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Periodontopathic bacteria detection method

A technology for periodontal disease and bacteria, which is applied in the field of detection of periodontal disease bacteria, can solve the problems of heavy burden on patients and the inability to check multiple patients at the same time, and achieve the effect of convenient analysis and determination

Pending Publication Date: 2021-12-03
ADTEC INC A OF DE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods require not only time, labor, and cost but also special facilities and skills, and thus cannot simultaneously examine multiple patients in an easy and quick manner, and the burden on patients is also great

Method used

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  • Periodontopathic bacteria detection method
  • Periodontopathic bacteria detection method
  • Periodontopathic bacteria detection method

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0156] (Preparation of the first compound and the second compound)

[0157] As the compound (first compound) having antioxidant effect, sampling L- ascorbic acid, L- cysteine ​​hydrochloride and glutathione each 62.5 mM, 125 mM, 250 mM, 500 mM and 1000 mM, and each a first compound samples were dissolved in 50 mM tris- maleate buffer pH 8.5, and thereafter the pH was adjusted to bring the pH became 8.5.

[0158] As the compound (a second compound) serves to protect SH group and disulfide bond cleavage of the DTT, thioglycolic acid, thioglycerol, mercaptoethanol and TCEP were each dissolved in 50 mM tris- maleate buffer solution pH 8.5 in to 62.6mM, 125 mM, 250 mM, 500 mM and 1000 mM concentration, and the pH adjusted thereafter so that the pH became 8.5.

[0159] (Preparation of Substrate)

[0160]As the substrate, N-α-benzoyl-DL-arginine-2-naphthamide hydrochloride (obtained from Sigma-Aldrich JapanLLC) is dissolved in 50 mm Tris-maleate buffer solution pH 8.5. The substrate conc...

Embodiment 1

[0168] (Example 1) Dry retention method of the first compound or the second compound alone

[0169] (A) Dry retaining of substrates and first compounds or second compounds

[0170] The preparative first compound or the second compound and the substrate were mixed in a ratio of 1: 9, and 10 mm of a tray (obtained from Advantec Co., Ltd.) was impregnated with 80 μL of the mixture / disc. The tray was then dried overnight at 25 ° C.

[0171] (B) Preparation of P.G

[0172] After 24 hours of culture, the bacterial culture fluid was diluted 10 times with 50 mm Tris-maleate buffer solution pH 8.5 to prepare 1.0 × 10 6 CFU / ml, 1.0 × 10 5 CFU / ml, 1.0 × 10 4 CFU / ML and 1.0 × 10 3 CFU / ml of bacteria suspension.

[0173] (C) test method

[0174] Two tray were prepared, which was impregnated with respective concentrations of the first compound or the second compound and the substrate and then equipped with 80 μL of the prepared pg bacteria suspension, and left for 2 minutes, 5 minutes...

Embodiment 2

[0182] (Example 2) Explanation of a solution method of a first compound or a second compound alone

[0183] (A) dry retention of substrate

[0184] The preparative substrate was mixed into 50 mm Tris-maleate buffer solution pH 8.5 in a volume ratio of 9: 1 and 10 mm (from Advantec Co., Ltd.) with 80 μl of mixture / disc. get). The tray was then dried overnight at 25 ° C.

[0185] (B) Preparation of P.G bacteria

[0186] The prepared first compound or the second compound was diluted 9 times with a 50 mM Tris-maleate buffer solution pH 8.5 to obtain a dilution solution of the compound.

[0187] After 24 hours of culture, the bacterial culture solution was continuously diluted 10 times with a diluted compound solution to prepare 1.0 × 10 6 CFU / ml, 1.0 × 10 5 CFU / ml, 1.0 × 10 4 CFU / ML and 1.0 × 10 3 CFU / ml of bacteria suspension.

[0188] (C) test method

[0189] Two trays were prepared, which was impregnated with a substrate and then equipped with 80 μL of the prepared Pg bac...

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Abstract

The present invention makes it possible to simply, rapidly, and with high sensitivity detect periodontopathic bacteria at room temperature and in under ten minutes (approximately 2-8 minutes). The arginine-specific peptidase activity of three strains of periodontopathic bacteria, i.e., Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, are analyzed at room temperature, in under ten minutes, and with high sensitivity. A first compound that has an anti-oxidizing effect and / or a second compound that protects SH groups (mercapto groups) and has a disulfide bond-cleaving effect are present during analysis of a sample. The first compound is at least one among L-ascorbic acid, L-cysteine hydrochloride, and glutathione, and the second compound is at least one among DTT, thioglycolic acid, thioglycerol, mercaptoethanol, and TCEP.

Description

Technical field [0001] Method for detecting periodontal disease bacteria present invention relates to, and more particularly relates to a method for detecting periodontal disease bacteria does not require special apparatus, which allows periodontal disease bacteria at high temperature measured at room temperature in less than 10 within minutes (approximately 2-8 minutes) are detected. Background technique [0002] It is said that about 80% of adults in Japan by the impact of periodontal disease. Periodontal disease causes tooth loss, and already it is clear that accompanied not only cause taste dysfunction and abnormal secretion of saliva, but also cause abnormal central nervous system and the autonomic nervous system. In addition, in recent years, periodontal disease has been noted that a variety of systemic diseases such as coronary heart disease risk factors, such as cerebral infarction. [0003] Periodontal bacterial infection and inspection items inflammation inspection meth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12Q1/37G01N33/48G01N21/77G01N21/78
CPCC12Q1/04C12Q1/37G01N2333/20G01N2333/952G01N33/56955G01N33/532G01N21/78
Inventor 山下友里板垣初惠森若专太小林薰小林行治
Owner ADTEC INC A OF DE
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