WO2004033716A1 - Methode d'examen et diagnostic d'examen pour les parodontopathies - Google Patents

Methode d'examen et diagnostic d'examen pour les parodontopathies Download PDF

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Publication number
WO2004033716A1
WO2004033716A1 PCT/JP2003/009493 JP0309493W WO2004033716A1 WO 2004033716 A1 WO2004033716 A1 WO 2004033716A1 JP 0309493 W JP0309493 W JP 0309493W WO 2004033716 A1 WO2004033716 A1 WO 2004033716A1
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WO
WIPO (PCT)
Prior art keywords
periodontal disease
gtp
examination
activity
diagnostic
Prior art date
Application number
PCT/JP2003/009493
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English (en)
Japanese (ja)
Inventor
Yasuyuki Ishizuka
Original Assignee
Applied Cell Biotechnologies, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication date
Application filed by Applied Cell Biotechnologies, Inc. filed Critical Applied Cell Biotechnologies, Inc.
Publication of WO2004033716A1 publication Critical patent/WO2004033716A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/91—Transferases (2.)
    • G01N2333/91045—Acyltransferases (2.3)
    • G01N2333/91074—Aminoacyltransferases (general) (2.3.2)
    • G01N2333/9108—Aminoacyltransferases (general) (2.3.2) with definite EC number (2.3.2.-)

Definitions

  • the present invention relates to a method for detecting periodontal disease and a diagnostic reagent. More specifically, the present invention relates to a biochemical test method for periodontal disease using y-glutamyl transpeptidase as a marker and a test reagent for periodontal disease.
  • Periodontal disease is a chronic disease in which the periodontal tissue around the teeth gradually becomes loom. Generally, except for apical periodontitis and tumors formed in the periodontal tissue, diseases related to other periodontal tissues are collectively called periodontal disease (or periodontal disease). .
  • Symptoms of periodontal disease include swelling of the gums, bleeding from the gums when eating or brushing teeth, increasing bad breath, and pus coming out around the teeth when pushing the gums.
  • periodontal diseases pass quietly without any subjective symptoms such as pain. Many have been lost.
  • periodontal disease is a slow but periodic, quiescent period of inactivity, followed by severe acute inflammation and destruction, resulting in cracks between the teeth and gums (generally referred to as periodontal pockets).
  • the connective tissue around the teeth is soiled and the alveolar bone is resorbed. Observation from the outside shows that the gums are thin and she evacuates, and the 'roots' appear, and eventually the teeth become loose and eventually fall out.
  • Periodontal disease testing method using enzyme activity measurement a sample collected from the oral cavity is allowed to react with a substrate for measuring the activity of lipophilic phosphatase. The degree of color development is visually observed, and gingivitis and periodontitis are observed.
  • a method for measuring the enzyme activity of collagenase contained in gingival crevicular fluid Japanese patent Publication No. 61-710000 publication has been proposed.
  • the present invention examines the degree of alveolar bone rupture that occurs with inflammation of periodontal disease by examining the activity of an enzyme having an activity of inducing osteoclast differentiation, that is, the activity of 0 / -daltamyl transpeptidase. Grasp by periodontal disease It is an object of the present invention to provide a method for examining periodontal disease and a diagnostic reagent capable of accurately predicting progression. Disclosure of the invention
  • the present invention employs the following solutions.
  • the present inventors express y-glutamyl transpeptidase enzyme (common name; hereinafter abbreviated as “ ⁇ -GT GT”) in the gingival crevicular fluid of a periodontal disease patient. Based on the newly discovered fact that it is present, a method for detecting periodontal disease by measuring the activity of ⁇ -GTGT is provided.
  • ⁇ -GT GT y-glutamyl transpeptidase enzyme
  • Gingival crevicular fluid is known to increase its outflow when gingival irritation occurs, and y-GTP hydrolyzes ⁇ -glutamyl peptide, including daltathione.
  • y-GTP hydrolyzes ⁇ -glutamyl peptide, including daltathione.
  • it is known as an enzyme that transfers a ⁇ -glutamyl group to another amino acid peptide.
  • the ⁇ / -GTP used as a test marker in the present invention is a ⁇ -GT ⁇ activity measurement method generally used in the test diagnosis of liver diseases and the like, and is specific to the anti-0 / -GTP antibody -GT ⁇ .
  • a method for confirming the expression of mRNA encoding ⁇ -GTP by a direct detection method utilizing the reaction between an antibody that binds specifically and GTP protein, or a PCR (polymerase cha in reaction: synthase chain reaction) method Based on a known method selected from such as above, it is possible to simply and reliably detect.
  • the present invention also provides a diagnostic reagent for periodontal disease that contains at least a substance that reacts (interacts) with ⁇ -GTP. '
  • ⁇ -GTP as a marker for periodontal disease can provide a biochemical test / diagnosis method for periodontal disease that can test, diagnose, and predict progression of periodontal disease. Significance, and there is technology that has already been established in the medical field, etc. for measuring or detecting ⁇ -GT ⁇ .
  • the biochemical examination and diagnosis method of periodontal disease can be easily and quickly applied to the medical field. It has the second technical significance that it can be spread.
  • Fig. 1 shows the experimental results (Table).
  • gingival crevicular fluid of a periodontal disease patient is collected from a patient, and the activity of ⁇ -GTP in this gingival crevicular fluid is determined.
  • a method of confirming by a known ⁇ -GTPP activity measurement method which is generally used in laboratory diagnosis of liver diseases and the like at present.
  • ⁇ -GT ⁇ activity is, for example, that ⁇ - (DL- ⁇ -glutamyl) aniline (I) is used as a substrate, amino acid (II) is added as an acceptor, and the azo dye of the reaction product is added. It can be implemented by using the sales kit as appropriate.
  • a specific antigen-antibody reaction between an anti- ⁇ -GTP antibody and a specific ⁇ -GTP protein was used. More preferably, an embodiment in which the expression of the ⁇ -GTPP protein is directly confirmed by a direct detection method (immunological method) and the examination and diagnosis of periodontal disease are performed reliably is performed. You can also.
  • ⁇ -GTPP protein expressed in relation to osteoclast-forming activity or bone destruction is considered to be suitable as a marker for periodontal disease examination.
  • the anti- ⁇ -GTP antibody is not particularly limited as long as it specifically binds to the ⁇ -GT ⁇ protein.
  • ⁇ -GTP was confirmed based on the method of confirming the expression of niRNA encoding ⁇ -GTP protein by a known RT-PCR (polymerase chain reaction) method.
  • RT-PCR polymerase chain reaction
  • any method other than the above can be adopted as an example of the present invention as long as the method can surely detect the ⁇ -GTP protein.
  • a reagent having a configuration containing at least an anti- ⁇ -GTP antibody specifically binding to ⁇ -GTP is used. Conceivable.
  • this reagent is widely interpreted, and furthermore, a test / diagnosis kit provided with this reagent, a biosensor chip having a detection surface provided with a ligand such as an anti-GTP antibody, etc. are also provided. be able to. Example.
  • comparison samples used were extracted teeth from a caries patient without periodontal disease (specimen number 11), and extracted teeth (sample numbers 12 to 14) extracted during orthodontic treatment when the teeth were healthy.
  • ⁇ -GTP activity was measured in the same manner as described above.
  • Figure 1 shows the measurement results.
  • the _GT ⁇ activity in the eluate of the extracted teeth of 10 patients with periodontal disease was high (average 17.79).
  • the ⁇ -GTP activity of the extract of the extracted tooth from a caries patient without periodontal disease has 0 ⁇ -GTP activity, and the extracted tooth was extracted during orthodontic treatment.
  • the average GTP activity was 0.2 U.
  • ⁇ -GTP activity contained in saliva of 10 patients with periodontal disease was measured, and it was confirmed that the activity was about 1 U. From this, it was confirmed that most of y-GTP in the eluate obtained from sample numbers 1 to 10 was derived from gingival crevicular fluid.
  • ⁇ -GTP as a marker for periodontal disease
  • the method for measuring or detecting ⁇ -GTP can be easily and quickly applied to biochemical testing methods and diagnostic reagents for periodontal disease in the medical field, using technologies already established in the medical field, etc. It can be spread. Furthermore, it can provide definitive biochemical examination and diagnosis techniques for periodontal disease, and can be useful for early diagnosis and early treatment of periodontal disease.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne une méthode d'examen et un diagnostic d'examen au moyen desquels on peut estimer de manière appropriée la progression d'une parodontopathie. De manière plus précise cette invention se rapporte à une méthode d'examen d'une parodontopathie dans laquelle on procède à l'évaluation de l'activité de la η-glutamyltranspeptidase qui participe à l'induction de la différenciation des ostéoclastes et qui apparaît dans le fluide parodontal et donne lieu à un flux accru du fait de la parodontopathie; et à un diagnostic pour l'examen de la parodontopathie qui contient au moins une substance qui réagit avec ladite η-glutamyltranspeptidase telle qu'un anticorps anti-η-GTP.
PCT/JP2003/009493 2002-10-11 2003-07-28 Methode d'examen et diagnostic d'examen pour les parodontopathies WO2004033716A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2002298177A JP2004129584A (ja) 2002-10-11 2002-10-11 歯周病の検査方法及び検査診断薬
JP2002-298177 2002-10-11

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WO2004033716A1 true WO2004033716A1 (fr) 2004-04-22

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7432071B2 (en) * 2003-06-30 2008-10-07 Applyed Cell Biotechnologies Inc. Method of diagnosing periodontitis by determining γ-glutamyltranspeptidase

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006121956A (ja) * 2004-10-28 2006-05-18 Ortho Corp 抗γ−ＧＴＰ抗体を有効成分とする機能性食品
JP2014222241A (ja) * 2014-07-10 2014-11-27 コルゲート・パーモリブ・カンパニーＣｏｌｇａｔｅ−Ｐａｌｍｏｌｉｖｅ Ｃｏｍｐａｎｙ 口の健康のための代謝産物およびその使用
WO2017195268A1 (fr) * 2016-05-10 2017-11-16 株式会社アイシーエレクトロニクス Kit de test de parodontite utilisant un miarn comme marqueur, et procédé de test

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990004787A1 (fr) * 1988-10-26 1990-05-03 The Board Of Trustees Of The University Of Illinois Procede de determination du type et de la gravite d'affections du parodonte
WO1990009589A1 (fr) * 1989-02-14 1990-08-23 Xytronyx, Inc. Procede permettant de diagnostiquer une paradontolyse par la presence d'une alanine-aminotransferane
JPH04229198A (ja) * 1990-12-26 1992-08-18 Lion Corp 歯周疾患検査方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990004787A1 (fr) * 1988-10-26 1990-05-03 The Board Of Trustees Of The University Of Illinois Procede de determination du type et de la gravite d'affections du parodonte
WO1990009589A1 (fr) * 1989-02-14 1990-08-23 Xytronyx, Inc. Procede permettant de diagnostiquer une paradontolyse par la presence d'une alanine-aminotransferane
JPH04229198A (ja) * 1990-12-26 1992-08-18 Lion Corp 歯周疾患検査方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CHU, L ET AL: "Role for Recombinant gamma-Glumamyltreansferase from Treponema denticola in Glutathione Metabolism", INFECTION AND IMMUNITY, vol. 71, no. 1, January 2003 (2003-01-01), pages 335 - 342, XP002975853 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7432071B2 (en) * 2003-06-30 2008-10-07 Applyed Cell Biotechnologies Inc. Method of diagnosing periodontitis by determining γ-glutamyltranspeptidase

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