WO2001073441A1 - Kit de detection d'infections fongiques de prelevements dans la cavite orale - Google Patents

Kit de detection d'infections fongiques de prelevements dans la cavite orale Download PDF

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Publication number
WO2001073441A1
WO2001073441A1 PCT/JP2001/002470 JP0102470W WO0173441A1 WO 2001073441 A1 WO2001073441 A1 WO 2001073441A1 JP 0102470 W JP0102470 W JP 0102470W WO 0173441 A1 WO0173441 A1 WO 0173441A1
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WO
WIPO (PCT)
Prior art keywords
oral
antigen
fungal
candida
derived
Prior art date
Application number
PCT/JP2001/002470
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English (en)
Japanese (ja)
Inventor
Tadakazu Yamauchi
Kamon Shirakawa
Original Assignee
Mochida Pharmaceutical Co., Ltd.
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.)
Filing date
Publication date
Application filed by Mochida Pharmaceutical Co., Ltd. filed Critical Mochida Pharmaceutical Co., Ltd.
Priority to AU42810/01A priority Critical patent/AU4281001A/en
Publication of WO2001073441A1 publication Critical patent/WO2001073441A1/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
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56961—Plant cells or fungi
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • Oral candidiasis is well known as a fungal infection occurring in the oral cavity.
  • Oral candidiasis is an infectious disease caused by Candida bacteria, and presents various clinical symptoms such as subjective symptoms such as oral pain, burning sensation, and dryness, and objective findings such as vitiligo, redness, and erosion of the oral mucosa. You.
  • kits or methods that solve the above-mentioned problems in the prior art, specifically, measuring the amount of antigens of oral fungal resident bacteria, being unaffected by therapeutic agents, rapid
  • An object of the present invention is to provide the kit or the method, which satisfies at least one of the properties, simplicity, and accuracy. Disclosure of the invention
  • a fungal-derived antigen in an oral specimen is quantified by using an immunological assay using an antibody specific to a fungal-derived antigen, and the oral fungal It is a kit for diagnosing oral mycosis, denture stomatitis or periodontal disease, characterized by quantifying the bacterial count or activity.
  • a pathological threshold value of an oral fungal-derived antigen is set in advance from a relationship between the number or activity of oral fungal bacteria and a quantitative value of an oral fungal-derived antigen, Using a quantitative value of an antigen derived from an oral fungus in an oral specimen of a subject as a pathological index, and comparing this value with the threshold value, wherein the disease is derived from an oral fungus in the subject. This is a method to determine the physical state.
  • fungus is a common name for the fungus Eumy cota, which is a common name that includes yeast. More specifically, fungi belonging to the genus Candida include Candida albicans, Candida glabura, Candida parapsilosis, Candida's tropicalis, Candida's crussey, Candida, Gilliermondi, and Candida lusitaniae. Fungi belonging to the genus Alpergillus include Alpergillus fumigas, Alpergillus niger, Aspergillus flavans, Alpergillus nidulans, Aspergillus nigerans and the like. Fungi belonging to the genus Cryptococcus include Cryptococcus Neoformans and the like.
  • Periodontal disease is a general term for lesions that cause periodontal tissue. Usually, lesions are broadly divided into gingivitis, which is limited to the marginal gingiva, and periodontitis, which involves resorption of alveolar bone. "Detection” refers to, for example, measuring the amount of an antigen derived from a fungus in an oral sample using the immunological measurement method of the present application, and comparing the measured value with a preset pathological threshold. Doctors, dentists, laboratory technicians, caregivers, patients themselves, etc., evaluate or evaluate the disease, understand the condition of the disease or the state of progression to the disease, and determine the healthy state Or to do.
  • a cut-off value is set, and if the antigen amount exceeds this set value, it is determined that the person is in a condition derived from oral fungi. If it is equal to or less than the set value, it can be determined that the person is healthy.
  • the threshold in this case is, for example, 100 to 1000 pg / mL, preferably 100 to 50 Q pg / mL, more preferably 100 to 300 pg / mL, and still more preferably 150 to 250 pg / mL.
  • the cutoff value is set, for example, it is 1000 pgZmL or less, preferably 500 pgZmL or less, and more preferably 300 pgZmL or less.
  • the previously set pathological threshold value fluctuates according to the oral specimen collecting method described above, and is desirably set for each oral specimen collecting method.
  • the obtained oral sample is heat-extracted, and the precipitate is removed by centrifugation.
  • the supernatant be used as a measurement sample after filtration and removal. This allows for highly sensitive measurements.
  • a pretreatment such as addition of a surfactant, addition of a pH adjusting agent such as a buffer, addition of a chelating agent, and deproteinization can also be performed.
  • a pretreatment such as addition of a surfactant, addition of a pH adjusting agent such as a buffer, addition of a chelating agent, and deproteinization can also be performed.
  • a pretreatment such as addition of a surfactant, addition of a pH adjusting agent such as a buffer, addition of a chelating agent, and deproteinization can also be performed. .
  • a method of storing the threshold value in the storage means of the computer retrieving the threshold value from the storage means into the calculation means, comparing the result with the measurement result, and judging a disease state derived from the oral fungus of the subject (in the storage means)
  • the threshold value may be stored as data that changes for each predetermined condition or may be stored as an appropriate input value.
  • a computer to determine disease state, monitor, administer medication, manage oral flora The method may be performed.
  • Those skilled in the art can use the immunological assay method used in the present invention as a measurement kit according to a conventional method.
  • the absorbance at 550 nm was measured and adjusted to 20 OD / mL, which was designated as 20 mgZmL.
  • Screening of the purified polyclonal antibody was performed using the reactivity of the antibody with purified mannan derived from various fungi as an index. After dispensing the purified mannan 100 from various fungi into a micro-tie Yuichi plate (Immu nomodu 1 e Maisorp, Nalge Nunc Internatinal, Rochester, NY, USA), 70 ° C, heated for 30 minutes to solidify. The plate was coated with 0.5% serum albumin (BSA) and washed to obtain a purified mannan-immobilized plate.
  • BSA serum albumin
  • each antibody solution (0.3 g / mL) to be screened is added to the purified mannan-immobilized plate and reacted at room temperature for 1.5 hours.
  • HRP-labeled antibody against polyclonal antibodies A giant immunoglobulin antibody (Sigma-Aldrich, St. Louis, MS, USA) was reacted for 1 hour at room temperature.
  • the sample processing solution used for heat treatment of oral samples is EDTA disodium 11 g And 0.5 mL of Tween 20 were prepared with 0.1 M phosphate buffer (pH 7.0) to make a total volume of 50 OmL.
  • the wells were washed 5 times with a washing solution, and a TMB substrate color developing solution (10 O LZ) was added, and the enzyme reaction was carried out at room temperature. After 15 minutes, the reaction was stopped by the addition of a stop solution (100 ⁇ l), and the absorbance at 450 nm was measured.
  • the amount of Candida mannan antigen was measured by the ELISA method described in 5. of the above example using saliva collected from an oral dental outpatient outpatient.
  • a score indicating the amount of colonies was obtained according to the protocol, in which a sample collected from an intraoral lesion using a cotton swab was examined by the CHROMag method (Kanto Chemical).
  • Figure 3 shows the Candida mannan antigen concentration and the results of the CHROM agar method. As a result, the Candida mannan antigen concentration showed a good correlation with the score of the CHROM agar method.
  • the use of the kit or method of the present invention enables the diagnosis of oral mycosis, denture stomatitis or periodontal disease.

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

Abstract

L'invention concerne un kit de dosage de prélèvements oraux par une méthode d'immunodosage à l'aide d'un anticorps spécifique à un antigène rencontré dans des champignons, lequel est utile à la détection précoce d'infections fongiques dans la cavité orale et au traitement rapide de celles-ci.
PCT/JP2001/002470 2000-03-27 2001-03-27 Kit de detection d'infections fongiques de prelevements dans la cavite orale WO2001073441A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU42810/01A AU4281001A (en) 2000-03-27 2001-03-27 Kit for detecting fungal infection of specimens in oral cavity

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2000-087443 2000-03-27
JP2000087443 2000-03-27

Publications (1)

Publication Number Publication Date
WO2001073441A1 true WO2001073441A1 (fr) 2001-10-04

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2001/002470 WO2001073441A1 (fr) 2000-03-27 2001-03-27 Kit de detection d'infections fongiques de prelevements dans la cavite orale

Country Status (2)

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AU (1) AU4281001A (fr)
WO (1) WO2001073441A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016070702A (ja) * 2014-09-26 2016-05-09 花王株式会社 口腔内の乾燥状態の評価方法
JP2017129456A (ja) * 2016-01-20 2017-07-27 国立大学法人信州大学 免疫力検査方法および免疫力検査キット
KR101987754B1 (ko) * 2018-12-24 2019-06-12 주식회사 덴오믹스 구강 질환 검사용 용기 및 이를 포함하는 검사 키트

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS62502193A (ja) * 1985-03-12 1987-08-27 コラル・ソシエダデ・ブラシレイラ・デ・ペスキサス・エ・デセンボルビメント・リミタダ 抗原、抗体およびその用途

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS62502193A (ja) * 1985-03-12 1987-08-27 コラル・ソシエダデ・ブラシレイラ・デ・ペスキサス・エ・デセンボルビメント・リミタダ 抗原、抗体およびその用途

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
MATTHEW P. HOFFMAN ET AL.: "Analysis of candida albicans adhesion to salivary mucin", INFECTION AND IMMUNITY, vol. 61, no. 5, 1993, pages 1940 - 1949, XP002942223 *
SEIKO NAGUMO: "Kou candida albicans koutai ni yoru candida shou no shindan", EISEI KENSA, vol. 38, no. 9, 1997, pages 1207 - 1210, XP002942224 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016070702A (ja) * 2014-09-26 2016-05-09 花王株式会社 口腔内の乾燥状態の評価方法
JP2017129456A (ja) * 2016-01-20 2017-07-27 国立大学法人信州大学 免疫力検査方法および免疫力検査キット
KR101987754B1 (ko) * 2018-12-24 2019-06-12 주식회사 덴오믹스 구강 질환 검사용 용기 및 이를 포함하는 검사 키트

Also Published As

Publication number Publication date
AU4281001A (en) 2001-10-08

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