Reagent detection method for high-frequency mutation sites of hereditary hearing loss

Abstract

The invention relates to the field of quality testing, and discloses a reagent detection method for a hereditary hearing loss high-frequency mutation site, which is used for providing a reliable measurement technical means for quality detection of a hereditary hearing loss high-frequency mutation site detection reagent. Comprising the following steps: acquiring original fluorescence physical signals of a reagent solution under a plurality of optical detection channels by using an optical detection instrument; performing photoelectric conversion and standardization on the repeated physical detection signals, and generating a weighted synthesis signal in combination with an expected response weight so as to evaluate the overall optical luminescence physical stability of the reagent solution; a correlation network between physical response signals of different detection channels is constructed, the luminescence dynamics consistency in a reagent solution system is quantified by calculating the global efficiency of the correlation network, and the fluctuation amplitude of the physical luminescence state of the reagent solution is accurately measured. According to the invention, comprehensive, sensitive and anti-interference closed-loop testing of real physical properties of the reagent materials can be realized.

Description

Technical Field

[0001] This invention relates to the field of quality testing, and more particularly to a reagent detection method for a high-frequency mutation site in hereditary deafness. Background Technology

[0002] Hereditary deafness, a common birth defect, places a heavy burden on patients and their families. Accurate detection of high-frequency mutation sites associated with hereditary deafness is crucial for early diagnosis, genetic counseling, and the development of preventative interventions. With the continuous development of gene testing technology, testing reagents targeting mutation sites in hereditary deafness have emerged and are widely used in clinical diagnosis and screening. To ensure the accuracy of diagnosis and screening, manufacturers need to test the composition of the solvents in the reagents before selling them to ensure that the solvent costs meet requirements.

[0003] Currently, the detection method used to detect solutions in reagents is based on a single light source. However, a single light source has limited dimensions, resulting in inaccurate detection results and serious bias in the measurement results of solution components. Existing detection instruments fail to adequately compensate for background fluctuations at the physical level when processing raw photoelectric signals. When faced with aging photoelectric devices, voltage noise, or minor environmental thermodynamic disturbances, the detection results are highly susceptible to external hardware interference.

[0004] Therefore, we propose a reagent detection method for high-frequency mutation sites in hereditary deafness to address the above-mentioned problems. Summary of the Invention

[0005] This invention provides a reagent detection method for high-frequency mutation sites in hereditary deafness, providing a reliable technical means for quality testing of reagents for detecting high-frequency mutation sites in hereditary deafness.

[0006] The first aspect of this invention provides a reagent detection method for high-frequency mutation sites in hereditary deafness. The method includes: repeatedly physical excitation and optical signal acquisition of a test reagent solution using an optical detection instrument to obtain the original fluorescence physical signals of the reagent solution under multiple optical detection channels, and processing them to form a standardized physical detection signal matrix; applying a phase weight vector to the standardized physical detection signal matrix for beamforming operations to generate a phase-weighted synthesized signal sequence to extract a stability index characterizing the overall optical luminescence physical stability of the test reagent solution; analyzing the standardized physical detection signal matrix and filtering it according to a preset significance threshold to construct a channel physical response correlation network, and calculating the global efficiency of the network to obtain a physical structure consistency index; analyzing the distribution probability of the initial physical detection signals of the test reagent solution under each optical detection channel, and calculating a luminescence state uncertainty index; calculating a comprehensive physical quality index based on the stability index of the phase-weighted synthesized signal sequence, the physical structure consistency index, and the luminescence state uncertainty index; and determining the quality of the reagent solution by comparing the comprehensive physical quality index with a preset threshold.

[0007] Optionally, in a first implementation of the first aspect of the present invention, the method includes: acquiring expected response characteristic data of each optical detection channel to form a priori information set; generating a phase weight vector based on the priori information set through mathematical transformation, wherein the mathematical transformation makes the expected response intensity negatively correlated with the weight value; generating an intermediate synthesis vector based on the phase weight vector and the standardized physical detection signal matrix; processing the intermediate synthesis vector to generate a phase-weighted synthesis signal sequence; and calculating the coefficient of variation of the phase-weighted synthesis signal sequence as a stability index characterizing the overall optical luminescence physical stability of the test reagent solution.

[0008] Optionally, in a second implementation of the first aspect of the present invention, the method includes: calculating the correlation coefficient between column vectors corresponding to any two different optical detection channels in the standardized physical detection signal matrix to form a physical response correlation coefficient matrix; comparing each element in the physical response correlation coefficient matrix with a preset significance threshold to select elements whose absolute value is greater than or equal to the significance threshold; based on the selection results, constructing an undirected weighted network with each optical detection channel as a node and the selected significant correlation relationships as connecting edges, as a channel physical response correlation network; and calculating the reciprocal of the harmonic mean of the shortest path lengths between all possible node pairs in the channel physical response correlation network as the global efficiency of the network to obtain a physical structure consistency index.

[0009] Optionally, in a third implementation of the first aspect of the present invention, based on the physical response correlation coefficient matrix, the average of the absolute values ​​of all elements is calculated to generate an initial correlation level reference value; based on the initial correlation level reference value, an initial candidate threshold is calculated and generated through a preset threshold adjustment rule; the physical response correlation coefficient matrix is ​​screened using the initial candidate threshold to construct an initial candidate network; the average number of shortest paths between nodes and the node clustering coefficient of the initial candidate network are calculated to generate network topology feature indicators; based on the degree of difference between the network topology feature indicators and the theoretical ideal network features, the initial candidate threshold is calibrated to generate a preset significance threshold.

[0010] Optionally, in the fourth implementation of the first aspect of the present invention, the method includes: based on the initial physical detection data matrix of the test reagent, counting the number of times the original fluorescence physical signal of each optical detection channel breaks through the preset reference emission threshold in all repeated physical excitation detections, and generating the effective emission frequency of each optical detection channel; dividing the effective emission frequency of each optical detection channel by the total number of detections to calculate the effective emission probability value of each optical detection channel; calculating the information content value corresponding to each optical detection channel according to the effective emission probability value of each optical detection channel through a preset information content calculation rule; and generating a comprehensive information entropy value based on the information content values ​​of all optical detection channels as an uncertainty index of the emission state.

[0011] Optionally, in the fifth implementation of the first aspect of the present invention, the effective emission probability value of each optical detection channel is logarithmically transformed to generate a corresponding first transformation value; the absolute difference between the effective emission probability value and the fixed reference probability value of each optical detection channel is calculated to generate a corresponding probability deviation value; the first transformation value and the probability deviation value of each optical detection channel are calculated to generate a second transformation value of the detection channel; a negative operation is applied to the second transformation value of each optical detection channel to generate the information value corresponding to the detection channel.

[0012] Optionally, in a sixth implementation of the first aspect of the present invention, the method includes: normalizing the stability index, the physical structure consistency index, and the luminescence state uncertainty index respectively to generate corresponding normalized stability index, normalized physical structure consistency index, and normalized uncertainty index; processing the normalized stability index, normalized physical structure consistency index, and normalized uncertainty index with a preset set of coefficients to obtain a comprehensive physical quality index; comparing the comprehensive physical quality index with a preset quality qualification threshold to generate a comparison result; and outputting the quality judgment result of the test reagent solution based on the comparison result.

[0013] Optionally, in a seventh implementation of the first aspect of the invention, an execution comprehensive physical quality index is introduced. : ; in, As a normalized stability index, , The coefficient of variation; This is a normalized physical structure consistency index; The normalized uncertainty index , For comprehensive information entropy; As a stability weight, For consistency weight, This represents the uncertainty weight.

[0014] Optionally, in an eighth implementation of the first aspect of the present invention, the method further includes: acquiring calibration and testing data of a standard reference reagent, wherein the standard reference reagent has known and qualified physicoluminescence properties; processing the calibration and testing data using the same physical detection and analysis steps as the test reagent solution to generate a baseline physical quality index of the standard reference reagent; comparing the baseline physical quality index of the standard reference reagent with the theoretical qualified index to obtain a system calibration coefficient to compensate for the physical attenuation error of the optical instrument; adjusting the comprehensive physical quality index of the test reagent solution using the system calibration coefficient to generate a calibrated comprehensive physical quality index; and outputting a calibrated quality judgment result based on the comparison result between the calibrated comprehensive physical quality index and the theoretical qualified index.

[0015] Optionally, in the ninth implementation of the first aspect of the present invention, obtaining the calibration and testing data of the standard reference reagent includes: performing repeated physical excitation and optical signal acquisition and testing on the standard reference reagent a predetermined number of times under the same optical detection instrument to obtain the original calibration fluorescence physical signal results for each optical detection channel; organizing the original calibration fluorescence physical signal results into a calibration initial data matrix, wherein the rows of the matrix correspond to the number of calibration tests and the columns correspond to each optical detection channel; calculating the calibration mean and calibration standard deviation of the corresponding column for each optical detection channel in the calibration initial data matrix; based on the calibration mean and calibration standard deviation of each optical detection channel, performing a standardized transformation on all the corresponding original calibration fluorescence physical signal results to generate a standardized physical calibration signal for each optical detection channel; and combining the standardized physical calibration signals of all optical detection channels to construct a calibration and testing data matrix of the standard reference reagent.

[0016] Beneficial effects: It overcomes the problem that traditional optical instruments are easily interfered with by photoelectric background noise when directly reading single-point fluorescence physical quantities. By using the prior information of the expected response characteristics of each optical detection channel to construct a phase weight vector, the original physical measurement signal after photoelectric conversion is directionally enhanced, effectively filtering random photoelectric noise, and realizing efficient and high signal-to-noise ratio physical signal extraction for reagent material quality assessment.

[0017] This approach transforms the complex quality assessment of multi-component reagents into the deep extraction of multi-dimensional optical physical signals and network topology measurement. It precisely quantifies the underlying optical luminescence physical stability of the reagent solution and the consistency of luminescence dynamics among different detection channels. By calculating the luminescence state uncertainty index, it accurately measures the fluctuation range of the physical luminescence state of the reagent solution, providing comprehensive and reliable pure physical material performance parameters for the research and development, quality control, and factory inspection of multi-channel fluorescence detection reagents.

[0018] By introducing a standard reference reagent with known and qualified luminescence properties, and calculating the system calibration coefficient based on it, the systemic physical measurement background error caused by the attenuation of the excitation light source and the decrease in the sensitivity of the optical sensor can be effectively compensated. This significantly improves the cross-device reusability and objective accuracy of the reagent physical performance judgment results, and ensures a high degree of consistency in quality assessment standards between different batches and different measuring instruments. Attached Figure Description

[0019] Figure 1 This is a schematic diagram of an embodiment of a reagent detection method for high-frequency mutation sites of hereditary deafness in this invention.

[0020] Figure 2 This is a schematic diagram illustrating the process of standardizing raw detection data from different optical detection channels.

[0021] Figure 3 This is a schematic diagram of another embodiment of the reagent detection method for high-frequency mutation sites of hereditary deafness in this invention. Detailed Implementation

[0022] This invention provides a reagent detection method for high-frequency mutation sites in hereditary deafness, offering a reliable technical means for quality testing of reagents used to detect high-frequency mutation sites in hereditary deafness. The terms "first," "second," "third," "fourth," etc. (if present) in the specification, claims, and accompanying drawings are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate so that the embodiments described herein can be implemented in orders other than those illustrated or described herein. Furthermore, the terms "comprising" or "having" and any variations thereof are intended to cover a non-exclusive inclusion; for example, a process, method, system, product, or apparatus that comprises a series of steps or units is not necessarily limited to those steps or units explicitly listed, but may include other steps or units not explicitly listed or inherent to such processes, methods, products, or apparatus.

[0023] For ease of understanding, the specific process of the embodiments of the present invention is described below. Please refer to [link / reference]. Figure 1 One embodiment of the reagent detection method for high-frequency mutation sites of hereditary deafness in this invention includes: 101. Using an optical detection instrument, the test reagent solution is subjected to repeated physical excitation and optical signal acquisition to obtain the original fluorescence physical signal of the reagent solution under multiple optical detection channels. The original fluorescence physical signal of each optical detection channel is subjected to photoelectric conversion and standardization processing to form a standardized physical detection signal matrix. It is understood that the executing entity of this invention can be a reagent detection device for high-frequency mutation sites in hereditary deafness, or it can be a terminal or a server; the specific implementation is not limited here. This embodiment of the invention will be described using a server as an example.

[0024] It should be noted that quality control personnel place the test reagent in a 96-well reaction plate and then place it in a multi-channel optical detection instrument equipped with a specific excitation light source and a high-sensitivity photomultiplier tube (PMT). Under the set thermodynamic temperature control cycle, the excitation light source irradiates the reagent solution to induce a physicochemical reaction. The optical sensor captures the original physical fluorescence photons of the reagent solution in the first optical detection channel (FAM fluorescence band), the second optical detection channel (VIC fluorescence band), and the third optical detection channel (ROX fluorescence band) in real time, converts them into voltage analog signals, and outputs them as original digital fluorescence signals after A / D conversion.

[0025] The raw fluorescence physical signal exported by the instrument exhibits natural photoelectric physical fluctuations. Standardization and division are performed based on the ideal full-scale reference value (5000) set by the instrument.

[0026] To demonstrate matrix operations, the standardized results of the first 5 physical detections are extracted: First time: the first optical detection channel is 0.90, the second optical detection channel is 0.64, and the third optical detection channel is 1.00.

[0027] Second time: The first optical detection channel is 0.92, the second optical detection channel is 0.65, and the third optical detection channel is 1.01.

[0028] The third time: the first optical detection channel is 0.88, the second optical detection channel is 0.62, and the third optical detection channel is 0.98.

[0029] Fourth time: The first optical detection channel is 0.91, the second optical detection channel is 0.66, and the third optical detection channel is 1.02.

[0030] Fifth time: The first optical detection channel is 0.89, the second optical detection channel is 0.63, and the third optical detection channel is 0.99.

[0031] The server combines these five tests to generate a standardized physical detection signal matrix of five rows and three columns. In reality, the backend generates a massive matrix of ninety-six rows and three columns.

[0032] 102. Apply the phase weight vector determined based on the prior information of the expected response characteristics of each optical detection channel to the standardized physical detection signal matrix to perform beamforming operation, generate a phase-weighted synthesized signal sequence, and extract a stability index characterizing the overall optical luminescence physical stability of the test reagent solution. It should be noted that, based on the prior information of the theoretical luminescence intensity ratio and concentration proportion of each luminescent component within the reagent, a weight of 0.5 is assigned to the first optical detection channel with the strongest physical response, a weight of 0.3 is assigned to the second optical detection channel, and a weight of 0.2 is assigned to the third optical detection channel. These three values ​​are arranged sequentially to form the phase weight vector.

[0033] Perform row-by-row beamforming on the five-row, three-column matrix shown in step 101: For the first test (first row), multiply 0.90 by 0.5, 0.64 by 0.3, and 1.00 by 0.2, and add them together to get 0.842.

[0034] For the second test (second row), multiply 0.92 by 0.5, 0.65 by 0.3, and 1.01 by 0.2, and add them together to get 0.857.

[0035] For the third test (third row), multiply 0.88 by 0.5, 0.62 by 0.3, and 0.98 by 0.2, and add them together to get 0.822.

[0036] For the fourth test (fourth row), multiply 0.91 by 0.5, 0.66 by 0.3, and 1.02 by 0.2, and add them together to get 0.857.

[0037] For the fifth test (the fifth row), multiply 0.89 by 0.5, 0.63 by 0.3, and 0.99 by 0.2, and add them together to get 0.832.

[0038] Perform this operation on all 96 rows of data to generate a one-dimensional phase-weighted synthetic signal sequence containing 96 values.

[0039] 103. Analyze the correlation between physical response signals of different optical detection channels in the standardized physical detection signal matrix and filter them according to the preset significance threshold. Construct a channel physical response correlation network and calculate the global efficiency of the network to obtain a physical structure consistency index that characterizes the consistency of luminescence dynamics inside the reagent solution. It should be noted that by calling the normalized matrix containing 96 complete tests, the complete column sequences of the three optical detection channels were extracted and the absolute correlation coefficients were calculated. Due to the small errors in the chemiluminescence kinetics of different luminescent components, these three sequences exhibit complex fluctuations that are not entirely linear.

[0040] The calculations show that the first and third optical detection channels have extremely high synchronicity in fluctuations, with a correlation coefficient of 0.98; the first and second optical detection channels have a correlation coefficient of 0.95; while the second and third optical detection channels have a relatively low correlation coefficient of 0.85.

[0041] The pre-set saliency threshold for the research and development was 0.90. Based on this, the network was constructed as follows: the connection edges between the "first channel and the third channel" and between the "first channel and the second channel" were retained; the direct connection between the "second channel and the third channel" was severed. The network exhibits a relay structure centered on the first optical detection channel.

[0042] Calculate the global efficiency of the network: For the first pair (from the first channel to the second channel), the shortest path is one, and the reciprocal is one.

[0043] For the second pair (from the first channel to the third channel), the shortest path is one, and the reciprocal is one.

[0044] The third pair (from the second channel to the third channel) requires a transfer in the first channel. The shortest path is two, and the reciprocal is 0.5.

[0045] Adding these three reciprocals gives 2.5, which, when divided by the logarithm of 3, yields an average of 0.833. This value represents the physical structure consistency index of the batch of reagents, reflecting the degree of chemical kinetic synchronization during the mixed reaction of multiple luminescent components within the detection system.

[0046] 104. Analyze the distribution probability of the initial physical detection signal of the test reagent solution under each optical detection channel, and calculate the luminescence state uncertainty index, which characterizes the fluctuation amplitude of the physical luminescence state of the reagent solution. It should be noted that, for the complete data from 96 tests, the signal mean of each independent optical detection channel was first calculated. Then, the data was divided into three intervals according to the degree of deviation: the central stable zone (deviation from the mean within 2 percent), the edge drift zone (deviation from 2 percent to 5 percent), and the abnormal fluctuation zone (deviation from more than 5 percent).

[0047] Simultaneously, the unstandardized initial 96-row physical detection data matrix is ​​retrieved. At this point, the instrument's preset baseline physical luminescence thresholds (i.e., the minimum photoelectric response level required for effective luminescence of solution components) for the first, second, and third detection channels are 2020, 1490, and 3120, respectively. The entire 96 data points are iterated through, and the number of effective luminescence frequencies exceeding these physical thresholds is counted. The first optical detection channel has a central stable region accounting for 80%, an edge drift region for 15%, and an abnormal fluctuation region for 5%. In 96 stimulated tests, it exceeded the physical threshold 86 times, resulting in an effective emission frequency of 86 and an effective emission probability of approximately 0.896. Using the information entropy rule, its uncertainty is calculated to be 0.883.

[0048] Second optical detection channel: The light emission performance of this channel is slightly unstable, with a stable central region accounting for 70%, a drifting edge region accounting for 20%, and an abnormal region accounting for 10%. The effective light emission frequency is 77, and the effective light emission probability value is approximately 0.802. The calculated information entropy is 1.156.

[0049] The third optical detection channel exhibits extremely stable performance, with a 90% stable central region, 8% at the edges, and 2% in the abnormal region; the effective emission frequency is 91, and the effective emission probability is approximately 0.948. The calculated information entropy is 0.529.

[0050] Adding the information entropy of these three channels and dividing by three yields an average uncertainty of 0.856. This value, serving as an overall uncertainty index for the luminescence state, precisely quantifies the background random fluctuation amplitude of the physicoluminescence state of the reagent solution upon excited excitation.

[0051] 105. Based on the stability index of the phase-weighted synthesized signal sequence, the physical structure consistency index, and the luminescence state uncertainty index, calculate the comprehensive physical quality index, and determine the quality of the reagent solution by comparing the comprehensive physical quality index with a preset threshold. It should be noted that variance statistics were performed on the 96-bit weighted synthetic sequence generated in step 102, and the "stability index" was calculated to be 0.940.

[0052] Retrieve the "Physical Structure Consistency Index" of 0.833 generated in step 103.

[0053] The "luminescence state uncertainty index" of 0.856 calculated in step 104 is retrieved.

[0054] In the quality control model, the weights are set as follows: stability index accounts for 40 points; physical structure consistency index accounts for 40 points; since uncertainty is usually in the range of 0 to 2 under the natural logarithm, it is set to be directly multiplied by 10 as a negative deduction item. The qualified factory exit threshold is set at 60.00 points.

[0055] Calculation process: Stability 0.940 multiplied by 40 equals 37.60 points; Consistency 0.833 multiplied by 40 equals 33.32 points; Uncertainty 0.856 multiplied by 10 equals 8.56 points to be deducted.

[0056] The total score is 37.60 plus 33.32, minus 8.56, resulting in a final comprehensive physical quality index of 62.36.

[0057] Since the score of 62.36 is greater than the passing threshold of 60.00, the reagent solution is deemed to be of acceptable quality, as shown in Table 1 below: Table 1

[0058] Please see Figures 2-3 ,in Figure 2 The three dot plots on the left represent the raw physical detection signals from three different optical detection channels, each with a different mean and standard deviation. The three dot plots on the right show the results of these signals after standardization; they have all been converted into a distribution with a mean of 0 and a standard deviation of 1. Another embodiment of the reagent detection method for high-frequency mutation sites in hereditary deafness, as described in this invention, includes: 201. Using optical detection instruments, the test reagent solution is subjected to repeated physical excitation and optical signal acquisition to obtain the original fluorescence signal reflecting the overall excited photophysical properties of the reagent solution. The signal is then converted into photoelectric signal and standardized to form a standardized physical detection signal matrix. Specifically, the test reagent solution is subjected to repeated physical excitation detections to obtain its original physical detection results in each optical detection channel, forming an initial physical detection data matrix with the number of detections as rows and the optical detection channels as columns. The statistical mean and statistical standard deviation of the corresponding column for each optical detection channel in the initial physical detection data matrix are calculated. Based on the statistical mean and statistical standard deviation of each optical detection channel, all the corresponding original detection results are standardized and transformed to generate standardized detection signals for each optical detection channel. The standardized detection signals of all optical detection channels are combined to construct a standardized physical detection signal matrix. Among them, the initial physical detection data matrix is ​​used to calculate the luminescence state uncertainty index, and the standardized physical detection signal matrix is ​​used to generate a phase-weighted synthetic signal sequence and construct a channel physical response correlation network.

[0059] It should be noted that the quality control personnel conducted 96 independent and repeated physical excitation tests using the test reagent solution and the standard luminescent reference matrix. Three optical detection channels were the focus of monitoring: the first optical detection channel (FAM channel), the second optical detection channel (VIC channel), and the third optical detection channel (ROX channel).

[0060] The instrument generated an "initial physical detection data matrix" consisting of 96 rows and 3 columns. To clearly illustrate the data extrapolation process, excerpts of the original data from the first five detections are provided below: First test: Channel 1 2100, Channel 2 1500, Channel 3 3200.

[0061] Second test: Channel 1 2000, Channel 2 1550, Channel 3 3100.

[0062] Third test: Channel 1 2200, Channel 2 1450, Channel 3 3300.

[0063] Fourth test: Channel 1 2050, Channel 2 1480, Channel 3 3150.

[0064] Fifth test: Channel 1 2150, Channel 2 1520, Channel 3 3250.

[0065] The statistics for each column of these 96 data points were calculated independently. The global mean of the first optical detection channel was 2100, and the standard deviation was approximately 79.05; the mean of the second optical detection channel was 1500, and the standard deviation was approximately 38.08; the mean of the third optical detection channel was 3200, and the standard deviation was approximately 79.05.

[0066] Then, perform standardization transformation one by one: subtract the mean of the column from the original physical value, and then divide by the standard deviation of the column.

[0067] In the first test, the raw values ​​of the three channels were exactly equal to the mean, and after conversion, they were all 0.00.

[0068] The second test yielded a value of -1.27 for the first channel (2000 minus 2100 divided by 79.05); a value of +1.31 for the second channel; and a value of -1.27 for the third channel.

[0069] The third test yielded positive values ​​of 1.27, negative values ​​of 1.31, and positive values ​​of 1.27 in sequence.

[0070] The fourth test yielded values ​​of -0.63, -0.53, and -0.63.

[0071] The fifth test yielded positive values ​​of 0.63, 0.53, and 0.63.

[0072] These 96 lines of converted data are assembled to generate a complete standardized physical detection signal matrix.

[0073] 202. Apply the phase weight vector determined based on the prior information of the expected response characteristics of each optical detection channel to the standardized physical detection signal matrix to perform beamforming operation and generate a phase-weighted synthesized signal sequence. Specifically, the expected response characteristic data of each optical detection channel are acquired to form a priori information set; based on the priori information set, a phase weight vector is generated through mathematical transformation, which makes the expected response intensity negatively correlated with the weight value; the phase weight vector is multiplied by a standardized physical detection signal matrix to generate an intermediate composite vector; the intermediate composite vector is normalized to generate a phase-weighted composite signal sequence; the coefficient of variation of the phase-weighted composite signal sequence is calculated as a stability index; wherein, the phase weight vector is used for matrix multiplication, and the coefficient of variation of the phase-weighted composite signal sequence is used to calculate the comprehensive physical quality index.

[0074] It should be noted that the theoretical physical ratio and luminescence characteristics of the luminescent components inside the extraction reagent are as follows: the expected response intensity ratio of the first optical detection channel is 20%, the second optical detection channel is 10%, and the third optical detection channel is 5%.

[0075] To achieve negative correlation, the reciprocals are calculated: five, ten, and twenty respectively, with the sum of the reciprocals being thirty-five. After calculating the proportions for each, the phase weight vectors are obtained: the weight of the first optical detection channel is approximately 0.143, the weight of the second optical detection channel is approximately 0.286, and the weight of the third optical detection channel is approximately 0.571.

[0076] Perform row-by-row matrix multiplication on this vector and a 96-row normalized matrix (multiplying each column element-wise and then summing the results): For the extracted fragment, the first row is all zeros, and the composite value is 0.000.

[0077] Second line: Multiply 1.27 by 0.143, 1.31 by 0.286, and 1.27 by 0.571, and add them together to get the midpoint 0.532.

[0078] The third line is calculated similarly, yielding a positive value of 0.532.

[0079] The result of the fourth line of calculation is negative 0.602.

[0080] The calculation in the fifth line yields a positive value of 0.602.

[0081] Perform magnitude normalization on the intermediate composite vector containing 96 values. Lock the global minimum to -0.602, the maximum to +0.602, and the range to 1.204. Subtract the minimum value from each value and divide by the range: the first row is converted to 0.500, the second to 0.058, the third to 0.942, the fourth to 0.000, and the fifth to 1.000.

[0082] A phase-weighted composite signal sequence of length 96 was generated. Statistical analysis showed that the arithmetic mean of this sequence was 0.500, and the sample standard deviation was 0.472. Dividing the standard deviation by the mean yielded a coefficient of variation of 0.944. This coefficient serves as an indicator of the overall stability of the test reagent solution.

[0083] 203. Analyze the correlation between response signals of different optical detection channels in the standardized physical detection signal matrix and filter them according to the preset significance threshold. Construct a channel physical response correlation network and calculate the global efficiency of the network as a physical structure consistency index. Specifically, the correlation coefficient between column vectors corresponding to any two different optical detection channels in the standardized physical detection signal matrix is ​​calculated to form a physical response correlation coefficient matrix. Each element in the physical response correlation coefficient matrix is ​​compared with a preset significance threshold, and elements with absolute values ​​greater than or equal to the threshold are selected. Based on the selection results, an undirected weighted network is constructed with each optical detection channel as a node and the selected significant correlations as connecting edges, serving as the channel physical response correlation network. The reciprocal of the harmonic mean of the shortest path lengths between all possible node pairs in the channel physical response correlation network is calculated as the global efficiency of the network, i.e., the physical structure consistency index. Among these, the physical response correlation coefficient matrix is ​​used for network construction, and the global efficiency of the network is used to calculate the comprehensive physical quality index.

[0084] Furthermore, the determination of the preset significance threshold includes: calculating the average of the absolute values ​​of all elements based on the physical response correlation coefficient matrix to generate an initial correlation level reference value; calculating and generating an initial candidate threshold based on the initial correlation level reference value and a preset threshold adjustment rule; using the initial candidate threshold to filter the physical response correlation coefficient matrix and construct an initial candidate network; calculating the average shortest path number between nodes and the node clustering coefficient of the initial candidate network to generate network topology feature indicators; and calibrating the initial candidate threshold based on the degree of difference between the network topology feature indicators and the theoretical ideal network features to generate the final preset significance threshold.

[0085] It should be noted that the 96-row standardized matrix from step 201 is retrieved, and its three column vectors are paired to calculate the absolute value of the Pearson correlation coefficient. Based on the physical fluctuation trend of the aforementioned standardized data, the following results are calculated: the absolute correlation coefficient between the first and second optical detection channels is approximately 0.66; the emission fluctuations of the first and third optical detection channels are completely synchronized, with a coefficient of 1.00; and the coefficient between the second and third optical detection channels is approximately 0.66.

[0086] Calculate the arithmetic mean of these three values ​​to obtain an initial reference value of 0.77 for the correlation level. Following a preset rule (increased by 10%), generate an initial candidate threshold of 0.85.

[0087] Using a threshold of 0.85 for exploratory screening, only the first and third channels (1.00) passed, while the second channel became an isolated node. The clustering coefficient of the nodes in this initial candidate network was calculated to be zero, indicating that it seriously deviated from the ideal network topology characteristic that "all luminescent components should be in the same stable chemiluminescent system".

[0088] After the calibration mechanism is triggered, the significance threshold is lowered and finally determined to be 0.60.

[0089] Re-screening using a final threshold of 0.60 resulted in all correlation coefficients meeting the requirement. An undirected weighted network containing all three optical detection channel nodes was constructed. The correlation coefficient was used as the weight, and its reciprocal was used as the shortest path length of the network. The path length from the first channel to the second channel is 1 divided by 0.66, approximately 1.51.

[0090] The path length from the first channel to the third channel is 1 divided by 1.00, which equals 1.00.

[0091] The path length from the second channel to the third channel is 1 divided by 0.66, approximately 1.51.

[0092] Calculate the reciprocal of the harmonic mean of the three shortest path lengths: take the reciprocals of 1.51, 1.00, and 1.51 again to restore them to 0.66, 1.00, and 0.66, add them together to get 2.32, divide by the total logarithm 3, and finally obtain the average value of 0.773.

[0093] This value of 0.773 represents the objective physical structural consistency index of this batch of reagents, reflecting the robustness of the luminescence dynamics of the multi-component detection system.

[0094] 204. Analyze the distribution probability of the initial physical detection signal of the test reagent solution under each optical detection channel, and calculate the luminescence state uncertainty index, which characterizes the fluctuation amplitude of the physical luminescence state of the reagent. Specifically, based on the initial physical detection data matrix of the test reagent, the number of times the original fluorescence physical signal of each optical detection channel breaks through the preset reference emission threshold in all repeated physical excitation detections is counted to generate the effective emission frequency of each optical detection channel; the effective emission frequency of each detection channel is divided by the total number of detections to calculate the effective emission probability value of each optical detection channel. Based on the effective emission probability value of each optical detection channel, the information value corresponding to each optical detection channel is calculated according to the preset information value calculation rules. The information value of all detection channels is then averaged to generate a comprehensive information entropy value, which serves as the uncertainty index of the emission state.

[0095] Furthermore, the preset information content calculation rules include: performing a logarithmic domain transformation on the effective emission probability value of each optical detection channel to generate a corresponding first transformation value; calculating the absolute difference between the effective emission probability value of each detection channel and a fixed reference probability value to generate a corresponding probability deviation value; multiplying the first transformation value and the probability deviation value of each detection channel to generate a second transformation value for that channel; and applying a negative sign operation to the second transformation value of each detection channel to generate the information content value corresponding to that optical detection channel.

[0096] It should be noted that the 96 lines of the "Initial Physical Detection Data Matrix" that were not standardized in step 201 are retrieved. At this time, the instrument's preset reference physical emission thresholds (i.e., the minimum photoelectric response level for determining that the solution components have generated effective light emission) for the first, second, and third optical detection channels are 2020, 1490, and 3120, respectively.

[0097] Iterate through all 96 data points and count the effective emission frequencies that exceed the above threshold: According to statistics, the first optical detection channel exceeded the physical threshold 86 times out of 96 stimulated tests, with an effective emission frequency of 86 and an effective emission probability of approximately 0.896.

[0098] The second optical detection channel showed slightly weaker light emission, exceeding the threshold 77 times, with an effective light emission frequency of 77 and an effective light emission probability of approximately 0.802.

[0099] The third optical detection channel performed best, exceeding the threshold 91 times, with an effective emission frequency of 91 and an effective emission probability of approximately 0.948.

[0100] The preset ideal fixed reference probability value is 1.0 (i.e., 100% stable stimulated response).

[0101] Calculate the information content of each optical detection channel one by one: For the first optical detection channel: Taking the natural logarithm of 0.896 yields a first conversion value of -0.110. Calculating the absolute difference between this and 1.0 yields a probability deviation of 0.104. Multiplying the two yields a second conversion value of -0.0114. Applying a negative sign transforms this to a positive value, resulting in an information content value of 0.0114 for the first channel.

[0102] For the second optical detection channel: taking the natural logarithm of 0.802 gives -0.221. The absolute difference is 0.198. Multiplying the two gives -0.0438. After applying the negative sign, the information content of the second channel is 0.0438 (large deviation, the entropy value caused by physical state fluctuations increases significantly).

[0103] For the third optical detection channel: taking the natural logarithm of 0.948 gives -0.053. The absolute difference is 0.052. Multiplying these gives -0.0028. After applying the negative sign, the information content of the third channel is 0.0028.

[0104] Adding these three information values ​​(0.0114, 0.0438, and 0.0028) gives 0.0580. Then, performing an arithmetic mean (dividing by 3) yields a comprehensive information entropy value of approximately 0.0193. This value is the uncertainty index for the luminescence state of this batch of reagent solutions.

[0105] 205. Based on the stability index, physical structure consistency index, and luminescence state uncertainty index of the phase-weighted synthesized signal sequence, calculate the comprehensive physical quality index, and determine the quality of the reagent solution based on the comparison result of the index with the preset threshold. Specifically, the coefficient of variation of the phase-weighted synthesized signal sequence is calculated as a stability index; the stability index, physical structure consistency index, and luminescence state uncertainty index are normalized to generate corresponding normalized stability index, normalized physical structure consistency index, and normalized uncertainty index; the above three normalized indices are weighted and summed with a preset set of coefficients to calculate the comprehensive physical quality index; the comprehensive physical quality index is compared with a preset quality qualification threshold to generate a comparison result; based on the comparison result, the quality judgment result of the test reagent solution is output.

[0106] It should be noted that the three core indicators generated in the preceding process are: stability indicator (coefficient of variation S). idx The physical structure consistency index (global efficiency C) is 0.944; idx The uncertainty index of luminescence state (comprehensive information entropy U) is 0.773; idx The value is 0.0193.

[0107] To satisfy the requirements of weighted summation, the following normalization transformation is performed: Normalized stability index (N) S ): Using a negative exponential mapping of the natural constant. Calculation That is, e -0.944 The normalized value is 0.389.

[0108] Normalized physical structure consistency index (N C The indicator itself is positive and between 0 and 1, so it is directly assigned an equivalent value, resulting in a normalized value of 0.773.

[0109] Normalized uncertainty index (N) U ): An exponential decay amplification difference mapping is used, with a decay coefficient set to 10. Calculation That is, e -0.193 The normalized value is 0.824.

[0110] The preset set of weighting coefficients is as follows: (Stability weight) accounts for 0.40. (Consistency weight) accounts for 0.40. (Uncertainty weight) accounts for 0.20.

[0111] Introducing the implementation of the Comprehensive Physical Quality Index (CQI) calculation: ; Substituting the data: 0.40 × 0.389 = 0.1556; 0.40 × 0.773 = 0.3092; 0.20 × 0.824 = 0.1648.

[0112] The sum of the three terms is 0.6296, which, when multiplied by 100, yields a comprehensive physical quality index of 62.96.

[0113] The quality pass threshold (60.00 points) was retrieved. Since 62.96 points is greater than 60.00 points, the reagent is preliminarily determined to be of acceptable quality, as shown in Table 2 below: Table 2

[0114] 206. Obtain calibration test data for the standard reference reagent, which has known and qualified physicoluminescence properties; process the calibration test data using the same physical testing and analysis steps as the test reagent solution to generate the baseline physical quality index of the standard reference reagent; compare the baseline physical quality index of the standard reference reagent with the theoretical qualified index to calculate the system calibration coefficient to compensate for the physical attenuation error of the optical instrument; adjust the comprehensive physical quality index of the test reagent solution using the system calibration coefficient to generate the calibrated comprehensive physical quality index; based on the comparison result between the calibrated comprehensive physical quality index and the theoretical qualified index, output the final calibrated quality judgment result. Furthermore, obtaining calibration and testing data for the standard reference reagent includes: performing a predetermined number of repeated physical excitation tests on the standard reference reagent using a standard testing procedure to obtain the original calibration fluorescence physical signal results for each optical detection channel; organizing the original calibration fluorescence physical signal results into an initial calibration data matrix, where the rows of the matrix correspond to the number of calibration tests and the columns correspond to each optical detection channel; calculating the calibration mean and calibration standard deviation for each column corresponding to each optical detection channel in the initial calibration data matrix; based on the calibration mean and calibration standard deviation for each channel, standardizing and transforming all corresponding original calibration results to generate standardized physical calibration signals for each optical detection channel; and combining the standardized physical calibration signals of all optical detection channels to construct the calibration and testing data matrix for the standard reference reagent.

[0115] It should be noted that the quality inspectors took out a batch of standard reference reagents that were certified by a national statutory body and had perfect physical luminescence performance, and performed 96 repeated excitation tests on the same multi-channel fluorescence detector according to the standard physical measurement procedure, generating the original calibration fluorescence physical signal results for each optical detection channel.

[0116] The results were organized into an initial calibration data matrix, and the aforementioned operational models 201 to 205 were fully reused (covering calculation of the mean and standard deviation of photoelectric signals, standardization, beamforming, construction of correlation network topology, and normalized weighting, etc.). The calculated "benchmark physical quality index" score of the standard reference reagent on that day was 80.00.

[0117] According to the physical properties specifications, the "theoretical qualification index" of the reference reagent under ideal conditions without photoelectric background interference is calibrated to 95.00 points.

[0118] The system calibration coefficient was calculated by dividing the theoretical pass index of 95.00 by the actual measured baseline physical quality index of 80.00, resulting in a coefficient of 1.1875. This indicates that the excitation source or sensor components inside the current optical detection instrument have a negative error due to overall physical signal attenuation.

[0119] The initial comprehensive physical quality index of the test reagent solution is 62.96 points. Hardware error compensation adjustment is performed using a coefficient: multiplying 62.96 by 1.1875 yields a theoretical value of 74.76 points after calibration. To ensure the consistency of the scoring system (full score limit), a maximum value truncation function is built-in (i.e., the maximum score does not exceed 100 points). Since 74.76 points does not exceed the limit, the "calibrated comprehensive physical quality index" is output as 74.76 points.

[0120] See Table 3 below. The index after physical error calibration (74.76 points) is compared again with the set passing threshold of 60.00 points to confirm that it is still firmly above the passing line. The final quality judgment result after calibration is then generated and output.

[0121] Table 3

[0122] The present invention also provides a reagent detection device for high-frequency mutation sites of hereditary deafness. The reagent detection device for high-frequency mutation sites of hereditary deafness includes a memory and a processor. The memory stores computer-readable instructions. When the computer-readable instructions are executed by the processor, the processor performs the steps of the reagent detection method for high-frequency mutation sites of hereditary deafness described in the above embodiments.

[0123] The present invention also provides a computer-readable storage medium, which may be a non-volatile computer-readable storage medium or a volatile computer-readable storage medium, wherein the computer-readable storage medium stores instructions that, when the instructions are executed on a computer, cause the computer to perform the steps of the reagent detection method for high-frequency mutation sites in hereditary deafness.

[0124] Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the specific working processes of the systems, devices, and units described above can be referred to the corresponding processes in the foregoing method embodiments, and will not be repeated here.

[0125] If the integrated unit is implemented as a software functional unit and sold or used as an independent product, it can be stored in a computer-readable storage medium. Based on this understanding, the technical solution of the present invention, in essence, or the part that contributes to the prior art, or all or part of the technical solution, can be embodied in the form of a software product. This computer software product is stored in a storage medium and includes several instructions to cause a computer device (which may be a personal computer, server, or network device, etc.) to execute all or part of the steps of the methods described in the various embodiments of the present invention. The aforementioned storage medium includes various media capable of storing program code, such as USB flash drives, portable hard drives, read-only memory (ROM), random access memory (RAM), magnetic disks, or optical disks.

[0126] The above-described embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A reagent detection method for high-frequency mutation sites in hereditary deafness, characterized in that, Includes the following steps: The test reagent solution is subjected to repeated physical excitation and optical signal acquisition using an optical detection instrument to obtain the original fluorescence physical signal of the reagent solution under multiple optical detection channels, and then processed to form a standardized physical detection signal matrix. The phase weight vector is applied to the standardized physical detection signal matrix to perform beamforming operation, generating a phase-weighted synthesized signal sequence to extract a stability index characterizing the overall optical luminescence physical stability of the test reagent solution. The standardized physical detection signal matrix is ​​analyzed and filtered according to a preset significance threshold to construct a channel physical response correlation network, and the global efficiency of the network is calculated to obtain a physical structure consistency index. Analyze the distribution probability of the initial physical detection signal of the test reagent solution under each optical detection channel, and calculate the luminescence state uncertainty index; Based on the stability index of the phase-weighted synthesized signal sequence, the physical structure consistency index, and the luminescence state uncertainty index, a comprehensive physical quality index is calculated, and the quality of the reagent solution is determined by comparing the comprehensive physical quality index with a preset threshold.

2. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 1, characterized in that, include: Acquire the expected response characteristic data of each optical detection channel to form a set of prior information; Based on the prior information set, a phase weight vector is generated through mathematical transformation, which makes the expected response intensity negatively correlated with the weight value. An intermediate composite vector is generated based on the phase weight vector and the standardized physical detection signal matrix; The intermediate synthesis vector is processed to generate a phase-weighted synthesized signal sequence; The coefficient of variation of the phase-weighted synthesized signal sequence is calculated as a stability index characterizing the overall optical luminescence physical stability of the test reagent solution.

3. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 1, characterized in that, include: Calculate the correlation coefficient between column vectors corresponding to any two different optical detection channels in the standardized physical detection signal matrix to form a physical response correlation coefficient matrix; Each element in the physical response correlation coefficient matrix is ​​compared with a preset significance threshold, and elements with an absolute value greater than or equal to the significance threshold are selected. Based on the screening results, an undirected weighted network is constructed with each optical detection channel as a node and the significant correlations retained by the screening as connecting edges, serving as the channel physical response correlation network; The reciprocal of the harmonic mean of the shortest path lengths between all possible node pairs in the channel physical response correlation network is calculated as the global efficiency of the network, thus obtaining the physical structure consistency index.

4. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 3, characterized in that, Based on the physical response correlation coefficient matrix, the average of the absolute values ​​of all elements is calculated to generate an initial correlation level reference value. Based on the initial correlation level reference value, an initial candidate threshold is calculated and generated using a preset threshold adjustment rule; The physical response correlation coefficient matrix is ​​filtered using an initial candidate threshold to construct an initial candidate network; Calculate the average shortest path number between nodes and the node clustering coefficient of the initial candidate network to generate network topology feature indicators; Based on the degree of difference between the network topology feature indicators and the theoretical ideal network features, the initial candidate thresholds are calibrated to generate a preset significance threshold.

5. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 1, characterized in that, include: Based on the initial physical detection data matrix of the test reagent, the number of times the original fluorescence physical signal of each optical detection channel breaks through the preset reference emission threshold in all repeated physical excitation detections is counted to generate the effective emission frequency of each optical detection channel. The effective emission frequency of each optical detection channel is divided by the total number of detections to calculate the effective emission probability value of each optical detection channel. Based on the effective light emission probability value of each optical detection channel, the information content value corresponding to each optical detection channel is calculated according to the preset information content calculation rules. A comprehensive information entropy value is generated based on the information values ​​of all optical detection channels, which serves as an index for the uncertainty of the luminescence state.

6. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 5, characterized in that, Logarithmic domain transformation is performed on the effective emission probability value of each optical detection channel to generate the corresponding first transformation value; Calculate the absolute difference between the effective emission probability value and the fixed reference probability value for each optical detection channel, and generate the corresponding probability deviation value. The second conversion value for each optical detection channel is generated by calculating the first conversion value and the probability deviation value. A negative operation is applied to the second conversion value of each optical detection channel to generate the information value corresponding to that detection channel.

7. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 1, characterized in that, include: The stability index, the physical structure consistency index, and the luminescence state uncertainty index are normalized respectively to generate corresponding normalized stability index, normalized physical structure consistency index, and normalized uncertainty index. The normalized stability index, normalized physical structure consistency index, and normalized uncertainty index are processed with a preset set of coefficients to obtain the comprehensive physical quality index. The comprehensive physical quality index is compared with a preset quality qualification threshold to generate a comparison result; Based on the comparison results, the quality determination result of the test reagent solution is output.

8. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 7, characterized in that, Introducing the implementation of the comprehensive physical quality index : ； in, As a normalized stability index, , The coefficient of variation; This is a normalized physical structure consistency index; The normalized uncertainty index , For comprehensive information entropy; As a stability weight, For consistency weight, This represents the uncertainty weight.

9. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 1, characterized in that, Also includes: Obtain calibration test data for a standard reference reagent, which has known and qualified physicoluminescent properties; The calibration test data are processed using the same physical detection and analysis steps as the test reagent solution to generate a baseline physical quality index for the standard reference reagent. The system calibration coefficient is obtained by comparing the reference physical quality index of the standard reference reagent with the theoretical qualification index to compensate for the physical attenuation error of the optical instrument. Using the system calibration coefficient, the comprehensive physical quality index of the test reagent solution is adjusted to generate the calibrated comprehensive physical quality index; Based on the comparison between the calibrated comprehensive physical quality index and the theoretical pass index, the calibrated quality judgment result is output.

10. The reagent detection method for high-frequency mutation sites in hereditary deafness according to claim 9, characterized in that, The calibration and testing data obtained from the standard reference reagent include: Under the same optical detection instrument, the standard reference reagent is subjected to repeated physical excitation and optical signal acquisition detection a predetermined number of times to obtain its original calibration fluorescence physical signal results for each optical detection channel; The original calibration fluorescence physical signal results are organized into a calibration initial data matrix, where the rows of the matrix correspond to the number of calibration tests and the columns correspond to each optical detection channel; Calculate the calibration mean and calibration standard deviation for each column corresponding to each optical detection channel in the initial calibration data matrix; Based on the calibration mean and calibration standard deviation of each optical detection channel, all the corresponding original calibration fluorescence physical signal results are standardized and transformed to generate standardized physical calibration signals for each optical detection channel. By aggregating the standardized physical calibration signals from all optical detection channels, a calibration detection data matrix for a standard reference reagent is constructed.

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