Application of biomarkers in the preparation of products for diagnosing patients with oral diseases and kits for detecting oral diseases

Abstract

The present application discloses an application of a biomarker in the preparation of a product for diagnosing patients with oral diseases and a kit for detecting oral diseases, wherein the oral diseases include at least one of periodontitis, gingivitis and caries, and the biomarkers include cytokines and / or oral microorganisms, wherein the cytokines include at least one of IFN-γ, IL-6, IL-10, IL-18 and IL-1β, and the oral microorganisms include at least one of Neisseria, Porphyromonas, Fusobacterium and Treponema. In the present application, auxiliary diagnosis of oral diseases can be achieved by detecting biomarkers, which is conducive to timely detection of the condition in the early stage of oral diseases or before the disease progresses for timely intervention.

Description

Technical Field

[0001] The present application relates to the field of oral medicine technology, and in particular to the use of a biomarker in the preparation of a product for diagnosing patients with oral diseases and a kit for detecting oral diseases. Background Art

[0002] Dental caries is a process in which bacteria on the surface of teeth metabolize and produce acid, leading to demineralization of enamel and dentin, and then forming cavities. If not treated in time, dental caries may further develop into pulpitis or apical periodontitis, leading to more serious complications. Gingivitis and periodontitis are chronic inflammatory diseases caused by microbial communities in dental plaque. Gingivitis is mainly manifested by symptoms such as redness, swelling and bleeding of the gums, which are usually limited to the gingival tissue; while periodontitis involves deeper periodontal tissues, including the gums, periodontal ligament, alveolar bone and cementum. Untreated periodontitis can lead to alveolar bone resorption, loose teeth and loss, seriously affecting the patient's oral health and quality of life.

[0003] At present, the diagnosis and analysis of traditional oral diseases are mostly based on the clinical diagnosis of doctors, mainly relying on clinical examination and radiological evaluation. This analysis method has certain limitations, such as the results rely on the subjective interpretation of experienced physicians, lack of objectivity, and the need to spend a lot of time and manpower costs. In addition, traditional clinical examination methods can often only detect lesions when the disease progresses to a certain extent, which may lead to missing the best time for treatment.

[0004] Therefore, it is necessary to develop biomarkers that can identify these oral diseases to detect early periodontitis and predict future periodontal destruction so that timely intervention can be carried out before advanced periodontal lesions occur. Summary of the invention

[0005] In order to solve the problem that traditional oral disease examinations lack objectivity and can only be detected when the disease is progressing, the present application provides an application of a biomarker in the preparation of a product for diagnosing oral diseases and a kit for detecting oral diseases.

[0006] This application adopts the following technical solutions:

[0007] The first aspect of the present application discloses the use of a biomarker in the preparation of a product for diagnosing patients with oral diseases, wherein the oral diseases include at least one of periodontitis, gingivitis and caries, and the biomarker includes cytokines and / or oral microorganisms, wherein the cytokines include at least one of IFN-γ, IL-6, IL-10, IL-18 and IL-1β, and the oral microorganisms include at least one of Neisseria, Porphyromonas, Fusobacterium and Treponema. It should be noted that the biomarkers of the present application have significant expression differences or abundance differences (P < 0.05) between patients with oral diseases and healthy people, and the detection of the biomarkers can help diagnose patients with oral diseases.

[0008] In one implementation of the present application, the oral disease patients include a first type, a second type, a third type, a fourth type and a fifth type. The first type is patients suffering from only gingivitis (abbreviated as HGH), the second type is patients suffering from only caries (abbreviated as HHC), the third type is patients suffering from gingivitis combined with caries (abbreviated as HGC), the fourth type is patients suffering from periodontitis combined with gingivitis (abbreviated as PGH), the fifth type is patients suffering from periodontitis combined with gingivitis and caries (abbreviated as PGC), and healthy people are those who do not suffer from periodontitis, gingivitis or caries (abbreviated as HHH).

[0009] In one implementation of the present application, the biomarker is IL-6 and / or IL-1β, and the biomarker is used to distinguish the oral disease patient from the healthy person. It should be noted that for patients with at least one of periodontitis, gingivitis and caries, IL-6 and / or IL-1β have good diagnostic performance. According to one embodiment, IL-6 as a biomarker has a sensitivity of 60% and a specificity of 81.5%; IL-1β as a biomarker has a sensitivity of 80.39% and a specificity of 76%; IL-6 combined with IL-1β as a biomarker has a sensitivity of 83.33% and a specificity of 83.33%.

[0010] In one implementation of the present application, the biomarkers are IL-6, IL-1β and IL-10, and the biomarkers are used to distinguish the oral disease patients from the healthy people. It should be noted that for patients with at least one of periodontitis, gingivitis and dental caries, IL-6 combined with IL-1β and IL-10 as biomarkers has good diagnostic performance. According to one embodiment, IL-6 combined with IL-1β and IL-10 as biomarkers has a sensitivity of 78.85% and a specificity of 66.67%.

[0011] In one implementation of the present application, the biomarker is IL-6, and the biomarker is used to distinguish the fifth type of patients from the healthy people (that is, the biomarker is used to distinguish PGCvs HHH). It should be noted that IL-6 has good diagnostic performance for patients with periodontitis combined with gingivitis and dental caries. According to one embodiment, IL-6 as a biomarker has a sensitivity of 60% and a specificity of 81.25%.

[0012] In one implementation of the present application, the biomarker is IL-1β, and the biomarker is used to distinguish the second type of patients / the healthy people from other types of patients (that is, the biomarker is used to distinguish (HHC and HHH) vs (HGH and HGC and PGH and PGC), that is, the biomarker is used to distinguish HHX vs X.GX). It should be noted that for patients in the two groups of HHX and XGX, IL-1β has good diagnostic performance. According to one embodiment, IL-1β as a biomarker has a sensitivity of 80.39% and a specificity of 76.47%.

[0013] The second aspect of the present application discloses an application of a biomarker in the preparation of a product for identifying patients with different types of oral diseases, wherein the oral disease patients include a first type, a second type, a third type, a fourth type and a fifth type, wherein the first type is patients suffering only from gingivitis (referred to as HGH), the second type is patients suffering only from caries (referred to as HHC), the third type is patients suffering from gingivitis combined with caries (referred to as HGC), the fourth type is patients suffering from periodontitis combined with gingivitis (referred to as PGH), and the fifth type is patients suffering from periodontitis combined with gingivitis and caries (referred to as PGC), the biomarkers include cytokines and / or oral microorganisms, the cytokines include at least one of IFN-γ, IL-6, IL-10, IL-18 and IL-1β, and the oral microorganisms include at least one of Neisseria, Porphyromonas, Fusobacterium and Treponema. It should be noted that the biomarkers of the present application have significant expression differences or abundance differences (P < 0.05) in patients with different types of oral diseases. Detection of the biomarkers can help distinguish patients with different types of oral diseases.

[0014] In one implementation of the present application, in the applications involved in the first aspect and the second aspect of the present application, the product includes a reagent or a kit for detecting the abundance or concentration of the biomarker.

[0015] In one implementation of the present application, in the applications involved in the first aspect and the second aspect of the present application, the sample type detected by the product is saliva.

[0016] The third aspect of the present application discloses a kit for detecting patients with oral diseases, the kit comprising reagents for detecting the abundance or content of biomarkers, the oral diseases comprising at least one of periodontitis, gingivitis and caries, the biomarkers comprising cytokines and / or oral microorganisms, the cytokines comprising at least one of IFN-γ, IL-6, IL-10, IL-18 and IL-1β, and the oral microorganisms comprising at least one of Neisseria, Porphyromonas, Fusobacterium and Treponema.

[0017] The beneficial effects of this application are:

[0018] The application of the biomarkers provided in the present application in the preparation of products for diagnosing patients with oral diseases and in the kits for detecting oral diseases can achieve auxiliary diagnosis of oral diseases through the detection of biomarkers, thereby facilitating timely detection of oral diseases in the early stages or before the disease progresses for timely intervention. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a graph showing the difference in species abundance between PGC patients and HHH healthy people.

[0020] Figure 2 This is a graph showing the difference in species abundance between HHC patients and HHH healthy people.

[0021] Figure 3 This is a graph showing the expression results of cytokines that are differentially expressed in different groups of people.

[0022] Figure 4 This is a graph showing the expression of various cytokines in PGC patients and HHH healthy subjects.

[0023] Figure 5 This is a graph showing the expression of various cytokines in HHC patients and PGC patients.

[0024] Figure 6 It is a graph showing the expression of various cytokines in HHC patients and HHH healthy subjects.

[0025] Figure 7 It is a graph showing the expression of various cytokines in HHC patients and HGC patients.

[0026] Figure 8This is a graph showing the expression of various cytokines in HHH healthy subjects and HGC patients. DETAILED DESCRIPTION

[0027] The present invention is further described in detail below by specific embodiments in conjunction with the accompanying drawings. In the following embodiments, many detailed descriptions are intended to enable the present application to be better understood. However, those skilled in the art can effortlessly recognize that some of the features can be omitted in different situations, or can be replaced by other materials and methods. In some cases, some operations related to the present application are not shown or described in the specification, in order to avoid the core part of the present application being overwhelmed by too much description, and for those skilled in the art, it is not necessary to describe these related operations in detail, and they can fully understand the related operations according to the description in the specification and the general technical knowledge in the art.

[0028] In addition, the features, operations or characteristics described in the specification can be combined in any appropriate manner to form various implementations. At the same time, the steps or actions in the method description can also be interchanged or adjusted in a manner that is obvious to those skilled in the art. Therefore, the various sequences in the specification and the drawings are only for the purpose of clearly describing a certain embodiment and are not meant to be a required sequence, unless otherwise specified that a certain sequence must be followed.

[0029] The serial numbers assigned to the components in this article, such as "first", "second", etc., are only used to distinguish the objects described and do not have any order or technical meaning.

[0030] The present application is further described in detail below by specific examples. The following examples are only further described for the present application and should not be construed as limiting the present application. It should be noted that the reagents, test kits, instruments, etc. used in the examples are all commercially available, and the operating steps are carried out according to the instructions or the general operating steps in the art.

[0031] [Example]

[0032] Detection and analysis:

[0033] (1) Subject inclusion. Among them, 61 patients with oral diseases treated in our hospital were selected as the observation group, and 29 healthy subjects without oral diseases were selected as the control group. Inclusion criteria: ① Red, swollen, shiny gums that bleed easily when touched; ② Plaque covering the tooth surface; ③ No implant restorations in the mouth and no history of orthodontic treatment; ④ Attachment loss of 1 to 2 mm can be seen clinically; ⑤ The depth of the periodontal pocket does not exceed 4 mm; ⑥ No periodontal treatment has been performed within 3 months; ⑦ The number of natural teeth in the mouth is greater than 20, the number of molars is greater than 4, and there are natural teeth remaining in each quadrant. Exclusion criteria: ① patients with severe systemic diseases such as diabetes, hypertension, cardiovascular disease, and immune system diseases; ② patients with severe organ dysfunction, infectious diseases, mental illness, impaired consciousness, and malignant tumors; ③ patients with other oral diseases except periodontitis; ④ patients with a history of using mouthwash within 3 months and a history of taking antibiotics or nonsteroidal anti-inflammatory drugs, immunosuppressants, or immunostimulants; ⑤ clinical attachment loss >2mm, or periodontal pocket depth exceeding 4mm.

[0034] (2) Saliva specimen collection. After stopping eating and drinking for 2 hours and 10 minutes before saliva collection, the subjects rinsed their mouths with distilled water, and then used sterile cotton balls to clean the soft plaque or food residues in the volunteers' mouths. During the baseline periodontal examination (before clinical evaluation), 5 ml of whole saliva secreted naturally without stimulation was collected by drainage method into sterile enzyme-free EP tubes, which were stored in liquid nitrogen tanks for later use. They were then transferred to the laboratory on dry ice and centrifuged at 4°C (15 minutes, 4000 rpm / min). The supernatant was divided into 2 ml EP tubes and stored at 1-80°C for subsequent testing (thawed and used within 3 months). To prevent the periodontal clinical examination from affecting saliva collection, saliva collection for all subjects was performed before the baseline periodontal examination.

[0035] (3) After collecting saliva, a baseline periodontal examination was performed to obtain the clinical examination results of oral diseases of each subject. Six teeth were selected for each subject (according to the FDI international tooth numbering system, the selected teeth were numbered 16, 11, 26, 31, 36, and 46, respectively) for clinical index evaluation. According to the specific conditions of the 61 patients with oral diseases, they were divided into patients with only gingivitis (HGH), patients with caries suffering from gingivitis (HGC), patients with periodontitis and gingivitis (PGH), patients with caries suffering from periodontitis and gingivitis (PGC), patients with caries without periodontitis or gingivitis (HHC), and those who did not suffer from periodontitis, gingivitis or caries were healthy people (HHH). In this embodiment, the naming rules of each group can be found in Table 1 below:

[0036] Table 1 Naming rules for subject groups

[0037] project Do you have periodontitis? Do you have gingivitis? Whether you have dental caries yes P G C no H H H Yes or No X X X

[0038] (4) Detect the differences in bacterial flora in the saliva samples of the subjects. The DNA extraction methods of the samples were strictly carried out in accordance with the specifications of the DNAStool Mini Kit, and the extracted DNA samples were subjected to genome integrity detection (agarose gel electrophoresis) and quality inspection (NanoDrop2000). Specifically, a tube of 1.5 ml of saliva was transferred to a 2 ml centrifuge tube, centrifuged at 8000 rpm for 4 minutes, the supernatant was discarded, and the precipitate was retained. 180 μL ATL buffer was added to the cell pellet and fully resuspended. 20 μL proteinase K was added to the resuspended solution and mixed. Incubate in a 56°C water bath for 30 minutes and gently mix to obtain a pretreated sample. Add 200 μL AL buffer to the pretreated sample and mix. Add 200 μL anhydrous ethanol and mix thoroughly. Transfer the mixture to a collection tube containing a DNA easy mini column, centrifuge at 8000 rpm for 1 minute, discard the effluent and retain the mini column. Add 500 μL AW1 buffer to the mini column, centrifuge at 8000 rpm for 1 minute, and discard the effluent. Add 500 μL AW2 buffer to the mini column, centrifuge at 14000 rpm for 3 minutes, discard the effluent and idle for 1 minute to remove residual ethanol. Transfer the mini column to a new 1.5 mL centrifuge tube. Add 100 μL AE buffer to the center of the mini column and leave at room temperature for 1 minute. Centrifuge at 8000 rpm for 1 minute and collect the DNA in the eluate. Use NanoDrop2000 to determine the DNA concentration and purity (260 / 280 and 260 / 230 ratios). The quality and integrity of the DNA were detected by agarose gel electrophoresis. For those whose DNA quality meets the requirements, the next experiment was carried out. Subsequently, Beijing Novogene Technology Co., Ltd. performed paired-end sequencing on the V4 variable region of the 16S r RNA gene of the saliva sample to obtain the types and abundance of microorganisms in the sample.

[0039] (5) Detect the expression of cytokines IL-1β, TNF-α, IFN-γ, IL-6, IL-8, IL-10, IL-18, and Fractalkine in the saliva samples of the subjects. MSD electrochemiluminescence detection technology was used, and SULFO-TAGTM markers were used. After the electrode surface of the MULTI-ARRAY and MULTI-SPOT microplates was energized, the electrochemical action stimulated the SULFO-TAGTM markers to emit strong light. A customized 8-factor detection 96-well plate was used. One person in the observation group and one person in the healthy group (negative control) took 1.5ml of saliva and thawed it for pretreatment, and diluted it according to 1:2 and 1:4. Different concentrations of standard samples were prepared with sample diluent, and the standard samples and negative controls were added to each well. The background value and dilution were determined and a standard curve was drawn. Subsequently, the samples were diluted according to a certain dilution, and 40ul of the diluted samples were added to the corresponding wells. The samples were shaken to cover the bottom of the wells and bind to the capture antibody. Then incubate and wash. Incubation: seal the plate and incubate at room temperature for 1 hour. Gently shake it on a shaker to ensure that the target protein in the sample is fully bound to the capture antibody. Washing: wash the plate wells 3-5 times with washing buffer to remove unbound samples and ensure the specificity of the detection. Read the electrochemical signal: add a dedicated reading buffer to promote the electrochemical reaction. Then put the plate into the MSD electrochemiluminescence analyzer and start the detection. The instrument excites SULFO-TAG to generate an electrochemiluminescence signal by applying current and records the luminescence intensity of each well. Data analysis: Use the standard data to generate a standard curve, use the instrument's own software or other statistical software for data fitting, and calculate the concentration of the target protein in the sample based on the standard curve.

[0040] (6) Data statistics: SPSS 18.0 software package was used for statistical analysis. Categorical numerical data were expressed as absolute values ​​and percentages (%). The t-test was used for inter-group comparison of parametric data; the Mann-Whitney test was used for inter-group comparison of non-parametric data; the χ2 test was used for inter-group comparison of categorical variables. P < 0.05 was considered statistically significant.

[0041] Test results:

[0042] (1) Figure 1 This is the result of the species abundance difference analysis between PGC patients and HHH healthy people. Figure 2 This is the result of the species abundance difference analysis between HHC patients and HHH healthy people, such as Figure 1 and Figure 2As shown in the figure, through pairwise comparison of species abundance among groups, it was found that Neisseria, Porphyromonas, Fusobacterium, and Treponema in caries patients with periodontitis and gingivitis (PGC) were significantly higher than those in completely healthy people (HHH) (P < 0.05), and Neisseria and Porphyromonas in caries patients without periodontitis and gingivitis (HHC) were significantly higher than those in completely healthy people (HHH) (P < 0.05).

[0043] (2) Analysis of cytokine detection results:

[0044] 1) The rank sum test was used to compare the expression of cytokines in each group, and it was found that the expression of IFN-γ and IL-10 in each group (including patients with gingivitis only (HGH), patients with caries with gingivitis (HGC), patients with periodontitis and gingivitis (PGH), patients with caries with periodontitis and gingivitis (PGC), patients with caries without periodontitis or gingivitis (HHC), and healthy people (HHH)) was statistically significant (P < 0.05, Figure 3 ), Figure 3 This is a graph showing the expression results of cytokines that are differentially expressed in different groups of people. The MFI on the vertical axis refers to the mean fluorescence intensity (Mean Fluorescence Intensity) detected.

[0045] 2) Compare the differences in cytokine expression between the two groups, Figure 4 This is a chart showing the expression of various cytokines in PGC patients and HHH healthy people. Figure 5 This is a chart showing the expression of various cytokines in HHC patients and PGC patients. Figure 6 This is a chart showing the expression of various cytokines in HHC patients and HHH healthy subjects. Figure 7 This is a chart showing the expression of various cytokines in HHC and HGC patients. Figure 8 This is a graph showing the expression of various cytokines in HHH healthy subjects and HGC patients. The results are as follows Figures 4 to 8There were significant differences in the expression of IFN-γ and IL-10 between caries patients without gingivitis (HHC) and caries patients with gingivitis (HGC) (P < 0.05); there were significant differences in the expression of IL-1β, IL-10 and IL-18 between caries patients without periodontitis and gingivitis (HHC) and caries patients with periodontitis and gingivitis (PGC); there were differences in the expression levels of IL-10 and IL-6 between completely healthy people (HHH) and caries patients with gingivitis (HGC), while there was only a difference in the expression level of IL-10 between completely healthy people (HHH) and caries patients with periodontitis and gingivitis (PGC); there was no difference in the cytokine levels of completely healthy people (HHH) and patients with only caries (HHC) in this screening.

[0046] (3) Diagnostic efficacy study:

[0047] Sensitivity refers to the ability of a detection method to correctly identify positive samples. That is, the proportion of positive samples detected among all truly positive samples. Tests with high sensitivity can effectively avoid missed diagnoses. The calculation formula for sensitivity is as follows: Sensitivity = True Positive / (True Positive + False Negative). High sensitivity means that the test can detect even a small amount of target substance in the sample, and low sensitivity may cause some positive samples to be missed (false negative). Specificity refers to the ability of a detection method to correctly identify negative samples. That is, the proportion of negative samples detected among all truly negative samples. Tests with high specificity can effectively avoid misdiagnosis. The calculation formula for specificity is as follows: Specificity = True Negative / (True Negative + False Positive). High specificity means that the test can effectively avoid false positives of negative samples, and low specificity may cause some negative samples to be misjudged as positive (false positive). Youden Index: It is a statistical indicator used to evaluate binary diagnostic tests. It can reflect both sensitivity and specificity and is usually used to measure the overall diagnostic effect of the test. The calculation formula is as follows: Youden Index = Sensitivity + Specificity - 1. The Youden Index ranges from 0 to 1. If the Youden Index is 1, it means that the test method has perfect sensitivity and specificity and can accurately distinguish all positive and negative samples. If the Youden Index is 0, it means that the diagnostic effect of the test method is no different from random guessing, and it is completely unable to distinguish between positive and negative samples. Generally speaking, the higher the Youden Index, the better the overall effect of the test method, which can effectively detect positive cases and effectively exclude negative cases.

[0048] According to the detection results of each cytokine, combined with the clinical diagnosis results of the subjects, the diagnostic efficacy of biomarkers was analyzed. The average concentrations of IL6, IL-10, IL-18 and IL-1β in the overall samples were 2.48pg / ml, 0.37pg / ml, 380.36pg / ml and 293.96pg / ml, respectively. When IL-6 took 2.05pg / ml as the threshold (IL-6 concentration greater than 2.05pg / ml was positive, and vice versa was negative), its sensitivity and specificity were 60% and 81.25%, respectively. When IL-10 took 0.34pg / ml as the threshold, its sensitivity and specificity were 60% and 81.25%, respectively. / ml as the threshold (IL-10 concentration less than 0.34pg / ml is positive, otherwise it is negative), the sensitivity is 76.47% and the specificity is 40.0%; when IL-18 takes 110pg / ml as the threshold (IL-18 concentration greater than 110pg / ml is positive, otherwise it is negative), its sensitivity and specificity are 74.5% and 44.4% respectively; when IL-1β takes 92.02pg / ml as the threshold (IL-1β concentration greater than 92.02pg / ml is positive, otherwise it is negative), its sensitivity and specificity are 80.39% and 76% respectively. When IL-6 and IL-1β are combined as biomarkers, the Youden index is 0.67, the sensitivity is 83.33%, and the specificity is 83.33%. When IL-6, IL-18 and IL-1β were used as biomarkers, the Youden index was 0.46, the sensitivity was 78.85%, and the specificity was 66.67%. The specific results are shown in Table 2 below.

[0049] Table 2 Diagnostic efficacy of various cytokines (oral disease patients vs healthy subjects)

[0050] Cytokines Youden Index Critical value (pg / ml) Sensitivity (%) Specificity (%) IL-10 0.14 0.34 76.47 40.00 IL-6 0.42 2.05 60 81.5 IL-18 0.19 110 74.5 44.4 IL-1β 0.56 92.02 80.39 76 IL-6 and IL-1β 0.67 - 83.33 83.33 IL-6, IL-10, and IL-1β 0.46 - 78.85 66.67

[0051] In Table 2, oral disease patients refer to patients suffering from at least one of periodontitis, gingivitis and caries. It can be seen from Table 2 that IL-6, IL-1β, IL-6 combined with IL-1β, IL-6 combined with IL-1β and IL-10 as biomarkers have good sensitivity and specificity for distinguishing oral disease patients (HGH or HGC or PGH or PGC or HHC) and healthy people (HHH), both of which are 60% or above. Among them, the sensitivity and specificity of IL-6 combined with IL-1β as a biomarker are both over 80%, which has excellent diagnostic efficacy.

[0052] In addition, different biomarkers also have good diagnostic efficacy in subdividing different types of patients, as shown in Tables 3 to 6 below.

[0053] Table 3 Diagnostic efficacy of IL-6 (HGC vs HHH)

[0054]

[0055] Table 4 Diagnostic efficacy of IL-10 (XGC vs HHH)

[0056]

[0057]

[0058] Table 5 Diagnostic efficacy of IL-18 (XGX vs HHC)

[0059]

[0060] Table 6 Diagnostic efficacy of IL-1β (XGX vs HHX)

[0061]

[0062] Among them, XGC includes PGC and HGC, XGX includes HGC, HGH, PGC and PGH, and HHX includes HHC and HHH. From Tables 3 to 6 above, it can be seen that IL-6, IL-10, IL-18 and IL-1β have good effects in distinguishing specific types of people. In particular, IL-6 has good sensitivity and specificity for distinguishing HGC and HHH people, and IL-1β has good sensitivity and specificity for distinguishing XGX and HHX people.

[0063] The above contents are further detailed descriptions of the present application in combination with specific implementation methods, and it cannot be determined that the specific implementation of the present application is limited to these descriptions. For ordinary technicians in the technical field to which the present application belongs, several simple deductions or substitutions can be made without departing from the concept of the present application.

Claims

1. Use of a biomarker in the preparation of a product for diagnosing patients with oral diseases, characterized in that: The oral diseases include at least one of periodontitis, gingivitis and caries, the biomarkers include cytokines and / or oral microorganisms, the cytokines include at least one of IFN-γ, IL-6, IL-10, IL-18 and IL-1β, and the oral microorganisms include at least one of Neisseria, Porphyromonas, Fusobacterium, Treponema.

2. The use according to claim 1, characterized in that: The oral disease patients include the first type, the second type, the third type, the fourth type and the fifth type. The first type is suffering from gingivitis only, the second type is suffering from caries only, the third type is suffering from gingivitis combined with caries, the fourth type is suffering from periodontitis combined with gingivitis, the fifth type is suffering from periodontitis combined with gingivitis and caries, and healthy people are those who do not suffer from periodontitis, gingivitis or caries.

3. The use according to claim 2, characterized in that The biomarker is IL-6 and / or IL-1β, and the biomarker is used to distinguish the oral disease patients from the healthy people.

4. The use according to claim 2, characterized in that The biomarkers are IL-6, IL-1β and IL-10, and the biomarkers are used to distinguish the oral disease patients from the healthy people.

5. The use according to claim 2, characterized in that: The biomarker is IL-6, and the biomarker is used to distinguish the fifth type of patients from the healthy persons.

6. The use according to claim 2, characterized in that: The biomarker is IL-1β, and the biomarker is used to distinguish the second type of patients / the healthy persons from other types of patients.

7. Use of a biomarker in the preparation of a product for identifying patients with different types of oral diseases, characterized in that: The oral disease patients include a first type, a second type, a third type, a fourth type and a fifth type. The first type is patients suffering only from gingivitis, the second type is patients suffering only from caries, the third type is patients suffering from gingivitis combined with caries, the fourth type is patients suffering from periodontitis combined with gingivitis, and the fifth type is patients suffering from caries with periodontitis combined with gingivitis. The biomarkers include cytokines and / or oral microorganisms, the cytokines include at least one of IFN-γ, IL-6, IL-10, IL-18 and IL-1β, and the oral microorganisms include at least one of Neisseria, Porphyromonas, Fusobacterium, and Treponema.

8. The use according to any one of claims 1 to 7, characterized in that The product includes a reagent or a kit for detecting the abundance or concentration of the biomarker.

9. The use according to claim 8, characterized in that The sample type tested by the product is saliva.

10. A kit for detecting oral diseases, characterized in that: The kit includes reagents for detecting the abundance or content of biomarkers, the oral diseases include at least one of periodontitis, gingivitis and caries, the biomarkers include cytokines and / or oral microorganisms, the cytokines include at least one of IFN-γ, IL-6, IL-10, IL-18 and IL-1β, and the oral microorganisms include at least one of Neisseria, Porphyromonas, Fusobacterium and Treponema.

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