Method for providing information for in vitro diagnosis of periodontal disease and method for detecting Pg bacteria

Abstract

An in vitro diagnosis method for periodontal diseases according to the present invention comprises: bringing a specimen derived from the oral cavity of a subject into contact with an electrode, and conducting electrochemical measurement in anaerobic environment in the presence of a glucogenic amino acid and an electron mediator; and providing, if generation of an electronic current is detected as a result of the electrochemical measurement, information for determining that a periodontal disease has advanced in the oral cavity of the subject. With the in vitro diagnosis method for periodontal diseases according to the present invention, information for determining the state of advance of a periodontal disease can be provided by a simple operation.

Description

[Technical Field]

[0001] The present invention relates to an in vitro diagnostic method for periodontal disease and a method for detecting Pg bacteria. [Background technology]

[0002] Periodontal disease is a general term for diseases of the periodontal tissues, which are the tissues that support the teeth, and is considered a multifactorial disease caused by a variety of factors. Among these many factors, one of the main ones is the infection of periodontal pockets by periodontal pathogenic bacteria. Such bacterial infection leads to the breakdown of periodontal tissues and ultimately to tooth loss.

[0003] It is known that there are many periodontal pathogenic bacteria. Among them, Porphyromonas gingivalis (hereinafter referred to as "Pg bacteria"), Tannerella forsythia (hereinafter referred to as "Tf bacteria"), and Treponema denticola (hereinafter referred to as "Td bacteria") are particularly likely to be the causative bacteria of periodontal disease. Furthermore, they are thought to be involved not only in periodontal disease but also in diseases of other organs. Therefore, research is underway into in vitro diagnostic methods for detecting the activity of these bacteria in the oral cavity of subjects.

[0004] One such method utilizes the trypsin-like enzyme activity of the aforementioned periodontal pathogenic bacteria (Pg, Tf, and Td). That is, this is a technology for diagnosing periodontal disease by detecting the enzyme activity. For example, Patent Document 1 describes a "periodontal disease detection method characterized by using a cysteine protease produced by Porphyromonas gingivalis as a periodontal disease marker for detecting periodontal disease." [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2004 / 106541 Summary of the Invention [Problem to be solved by the invention]

[0006] The periodontal disease detection method described in Patent Document 1 measures enzyme activity, but the activity varies greatly depending on the reaction temperature, the measurement procedure is complicated, and the measurement requires skill, leaving room for improvement.

[0007] Therefore, an object of the present invention is to provide an in vitro diagnostic method for periodontal disease that can provide information for determining the progression of periodontal disease with a simple operation, and a method for detecting Pg bacteria. [Means for solving the problem]

[0008] As a result of extensive research into achieving the above object, the present inventors have found that the above object can be achieved by the following configuration.

[0009] [1] A method for in vitro diagnosis of periodontal disease, comprising: contacting a sample derived from a subject's oral cavity with an electrode; performing electrochemical measurement in an anaerobic environment in the presence of a glycogenic amino acid and an electron mediator; and, if current generation is detected as a result of the electrochemical measurement, providing information for determining that periodontal disease is progressing in the subject's oral cavity. [2] The in vitro diagnostic method for periodontal disease according to [1], wherein the glucogenic amino acid comprises at least one selected from the group consisting of arginine, histidine, aspartic acid, and glutamic acid. [3] The in vitro diagnostic method for periodontal disease described in [1] or [2], wherein the electron mediator comprises at least one selected from the group consisting of flavin mononucleotide, riboflavin, and 2-hydroxy-1,4-napthoquinone. [4] The in vitro diagnostic method for periodontal disease according to any one of [1] to [3], wherein the electron mediator is 2-hydroxy-1,4-naphthoquinone. [5] An in vitro diagnostic method for periodontal disease described in any of [1] to [4], wherein the electrochemical measurement method is a method of controlling the potential of the electrode and measuring the current flowing through the electrode as a function of time. [6] The in vitro diagnostic method for periodontal disease according to [5], wherein the potential is in the range of more than 0 V with respect to a silver-silver chloride electrode and less than the upper limit of the potential window. [7] An in vitro diagnostic method for periodontal disease described in any of [1] to [6], wherein the information for making the judgment includes at least one measurement result selected from the group consisting of the time from the start of the electrochemical measurement to the detection of the current generation, and the maximum current density of the generated current. [8] The in vitro diagnostic method for periodontal disease described in [7], wherein the information for making the judgment includes comparison information between a predetermined reference value and the measurement result. [9] The in vitro diagnostic method for periodontal disease according to any one of [1] to [8], wherein the causative bacteria of periodontal disease include Porphyromonas gingivalis.

[10] A method for detecting Porphyromonas gingivalis, comprising: bringing a sample into contact with an electrode; performing electrochemical measurement in an anaerobic environment in the presence of a glycogenic amino acid and 2-hydroxy-1,4-napthoquinone; and, if the generation of a current is detected as a result of the electrochemical measurement, providing information for determining that the sample contains Porphyromonas gingivalis.

[11] The method for detecting Pg bacteria according to

[10] , wherein the electrochemical measurement method is a method of controlling the potential of the electrode and measuring the current flowing through the electrode as a function of time.

[12] The Pg bacteria detection method according to

[11] , wherein the potential is in the range of more than 0 V versus a silver-silver chloride electrode and less than the upper limit of the potential window.

[13] A Pg bacteria detection method according to any one of

[10] to

[12] , wherein the information for making the judgment includes at least one measurement result selected from the group consisting of the time from the start of the electrochemical measurement to the detection of the current generation, and the maximum current density of the generated current.

[14] The Pg bacteria detection method described in

[13] , wherein the information for making the judgment includes information for comparing the measurement result with a predetermined reference value. [Effects of the Invention]

[0010] The in vitro diagnostic method of the present invention includes contacting a sample derived from the oral cavity of a subject with an electrode, performing electrochemical measurement in an anaerobic environment in the presence of a glucogenic amino acid and an electron mediator, and, if the generation of a current is detected as a result of the electrochemical measurement, providing information for determining whether periodontal disease is progressing in the oral cavity of the subject. According to this method, information for determining the progression of periodontal disease can be obtained by the simple procedure of electrochemically measuring the sample in the presence of a glucogenic amino acid and an electron mediator.

[0011] Furthermore, when the glucogenic amino acid contains at least one selected from the group consisting of arginine, histidine, aspartic acid, and glutamic acid, the resulting generated current tends to be larger, which allows for more sensitive measurements and, as a result, more accurate information can be obtained.

[0012] Furthermore, when the electron mediator contains at least one selected from the group consisting of flavin mononucleotide (FMN), riboflavin (RF), and 2-hydroxy-1,4-naphthoquinone (hereinafter also referred to as "HNQ"), the flow of electrons transferred to the electrode by the metabolism of periodontal pathogenic bacteria tends to be smoother. In other words, the specific electron mediator makes it easier for electrons to be extracted from periodontal pathogenic bacteria. As a result, a larger current is generated and the dynamic range is increased.

[0013] The above tendency is particularly pronounced when the electron mediator contains HNQ. The present inventors have experimentally confirmed that, surprisingly, when the electron mediator contains HNQ, the time until current generation is detected is also shorter. In other words, the time until information is provided can be shortened.

[0014] Additionally, if the electrochemical measurement method is one in which the potential of an electrode is controlled and the current flowing through said electrode is measured as a function of time, the current generation is more easily detected and, as a result, more accurate information is more likely to be obtained.

[0015] Furthermore, when the potential of the controlled electrode is above 0 V (vs. Ag / AgCl: silver-silver chloride electrode) and below the upper limit of the potential window, the flow of electrons from periodontal pathogenic bacteria to the electrode via the electron mediator becomes smoother, enabling more accurate measurements.

[0016] Furthermore, it is preferable that the information for determination includes at least one measurement result selected from the group consisting of the time from the start of electrochemical measurement to the detection of current generation and the maximum current density of the generated current, since quantitative evaluation is easier. For example, when samples are collected from the same subject multiple times at intervals, the progress of periodontal disease can be determined over time by comparing the obtained information.

[0017] Furthermore, when the information for the above judgment includes information comparing the measurement results with predetermined reference values, for example, by using reference values determined for each subject, it becomes easier to judge the progression of periodontal disease. Furthermore, by using reference values determined for each subject's attributes, such as age and gender, the comparison target becomes clear even with a single measurement, making it easier to judge the progression of periodontal disease from the obtained information.

[0018] The Pg detection method of the present invention includes contacting a sample with an electrode and performing electrochemical measurement in an anaerobic environment in the presence of a glucogenic amino acid and HNQ, and providing information for determining that the sample contains Pg if current generation is detected as a result of the electrochemical measurement. The above method uses HNQ as an electron mediator, allowing for specific detection of Pg bacteria with a simple procedure even when the sample contains contaminants and / or other bacteria. Because Pg bacteria, which are periodontal pathogens, can be detected without culturing or other procedures, the method can be easily applied to determining dental treatment strategies.

[0019] Furthermore, if the electrochemical measurement method involves controlling the potential of the electrode and measuring the current flowing through the electrode as a function of time, it is easier to detect the current generated by Pg bacteria, and as a result, more accurate information can be obtained.

[0020] Furthermore, when the controlled potential of the electrode is within a range exceeding 0 V (vs. Ag / AgCl) and less than the upper limit of the potential window, the flow of electrons from the Pg bacteria to the electrode via HNQ becomes smoother, enabling more accurate measurements.

[0021] Furthermore, it is preferable that the information for the determination includes at least one measurement result selected from the group consisting of the time from the start of the electrochemical measurement to the detection of the current generation and the maximum current density of the generated current, since this makes quantitative evaluation easier, and for example, makes it possible to compare the number of Pg bacteria between samples.

[0022] Furthermore, when the information for making a judgment includes information for comparing a predetermined reference value with the measurement result, more accurate measurements can be achieved by creating the reference value based on, for example, a blank sample. [Brief explanation of the drawings]

[0023] [Figure 1] 1 is a flowchart of an in vitro diagnostic method according to an embodiment of the present invention. [Figure 2]1 is a chronoamperogram showing the effect of histidine on the generated current. [Figure 3] 1 is a chronoamperogram showing the effect of aspartic acid on the generated current. [Figure 4] 1 is a chronoamperogram showing the effect of glutamate on the generated current. [Figure 5] FIG. 10 is a diagram showing Pg-specific current generation when aspartic acid is used as a glycogenic amino acid. [Figure 6] These are the experimental results of investigating the effect of HNQ on current generation. [Figure 7] These are the experimental results of investigating the effect of FMN on current generation. [Figure 8] These are the results of an experiment investigating the effect of RF on current generation. DETAILED DESCRIPTION OF THE INVENTION

[0024] The present invention will be described in detail below. The following description of the components may be based on a representative embodiment of the present invention, but the present invention is not limited to such an embodiment. In this specification, a numerical range expressed using "to" means a range that includes the numerical values before and after "to" as the lower and upper limits.

[0025] [In vitro diagnostic methods] FIG. 1 is a flowchart of an in vitro diagnostic method according to an embodiment of the present invention. First, a sample derived from the oral cavity of a subject is brought into contact with an electrode, and electrochemical measurement is performed in the presence of a glucogenic amino acid and an electron mediator in an anaerobic environment (step S11).

[0026] The specimen is not particularly limited as long as it is derived from the oral cavity of the subject, but preferably includes saliva, plaque, blood, pus, a mixture of these, etc. Among these, a specimen containing saliva is preferred because it is more non-invasive and easier to collect.

[0027] The specimen may contain water, electrolytes, and the like as components other than those mentioned above. The electrolyte is not particularly limited, and any known electrolyte can be used, provided that the electrolyte does not preferably contain any organic substance other than the glucogenic amino acids described below.

[0028] The electrochemical measurement is carried out in the presence of a glucogenic amino acid. The electrochemical measurement in the presence of a glucogenic amino acid typically includes adding a glucogenic amino acid to a sample. Examples of glycogenic amino acids include alanine, glycine, serine, threonine, cysteine, tryptophan, isoleucine, methionine, valine, aspartic acid, arginine, glutamic acid, histidine, proline, tyrosine, and phenylalanine. Among these, from the viewpoint of increasing the generated current (density), at least one selected from the group consisting of arginine, histidine, aspartic acid, and glutamic acid is preferred, and at least one selected from the group consisting of arginine and histidine is more preferred.

[0029] Histidine is preferred over aspartic acid and glutamic acid because it has higher water solubility and tends to be more uniform when added to a sample. High water solubility also makes it unnecessary to add an acid or the like to the sample, which tends to reduce background current during measurement. Furthermore, the present inventors have experimentally confirmed that when the glycogenic amino acid is histidine, the generated current derived from Pg bacteria tends to be larger (see Examples).

[0030] When aspartic acid or glutamic acid is used as the glucogenic amino acid, a more uniform sample can be obtained by adding an acid (e.g., hydrochloric acid) to the sample to adjust the pH to 6.0 or less (e.g., about 5.2). The content of glucogenic amino acids in a sample is not particularly limited, but is generally preferably 0.1 to 1000 mM.

[0031] Furthermore, electrochemical measurements are performed in the presence of an electron mediator. Although some periodontal pathogenic bacteria generate current in an anaerobic environment and transfer that current to an extracellular electron acceptor (e.g., an anode electrode), electrochemical measurements are preferably performed in the presence of an electron mediator, as this allows for smoother electron transfer. As in the case of the glucogenic amino acids, electrochemical measurement in the presence of an electron mediator typically involves adding the electron mediator to the sample.

[0032] The electron mediator that can be used is not particularly limited, and known electron mediators can be used. The electron mediator is preferably water-soluble, and in particular, when the electron mediator contains at least one selected from the group consisting of flavin mononucleotide, riboflavin, and 2-hydroxy-1,4-napthoquinone (HNQ), a larger generated current (density) can be obtained.

[0033] In particular, the present inventors' research has surprisingly revealed that when the electron mediator contains HNQ, the generated current derived from Pg bacteria, among oral bacteria, is significantly increased. This suggests that the generated current can be specifically detected from Pg bacteria even when a sample contains Porphyromonas gingivalis (Pg bacteria), Streptococcus mutans (Sm bacteria), and Capnocytophaga ochracea (Co bacteria).

[0034] Although the above description has been given of the case where the sample contains a glucogenic amino acid and an electron mediator, in the extracorporeal diagnostic method, it is sufficient that the electrochemical measurement is performed in the presence of the glucogenic amino acid and the electron mediator, and the glucogenic amino acid and the electron mediator do not necessarily need to be added to the sample. In such a case, for example, the glucogenic amino acid may be immobilized on the surface of an electrode.

[0035] The electrochemical measurement method is preferably a method in which the potential of the electrode is controlled and the current is measured as a function of time, such as amperometry, cyclic voltammetry, linear sweep voltammetry, and square wave voltammetry.

[0036] The material of the electrode is not particularly limited, and known electrodes for electrochemical measurement can be used.The material of the electrode can be, for example, ITO (indium tin oxide), noble metals (gold (Au), silver (Ag), platinum (Pt), palladium (Pd), rhodium (Rh), iridium (Ir), ruthenium (Ru), etc.), copper (Cu), aluminum (Al), tungsten (W), molybdenum (Mo), chromium (Cr), titanium (Ti), nickel (Ni), etc. Also, carbon materials, such as carbon and graphite (graphene), etc., can be used.In addition, boron-doped diamond electrodes are also preferred because of their wide potential window.

[0037] For the electrochemical measurement, a general electrochemical measurement device can be used, for example, a three-electrode electrochemical measurement device in which a working electrode, a counter electrode, and a reference electrode are housed in a cell. The temperature at which the electrochemical measurement is performed is not particularly limited, but it can be performed at a temperature similar to that at the location where the sample is collected, or the temperature may be controlled. In this case, the sample temperature is preferably 10 to 40°C. The measurement may be carried out in a general anaerobic glove box (anaerobic chamber), preferably in an oxygen-free state.

[0038] In order to detect a current by receiving electrons generated by the anaerobic metabolism of periodontal pathogenic bacteria at the electrode, the potential of the electrode (working electrode) is preferably controlled within a range exceeding 0 V (vs. Ag / AgCl) and below the upper limit of the potential window. From the viewpoint of easily obtaining a larger generated current or easily generating a current in a shorter time, the potential of the electrode is preferably set to 0 to +0.6 V (vs. Ag / AgCl).

[0039] If the electrochemical measurement detects the generation of a current (step S12: Yes), the presence of periodontal pathogenic bacteria in the sample is suggested, and information is provided for determining whether periodontal disease is progressing in the subject's oral cavity (step S13).

[0040] On the other hand, if no current generation is detected by this electrochemical measurement (step S12: No), the diagnosis ends and no information is provided for determining whether periodontal disease is progressing.

[0041] The above information is based on measurement results and its form is not particularly limited, but if it is information (hereinafter also referred to as "specific information") that includes at least one measurement result selected from the group consisting of the time from the start of electrochemical measurement to the detection of current generation and the maximum current density of the generated current, it is more preferable in that it is more likely to be information that contributes to quantitative evaluation of the progression of periodontal disease.

[0042] The above-mentioned specific information is a numerical value related to the content of periodontal pathogenic bacteria in the sample, and by comparing this with, for example, values from healthy individuals or with past values from the same subject, it is likely to provide information that will be useful in determining the degree of periodontal disease occurring in the subject's oral cavity and its progression over time.

[0043] (Variation) A variation of the in vitro diagnostic method according to the above embodiment is an in vitro diagnostic method in which, when current generation is detected, the obtained measurement result is compared with a reference value, and information including the comparison information is provided as information for determining whether periodontal disease is progressing in the subject's oral cavity.

[0044] Examples of reference values in ex vivo diagnostic methods include the results of measurements made by the same method using samples from healthy subjects, and past test results of the same subjects. Examples of comparative information include the difference between specific information measured by the same method using a sample from a healthy subject and the specific information of the sample.

[0045] More specifically, examples of such information include the difference between the time from the start of electrochemical measurement until current generation is detected when a sample from a healthy individual is measured using the same method, and the time measured using the same method on a sample obtained from a subject. If the time measured on the sample is shorter and the difference is larger, this can be used as information for determining that the sample contains a higher amount of periodontal pathogenic bacteria. Note that the above comparison information is just an example, and other information may also be used.

[0046] [Pg bacteria detection method] A method for detecting Pg bacteria according to an embodiment of the present invention includes contacting a sample with an electrode, performing electrochemical measurement in an anaerobic environment in the presence of a glycogenic amino acid and 2-hydroxy-1,4-napthoquinone, and providing information for determining that the sample contains Porphyromonas gingivalis if current generation is detected as a result of the electrochemical measurement.

[0047] As described in the Examples below, the present inventors have experimentally confirmed that when electrochemical measurements are performed in the presence of a glucogenic amino acid and HNQ in an anaerobic environment, a large generated current can be detected specifically for Pg bacteria.

[0048] For example, in the field of periodontal disease prevention, there is a strong demand for a simple method to detect the proliferation of the particularly influential periodontal pathogenic bacteria known as the "red complex." As already explained, the enzymatic method is not simple due to its complicated procedures and large variability. However, the above method is very simple and can detect Pg bacteria almost specifically.

[0049] The conditions for electrochemical measurement, etc., can be the same as those already described as the electrochemical measurement method in the in vitro diagnostic method according to the first embodiment, and the preferred embodiments are also the same, so description thereof will be omitted.

[0050] The specimen in this detection method is not limited to specimens derived from the oral cavity of a subject, but may also be specimens cultured for research purposes, environmental specimens (water and garbage), etc. If the specimen is not liquid, it can be extracted and purified using water or the like before use. [Example]

[0051] The present invention will be described in more detail below with reference to examples. The materials, amounts used, ratios, treatment contents, treatment procedures, etc. shown in the following examples can be changed as appropriate without departing from the spirit of the present invention. Therefore, the scope of the present invention should not be construed as being limited by the examples shown below.

[0052] [Effect of glycogenic amino acids on the generated current] (Experiment 1: Reference Example) The effect of glycogenic amino acids on current generation was confirmed using Pg bacteria. A three-electrode electrochemical cell (working electrode: ITO, counter electrode: platinum, reference electrode: Ag / AgCl) was used to measure a total of 5 mL of sample. The temperature was 37°C. The specimen was prepared by adding 10 mM histidine or glucose to DM liquid medium without yeast extract, and then adding Pg bacterial solution (OD 600 The measurement was carried out by chronoamperometry in a COY anaerobic chamber filled with 100% nitrogen.

[0053] Figure 2 shows chronoamperograms showing the effect of histidine on the current generated. "PG-w / Histidine" indicates the results for a sample containing the glycogenic amino acid histidine, while "PG-w / Glucose" indicates the results for a sample containing glucose instead of histidine. As shown in Figure 2, current generation was detected immediately after the addition of the Pg bacterial solution in samples containing histidine. On the other hand, current generation was detected in samples containing glucose, but the magnitude was smaller and it took longer for the current to be generated.

[0054] (Experiment 2: Reference Example) Next, we tested using aspartic acid and glutamic acid instead of histidine as glucogenic amino acids. Because aspartic acid and glutamic acid have low solubility in water, they were dissolved in 0.5 M HCl. Therefore, the pH of the DM liquid medium was adjusted to approximately 5.2.

[0055] Figure 3 shows chronoamperograms illustrating the effect of aspartic acid on the generated current. Figure 4 shows chronoamperograms illustrating the effect of glutamic acid on the generated current. In both cases, current generation was detected immediately after the addition of the Pg bacterial solution.

[0056] The results in Figures 2 to 4 indicate that when the sample contains a glucogenic amino acid, current generation is rapidly detected upon addition of the Pg bacterial solution. Furthermore, when the glucogenic amino acid is histidine, the generated current is larger and the time until current generation is shorter than when the glucogenic amino acid is aspartic acid or glutamic acid.

[0057] [Histidine-specific current generation by Pg bacteria] (Experiment 3: Reference Example) Pg bacterial solution (OD 600 :0.1) instead of Pg bacterial solution (OD 600 :0.5), Streptococcus mutans bacterial solution (OD 600 :0.5), and Capnocytophaga ochracea (OD 600 The generated current was investigated in the same manner as in Experiment 1, except that a 0.5 (.0.5) was used.

[0058] The results are shown in Figure 5. Figure 5 shows that current generation specific to Pg bacteria can be detected in samples containing aspartic acid.

[0059] [Effect of electron mediators on current generation] (Experiment 4: Example) Instead of a three-electrode electrochemical cell (working electrode: ITO, counter electrode: platinum, reference electrode: Ag / AgCl), an electrochemical measurement plate with three electrodes printed on the bottom of a 96-well plate was used to investigate the effect of the electron mediator on current generation. The electron mediators used were HNQ, FMN, and RF, with concentrations of 10 μM, 50 μM, and 100 μM, respectively. 600 :0.5), and other conditions were the same as in Experiment 1.

[0060] Figure 6 shows the experimental results when HNQ was used, Figure 7 shows FMN, and Figure 8 shows RF. The results in Figures 6 to 8 show that when any of the electron mediators was used, a larger current was obtained quickly after the addition of the bacterial solution (compare with Figure 2). In particular, when HNQ was used, a larger current was obtained than when FMN and RF were used, and this tendency was particularly noticeable at concentrations exceeding 10 μM. [Industrial Applicability]

[0061] According to the in vitro diagnostic method for periodontal disease of the present invention, saliva can be used as a sample, and information for determining the progression of periodontal disease can be obtained in a simple manner. The information provided by the in vitro diagnostic method of the present invention can be used not only for dentists to decide on treatment plans, but also for patients to manage their own oral health at home or in remote locations.

[0062] Furthermore, the Pg detection method of the present invention enables the detection of Pg-specific current generation by using a combination of a glycogenic amino acid and HNQ. This method is useful not only for formulating dental treatment strategies but also in experimental research.

Claims

1. Adding a glucogenic amino acid and an electron mediator to a sample derived from the oral cavity of a subject, bringing the sample into contact with an electrode, and performing electrochemical measurement in an anaerobic environment; A method for providing information for in vitro diagnosis of periodontal disease, comprising: providing information for determining that periodontal disease is progressing in the subject's oral cavity if current generation is detected as a result of the electrochemical measurement.

2. A method for providing information for in vitro diagnosis of periodontal disease as described in claim 1, wherein the glycogenic amino acid includes at least one selected from the group consisting of arginine, histidine, aspartic acid, and glutamic acid.

3. The method for providing information for in vitro diagnosis of periodontal disease according to claim 1, wherein the electron mediator comprises at least one selected from the group consisting of flavin mononucleotide, riboflavin, and 2-hydroxy-1,4-napthoquinone.

4. The method for providing information for in vitro diagnosis of periodontal disease according to any one of claims 1 to 3, wherein the electron mediator is 2-hydroxy-1,4-naphthoquinone.

5. 2. A method for providing information for in vitro diagnosis of periodontal disease as described in claim 1, wherein the electrochemical measurement method is a method of controlling the potential of the electrode and measuring the current flowing through the electrode as a function of time.

6. 6. The method for providing information for in vitro diagnosis of periodontal disease according to claim 5, wherein the potential is in the range of more than 0 V versus a silver-silver chloride electrode and less than the upper limit of the potential window.

7. A method for providing information for in vitro diagnosis of periodontal disease according to any one of claims 1 to 3, wherein the information for making the judgment includes at least one measurement result selected from the group consisting of the time from the start of the electrochemical measurement to the detection of the current generation, and the maximum current density of the generated current.

8. The method for providing information for in vitro diagnosis of periodontal disease according to claim 7 , wherein the information for making the judgment includes information for comparing the measurement results with a predetermined reference value.

9. The method for providing information for in vitro diagnosis of periodontal disease according to any one of claims 1 to 3, wherein the causative bacteria of periodontal disease include Porphyromonas gingivalis.

10. adding a glucogenic amino acid and 2-hydroxy-1,4-naphthoquinone to a sample, bringing the sample into contact with an electrode, and performing electrochemical measurement in an anaerobic environment; and providing information for determining that the sample contains Porphyromonas gingivalis when current generation is detected as a result of the electrochemical measurement.

11. 11. The method for detecting Pg bacteria according to claim 10, wherein the electrochemical measurement method is a method in which the potential of the electrode is controlled and the current flowing through the electrode is measured as a function of time.

12. The Pg bacteria detection method according to claim 11, wherein the potential is in the range of more than 0 V versus a silver-silver chloride electrode and less than the upper limit of the potential window.

13. The Pg bacteria detection method according to any one of claims 10 to 12, wherein the information for making the determination includes at least one measurement result selected from the group consisting of the time from the start of the electrochemical measurement to the detection of the current generation, and the maximum current density of the generated current.

14. The Pg bacteria detection method according to claim 13 , wherein the information for making the determination includes information for comparing the measurement result with a predetermined reference value.

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