A set of noise-induced hearing loss markers and its detection kit and application

Abstract

The present invention discloses a group of noise-induced hearing loss marker combinations and a detection kit and application thereof, wherein the marker combination includes rs2394795, rs2862833, rs3751385 and rs1053023. When the rs2394795 is of TT or TC genotype, rs2862833 is of TT or CC genotype, rs3751385 is of TT genotype and rs1053023 is of AG genotype, the noise-induced hearing loss is susceptible; if not, the noise-induced hearing loss is tolerant. The present invention firstly discovers the SNPs sites associated with ONID and finds that the above-mentioned marker combination has the correlation with the onset of noise-induced hearing loss, which can be used for the effective assessment of the risk of occupational noise-induced hearing loss, thereby realizing the effective identification and screening of the population susceptible to noise-induced hearing loss, thereby having a primary preventive effect on occupational noise-induced hearing loss.

Description

Technical Field

[0001] The present invention belongs to the field of gene detection, and specifically relates to a group of noise-induced hearing loss marker combinations and a detection kit and application thereof. Background Art

[0002] Occupational noise-induced deafness (ONID), also known as noise-induced hearing loss, is one of the most common and important occupational diseases in the world. The occurrence of noise-induced hearing loss is the result of the combined effects of environmental and genetic factors, and noise-induced hearing loss (NIHL) is an early manifestation of noise-induced hearing loss. In related technologies, more than 60 single nucleotide polymorphisms (SNPs) of more than 30 genes have been found to be associated with the susceptibility to noise-induced hearing loss through studies on occupational populations and animal experiments. The discovery of these susceptibility-related genes is mainly based on candidate genes in the NIHL pathogenesis process through animal experiments of gene knockout and verification experiments of some noise-exposed populations. These studies are all aimed at detecting a certain candidate gene, and the research methods (selection of research subjects, gene detection methods, statistical methods, etc.) are different, which are very accidental and blind, lack of systematicity, lack of unified standards for research conclusions, and the accuracy needs to be further studied. Currently, there is still a lack of accurate and sensitive detection methods and markers for the prediction and diagnosis of noise-induced hearing loss.

[0003] Therefore, developing a prediction and diagnosis product for noise-induced hearing loss with high specificity and good sensitivity is of great significance for the screening and early warning of people who are prone to noise-induced hearing loss in clinical practice. Summary of the invention

[0004] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention proposes a set of noise-induced hearing loss marker combinations and a detection kit and application thereof, through which the susceptibility of a subject to noise-induced hearing loss can be accurately determined, thereby effectively providing an early warning for the subject and carrying out relevant preventive work in advance, which is of great significance for reducing the occurrence of clinical noise-induced hearing loss.

[0005] The first aspect of the present invention provides a set of SNP marker combinations associated with susceptibility to noise-induced hearing loss, which includes rs2394795, rs2862833, rs3751385 and rs1053023.

[0006] Among them, the site information of rs2394795, rs2862833, rs3751385 and rs1053023 is as follows.

[0007]

[0008]

[0009] The SNP of the susceptibility gene for noise-induced hearing loss is not simply caused by a single independent gene mutation. Its genetic characteristics are realized by the interaction of multiple susceptibility gene mutations. It may be a set (group) of SNPs, but there have been no public reports on this in the prior art. In the embodiments of the present invention, the inventors found through research and actual verification that eight SNPs sites, CDH23-rs2394795, FAS-rs1468063, FAS-rs2862833, GJB2-rs3751385, PTPRN2-rs10081191, SIK3-rs6589574, STST3-rs1053023, and STST3-rs1053005, may be related to ONID susceptibility, among which the combination of four SNPs sites, CDH23-rs2394795, FAS-rs2862833, GJB2-rs3751385, and STST3-rs1053023, has actual relevance to the substantial occurrence of ONID susceptibility, and is statistically significant.

[0010] In the embodiment of the present invention, the inventors found that for the combination of the above SNPs, the ROC curve was used for verification, and it was found that compared with FAS-rs2862833 (AA / BB) alone for diagnosis, the AUC of FAS-rs2862833 (AA / BB) + STAT3-rs1053023 (AB) combined diagnosis was 0.593 (P = 0.008), and FAS-rs2862833 (AA / BB) + STAT3-rs1053023 (AB) + CDH23-rs1053023 (AB) combined diagnosis was 0.593 (P = 0.008). The AUC of the combined diagnosis of s2394795 (AA / AB) was 0.601 (P=0.004), and the AUC of the combined diagnosis of FAS-rs2862833 (AA / BB) + STAT3-rs1053023 (AB) + CDH23-rs2394795 (AA / AB) + GJB2-rs3751385 (BB) was 0.602 (P=0.004), indicating that these four SNPs loci as combined diagnostic indicators can improve the diagnostic accuracy of ONID susceptible people. The composition and structure of this set will help to judge the susceptible population of noise-induced hearing loss and have diagnostic value for noise-induced hearing loss susceptible people.

[0011] In the embodiments of the present invention, the inventors found through various statistical analyses and 268 clinical verifications that the combination of four SNPs sites, CDH23-rs2394795, FAS-rs2862833, GJB2-rs3751385, and STST3-rs1053023, has a practical relevance to the substantial occurrence of ONID susceptibility, and when the rs2394795 is TT or TC genotype, rs2862833 is TT or CC genotype, rs3751385 is TT genotype, and rs1053023 is AG genotype, it can be considered that there is a risk of ONID, and it is statistically significant.

[0012] The second aspect of the present invention provides Taqman primers for detecting the SNP marker combination described in the first aspect of the present invention, wherein the nucleotide sequence of the primers is:

[0013] rs2394795: SEQ ID NO: 187 and SEQ ID NO: 189;

[0014] rs2862833: SEQ ID NO: 191 and SEQ ID NO: 193;

[0015] rs3751385: SEQ ID NO: 195 and SEQ ID NO: 197;

[0016] rs1053023: SEQ ID NO:199 and SEQ ID NO:201.

[0017] Of course, those skilled in the art can also reasonably design other Taqman primers with high specificity to amplify the above-mentioned SNP marker combination according to actual use requirements, including but not limited to the Taqman primers listed above.

[0018] The third aspect of the present invention provides a probe for detecting the SNP marker combination described in the first aspect of the present invention, wherein the nucleotide sequence of the probe is:

[0019] rs2394795:

[0020] C-type probe: SEQ ID NO: 188, T-type probe: SEQ ID NO: 190;

[0021] rs2862833:

[0022] C-type probe: SEQ ID NO: 192, T-type probe: SEQ ID NO: 194;

[0023] rs3751385:

[0024] C-type probe: SEQ ID NO: 196, T-type probe: SEQ ID NO: 198;

[0025] rs1053023:

[0026] C-type probe: SEQ ID NO: 200, T-type probe: SEQ ID NO: 202.

[0027] Of course, those skilled in the art can also reasonably design other probes to identify the above SNP marker combination according to actual use requirements, including but not limited to the C-type probes and T-type probes listed above.

[0028] A fourth aspect of the present invention provides a detection kit for detecting susceptibility to noise-induced hearing loss, the kit comprising the Taqman primers described in the second aspect of the present invention and the probe described in the third aspect of the present invention.

[0029] In the embodiment of the present invention, the inventors can accurately identify the specific base mutation of each SNP in the above SNP combination based on the above Taqman primers and probes, thereby effectively analyzing its genotype and judging its susceptibility to noise-induced hearing loss based on this.

[0030] In some embodiments of the present invention, when the rs2394795 is of TT or TC genotype, rs2862833 is of TT or CC genotype, rs3751385 is of TT genotype and rs1053023 is of AG genotype, the noise-induced hearing loss is susceptible; otherwise, the noise-induced hearing loss is tolerant.

[0031] In some embodiments of the present invention, the kit further includes other auxiliary agents, including buffer, amplification reagents, and enzymes.

[0032] Of course, those skilled in the art can also reasonably add or replace auxiliary agents in the kit according to actual use requirements to complete the corresponding detection work, including but not limited to the above-mentioned buffer, amplification reagents, and enzymes.

[0033] The fifth aspect of the present invention provides the use of the Taqman primers described in the second aspect of the present invention and the probes described in the third aspect of the present invention in the preparation of a noise-induced hearing loss susceptibility detection product.

[0034] In the embodiment of the present invention, the inventors can accurately identify the specific base mutation of each SNP in the above SNP combination based on the above Taqman primers and probes, thereby effectively analyzing its genotype and judging its susceptibility to noise-induced hearing loss based on this.

[0035] In some embodiments of the present invention, the product includes a detection reagent, a detection kit, and a gene chip.

[0036] Of course, those skilled in the art can also prepare other forms of products using the above-mentioned Taqman primers and probes according to actual use requirements, so as to achieve the effect of determining the susceptibility of the subject to noise-induced hearing loss by detecting the above-mentioned SNPs combination.

[0037] A sixth aspect of the present invention provides a method for distinguishing a population susceptible to noise-induced hearing loss from a population tolerant to noise-induced hearing loss, comprising the following steps:

[0038] The Taqman primers described in the second aspect of the present invention and the probes described in the third aspect of the present invention are used to detect the DNA sample to be tested, and the susceptibility and tolerance to noise-induced deafness are judged according to the genotype; when the rs2394795 is a TT or TC genotype, the rs2862833 is a TT or CC genotype, the rs3751385 is a TT genotype, and the rs1053023 is an AG genotype, the noise-induced deafness is susceptible; if not, the noise-induced deafness is tolerant.

[0039] There is no research on the correlation between the interaction of the combination of the above four susceptible gene variation sites and noise-induced hearing loss in the prior art. The present invention uses case-control studies, SnaPshot experiments, and Taqman probe methods to verify and discover for the first time eight SNPs sites related to ONID, and evaluates the cumulative risk of ONID caused by the risk genes of risk SNPs based on the genetic risk score (GRS) method. Then, according to the CART classification results, Logistic regression analysis is performed to explore the SNP-SNP interaction and the interaction between SNP-environmental multi-level variables, and finally determines that the SNP combination of the four genes CDH23-rs2394795, FAS-rs2862833, GJB2-rs3751385 and STAT3-rs1053023 is associated with the onset of noise-induced hearing loss.

[0040] In the embodiments of the present invention, the inventors found that the combination of variant sites of four genes (CDH23-rs2394795, FAS-rs2862833, GJB2-rs3751385 and STAT3-rs1053023) has diagnostic value for susceptibility to noise-induced hearing loss based on a case-control study of occupational noise-induced hearing loss. The Taqman probe kit constructed based on the combination of variant sites of these four susceptible genes can be used for the identification and screening of people susceptible to noise-induced hearing loss, and has a primary preventive effect on occupational noise-induced hearing loss.

[0041] In some embodiments of the present invention, the detection system is:

[0042] Components content 2×ChamQ Geno-SNP Probe Master Mix 10μL DNA to be tested 1μL 10μmol / L upstream primer 1.8μL 10μmol / L downstream primer 1.8μL C-type probe 0.4μL T-type probe 0.4μL <![CDATA[ddH2O]]> Make up to 20 μL

[0043] The detection conditions were as follows: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 10 sec, annealing at 60°C for 30 sec, extension at 60°C for 30 sec, and 40 cycles.

[0044] The beneficial effects of the present invention are:

[0045] 1. The present invention discovered for the first time the SNPs sites associated with ONID, and verified through experiments that the SNPs combination of the four genes CDH23-rs2394795, FAS-rs2862833, GJB2-rs3751385 and STAT3-rs1053023 is associated with the onset of noise-induced hearing loss, and can be used as an effective marker combination for the risk of occupational noise-induced hearing loss.

[0046] 2. The present invention provides a kit that can be used for assessing susceptibility to occupational noise-induced hearing loss. The kit can accurately assess the risk of occupational noise-induced hearing loss in a subject, thereby enabling effective identification and screening of people susceptible to noise-induced hearing loss, thereby having a primary preventive effect on occupational noise-induced hearing loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 This is a representative peak diagram of the extension product in the example of the present invention, green represents A, red represents T, black represents C, and green represents G.

[0048] Figure 2 is the GRS distribution of NIHL subjects and control subjects, where A is the different distribution of GRS in NIHL subjects and control subjects, and B is the dose-response relationship between GRS level and NIHL risk.

[0049] Figure 3 This is the classification and regression tree analysis diagram of genetic polymorphism.

[0050] Figure 4 This is the ROC curve analysis diagram for different SNP combinations. DETAILED DESCRIPTION

[0051] In order to make the invention purpose, technical solution and technical effect of the present invention clearer, the present invention is further described in detail below in conjunction with specific implementation methods. It should be understood that the specific implementation methods described in this specification are only for explaining the present invention, not for limiting the present invention.

[0052] Unless otherwise specified, the experimental materials and reagents used are consumables and reagents that can be obtained from conventional commercial channels.

[0053] Principles for sample population inclusion

[0054] In the embodiment of the present invention, the sample population included in the experiment was selected from Chinese Han workers who were exposed to occupational noise in Guangdong Province, and their working experience of noise exposure was more than 3 years.

[0055] At the same time, questionnaires were used to collect the subjects' general personal information, occupational history (especially the length of noise exposure time), health status, past history, family ear disease history, whether effective noise prevention measures were taken, etc., in order to reduce the interference of confounding factors on the experimental results. For case-control studies, the biggest problem they face is the influence of bias factors. Since workers exposed to occupational noise come from different companies, confounding factors such as type of work, length of service, gender, age, noise exposure intensity, noise exposure working hours, and protective measures can all become bias factors in occupational epidemiological surveys. Doing a good job in quality control of the survey population is the key to achieving survey and research accuracy.

[0056] The gender of the subjects was not controlled. The inventors found in previous experiments that there was no statistical difference between the noise-induced hearing loss susceptible group and the noise-induced hearing loss tolerance group when the gender factor was controlled.

[0057] Earplug protection is an unavoidable confounding factor. When investigating individuals or subjects, the wearing rate of earplugs must be investigated (expressed as the ratio of earplug wearing time to the entire noise exposure period), so that the survey population only has the following three controllable variables: cumulative noise exposure (CNE), hearing test indicators and earplug wearing rate. By referring to the age factor, the quality control problem of the survey sample can be solved.

[0058] Conduct an on-site noise survey of each subject's work environment, including measuring the equivalent continuous A-weighted sound pressure level at each subject's work station and calculating each subject's CNE.

[0059] Cumulative noise exposure (CNE) can be used as an evaluation index for the external dose level of noise exposure, and its calculation method is:

[0060] CNE=10×log10 SPL ×T;

[0061] Where SPL is the noise exposure intensity, in dB(A); T is the noise exposure time, in years.

[0062] CNE is an evaluation parameter derived from the theory of equal energy exposure and equal biological effects. It provides comparable noise exposure indicators for individuals with different noise exposure intensities and exposure times.

[0063] The audiometry method is as follows: measure the 125Hz-8KHz pure tone air conduction hearing threshold of the left and right ears of each subject 16 hours after the noise removal operation, and make a clinical diagnosis in accordance with the Diagnostic Standards for Occupational Noise-Induced Hearing Loss (GBZ49-2014).

[0064] According to the above method, a total of 200 patients with confirmed occupational noise-induced deafness (susceptible population to occupational noise-induced deafness) were finally included in the trial. Then, according to the principle of case-control study, 200 noise-exposed non-occupational noise-induced deafness subjects were randomly selected as the tolerant population (as control) based on the similarity of CNE, and 2 mL of EDTA anticoagulated whole blood was drawn from the susceptible and tolerant populations as research samples.

[0065] Analysis of SNPs in susceptible genes of noise-induced hearing loss

[0066] Blood samples from the susceptible and tolerant populations obtained above were taken, and the SNaPshot technology was used to perform typing tests on 62 SNPs of 30 NIHL candidate susceptible genes (SNPs are shown in Table 1). Logistic multivariate regression analysis was performed with the CNE and hearing loss levels of noise-induced hearing loss individuals as dependent variables and the frequencies of differential SNP genotypes or alleles as independent variables to determine the SNP genotypes or alleles associated with occupational noise-induced hearing loss and screen out susceptible SNP sets (groups).

[0067] Table 1 62 SNPs of noise-induced hearing loss susceptibility genes

[0068]

[0069]

[0070] Among them, in this embodiment, the specific operation of the SNaPshot technology used by the inventor is:

[0071] (1) Design specific primers for the above genes:

[0072] The DNA sequences of the SNP sites in Table 1 above were searched from NCBI, and amplification primers and SNaPshot primers for each SNP site were designed. The designed primers were synthesized.

[0073] Among them, the designed amplification primers and SNaPshot primers are shown in Table 2.

[0074] Table 2 Amplification primers of SNPs loci and SNaPshot primers

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082] (2) PCR amplification and purification:

[0083] Amplification was performed based on the above PCR primers, and the amplification reaction system is shown in Table 3.

[0084] Table 3 PCR amplification reaction system

[0085]

[0086]

[0087] PCR amplification conditions were as follows: pre-denaturation at 94°C for 3 min; denaturation at 94°C for 15 sec, annealing at 55°C for 15 sec, extension at 72°C for 30 sec, 35 cycles; final extension at 72°C for 3 min.

[0088] After amplification, agarose gel electrophoresis was used to verify whether the electrophoresis band was the target band and the PCR amplification product was recovered.

[0089] The recovered PCR amplification products were purified with ExoI enzyme and FastAP purification enzyme to remove excess primers, Taq enzyme and dNTP to ensure the normal subsequent sequencing reaction.

[0090] The specific reaction steps are as follows: add 0.8 μL FastAP purified enzyme (1 μg / μL), 0.2 μL ExoI enzyme (20 μg / μL), and 0.7 μL ExoI buffer to 3 μL PCR amplification product, and then add ddH2O to 7 μL. Mix well, put on the machine, incubate at 37℃ for 15 minutes, inactivate at 80℃ for 15 minutes, and store at 4℃.

[0091] (3) SNaPshot single base extension reaction:

[0092] The purified PCR amplification product was taken for extension reaction.

[0093] The SNaPshot single base extension reaction system is shown in Table 4.

[0094] Table 4 SNaPshot single base extension reaction system

[0095] Components content PCR amplification products 2μL each 10 μmol SNaPshot primer 0.2μL / strip Snapshot Mix Reagents 1μL <![CDATA[ddH2O]]> Make up to 6 μL

[0096] SNaPshot single base extension sequencing reaction conditions: 96°C, 1 min; 96°C, 10 sec, 52°C, 5 sec, 60°C, 30 sec, 30 cycles. The extension product was obtained and stored at 4°C for future use.

[0097] (4) Genotype detection:

[0098] 1 μL of the extension product obtained in the above example was placed in a 96-well plate, and then 9 μL of HIDI (highly deionized-formamide) was added to each well, denatured at 95°C for 3 min, placed in an ice-water bath, and detected using an ABI-3730XL sequencer. The sequencing results were analyzed using Gene Mapper 5.0 software.

[0099] Genotyping results such as Figure 1 As shown. Among them, Figure 1 This is a representative peak diagram of the extension product in the example of the present invention, green represents A, red represents T, black represents C, and green represents G.

[0100] The allele and genotype data of a single SNP locus were statistically analyzed, and the allele and three genotype frequencies in the case-control group were obtained to determine whether there were statistical differences (OR and P value). The chi-square test was used to analyze whether there were differences between the heterozygous mutant genotype (AB), homozygous mutant genotype (BB) and wild genotype (AA) in the ONID group and the control group under different models. The allele model was (A vs.B), the dominant model was (AA vs.BB+AB), the recessive model was (AA+AB vs.BB), the overdominant model was (AA+BB vs.AB), and the additive model was (AAvs.BB). The above different models were selected to perform logistic multiple regression analysis on the analysis results.

[0101] Logistic regression analysis was performed on the sites with different genotypes / alleles in the univariate analysis under the genetic model. After analysis, eight SNPs sites, including CDH23-rs2394795, FAS-rs1468063, FAS-rs2862833, GJB2-rs3751385, PTPRN2-rs10081191, SIK3-rs6589574, STST3-rs1053023, and STST3-rs1053005, were screened out and may be related to ONID susceptibility.

[0102] Effect verification of SNP combination of candidate susceptibility genes for occupational noise-induced hearing loss

[0103] The validation cohort consisted of 268 subjects, including 125 in the ONID group and 143 in the control group, to validate the screened SNPs associated with ONID.

[0104] The basic characteristics of the validation population were the same as those in the above examples. There were no statistically significant differences between the ONID group and the control group in CNE, gender, ethnicity, education level, use of protective measures, and drinking status (P>0.05). There were significant differences between the ONID group and the control group in age, length of service in noise exposure, high-frequency hearing threshold shift, and smoking status (P<0.05).

[0105] The mean age of the ONID group was 47.97±6.83 (± standard deviation), which was significantly higher than that of the control group (44.10±8.73 (P<0.001). The mean length of time of working in the ONID group was 12.65±6.18 (± standard deviation), which was significantly higher than that of the control group (9.91±5.69 (P<0.001). The high-frequency hearing threshold shift of the ONID group was 56.33±10.79 (± standard deviation), which was significantly higher than that of the control group (26.76±7.14 (P<0.001).

[0106] The 8 SNPs sites selected in the above example were verified using the Taqman probe method, and positive results were determined by fluorescence excitation. The Taqman probe sequences and primers of some SNPs sites are shown in Table 6.

[0107] Table 6 Taqman probe sequences and primer sequences of some SNPs sites

[0108]

[0109]

[0110] Wherein, the detection system of the above-mentioned Taqman probe method is:

[0111] Table 7 Detection system of Taqman probe method

[0112] Components content 2×ChamQ Geno-SNP Probe Master Mix 10μL DNA to be tested 1μL 10μmol / L upstream primer 1.8μL 10μmol / L downstream primer 1.8μL C-type probe 0.4μL T-type probe 0.4μL <![CDATA[ddH2O]]> Make up to 20 μL

[0113] The above-mentioned ChamQ Geno-SNP Probe Master Mix was purchased from Nanjing Novoprotein Scientific Co., Ltd.

[0114] The reaction procedure was as follows: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 10 sec, annealing at 60°C for 30 sec, extension at 60°C for 30 sec, with 40 cycles.

[0115] Using the results of Logistic regression analysis, it was found that seven SNPs loci, namely CDH23-rs2394795, FAS-rs1468063, FAS-rs2862833, GJB2-rs3751385, PTPRN2-rs10081191, STAT3-rs1053023, and STAT3-rs1053005, were associated with the susceptibility to ONID (P<0.05).

[0116] Further analysis was performed using the gene-gene interaction analysis method. The SNPs with the strongest association with ONID and the corresponding genotypes increasing the risk of ONID onset, that is, the risk genotypes, were obtained by Logistic multiple regression analysis. The cumulative onset risk of ONID caused by the risk genes of the risk SNPs was evaluated using the genetic risk score (GRS).

[0117] RS was defined as the sum of the numbers of the risk genotypes of the SNPs related to ONID (the risk genotype was 2, the heterozygote was 1, and the non-risk genotype was 0). The calculation formula was as follows:

[0118]

[0119] where, n i was the number of the risk genotypes in SNPi; k was the number of the SNPs related to ONID.

[0120] P<0.05 was selected as having significant statistical differences, and 0.05<P<0.10 was selected as having marginal significant statistical differences. The dose-response relationship between GRS and OR was tested for trends among different GRS levels using the χ2 test.

[0121] The GRS analysis results were as Figure 2 shown in Table 8. It was found that the distribution of GRS between the ONID group and the control group was different, and the median of the ONID group (P 25 、P 75) GRS was 8.0 (6.0, 10.0), which was significantly higher than 6.0 (5.0, 9.0) in the control group (P<0.001). Moreover, the GRS level in the ONID group was higher than that in the control group. When further grading according to the GRS level, the subjects with the lowest GRS level (≤5) were used as the reference, and ORs (95% CIs) were calculated. It was found that ORs were marginally correlated with GRS (P trend <0.10). If the subjects were further divided into two groups (high GRS or low GRS) according to whether the GRS value was higher than 8, the risk of ONID in subjects with GRS ≧ 8 was significantly increased compared with that in subjects with GRS < 8, with an OR of 1.748 (95% CI = 1.079, 2.849).

[0122] Table 8 Relationship between GRS level and ONID susceptibility

[0123]

[0124] Among them, * means P < 0.05, # means 0.05 <P<0.10。

[0125] At the same time, CART analysis was performed on the above results using the rpart package in R 4.1.2 software, with the termination node being the binary variable of ONID or not. CART constructs a decision tree through recursive partitioning to analyze the prediction of outcomes for each genotype (AA, AB, BB) of susceptible SNPs or environmental factor variables (such as age, gender, CNE, smoking and drinking status, etc.). Logistic regression analysis was performed based on the CART classification results to explore SNP-SNP interactions and interactions between SNP-environmental multi-level variables.

[0126] The results of CART analysis are as follows Figure 3As shown in Table 9. According to the analysis results of CART, there were interactions among the four SNPs sites CDH23-rs2394795, FAS-rs2862833, GJB2-rs3751385 and STAT3-rs1053023. Compared with the subjects who simultaneously occurred FAS-rs2862833(AA / BB) and GJB2-rs3751385(AB), when CDH23-rs2394795(AA / AB), FAS-rs2862833(AA / BB), GJB2-rs3751385(AA / AB) and STAT3-rs1053023(AB) occurred simultaneously, the risk of ONID in the subjects would increase by 5.950 times, with 95% CIs of 2.563-14.600(P<0.001); and when GJB2-rs3751385(BB) appeared, the risk of ONID in the subjects would increase by 6.023 times, with 95% CIs of 3.077-12.187(P<0.001). The above results show that the influence of these four SNPs variant sites on the risk of ONID is as follows: FAS-rs2862833 (AA / BB) <STAT3-rs1053023(AB)<CDH23-rs2394795(AA / AB)<GJB2-rs3751385(BB)。

[0127] Table 9 Classification of gene interactions and risk estimation results of decision tree terminal nodes

[0128]

[0129] Wherein, a indicates adjustment for age, total working years, CNE, gender, nationality, drinking, smoking, and protective measures. * indicates P < 0.05.

[0130] Receiver operating characteristic curve (ROC) analysis was further performed based on these four SNP sites to reflect the classification effect of CART and evaluate the diagnostic ability of these four variant sites as evaluation indicators for ONID susceptibility.

[0131] The results are as follows Figure 4 As shown in Table 10. It can be seen that the combined diagnosis of FAS-rs2862833 (AA / BB) + STAT3-rs1053023 (AB) + CDH23-rs2394795 (AA / AB) + GJB2-rs3751385 (BB) is more accurate than the diagnosis of FAS-rs2862833 (AA / BB) alone.

[0132] Table 10 Area under the ROC curve and its trend analysis

[0133]

[0134] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention. SEQUENCE LISTING <110> Guangdong Pharmaceutical University <120> A combination of noise-induced hearing loss markers and its detection kit and application <130> <160> 202 <170> PatentIn version 3.5 <210> 1 <211> 20 <212> DNA <213> Artificial sequence <400> 1 agccaatcag aaggcagtcc 20 <210> 2 <211> 20 <212> DNA <213> Artificial sequence <400> 2 gctttctaaa cggaccttcg 20 <210> 3 <211> 35 <212> DNA <213> Artificial sequence <400> 3 tttttttttt tttgctgagc ctgaagtcgc cacgg 35 <210> 4 <211> 20 <212> DNA <213> Artificial sequence <400> 4 cctgaggagg tgtagaaatc 20 <210> 5 <211> 20 <212> DNA <213> Artificial sequence <400> 5 gatgggtgtt gatttcctcc 20 <210> 6 <211> twenty two <212> DNA <213> Artificial sequence <400> 6 tcaaaattcc tgcttacctg gg 22 <210> 7 <211> 20 <212> DNA <213> Artificial sequence <400> 7 ttgcctatcc tgacactcac 20 <210> 8 <211> 20 <212> DNA <213> Artificial sequence <400> 8 aaaaagcgcc cagttagcag 20 <210> 9 <211> 42 <212> DNA <213> Artificial sequence <400> 9 tttttttttt tttttttttt gagtggccaa ctaccagcgt ga 42 <210> 10 <211> twenty one <212> DNA <213> Artificial sequence <400> 10 ttaagggccc agattttgga g 21 <210> 11 <211> twenty one <212> DNA <213> Artificial sequence <400> 11 ccctctttct tctaggtctt g 21 <210> 12 <211> twenty two <212> DNA <213> Artificial sequence <400> 12 aaattggggt aaggctaccc ac 22 <210> 13 <211> twenty one <212> DNA <213> Artificial sequence <400> 13 tgacctcact tggaagagga g 21 <210> 14 <211> twenty two <212> DNA <213> Artificial sequence <400> 14 tcaggtaagt aggaggcgtg tg 22 <210> 15 <211> 32 <212> DNA <213> Artificial sequence <400> 15 tttttttttt atgccactca cctaaacgcc ag 32 <210> 16 <211> twenty one <212> DNA <213> Artificial sequence <400> 16 ttaagggccc agattttgga g 21 <210> 17 <211> twenty one <212> DNA <213> Artificial sequence <400> 17 ccctctttct tctaggtctt g 21 <210> 18 <211> twenty two <212> DNA <213> Artificial sequence <400> 18 cctccccctc caacctggtg tt 22 <210> 19 <211> twenty one <212> DNA <213> Artificial sequence <400> 19 ggttgcaaac atcttctgct c 21 <210> 20 <211> twenty one <212> DNA <213> Artificial sequence <400> 20 gagagatact tttcacgtgc c 21 <210> twenty one <211> 32 <212> DNA <213> Artificial sequence <400> twenty one tttttttttt aattattcca tcctacccaa gc 32 <210> twenty two <211> twenty one <212> DNA <213> Artificial sequence <400> twenty two taccacaaca tgctgtaatc g 21 <210> twenty three <211> twenty one <212> DNA <213> Artificial sequence <400> twenty three tgaagtcttc tctgggcaat g 21 <210> twenty four <211> twenty two <212> DNA <213> Artificial sequence <400> twenty four acataaaaat cgttgattcc ac 22 <210> 25 <211> 20 <212> DNA <213> Artificial sequence <400> 25 acgcattgcc cagttgttag 20 <210> 26 <211> 20 <212> DNA <213> Artificial sequence <400> 26 accaataacc cctaacagcc 20 <210> 27 <211> 35 <212> DNA <213> Artificial sequence <400> 27 tttttttttt tttaatcttt gtgttgggaa atgct 35 <210> 28 <211> twenty one <212> DNA <213> Artificial sequence <400> 28 ggattcttcg gcttcttcac c 21 <210> 29 <211> twenty one <212> DNA <213> Artificial sequence <400> 29 gccagatggt tttcaacgac a 21 <210> 30 <211> twenty two <212> DNA <213> Artificial sequence <400> 30 attcaacgtc tacatcgagt cc 22 <210> 31 <211> twenty one <212> DNA <213> Artificial sequence <400> 31 ctttccttat tcccaagtcc g 21 <210> 32 <211> twenty one <212> DNA <213> Artificial sequence <400> 32 tctggcttca catacacaga g 21 <210> 33 <211> twenty two <212> DNA <213> Artificial sequence <400> 33 gttgtcctag ctaatgaatg ca 22 <210> 34 <211> twenty one <212> DNA <213> Artificial sequence <400> 34 tgcagcagtg gaaccttaat g 21 <210> 35 <211> twenty one <212> DNA <213> Artificial sequence <400> 35 ctcgatgtag acgttgaatg g 21 <210> 36 <211> 32 <212> DNA <213> Artificial sequence <400> 36 tttttttttt ggcccgcagg tccccccgca gc 32 <210> 37 <211> 20 <212> DNA <213> Artificial sequence <400> 37 ctgagaaaga aaagcgagcc 20 <210> 38 <211> 20 <212> DNA <213> Artificial sequence <400> 38 agccctacaa agaccctcac 20 <210> 39 <211> twenty two <212> DNA <213> Artificial sequence <400> 39 cctccttcct ggtctacctg gc 22 <210> 40 <211> twenty one <212> DNA <213> Artificial sequence <400> 40 gacagaagga gcagaatctt g 21 <210> 41 <211> twenty one <212> DNA <213> Artificial sequence <400> 41 acccagaaac caaagccatt c 21 <210> 42 <211> 32 <212> DNA <213> Artificial sequence <400> 42 tttttttttt gccatttttg gcctcaaacc ca 32 <210> 43 <211> twenty one <212> DNA <213> Artificial sequence <400> 43 cccactatca ctgtttttag c 21 <210> 44 <211> twenty one <212> DNA <213> Artificial sequence <400> 44 gacctagtag ttataggata c 21 <210> 45 <211> 42 <212> DNA <213> Artificial sequence <400> 45 tttttttttt tttttttttt aggatataat caggttgatt ta 42 <210> 46 <211> twenty one <212> DNA <213> Artificial sequence <400> 46 gatgaggatc agtatggctt g 21 <210> 47 <211> twenty one <212> DNA <213> Artificial sequence <400> 47 gggattacta gtgctgttgt c 21 <210> 48 <211> 32 <212> DNA <213> Artificial sequence <400> 48 tttttttttt caaacagaaa gccattctga aa 32 <210> 49 <211> 20 <212> DNA <213> Artificial sequence <400> 49 acttccagtc aggtagttgc 20 <210> 50 <211> 20 <212> DNA <213> Artificial sequence <400> 50 tttgagcctt tgaccatgcc 20 <210> 51 <211> twenty two <212> DNA <213> Artificial sequence <400> 51 aaaaagttaa acattgaagt aa 22 <210> 52 <211> 20 <212> DNA <213> Artificial sequence <400> 52 tgttctaaag gtggtgaggc 20 <210> 53 <211> 20 <212> DNA <213> Artificial sequence <400> 53 gcaacaagaa atctcccgtg 20 <210> 54 <211> 40 <212> DNA <213> Artificial sequence <400> 54 ttttttttttttttttttca gccggaggcc agagctgagc 40 <210> 55 <211> twenty one <212> DNA <213> Artificial sequence <400> 55 cctttctttc ctgttgaagg c 21 <210> 56 <211> twenty one <212> DNA <213> Artificial sequence <400> 56 gtaaatatcc ccttggctgt g 21 <210> 57 <211> 32 <212> DNA <213> Artificial sequence <400> 57 tttttttttt tcctggagga gcacggaaaa ga 32 <210> 58 <211> twenty one <212> DNA <213> Artificial sequence <400> 58 cctttctttc ctgttgaagg c 21 <210> 59 <211> twenty one <212> DNA <213> Artificial sequence <400> 59 gtaaatatcc ccttggctgt g 21 <210> 60 <211> twenty two <212> DNA <213> Artificial sequence <400> 60 acagccatga gctttctcca ga 22 <210> 61 <211> twenty one <212> DNA <213> Artificial sequence <400> 61 tcatgtgctg tttggaagag g 21 <210> 62 <211> twenty one <212> DNA <213> Artificial sequence <400> 62 ttcttatttt tcccccaccc c 21 <210> 63 <211> 32 <212> DNA <213> Artificial sequence <400> 63 tttttttttt aatcttaaat cttagaaact tg 32 <210> 64 <211> twenty one <212> DNA <213> Artificial sequence <400> 64 tcatgtgctg tttggaagag g 21 <210> 65 <211> twenty one <212> DNA <213> Artificial sequence <400> 65 ttcttatttt tcccccaccc c 21 <210> 66 <211> 32 <212> DNA <213> Artificial sequence <400> 66 ttttttttttttctaaggga tccaagaagc at 32 <210> 67 <211> 20 <212> DNA <213> Artificial sequence <400> 67 acactgtcac tagtctcacc 20 <210> 68 <211> 20 <212> DNA <213> Artificial sequence <400> 68 tttgaggtag tcacagggag 20 <210> 69 <211> 45 <212> DNA <213> Artificial sequence <400> 69 tttttttttt tttttttttt tttttcagtg ccgacctgcg ccaat 45 <210> 70 <211> 20 <212> DNA <213> Artificial sequence <400> 70 acggcatagg tgagacccta 20 <210> 71 <211> 20 <212> DNA <213> Artificial sequence <400> 71 gctgttacca gcacaaacga 20 <210> 72 <211> 42 <212> DNA <213> Artificial sequence <400> 72 tttttttttt tttttttttt ggccttcctg atcatgctcc tc 42 <210> 73 <211> 20 <212> DNA <213> Artificial sequence <400> 73 aggacactta ggtccccaac 20 <210> 74 <211> 20 <212> DNA <213> Artificial sequence <400> 74 cagatagcac tgggaaagac 20 <210> 75 <211> 35 <212> DNA <213> Artificial sequence <400> 75 tttttttttt tttacactga cttaagattc taact 35 <210> 76 <211> twenty one <212> DNA <213> Artificial sequence <400> 76 cagacaaaga tgaggcagag g 21 <210> 77 <211> 20 <212> DNA <213> Artificial sequence <400> 77 tcagacccag gaatctgagc 20 <210> 78 <211> 42 <212> DNA <213> Artificial sequence <400> 78 tttttttttt tttttttttt gccgggggct ctcttcttca gc 42 <210> 79 <211> 20 <212> DNA <213> Artificial sequence <400> 79 tggcctgaag caaatggaag 20 <210> 80 <211> 20 <212> DNA <213> Artificial sequence <400> 80 ggtgtatttt ggagttctgg 20 <210> 81 <211> twenty two <212> DNA <213> Artificial sequence <400> 81 tatagtcata aacttttgct ga 22 <210> 82 <211> 20 <212> DNA <213> Artificial sequence <400> 82 accaacctgt aacccaatcc 20 <210> 83 <211> 20 <212> DNA <213> Artificial sequence <400> 83 tgaggtagga gaatcccttg 20 <210> 84 <211> twenty two <212> DNA <213> Artificial sequence <400> 84 ttgcactcca gcctgggcta ca 22 <210> 85 <211> 20 <212> DNA <213> Artificial sequence <400> 85 accaggttga tgtagtaggg 20 <210> 86 <211> 20 <212> DNA <213> Artificial sequence <400> 86 caacactgga actctctctc 20 <210> 87 <211> twenty two <212> DNA <213> Artificial sequence <400> 87 ccttccacgt ggggtggttg tc 22 <210> 88 <211> twenty one <212> DNA <213> Artificial sequence <400> 88 attagcagcc ctaatggtgt c 21 <210> 89 <211> twenty one <212> DNA <213> Artificial sequence <400> 89 gacatgagag aatagagcag g 21 <210> 90 <211> twenty two <212> DNA <213> Artificial sequence <400> 90 gtgaataatc accaaattgt at 22 <210> 91 <211> twenty one <212> DNA <213> Artificial sequence <400> 91 tacaaccact tgccgtcatt c 21 <210> 92 <211> twenty one <212> DNA <213> Artificial sequence <400> 92 gcatcactaa gtcaagacac c 21 <210> 93 <211> 32 <212> DNA <213> Artificial sequence <400> 93 tttttttttt tggtgacaat gacaacacca ca 32 <210> 94 <211> 20 <212> DNA <213> Artificial sequence <400> 94 caggattgcc tgatctgaac 20 <210> 95 <211> 20 <212> DNA <213> Artificial sequence <400> 95 gcataattcc tcgtagcacc 20 <210> 96 <211> twenty two <212> DNA <213> Artificial sequence <400> 96 ttgatttaat tctcataagc aa 22 <210> 97 <211> 20 <212> DNA <213> Artificial sequence <400> 97 tacagcaaca ggtcaacagg 20 <210> 98 <211> 20 <212> DNA <213> Artificial sequence <400> 98 tgtgtacagg cagcaagaag 20 <210> 99 <211> 45 <212> DNA <213> Artificial sequence <400> 99 tttttttttt tttttttttt ttttcatggc aggttaatgg gttgg 45 <210> 100 <211> 20 <212> DNA <213> Artificial sequence <400> 100 atgaaggact aagcggtctg 20 <210> 101 <211> 20 <212> DNA <213> Artificial sequence <400> 101 ttccgtctac ctttgctgtg 20 <210> 102 <211> 35 <212> DNA <213> Artificial sequence <400> 102 tttttttttt tttccttctg cgcatgcact acgcg 35 <210> 103 <211> 20 <212> DNA <213> Artificial sequence <400> 103 ggttatcggg aactccaagg 20 <210> 104 <211> 19 <212> DNA <213> Artificial sequence <400> 104 ggctcctgct ctgaagtcg 19 <210> 105 <211> 32 <212> DNA <213> Artificial sequence <400> 105 tttttttttt tgggaaaata ttgcagaagg gc 32 <210> 106 <211> 20 <212> DNA <213> Artificial sequence <400> 106 aaactgaggc taacagctgg 20 <210> 107 <211> 20 <212> DNA <213> Artificial sequence <400> 107 atgctcctca acgaaaccac 20 <210> 108 <211> 35 <212> DNA <213> Artificial sequence <400> 108 ttttttttttttttcacaag agctcagggc tggga 35 <210> 109 <211> 20 <212> DNA <213> Artificial sequence <400> 109 ttgcttcacc ttgtccttgc 20 <210> 110 <211> 20 <212> DNA <213> Artificial sequence <400> 110 ggaacagtcc ttccatcttg 20 <210> 111 <211> 45 <212> DNA <213> Artificial sequence <400> 111 tttttttttt tttttttttt tttgttcagc tgtgtttgcc cctca 45 <210> 112 <211> twenty one <212> DNA <213> Artificial sequence <400> 112 gacacctcca tggaagaaaa c 21 <210> 113 <211> twenty three <212> DNA <213> Artificial sequence <400> 113 ccctgtgttt ctgagaccct ttg 23 <210> 114 <211> 32 <212> DNA <213> Artificial sequence <400> 114 tttttttttt tgacactcag gaacagcacg ga 32 <210> 115 <211> twenty one <212> DNA <213> Artificial sequence <400> 115 agcctagacc aatagaaagc c 21 <210> 116 <211> twenty one <212> DNA <213> Artificial sequence <400> 116 acacctattc ttctgccaga g 21 <210> 117 <211> twenty two <212> DNA <213> Artificial sequence <400> 117 gagtgtttta atgggaaaga ag 22 <210> 118 <211> 20 <212> DNA <213> Artificial sequence <400> 118 ccagtttctt ggctcctatg 20 <210> 119 <211> 19 <212> DNA <213> Artificial sequence <400> 119 atctgtccag tcgctcgtc 19 <210> 120 <211> 35 <212> DNA <213> Artificial sequence <400> 120 ttttttttttttttcagactccagccgctcttgga 35 <210> 121 <211> 20 <212> DNA <213> Artificial sequence <400> 121 gacgccgcca tattttacag 20 <210> 122 <211> 20 <212> DNA <213> Artificial sequence <400> 122 cagagaacgc tggaaaaacc 20 <210> 123 <211> 35 <212> DNA <213> Artificial sequence <400> 123 tttttttttttttcggtgtg gggcagtggc tgacc 35 <210> 124 <211> 20 <212> DNA <213> Artificial sequence <400> 124 actcgttgat gttggtctcg 20 <210> 125 <211> 20 <212> DNA <213> Artificial sequence <400> 125 atcctgagta ggtgggaatg 20 <210> 126 <211> 35 <212> DNA <213> Artificial sequence <400> 126 tttttttttt tttcgtcccc gtgtaccctg gaccg 35 <210> 127 <211> twenty one <212> DNA <213> Artificial sequence <400> 127 tttgaaacca tgaccttggg c 21 <210> 128 <211> twenty one <212> DNA <213> Artificial sequence <400> 128 cacttctgcc cattgttcta c 21 <210> 129 <211> twenty two <212> DNA <213> Artificial sequence <400> 129 gtgaagggct atcacagcac ac 22 <210> 130 <211> twenty one <212> DNA <213> Artificial sequence <400> 130 gagggaaaac agagagaatg c 21 <210> 131 <211> twenty one <212> DNA <213> Artificial sequence <400> 131 ttcctcatgt accagcatca g 21 <210> 132 <211> twenty two <212> DNA <213> Artificial sequence <400> 132 ttcctaggac aggctgaccc ca 22 <210> 133 <211> twenty one <212> DNA <213> Artificial sequence <400> 133 tatgtgccct ctttctcttg g 21 <210> 134 <211> twenty one <212> DNA <213> Artificial sequence <400> 134 cagtaaacca gtcagttgct c 21 <210> 135 <211> 42 <212> DNA <213> Artificial sequence <400> 135 tttttttttt tttttttttt atgtagcctg ttggcattat gt 42 <210> 136 <211> 20 <212> DNA <213> Artificial sequence <400> 136 gggcccctaa gaatttgtgt 20 <210> 137 <211> 20 <212> DNA <213> Artificial sequence <400> 137 ccaccacacc cagctaattt 20 <210> 138 <211> twenty two <212> DNA <213> Artificial sequence <400> 138 gcagtgcttc ttgaactttc ac 22 <210> 139 <211> twenty one <212> DNA <213> Artificial sequence <400> 139 aaaagctagg tgatgctcca g 21 <210> 140 <211> twenty one <212> DNA <213> Artificial sequence <400> 140 tccggtatag catcgtgtat c 21 <210> 141 <211> 45 <212> DNA <213> Artificial sequence <400> 141 tttttttttt tttttttttt tttaaagg aagaaaatcc ttaga 45 <210> 142 <211> twenty two <212> DNA <213> Artificial sequence <400> 142 gtgcaaagtg tgtggatgta tg 22 <210> 143 <211> twenty one <212> DNA <213> Artificial sequence <400> 143 atcagaaagc tgagtctacc c 21 <210> 144 <211> twenty two <212> DNA <213> Artificial sequence <400> 144 gtattcacat gtgagtgtaa at 22 <210> 145 <211> twenty one <212> DNA <213> Artificial sequence <400> 145 tttgggcaag aatctggtca c 21 <210> 146 <211> twenty one <212> DNA <213> Artificial sequence <400> 146 acagatgtag cctagttggt c 21 <210> 147 <211> 32 <212> DNA <213> Artificial sequence <400> 147 tttttttttt acctccaagg accagggctg gg 32 <210> 148 <211> twenty one <212> DNA <213> Artificial sequence <400> 148 gcctttgaaa gaaagaaagg g 21 <210> 149 <211> twenty two <212> DNA <213> Artificial sequence <400> 149 tgggcacgca ccgagcgtgt gg 22 <210> 150 <211> 32 <212> DNA <213> Artificial sequence <400> 150 tttttttttt atgaaacagg aggactttgg ga 32 <210> 151 <211> twenty two <212> DNA <213> Artificial sequence <400> 151 ttccctctcc tttcctctag ac 22 <210> 152 <211> twenty one <212> DNA <213> Artificial sequence <400> 152 tatcccaatc cacggacaat c 21 <210> 153 <211> 32 <212> DNA <213> Artificial sequence <400> 153 tttttttttt cactttttag ggaatgctga ga 32 <210> 154 <211> twenty one <212> DNA <213> Artificial sequence <400> 154 caatgtggat gtcgagtagt g 21 <210> 155 <211> twenty one <212> DNA <213> Artificial sequence <400> 155 ctcctgttgg tcacaaattc c 21 <210> 156 <211> twenty two <212> DNA <213> Artificial sequence <400> 156 aactgtttac attttctatc tg 22 <210> 157 <211> twenty two <212> DNA <213> Artificial sequence <400> 157 caaaccaaac atttgacctc cc 22 <210> 158 <211> twenty one <212> DNA <213> Artificial sequence <400> 158 gttcagcctg gattatatgg g 21 <210> 159 <211> 32 <212> DNA <213> Artificial sequence <400> 159 tttttttttt acatctgtgg tatatcatta ca 32 <210> 160 <211> twenty one <212> DNA <213> Artificial sequence <400> 160 ggaggtggag tcttccaaat g 21 <210> 161 <211> twenty two <212> DNA <213> Artificial sequence <400> 161 acttccccca gttctcctac tc 22 <210> 162 <211> 40 <212> DNA <213> Artificial sequence <400> 162 ttttttttttttttttttag gctaagtaaa aagttagggg 40 <210> 163 <211> twenty two <212> DNA <213> Artificial sequence <400> 163 attctgggct acccagatct tc 22 <210> 164 <211> twenty two <212> DNA <213> Artificial sequence <400> 164 tttgccttgg tgatggagtc ag 22 <210> 165 <211> 48 <212> DNA <213> Artificial sequence <400> 165 tttttttttt tttttttttt ttttttccag cctgagctcc tgcacaca 48 <210> 166 <211> twenty one <212> DNA <213> Artificial sequence <400> 166 ttgtagagat gaaggtctcc c 21 <210> 167 <211> twenty one <212> DNA <213> Artificial sequence <400> 167 tgagcatgtg tggttatcct g 21 <210> 168 <211> 48 <212> DNA <213> Artificial sequence <400> 168 tttttttttt tttttttttt ttttttatct taccaaataa attttgag 48 <210> 169 <211> twenty one <212> DNA <213> Artificial sequence <400> 169 caacagccaa agccaaatca g 21 <210> 170 <211> twenty two <212> DNA <213> Artificial sequence <400> 170 cgcactgtgc ctgaatttta ac 22 <210> 171 <211> 32 <212> DNA <213> Artificial sequence <400> 171 tttttttttt cttatgaatg aaataaggtt tc 32 <210> 172 <211> twenty one <212> DNA <213> Artificial sequence <400> 172 agttccaggc actaagtaga c 21 <210> 173 <211> twenty one <212> DNA <213> Artificial sequence <400> 173 catgatcact gtgtgcctta g 21 <210> 174 <211> twenty two <212> DNA <213> Artificial sequence <400> 174 cccatagcaa tttgcctaca ct 22 <210> 175 <211> twenty one <212> DNA <213> Artificial sequence <400> 175 agctgatcag agtttctgtg g 21 <210> 176 <211> twenty one <212> DNA <213> Artificial sequence <400> 176 ccaccttata ggtaggtaag c 21 <210> 177 <211> twenty two <212> DNA <213> Artificial sequence <400> 177 gagctgagcc ctgttgtggc cc 22 <210> 178 <211> twenty one <212> DNA <213> Artificial sequence <400> 178 agctgatcag agtttctgtg g 21 <210> 179 <211> twenty one <212> DNA <213> Artificial sequence <400> 179 ccaccttata ggtaggtaag c 21 <210> 180 <211> 42 <212> DNA <213> Artificial sequence <400> 180 tttttttttt tttttttttt aagggggaga cgaccttctc ta 42 <210> 181 <211> twenty one <212> DNA <213> Artificial sequence <400> 181 actagaatgg tgagcttgtg c 21 <210> 182 <211> twenty one <212> DNA <213> Artificial sequence <400> 182 tgctcacttt gctttctcgt c 21 <210> 183 <211> 32 <212> DNA <213> Artificial sequence <400> 183 tttttttttt ggccttacca cggacacatc tg 32 <210> 184 <211> twenty one <212> DNA <213> Artificial sequence <400> 184 taccgctaat ggctactttg g 21 <210> 185 <211> twenty one <212> DNA <213> Artificial sequence <400> 185 caggagttca aggatgtagt g 21 <210> 186 <211> 42 <212> DNA <213> Artificial sequence <400> 186 tttttttttt tttttttttt ctagggcggt tgagaagtga ac 42 <210> 187 <211> twenty two <212> DNA <213> Artificial sequence <400> 187 tctctgtgtg tagcagtggt ga 22 <210> 188 <211> 17 <212> DNA <213> Artificial sequence <400> 188 ttgtatctga tttctgc 17 <210> 189 <211> twenty four <212> DNA <213> Artificial sequence <400> 189 atcagatgca ctcaactttg aata 24 <210> 190 <211> 17 <212> DNA <213> Artificial sequence <400> 190 ttgtatttga tttctgc 17 <210> 191 <211> 20 <212> DNA <213> Artificial sequence <400> 191 ttttttggta ggggcttgct 20 <210> 192 <211> 16 <212> DNA <213> Artificial sequence <400> 192 ctttgatgct tcttgg 16 <210> 193 <211> 27 <212> DNA <213> Artificial sequence <400> 193 aggggttaagg ttaaaaagga agtacct 27 <210> 194 <211> 18 <212> DNA <213> Artificial sequence <400> 194 tgtctttaat gcttcttg 18 <210> 195 <211> 19 <212> DNA <213> Artificial sequence <400> 195 cccgtgctca gctgtcaag 19 <210> 196 <211> 15 <212> DNA <213> Artificial sequence <400> 196 ctcagtcgcc agcat 15 <210> 197 <211> twenty two <212> DNA <213> Artificial sequence <400> 197 ggagtttcac ctgaggccta ca 22 <210> 198 <211> 16 <212> DNA <213> Artificial sequence <400> 198 ctcagtcgct agcatt 16 <210> 199 <211> 19 <212> DNA <213> Artificial sequence <400> 199 agggcctgag gaccctgtt 19 <210> 200 <211> 18 <212> DNA <213> Artificial sequence <400> 200 ctttaatggg ccacaaca 18 <210> 201 <211> twenty three <212> DNA <213> Artificial sequence <400> 201 aaagttctga gaggagctga gcc 23 <210> 202 <211> 17 <212> DNA <213> Artificial sequence <400> 202 ctttaacggg ccacaac 17

Claims

1. Use of a primer probe for detecting a SNP marker combination associated with noise-induced deafness susceptibility in the preparation of a noise-induced deafness susceptibility detection product, characterized in that: The SNP marker combination is rs2394795, rs2862833, rs3751385 and rs1053023; When the rs2394795 is of TT or TC genotype, the rs2862833 is of TT or CC genotype, the rs3751385 is of TT genotype, and the rs1053023 is of AG genotype, the mouse is susceptible to noise-induced hearing loss.

2. The use according to claim 1, characterized in that: The nucleotide sequence of the primer is: rs2394795: upstream primer SEQ ID NO: 187 and downstream primer SEQ ID NO: 189; rs2862833: upstream primer SEQ ID NO: 191 and downstream primer SEQ ID NO: 193; rs3751385: upstream primer SEQ ID NO: 195 and downstream primer SEQ ID NO: 197; rs1053023: upstream primer SEQ ID NO:199 and downstream primer SEQ ID NO:

201.

3. The use according to claim 1, characterized in that: The nucleotide sequence of the probe is: rs2394795： C-type probe: SEQ ID NO: 188, T-type probe: SEQ ID NO: 190; rs2862833： C-type probe: SEQ ID NO: 192, T-type probe: SEQ ID NO: 194; rs3751385： C-type probe: SEQ ID NO: 196, T-type probe: SEQ ID NO: 198; rs1053023： C-type probe: SEQ ID NO: 200, T-type probe: SEQ ID NO:

202.

4. The use according to claim 1, characterized in that: The products include detection kits and gene chips.

5. The use according to claim 4, characterized in that: The detection kit also includes other auxiliary agents, including buffer, amplification reagents, and enzymes.

Images