TW200948966A - Oral care methods and systems - Google Patents

Abstract

This invention relates to methods of assessing the bioflora of the mouth and of providing appropriate treatment utilizing a basic amino acid in accordance with the assessment.

Description

200948966 VI. INSTRUCTIONS: This application claims the priority of the US Patent Application No. 61/〇27,437 filed on February 9th, 2008. It also requests the application on February 9, 2008.

The contents of the applications of 61/027, 431, 61/027, 420, and 61/027, 435, are incorporated herein by reference in their entirety. TECHNICAL FIELD OF THE INVENTION The present invention relates to (d) a method for measuring the relative amount of caries and bacteria decomposing arginine in a mouthpiece containing a composition of _ or a salt-based amino acid (for example, as part of a dental care method) . 4 [Prior Art] Arginine and other amidine acids have been suggested for oral health care and are of great benefit to the limbs. Available in the market » The arginine toothpaste is DenClude8 with CaviStat8 and

ProClude® (which contains arginine and hydrogencarbonate). The type of biological flora in the mouth usually plays an important role in the formation of cavities and oral hygiene. For example, it is generally assumed that an important factor in the beneficial effects of arginine is that it can be metabolized with other basic amino acids by specific species of bacteria [eg blood chain 20 _ (& face)], and these bacteria do not produce Contact with the crane tooth bacteria [eg Streptococcus mutans (& compete with the teeth and the position in the mouth. The bacteria that break down arginine can use ammonia and other basic amino acids 200948966 to produce ammonia: thus improving their environment The pH value; and the lignin metabolism of sugar produces lactic acid, which tends to reduce the minerals of the teeth.

Quality 'final guide ship? . It is useful to have P 5 10 15 f T to be effective in monitoring the smear biopsy. For example, it is useful to determine the effectiveness of the treatment and the treatment. SUMMARY OF INVENTION The present invention provides a method for assessing the fascination of the biological flora in the mouth and is simple in the first method of the body tube 1 > (Decomposition of arginine fine 5, the present invention determines the plaque nitrogen production amount to confirm In another specific example, φ, the sputum bacteria lacks 4 τ '. The present invention determines the plaque lactic acid amount to determine the ff group, in the other example (PCR) (for example) Quantification g of the present invention using a polymerase chain backwash (in the case of PCR) 1) in the mouth (for example, plaque or in another 'example to identify the mouth (for example, the use of reverse transcriptase in the present invention) PCR (RT-PCB in another 'plaque or saliva) of the biological flora. In the body example, the post-probe) to use the antibody probe (for example, fluorescent anti-Chu, for example, The abundance of the abundance of at least a species of cariogenic bacteria (for example, the genus of serotonin bacteria (for example, bacteria): 20 200948966

In another embodiment, and in accordance with the indication of treatment, one of the methods described above is used to assess the method of the invention, and the treatment of the tooth is continued. In particular, it is used to detect the potentially harmful changes in the plaque ecology. There is a residual amount or significant demineralization or damage before entering.

Ίο 15 & months thus provide methods for improving oral health, such as a. reducing or inhibiting the formation of dental caries; b. reducing or inhibiting demineralization of teeth and promoting mineralization; c. treatment 'reducing or inhibiting early dental shafts D. Reduce allergic reactions to teeth; e• Reduce or inhibit gingivitis; f· Promote healing of ulcers or wounds in the mouth; g. Reduce the amount of acid-producing bacteria; h. Increase the relative amount of bacteria that break down arginine; To inhibit the formation of microbial biofilms in the oral cavity; j. to promote and/or maintain plaque at a pH of at least 5.5 after sugar stimulation; k. reduce plaque build-up; l. treat, alleviate or reduce dry mouth; 20 200948966 m. Whitening teeth; η. Enhancing general health, including cardiovascular health (eg 'utilizing the possibility of reducing systemic infections through the oral tissues'); 〇 making the teeth immune to caries bacteria and their effects; Tooth and mouth and/or q. reduce tooth erosion; the method comprises measuring the bacterial flora of the mouth (eg, using any of the methods described above), if necessary, administering an effective amount of an amino acid (eg, Arginine) or salt thereof in oral care products. The invention further provides for the manufacture of a medicament using a basic amino acid which is free or salt-formed, as measured by the method according to the invention, wherein the oral biota contains an increase in the amount of ore-producing bacteria and/or an increase in the amount of lactic acid, and/or decomposition Patients with low levels of arginine bacteria and/or low plaque ammonia production improve oral health. The invention further provides a method of enhancing the aesthetics of the mouth (wherein enhancing aesthetics may include, for example, making the teeth whiter and/or reducing bad breath), the method comprising measuring the oral flora using the method according to the invention, if indicated = causing the tooth If the amount of bacteria is increased and/or the amount of lactic acid is increased, and/or the amount of bacteria decomposing arginine is decreased and/or the amount of plaque ammonia is decreased, the oral cavity containing the free or salt-type amino acid is administered. Health Care Products. 200948966 [Embodiment] Plaque metabolism-ammonia production The ability of arginine to ammonia is a factor for decomposing arginine activity 5 ❹ 10

; will: the ability of diterpene arginine to ammonia, just as some fine sputum will transform sugar into ^. Increasing the bacteria that decomposes the 8 strains produces conditions that are unfavorable for the proliferation of _ bacteria (this proliferation: love acid = increase the risk of tooth decay). The use of arginine per day can be similar to that of sucrose-producing bacteria in the form of conditions that favor the production of acid-producing bacteria. Neutral plaque. Neutral Non-disease = fine as a benefit. Depending on the concentration of the selected arginine-degrading bacteria selected without the distinction between metabolically active (live) and inactivated (dead) bacteria (as described further below), the amount of ammonia produced can be measured to determine the conversion of spermine The contribution of all bacteria to which the acid is ammonia. Ammonia test kits are commercially available, for example, from Diagn〇stic Chemicals Limited (Oxford, CT), which can measure the amount of ammonia produced. The principle of quantification and determination is known as ammonia and alpha-ketoglutaric acid and reduction. Type nicotine indoleamine adenine dinucleotide phosphate (NADPH) reacts to form L-glutamic acid and NADp; this reaction is catalyzed by alanine dehydrogenase (GLDH); the absorbance at 340 nm is reduced due to NADPH oxidation It is proportional to the ammonia concentration. The plaque samples were collected after a predetermined treatment procedure. In several applications, plaque is obtained from enamel or HAP samples mounted on a retainer. In other applications, plaque is obtained directly from the teeth. 20 200948966 The plaque ecology based on the amount of lactic acid Just as the proxy for measuring the amount of arginine bacteria as a proxy for measuring the amount of ammonia, the lactic acid system is a proxy for the amount of caries caused by caries. Patients who were sampled by plaque stopped eating or drinking water the night before, and did not take 5 10 15 teeth in the morning; rinsed with 10% sucrose solution for 2 minutes; after 8 minutes, scraped the teeth and collected plaque. The plaque samples were collected in pre-weighted organs placed on ice and the plaque weight was measured. The analysis consisted of adding ice-cold water to a known plaque sample of 0, then heating the sample to 8 ° C for 5 minutes to kill the bacteria and release all the acid, then cooling the sample in ice water for 5 minutes. . The sample was then centrifuged and the supernatant was filtered. The lactic acid wave length was measured by capillary electrophoresis. Quantitative real-time PCR (polymerase chain reaction) based on quantitative real-time PCR is a method for quantifying DNa. Since the amount of DNA is directly related to the amount of bacteria present, the total bacterial amount is quantified using bacterial DNA isolated from dental plaque. Instant pcR is recognized as a powerful and sensitive technology by government agencies such as the Center for Disease Control and the FDA. Probes are designed to detect oral bacteria or specific bacteria, such as the total amount of Streptococcus mutans or Streptococcus sanguis, using known gene sequences of many oral bacteria. The DNA isolated from the dental plaque or saliva sample is amplified by polymerase chain reaction. The amount of DNA increases exponentially with each pCR reaction cycle. Since the reaction is carried out immediately after the use of fluorescent reporter molecules, the technique is referred to as "instant," and in one specific embodiment of the invention, SYBR Green is used as a reporter molecule. This molecule is a double strand 20 200948966

❹ ίο 15 Intensely fluorescing after DNA is combined. Quantification is achieved by setting the fluorescence threshold (thresh〇ld) and using the DNA standards of various concentrations to determine the number of cycles required to reach this threshold. The more DNA that is present, the less the number of DNA cycles required to reach a critical value. Commercially available real-time PCR instruments are commercially available from a number of manufacturers, such as Roche Diagnostics. Plaque samples were obtained from enamel or base-based apatite samples of known and fixed surface area. The standardization of plaque collection is critical because the amount of DNA present is directly related to the amount of plaque collected. The use of plaque amount As a method of standardizing the total bacterial amount measured by real-time PCR, it is not appropriate, and as a result, it is significantly correlated. The results were recorded as micrograms of DNA per milliliter. Statistical analysis can be performed on DNA concentration or Ln (DNA concentration). The total bacterial count was analyzed by a two-factor variance analysis (ANOVA) using the patient and the treatment as a factor. If the difference detects a 95% confidence criterion, then they are considered to be significant. For a specific bacterium such as Streptococcus mutans or ▲ bond ball, the total bacterial amount is used as a cross-variable to analyze the two-factor variation: the total amount of bacteria is related to the total bacterial population, so it is a tooth; Indicators. Spoon 于 In the specific embodiment of the present invention, the index of caries is measured; Streptococcus mutans is selected as a recognized risk factor. Although there is 1, , /, mouth and mouth, there are mouths, although there are other acid-producing bacteria involved in the sputum tooth sweep, generally known as the deformation chain pure + Hanban process, but the role. At the beginning and early stage of the process, the author played an important role in the present invention. In the specific example, Dong wants a healthier tooth g-ecosphere, and the chain 19 is transferred to the 玍~, the buckle', because the blood is known as 9 20 200948966 Amino acid (the ability to convert sperm into ammonia). RNA transcripts in samples were determined by plaque ecological reverse transcription PCR by RT-PCR. RNA is isolated, reverse transcriptase is used to convert transcripts into cDNA, and pcR is used to expand 5 fNA R11PC: R < Advantages are that the DNA system σ cavity fine sputum detection method cannot detect the viability of these strains. Because oral bacteria are most commonly found in biofilm communities, the DNA of dead bacteria can remain in the structure for a long time after death. Other methods, such as the glory viability assay (UveDeadi; Molecular Probes), can detect whether a organism has a damaged membrane 10 (C〇mpr〇mised membranes), but cannot directly detect a particular species. Therefore, the reverse transcription real-time PCR method is a method for quantifying the presence of a living organism in a complex colony of a fine g g strain. mRNA has a fairly short sound period and is therefore an indicator of newly viable bacteria. Applicants have issued a primer for the specificity of the elongatum factor. The 15th phase does not significantly control the growth stage, medium or environmental conditions, thus reducing the virtual impact of the number of bacteria detected. To the minimum. Use the symbiosis to release the line 3 You r (10) / Qing less her & & 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜 娜Survival differences were detected in the mixed population. 20 The secret phase containing up to six different (four) _ groups =; the identification of the presence of actinomycetes. The calculated fine ':y is based on the same culture as D6I. The estimated values are closely related to M〇; = agronomy and heterosexuality <1%). This assay represents a method for studying specific organisms in the complex environment of the oral cavity = 200948966. Cencus can develop a step-by-step gene sequence data available, that is, x see a wide variety of oral bacteria primers. The amount of bacteria according to the fluorescent antibody probe 5

10 Use the worm main tooth 4 break kit to detect the amount of ore-producing bacteria (4), Streptococcus mutans, and/or non-lying cleavage bacteria (eg, Streptococcus sanguis) in saliva by using monoclonal antibodies. . The specific antibody used is specific to the bacterial species and has fluorescence attached to the antibody. The amount of bacteria can be detected by measuring the amount of fluorescence emitted. Example 1

The total amount of plaque bacteria (microgram bacterial DNA/ml) in real-time PCR patients measuring total plaque bacteria was measured using various toothpaste formulations, as described previously: 250 ppm fluoride formulation (control) Total bacterial DNA 6.091 Streptococcus mutans DNA 0.09622 Streptococcus sanguis DNA 1.126 1450 ppm Fluoride formulation 6.018 0.099〇3 1.107 Formulation with 2% arginine bicarbonate and 1450 ppm fluoride 3.78 0.05998 1.291 10 11 200948966 Spermine The acid-fluoride blending agent effectively reduces the total bacterial plaque load and the plaque load of Streptococcus mutans (striate) and increases the load of Streptococcus sanguis (decomposed arginine). Example 2 - Ammonia Production Amount of ammonia produced using various toothpaste formulations, measured as described above: Ammonia (ppm) 250 ppm Topical Formulation 1.97 (Control) 1450 ppm Fluoride 1.79 The formulation has 2% arginine carbonate 2.77 hydrogen salt and 1450 ppm fluoride. The ammonia production in the plaque of patients with arginine formula is significantly higher. Example 3 - Amount of lactic acid 10 The plaque lactic acid amount of the patient was measured by capillary electrophoresis as described above, and the results showed that lactic acid was significantly increased in the presence of sucrose. Plaque sucrose-stimulated teeth plaque lactic acid (namol/mg) 1.87 7.82 ± 0.37 12 200948966

Shell Example 4 · Real-time PCR / RT-PCR refers to the principle of instant PCR detection of intracellular bioactive bacteria. From the pure environment of the bacterial sample, i ^ uses the reverse transcription instant P C R to detect the amount of simple or complex legs and its application. The sequence of ounces and

Ίο 15 ❿ One of the codes in the heart of the ship is encoded to synthesize proteins. The DNA South and H" calendar stocks are then used as a reminder of the final protein). Different households have a very short half-life (several seconds to several points in the cell, = live or fresh cells; DNA with a fixed number of different eggs (four) ❿ calendar I often reflect the conditions of cell survival. Qualitative performance may Changes in reaction temperature, growth medium, growth, and other environmental conditions are adjusted upwards or downwards. Therefore, changes in environmental conditions are likely to be misinterpreted as changes in the system. The invention uses extended u / gene #' as the stem sequence. Since there are few or no changes in different experimental conditions, this sequence has previously been used as a private brother. Instant PCR enables limbic genetic material polymerase chain reaction (pCR) ) The basic chemistry for amplification 'and the simultaneous detection of fluorescent markers (which is the mechanism for quantifying the number of sets of silk due to the number of strands after the cycle). The method of using these methods is to use SYBR Green! Fluorescent probes specifically embedded in double-stranded DNA (dsDNA); therefore, the amount of SYBR glory increased by 13 20 200948966 is related to higher dsDNA concentrations. When this dye is incorporated, the specific gene sequence is used. In the PCR reaction, the increase in fluorescence corresponds to an increase in the number of sets of target genes. Subsequently, the number of cycles in which the signal crosses the predetermined intensity threshold is related to the concentration of the gene sequence in the starting material. The development of real-time PCR technology has made it possible Rapid and highly accurate detection and measurement of specific biological species. The conventional methods for quantifying bacterial species rely on the development of primers encoding the variable region of the 16s ribosomal subunit DNA. This human unit is critical for bacterial replication. Sexuality, therefore, its sequence is not easily mutated. The detection of the 16s rDNA sequence specific to the species of the genus Diptera is helpful for the detection and enumeration of single bacterial species in complex environments. The design of the primer is based on a publicly available database. (Nati〇nal Center for Biotechnology Information and the Los Alamos Oral Pathogens Database) sequences such as/genes. Sequences are arranged using the DNA Star Lasergene program MegAlign standard elements. This arrangement is used to select regions with greater tillering properties, The similarity of strain specificity is maximized. The selection of primer sequences is based on the selection of R〇che Diagnostics LightCycl. Information analysis of er Probe Design software. The primers covered by this invention include not only those who have been designed and tested, but also all primers for this gene part of oral pathogens. Use appropriate RNA detachment kit or other rna detachment method from sample Any single RNA can be isolated; any preferred method of isolation from rnA can be used. The purified RNA is treated twice with the appropriate DNase treatment reagent; this step degrades any contaminating DNA in the RNA preparation and prevents false positive junctions from being obtained. . The transcribed RNA' was then reverse transcribed to produce a complementary dna (cDNA) molecule' amplified cDNA' was detected using SYBR Green. An immediate PCR reaction without a reverse transcription step can be performed as a quality control for complete removal of DNA; PCR products obtained without a reverse transcription reaction must be the result of contamination of DNA 5. An immediate reverse transcription PCR reaction was performed on RNA samples isolated from cultures containing known amounts of live bacteria to generate a standard curve. The known concentration of the second derivative of each known sample 作 is plotted against its known concentration of bacterial cells. One of the RNA amplification curves isolated from the unknown bacterial sample can then be compared from the maximum value of the v number to the standard curve of the shai to determine the concentration of living organisms in the sample population. This data will be valuable information to curb the effects of antibacterial and active molecules on the environmental ecology of the oral cavity. The following primer pairs were designed to amplify a 228 base pair region derived from the actinomycete A. faecalis plus/gene:

Tm %GC AG Bonding (C) (kcal/temperature Moh) 49.05 56.25 -27.56 55°C φ Initiator Sequence Name Front 5' A AAGCGCGTGGTATCAC 3,

31.52 40.00 -20.15 55 °C Reverse 5' TGTAAGGAACACCTA -3, I5 Table 1 · Designed to quantify the nature of the symbiosis release _ _ such as the ruler to make Wei. 15 200948966 These primers are used to amplify RNA from both pure cultures of Actinobacillus compositae and mixed populations with or without A. faecalis; the results, especially fluorescence (F1) and circulation The relationship between the numbers is shown in Figure 1. In Fig. 1, "water" represents the negative control group of purely symbiotic A. faecalis culture and "Aa" is its positive control group; "Mix 1" is self-contained by Prevohe Ua intermedia, distant Strept〇c〇ccus, Streptococcus faecalis (and (4), and actinomycetes are purified, so amplification using their primers should be negative; "Mix 3" is derived from Actinobacillus actinomycetes, gingiva Streptococcus mutans g/mail να/, Streptococcus mutans (& Γ印幻(10) Streptococcus mutans, and the group of Streptococcus sanguis, the amplification of Actinobacillus actinomycetes should be positive. This chart proves that At the same time that the mixed population containing Actinobacillus actinomycetes was amplified in a similar curve to the positive control group, the mixed population lacking Actinomyces faecalis followed the same amplification curve as the water control group; Accurate detection of Actinobacillus elegans in the RNA mixture. ^ The ability of the _# primer to accurately detect and quantify the live symbiotic A. faecalis organisms can be determined as described below. The bacillus cells were suspended in 8〇% ethanol for 15 minutes for sterilization, then centrifuged, and the rice was read/child; the sera was resuspended in fresh Brain Heart Infusion liquid growth medium. The ethanol-killed bacteria were cultured at 370C. Overnight, check the growth to confirm the retention of inactive organisms. Then, the ethanol-killed fine 200948966 5 ❹ 10 bacteria were mixed with living organisms at the same ratio, and reverse transcription A was followed by real-time PCR. The amplification is shown in Figure 2, RN A < ^ # V of the mixed population of the live and dead line symbiotic Actinobacillus, although the genus of the reaction model (4) contains the organisms of the genus _ _, but contains more living organisms The sample of the body indicates that the assay can amplify the living and dead fine g-mixing earlier, and the melting curve of the samples is further, ^ 3 = single- and identical products in the living biological sample. It was amplified and proved to be C. Figure 3 shows the peak analysis of the products from the purely symbiotic A. actinomycetes and the current products. The (four) line overlap table. The nutrient amplification-products were amplified. Table 2 shows the selection. Standard; = comparison of single measured value and calculated value ^ ~ in the image Bionumber predictions CFU calculated by CFU 100% 5.00 X 107 5.34 X 1〇7 50% 2.50 X 107 2.28 X 1〇7 40% 2.00 X 1〇7 2.54 X 1〇7 30% 1.50x 107 1.18 X 1〇7 20% 1.00 X 107 1.07 X 1〇7 10% 5.00 X 106 4.06 X 1〇6 5% 2.50 X 106 2.96 X 1〇6 Predicted value of biological number in the selected standard curve sample And the known thick sound of the cockroach culture that was killed, ▲~ the approximate number of living organisms in the group, combined with the expansion of the curve a 'calculation of the second derivative of each family 17 expansion curve maximum 15 200948966 value, The standard curve was generated; the results are shown in Figure 4, which illustrates the standard curve generated from the amplification of the known concentration live and dead line A. faecalis and the linear regression of the standard curve; the r2 value of the regression line is 0.96. 5 10 15 The r2 value of the linear regression line indicates the proximity of the regression equation to the observed value. A r2 close to 1.00 indicates that the observed value closely matches the regression line. With respect to the above example, the r2 value of the standard curve is 0.96, indicating that approximately 96% of the total variation observed in this line is due to the actual measured variation in the sample, so this standard curve can be used to calculate the concentration of living organisms in the unknown population. In fact, in a single experiment, the concentration of living organisms calculated from this standard curve was not significantly different from the actual concentration added prior to RNA detachment, and this difference was <20%. According to their data, this assay represents a rapid and accurate method for detecting and quantifying living organisms in specific strains in complex biota; a potential tool for analyzing the effects of various treatments on oral microbial ecology. [Simple description of the diagram] Figure 1 is obtained from the amplification of the symbiotic symbiotic bacillus of the pure culture and the mixed genus. Figure 2 shows the amplification of RNA from a mixed population of live and dead line symbiosis A. faecalis. Figure 3 is a graph showing the melting peaks of products amplified from pure line A. faecalis and its mixed cultures. Figure 4 is a standard curve generated from amplification of a known concentration of live and dead line A. faecalis and a linear regression of the standard curve. 18 20

Claims (1)

200948966 VII. Scope of application for patents: 1. A method for assessing oral flora, the amount of bacteria. The sputum method includes measuring the decomposition of arginine. 2. The method of the ninth aspect of the patent application is also used to measure the bacterial activity of the caries. 3. If the method of claim No. 2 or 2 is applied, the active system is used to decompose arginine bacteria by measuring the amount of plaque ammonia produced. ί ίο 鲁 15 4. The method of measuring the amount of lactic acid produced is evaluated as in the second or third aspect of the patent application. VIII. The bacterial activity of the genus 5. The range of benefits, including the use of quantitative = day-to-day, quantitative RT_PCR, and / or $ light antibody probes to quantify at least one cariogenic bacteria and / or at least A quantity that decomposes arginine bacteria. The above-mentioned application is a method of the term "the term", wherein the decomposition of arginine, and the scorpion includes Streptococcus sanguis S. 7. The method according to any one of the preceding claims, wherein the squamous sputum, the bacterium, the bacterium, the bacterium, the bacterium of the genus Streptococcus mutans (s-like). A method of any of the claims, wherein the method is for detecting potentially harmful changes in the plaque ecology prior to measurable or significant demineralization or injury to the teeth. 9. A method for improving oral health, for example, a. reducing or inhibiting the formation of dental caries; b. reducing or inhibiting demineralization of teeth and promoting mineralization 19 200948966 C. treating, reducing or inhibiting early enamel lesions d. Reduce allergic reactions to teeth; e. Reduce or inhibit gingivitis; f. Promote healing of ulcers or wounds in the mouth; 5 g. Reduce the amount of acid-producing bacteria; h. Increase the relative amount of bacteria that break down arginine; To inhibit the formation of microbial biofilms in the oral cavity; j. to increase and/or maintain the plaque pH to at least pH 5.5 after sugar stimulation; 10 k. reduce plaque accumulation; l. treat, alleviate or reduce dry mouth; m. whitening teeth; η. improve overall health, including cardiovascular health; 〇 protect teeth against caries bacteria or immunize them; 15 清洁 clean teeth and mouth and / or q. reduce tooth decay I insects; The use of the method according to any one of the preceding claims to measure the bacterial flora of the oral cavity, if the indication indicates an increase in the amount of bacteria and/or an increase in the amount of lactic acid, and/or the decomposition of 20 9200948966 Amine acid bacteria containing an effective amount of the administration of the test group decreased and / or decrease the amount of plaque acid produced ammonia or salt thereof in oral care products. 10. If you apply for patent scope 9 or its salt. The method of the invention wherein the basic amino acid is a succinic acid hydrazine/I or a salt-based basic amino acid, and the method of the method of measuring any one of the V π patent ranges It is used for oral health of the patient. The oral biog plexus of the patient contains an increase in the amount of bacteria and/or an increase in sputum sputum, and/or a low decomposition of arginine bacteria and/or plaque ammonia production. 10 12.- A method for improving the aesthetics of the mouth, the method comprising measuring the oral flora with the method according to any one of the items 1 to 10 of the patent application, if the amount of bacteria causing caries is increased and/or the amount of lactic acid is indicated If the amount of bacteria that degrades arginine is decreased and/or the amount of plaque ammonia is decreased, an oral health care product containing a free or salt-type basic amino acid is administered. twenty one