sample absorption part (X ₃ ) is disposed at an end opposite to the sample dropping part (X ₁ ) across the detection part (X ₂ ),
The porous membrane piece (Y) is a specimen in which an antibody (B) that specifically binds to SIgA secreted into human saliva is attached to one end thereof with blue colored particles (Pb) attached thereto. A dropping part (Y ₁ ) is arranged, and a detecting part (Y ₂ ) containing an antigen (S) that specifically binds to SIgA secreted in human saliva is arranged in the middle, The dental caries risk test tool is characterized in that a specimen absorption part (Y ₃ ) is arranged at an end opposite to the specimen dropping part (Y ₁ ) across the detection part (Y ₂ ).

  The caries risk test tool according to the present invention is capable of comprehensively judging the risk from the two aspects of oral bacteria and host with one caries risk test tool, and further exists in human saliva. If the number of Streptococcus mutans is high and the risk of dental caries is high, the red stain, and if there is a large amount of SIgA secreted in human saliva and the caries risk is low, a blue stain appears. Is a caries risk test tool that makes it easy to visually understand the level of caries risk.

  Hereinafter, although the caries risk test | inspection tool based on this invention is demonstrated in detail using drawing, this invention is not limited to a present Example.

  FIG. 1 schematically shows an embodiment of a caries risk inspection tool in which two types of porous membrane pieces X and porous membrane pieces Y are fixed to a base material Z made of a non-absorbable material. FIG.

  In FIG. 1, Z is a base material on which two kinds of porous membrane pieces X and porous membrane pieces Y are fixed. The substrate Z is made of a non-absorbable material, and for example, a material formed from polystyrene, polyester, cellulose acetate or the like can be used. The shape may be any of a film shape, a plate shape, a strip shape, a strip shape, a disc shape, and the like, but a plate shape having a thickness of about 0.1 to 5 mm is preferable for stably installing the porous membrane.

  In the drawing of FIG. 1, X and Y are porous membrane pieces, and the shape of the porous membrane pieces is preferably a shape having a rectangular equal length in order for saliva to be developed in the porous membrane by capillary action. However, it may be an ellipse with rounded corners, or the periphery may be a curved line instead of a straight line. As the material, for example, a cellulose membrane, a cellulose acetate membrane or a cellulose membrane derivative membrane such as nitrocellulose, a glass filter, a filter paper or the like can be used. The porous membrane piece X and the porous membrane piece Y may be the same or different in shape and material, and can be fixed to the substrate Z by a conventional method. For example, a double-sided tape or an adhesive is used. Use and fix. The arrangement of the porous membrane piece X and the porous membrane piece Y at the time of fixing is not particularly limited, but the porous membrane piece X and the porous membrane piece Y are parallel to the longitudinal direction as shown in FIG. There may be a sense of stability when arranged in a positional relationship. Further, there may be a plurality of porous films X and Y. In that case, improvement in inspection accuracy can be expected, and improvement in inspection efficiency can be expected.

In FIG. 1, X ₁ is a specimen dropping part, X ₂ is a detection part, X ₃ is a specimen absorption part, and in the porous membrane piece X, the specimen dropping part X ₁ and the specimen absorption part X ₃ are the detection part X ₂ . What is necessary is just to fix to the edge on the opposite side by using a clip or the like. Sample absorbing section X ₃ particularly limited without example filter paper or the like as long as material that easily absorbs dropped saliva can be used.

The specimen dropping part X ₁ contains the antibody A ′, and the detection part X ₂ contains the antibody A. The antibody A ′ and the antibody A are not particularly limited as long as they specifically bind to Streptococcus mutans prepared by a method commonly used in dental diagnosis and research bases. 'Need different types. The antibody A 'included in the sample dropping portion X ₁ reddish colored particle Pr such as gold colloidal particles (manufactured by British Biocell International, Inc.) is attached.

  The red colored particle Pr is not particularly limited as long as it is a substance exhibiting a red color that does not interfere with the activity of the antibody among those usually used in immunochromatography, and the red color is a commercially available product such as a 5RP 10RP 5R 10R 5YR and the like, and among them, particles exhibiting a color tone such as 10RP 5R 10R are preferable.

Antibodies A included in the detection section X ₂ are optionally diluted with bovine serum albumin-containing phosphate buffer or the like, for example, coating a porous membrane X longitudinal direction micropipette so that the vertical as shown in Figure 1 Thus, it is used by being fixed to the porous membrane X.

In FIG. 1, Y ₁ is a specimen dropping part, Y ₂ is a detection part, and Y ₃ is a specimen absorption part. Like the porous membrane piece X, the specimen dropping part Y ₁ and the specimen absorption part Y ₃ are the detection part Y. _What is necessary is just to fix to the opposite end on both sides of ₂ using a clip or the like. Sample absorbing section Y ₃ is particularly limited without example filter paper or the like as long as material that easily absorbs dropped saliva can be used.

The sample dripping section Y ₁ antibody B which SIgA specifically binds secreted in human saliva, the detection unit Y ₂ contains antigen S of SIgA secreted in human saliva. The antibody B that specifically binds to SIgA secreted in human saliva can be used without particular limitation as long as it is an antibody prepared by a method commonly used for dental diagnosis and research, and is colored blue Particles Pb are attached. In the case of the caries risk test tool shown in FIG. 1, blue particles are used for antibody B (MONOCLONAL ANTI-HUMAN IGA (manufactured by SIGMA)) that specifically binds to SIgA secreted in human saliva. Blue latex particles (manufactured by Merck Chimie SAS) are attached as Pb using 1-Ethyl-3- (3-dimethylaminopropyl) carbodiimide Hydrochloride.

Antigens S of SIgA included in the detection section X _2, for example, can use a protein called PAc (Protein Antigen cerotype C) ( 361-386), the use is not limited particularly if functions as an antigen SIgA Can do. Antigen S is diluted with a phosphate buffer or the like as necessary, and applied with a micropipette so as to be perpendicular to the longitudinal direction of porous membrane Y as shown in FIG. Is done.

  The blue colored particles Pb are not particularly limited as long as they are colored particles that are usually used in immunochromatography and do not interfere with the activity of the antibody. The blue colored particles are 10P 5P 10PB 5PB 10B 5B 10BG 5BG 10G 5G 10GY 5GY etc. are shown, and among them, particles having a color tone such as 5PB 10B 5B 10BG 5BG 10G 5G 10GY are preferable.

The caries risk is inspected using the caries risk inspection tool in which the two kinds of porous film pieces X and porous film pieces Y shown in FIG. 1 are fixed to the base material Z made of a non-absorbable material. A method will be described. Added dropwise and the porous membrane pieces saliva sample dropping portion X ₁ of the human X, if there is Streptococcus mutans in human saliva, Streptococcus mutans is included in the sample dropping portion X ₁ red colored particles Pr is been combine with antibody a 'attachment, moves the porous membrane piece in X in its associated state from the specimen dropping unit X ₁ to the sample absorbing section X ₃ by a capillary phenomenon. Streptococcus mutans bacteria when passing through the detecting section X ₂ in the middle of the movement is supplemented by antibody A which is fixed to the porous membrane strip X, greater the number of Streptococcus mutans present in human saliva if (if caries risk is high) appears as a red stain detector X ₂ on the porous membrane strip X.

Similarly, dropping the porous membrane pieces saliva sample dropping portion Y ₁ of the human Y, if present SIgA in human saliva, the blue SIgA is contained in the sample dropping portion X ₁ colored particles Pb ties and attached antibody B, and porous membrane strip in Y in a state bound by navigating from the sample dropping portion Y ₁ by capillary action to the sample absorbing section Y _3, the detection unit Y ₂ in the middle of the movement When passing, SIgA is captured by the immobilized antigen S, and if SIgA present in human saliva is large (if the caries risk is low), it appears as a blue stain on the porous membrane piece Y. In this way, the color of the stain that appears on each of them is visually judged to determine the caries risk of the patient.

  The saliva used in the examination using the caries risk examination tool of the present invention is usually prepared by the same method as the examination of saliva in a dental diagnosis, and can also be prepared as necessary. For example, when measuring the number of Streptococcus mutans bacteria, stimulated saliva is treated with a solution containing 1 mol / L NaOH and a solution containing 0.5 mol / L citric acid containing 5% polyoxyethylene (10) octylphenyl ether. In the measurement of SIgA antibody titer, the stimulated saliva can be diluted 10 times with PBS containing 1% Tween20-0.5% skim milk.

Further, when the saliva is dropped to the sample dropping portion Y ₁ of the sample dropping portion X ₁ of the porous membrane strip X and the porous membrane pieces Y are each different preparation, sample dropping portion X ₁ and sample dropping or a part Y ₁ in different shapes, the previously equal mark the different symbols, numbers, and more preferably no fear that wrong saliva dripping.

  When a caries risk is determined from the result of visual determination, it is more preferable that a risk determination table as shown in Table 1 is prepared in advance because the explanation to the patient is easy. In Table 1, Streptococcus mutans bacteria are expressed as caries.

As an application example of caries risk test device according to the present invention is to include antibodies A 'which binds to the analyte dropping unit X ₁ Streptococcus mutans specifically in the state where the colored particles Pr is deposited reddish instead keep may method of dropping from a mixture of the saliva of the solution and human containing antibody a 'which is deposited colored particles Pr for red outside caries risk test device to the sample dropping portion X _1. Similarly, instead of including the antibody B that specifically binds to SIgA secreted in the blue human saliva in the specimen dropping part Y ₁ with the blue colored particles Pb attached thereto, it is also possible to inspect the caries risk dropwise after mixing the saliva solution and human containing antibody B which is deposited colored particles Pb for blue in the sample dropping portion Y ₁ in corrosion risk testing tool out.

  Furthermore, as an application example of the caries risk inspection tool according to the present invention, a porous membrane piece fixed to the substrate Z is used as one piece, and the specimen dropping part at one end thereof is specifically different from Streptococcus mutans. The antibody A ′ to which the red colored particles Pr to be bound are attached and the antibody B to which blue colored particles (Pb) that specifically bind to SIgA secreted in human saliva are attached are simultaneously included. A caries of a form in which an antibody A different from antibody A ′ that specifically binds to Streptococcus mutans and an antigen S of SIgA secreted in human saliva are contained at intervals. Risk testing tools are also conceivable.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective explanatory view schematically showing one embodiment of a caries risk inspection tool according to the present invention.

Explanation of symbols

X: Porous membrane X
X ₁ : Sample dropping part Pr: Red colored particles
A ′: an antibody that specifically binds to Streptococcus mutans bacteria X ₂ : detection unit
A: Antibody that specifically binds to Streptococcus mutans X ₃ : Specimen absorption part Y: Porous membrane Y
Y ₁ : Sample dropping part Pb: Blue colored particles
B: Antibody that specifically binds to SIgA secreted in human saliva Y ₂ : Detection unit
S: Antigen of SIgA secreted into human saliva Y ₃ : Sample absorption part Z: Base material

Claims (1)

Two types of porous membrane pieces (X) having a length and a porous membrane piece (Y) are caries risk inspection tools fixed to a base material (Z) made of a non-absorbable material,
The porous membrane piece (X) has a specimen dropping part (Red) in which an antibody (A ′) that specifically binds to Streptococcus mutans bacteria is attached to one end of the porous membrane piece (X). X ₁ ) is arranged, and a detection part (X ₂ ) containing an antibody (A) different from the antibody (A ′) that specifically binds to Streptococcus mutans is arranged in the middle. , The specimen absorption section (X ₃ ) is disposed at the end opposite to the specimen dropping section (X ₁ ) across the detection section (X ₂ ),
The porous membrane piece (Y) is a specimen in which an antibody (B) that specifically binds to SIgA secreted into human saliva is attached to one end thereof with blue colored particles (Pb) attached thereto. A dropping part (Y ₁ ) is arranged, and a detecting part (Y ₂ ) containing an antigen (S) that specifically binds to SIgA secreted in human saliva is arranged in the middle, A caries risk inspection tool, characterized in that a specimen absorption part (Y ₃ ) is arranged at an end opposite to the specimen dropping part (Y ₁ ) across the detection part (Y ₂ ).

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