Tumor marker detection method based on surface enhanced Raman spectroscopy

Abstract

The invention relates to the technical field of optical testing, in particular to a tumor marker detection method based on a surface enhanced Raman spectroscopy technology, which comprises the following steps: acquiring Raman spectrograms of tumor markers at different temperatures, determining sample molecular adsorption parameters at different collection temperatures according to the variation trend of spectrum curves in the Raman spectrograms, and determining the molecular adsorption parameters of the tumor markers according to the sample molecular adsorption parameters. Constructing the characteristic significance degree of the adsorption equilibrium spectrum curve; constructing a stability index during tumor marker detection by utilizing the characteristic significance degree of a sample molecule adsorption parameter and an adsorption equilibrium spectrum curve; the optimal temperature for detecting the tumor marker is obtained by comparing the stability indexes of the tumor marker at a plurality of collection temperatures during detection, and the tumor marker is detected. The optimal temperature for detecting the tumor marker is determined by analyzing the change condition of the spectrum curve at different temperatures, so that a more accurate detection result is obtained.

Description

Technical Field

[0001] The present invention relates to the field of optical testing technology, and in particular to a tumor marker detection method based on surface enhanced Raman spectroscopy technology. Background Art

[0002] Tumor markers refer to substances that are characteristically present in malignant tumor cells or abnormally produced by malignant tumor cells, including metabolites of tumor cells, gene products of cells with disordered differentiation, substances released into the blood circulation by tumor cell necrosis and collapse, and cell-reactive products of tumor host cells. Tumor markers exist in the tissues, body fluids, and excretions of tumor patients and can be detected by immunological, biological, or chemical methods.

[0003] For rapid and accurate diagnosis and personalized treatment of cancer, blood tests can provide rich molecular information of tumor markers, while Raman spectroscopy can achieve label-free, non-destructive, and multi-index molecular information. Surface-enhanced Raman spectroscopy (SERS) uses the surface plasmon resonance effect of metal nanostructures (such as gold and silver nanoparticles). When these nanostructures are excited by lasers, they generate strong local electromagnetic fields, significantly enhancing the Raman scattering signals of molecules adsorbed on their surfaces, thereby improving detection sensitivity. It has important application value in analytical chemistry, biomedicine, and materials science. However, changes in temperature can affect the vibration state of molecules in the test sample and the stability of instrument performance, thereby interfering with the accuracy of the test results. Summary of the invention

[0004] The present invention provides a tumor marker detection method based on surface enhanced Raman spectroscopy technology to solve the existing problems.

[0005] The tumor marker detection method based on surface enhanced Raman spectroscopy technology of the present invention adopts the following technical scheme: One embodiment of the present invention provides a method for detecting tumor markers based on surface enhanced Raman spectroscopy technology, the method comprising the following steps: Obtaining Raman spectra of tumor markers at different temperatures, wherein the Raman spectra have Raman frequency shift as the horizontal axis and Raman scattering intensity as the vertical axis; According to the changing trend of the spectral curve in the Raman spectrum, the adsorption parameters of the sample molecules at different collection temperatures are determined; according to the performance of the Raman peak in the adsorption equilibrium spectral curve at different collection temperatures, the characteristic significance of the adsorption equilibrium spectral curve is constructed; using the sample molecule adsorption parameters and the characteristic significance of the adsorption equilibrium spectral curve, the stability index for tumor marker detection is constructed; By comparing the stability indicators of tumor markers detected at several collection temperatures, the optimal temperature for detecting tumor markers is obtained, and tumor marker detection is performed.

[0006] Furthermore, the Raman spectra of tumor markers at different temperatures are obtained by: A confocal micro-Raman spectrometer is used to detect the blood sample, and several groups of Raman spectral data are obtained at any collection temperature with a preset collection frequency and time interval; An image of several spectral curves formed by several groups of Raman spectral data at the same collection temperature is recorded as a Raman spectrum graph, and several Raman spectrum graphs at different collection temperatures are obtained within a set temperature range with a set change step.

[0007] Furthermore, the specific method of determining the sample molecule adsorption parameters at different collection temperatures according to the change trend of the spectral curve in the Raman spectrum diagram includes: For several spectral curves in the Raman spectrum at any collection temperature, the Raman scattering intensity changes of the sample molecules are analyzed to obtain the adsorption equilibrium spectral curve; and then the adsorption parameters of the sample molecules at different collection temperatures are determined in combination with the adsorption time; in, Indicates The sample molecular adsorption parameters at the acquisition temperature; Indicates The integral area of ​​the adsorption equilibrium spectrum curve at each collection temperature; Indicates The collection frequency of the adsorption equilibrium spectrum curve corresponds to the time at each collection temperature; Indicates The acquisition frequency corresponding to the time when the integrated area of ​​the first spectrum curve reaches the maximum value at the acquisition temperature; is the preset hyperparameter, Represents the absolute value symbol.

[0008] Furthermore, the characteristic significance of the adsorption equilibrium spectrum curve is constructed according to the performance of the Raman peak in the adsorption equilibrium spectrum curve at different collection temperatures, and the specific method includes: The degree of change of the adsorption equilibrium spectrum curve is calculated, and the overlap parameters of the Raman peaks in the adsorption equilibrium spectrum curve are determined. The characteristic significance of the adsorption equilibrium spectrum curve is constructed through the degree of change of the adsorption equilibrium spectrum curve and the overlap parameters of the Raman peaks.

[0009] Furthermore, the specific method for calculating the degree of change of the adsorption equilibrium spectrum curve includes: Determine the adsorption equilibrium spectral curves at several acquisition temperatures, and calculate the degree of change of the adsorption equilibrium spectral curve according to the position and peak width of the Raman peak in the adsorption equilibrium spectral curve; in, Indicates The degree of change of the adsorption equilibrium spectrum curve at each collection temperature; Indicates At the collection temperature, the adsorption equilibrium spectrum curve The Raman frequency shift corresponding to the peak; Indicates that at several acquisition temperatures, the adsorption equilibrium spectrum curve The mean of the Raman frequency shifts corresponding to the peaks; Indicates At the collection temperature, the adsorption equilibrium spectrum curve The peak width corresponding to the peak; Indicates that at several acquisition temperatures, the adsorption equilibrium spectrum curve The mean of the peak widths corresponding to the peaks; Represents the number of Raman peaks in the adsorption equilibrium spectrum curve; Represents the absolute value symbol.

[0010] Furthermore, the determination of the overlap parameters of the Raman peaks in the adsorption equilibrium spectrum curve includes the following specific methods: For the adsorption equilibrium spectral curves at several acquisition temperatures, the overlap factor of the overlapping peaks is constructed by using the differences in the Raman frequency shifts corresponding to several peaks in the overlapping peaks and the peak widths of the overlapping peaks; Determining the overlap parameter of the Raman peaks in the adsorption equilibrium spectrum curve according to the number of overlapping peaks and the overlapping factors of the overlapping peaks in the adsorption equilibrium spectrum curve; in, Indicates Overlap parameters of Raman peaks in the adsorption equilibrium spectrum curve at each acquisition temperature; Indicates The number of overlapping peaks in the adsorption equilibrium spectrum curve at each acquisition temperature; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, The peak width of overlapping peaks; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, Overlap factor of overlapping peaks.

[0011] Furthermore, the overlapping factor of the overlapping peaks is constructed by utilizing the difference in Raman frequency shifts corresponding to several peaks in the overlapping peaks and the peak width of the overlapping peaks, including the specific method of: The absolute value of the difference in Raman frequency shifts corresponding to the peak values ​​of all adjacent Raman peaks in the same overlapping peak is calculated, and the difference between the peak width of the overlapping peak and the absolute value of the difference in Raman frequency shifts corresponding to the peak values ​​of the adjacent Raman peaks is recorded as the overlapping factor of the overlapping peak.

[0012] Furthermore, the characteristic significance degree of the adsorption equilibrium spectrum curve is constructed by the variation degree of the adsorption equilibrium spectrum curve and the overlap parameter of the Raman peak, wherein the variation degree of the adsorption equilibrium spectrum curve and the overlap parameter of the Raman peak are negatively correlated with the characteristic significance degree of the adsorption equilibrium spectrum curve.

[0013] Furthermore, the method of constructing a stability index for tumor marker detection by using the characteristic significance of the sample molecule adsorption parameters and the adsorption equilibrium spectral curve includes the following specific methods: in, Indicates The stability index of tumor marker detection at each collection temperature. Indicates The characteristic significance of the adsorption equilibrium spectrum curve at each collection temperature is Indicates The sample molecular adsorption parameters at the acquisition temperature are: Represents the normalization function.

[0014] Furthermore, the optimal temperature for detecting tumor markers is obtained by comparing the stability indexes of tumor markers detected at several collection temperatures, and the tumor markers are detected, and the specific method includes: Obtaining stability indexes of tumor markers detected at several collection temperatures within the collection temperature range, and recording the collection temperature at which the stability index value is the highest as the optimal temperature for detecting tumor markers using surface enhanced Raman spectroscopy technology; Molecular dynamics simulation was used to analyze the equilibrium adsorption spectrum curve at the optimal temperature to obtain the concentration of tumor markers in blood samples.

[0015] The beneficial effects of the technical solution of the present invention are as follows: compared with the traditional tumor marker detection method, the embodiment of the present invention takes into account the influence of temperature on surface enhanced Raman spectroscopy technology, and therefore more accurately reflects the adsorption degree of sample molecules at different collection temperatures by determining the sample molecule adsorption parameters; and determines the characteristic significance of the adsorption equilibrium spectrum curve according to the performance of the Raman peak in the adsorption equilibrium spectrum curve at different collection temperatures, and more accurately reflects the resolution of the sample molecules by the Raman spectrum at different collection temperatures, so as to further construct a stability index during tumor marker detection, clearly reflect the stability of tumor marker detection at different collection temperatures, thereby obtaining the optimal temperature for detecting tumor markers, and performing tumor marker detection, thereby reducing the influence of temperature on the metal nanostructure and the diffusion rate of molecules, and improving the accuracy of the detection results. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0017] Figure 1 The present invention is a flow chart of the steps of the tumor marker detection method based on surface enhanced Raman spectroscopy technology. DETAILED DESCRIPTION

[0018] In order to further explain the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the following is a detailed description of the tumor marker detection method based on surface enhanced Raman spectroscopy technology proposed by the present invention, its specific implementation, structure, characteristics and effects, in combination with the accompanying drawings and preferred embodiments. In the following description, different "one embodiment" or "another embodiment" does not necessarily refer to the same embodiment. In addition, specific features, structures or characteristics in one or more embodiments may be combined in any suitable form.

[0019] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0020] The specific scheme of the tumor marker detection method based on surface enhanced Raman spectroscopy technology provided by the present invention is described in detail below with reference to the accompanying drawings.

[0021] See also Figure 1 , which shows a flow chart of the steps of a tumor marker detection method based on surface enhanced Raman spectroscopy technology provided by an embodiment of the present invention, the method comprising the following steps: Step S001: Obtaining Raman spectra of tumor markers at different temperatures.

[0022] It should be noted that understanding the spectral characteristics of tumor markers under different temperature conditions can provide important information for early cancer diagnosis, monitoring of treatment effects and disease physiology research. Surface-enhanced Raman spectroscopy technology enhances Raman signals, allowing the detection of molecules in samples even when the tumor marker content in blood samples is extremely low. It has the advantages of high sensitivity, high selectivity, rapid response and no labeling.

[0023] Specifically, in order to implement the tumor marker detection method based on surface enhanced Raman spectroscopy technology proposed in this embodiment, it is necessary to first obtain a Raman spectrum. The specific process is as follows: The embodiment of the present invention uses a confocal micro-Raman spectrometer to detect the blood sample, and obtains several groups of Raman spectrum data at any collection temperature, collection frequency and time interval.

[0024] With Raman frequency shift as the horizontal axis and Raman scattering intensity as the vertical axis, an image of several spectral curves formed by several groups of Raman spectral data at the same acquisition temperature is recorded as a Raman spectrum graph. Within a set temperature range and with a set change step size, several Raman spectra at different acquisition temperatures are obtained.

[0025] It should be noted that the preset temperature range in the embodiment of the present invention is , the change step size is The collection frequency is 30 seconds / time, and the collection frequency is 60 times.

[0026] It should be noted that the horizontal axis of the Raman spectrum is the Raman frequency shift, which reflects the vibration mode of the sample molecules; the vertical axis is the Raman scattering intensity, which reflects the relative intensity of the sample molecules in different vibration modes. The position and intensity of each Raman peak can provide information about the sample molecular structure, crystal structure, and their vibration and rotation modes. Therefore, the peaks in the Raman spectrum can be used to identify substances, analyze sample composition, and study molecular structures and their interactions.

[0027] So far, Raman spectra at different temperatures are obtained by the above method.

[0028] Step S002: Determine the sample molecule adsorption parameters at different acquisition temperatures according to the variation trend of the spectral curve in the Raman spectrum diagram; construct the characteristic significance of the adsorption equilibrium spectral curve according to the performance of the Raman peak in the adsorption equilibrium spectral curve at different acquisition temperatures; construct the stability index for tumor marker detection using the sample molecule adsorption parameters and the characteristic significance of the adsorption equilibrium spectral curve.

[0029] It should be noted that since surface enhanced Raman spectroscopy is a label-free method that relies on the random adsorption of sample molecules on nanomaterials to achieve signal enhancement, the adsorption sites of the sample molecules are random and there are adsorption and desorption processes. The adsorption and desorption will eventually reach a dynamic equilibrium on the SERS active surface. The overall process can be determined by analyzing the distribution density of the sample molecules, and the stability of the adsorption equilibrium at different temperatures can be obtained; then, the stability at the current temperature can be determined based on the distribution of the sample molecules in the spectral curve of the adsorption equilibrium.

[0030] Specifically, in step 2.1, the adsorption parameters of sample molecules at different collection temperatures are determined according to the change trend of the spectral curve in the Raman spectrum.

[0031] It should be noted that in surface enhanced Raman spectroscopy, in the early stage of adsorption, due to the large number of adsorption sites on the surface of the metal nanostructure, the adsorption rate of the sample molecules is fast, resulting in a rapid increase in the Raman scattering intensity. At this time, the adsorption curve shows a rapid upward trend. As time goes by, the adsorption sites are gradually occupied, the adsorption rate begins to slow down, and finally reaches adsorption equilibrium. When the temperature decreases, the thermal motion of the sample molecules will slow down, and the collision frequency between the sample molecules and the metal surface will decrease, resulting in a decrease in the adsorption amount; while the temperature gradually increases, the equilibrium adsorption amount will increase, and the metal surface will adsorb more sample molecules; but if the temperature is too high, the sample molecules will desorb from the metal surface, which will reduce the equilibrium adsorption amount.

[0032] For several spectral curves in the Raman spectrum at any collection temperature, the Raman scattering intensity changes of the sample molecules are analyzed to obtain the adsorption equilibrium spectral curve, and then the adsorption parameters of the sample molecules at different collection temperatures are determined in combination with the adsorption time.

[0033] It should be noted that in the initial stage, when the sample molecules just begin to adsorb, the Raman scattering intensity rises rapidly and reaches a maximum value. Subsequently, a small amount of sample molecules will desorb, and finally a dynamic equilibrium between desorption and adsorption is reached. Therefore, the distribution density of molecules in the blood sample can be expressed by the integral area of ​​the Raman spectrum curve; due to the influence of temperature, the sample molecules cannot quickly reach the equilibrium of desorption and adsorption, and the stability of molecular adsorption can be determined based on the time difference between the first appearance of the maximum integral area and the adsorption equilibrium.

[0034] As an optional embodiment, according to the change trend of the spectral curve in the Raman spectrum, the adsorption parameters of the sample molecules at different collection temperatures are determined, including the following method: Get the The average slope of all data points of the adsorption equilibrium spectrum curve at the acquisition temperature is obtained according to the average slope. The average slope is negatively correlated with the sample molecule adsorption parameter at the sample temperature.

[0035] As another optional embodiment, the adsorption parameters of sample molecules at different collection temperatures are determined according to the variation trend of the spectral curve in the Raman spectrum, including the following method: Get the At each collection temperature, the collection frequency of the adsorption equilibrium spectrum curve corresponds to the time , No. At the acquisition temperature, the acquisition frequency corresponding to the time when the integral area of ​​the first spectrum curve reaches the maximum value ,Will As the The adsorption parameters of sample molecules at different collection temperatures.

[0036] As a preferred embodiment, the specific calculation method of the sample molecule adsorption parameter is: in, Indicates The sample molecular adsorption parameters at the acquisition temperature; Indicates The integral area of ​​the adsorption equilibrium spectrum curve at each collection temperature; Indicates The collection frequency of the adsorption equilibrium spectrum curve corresponds to the time at each collection temperature; Indicates The acquisition frequency corresponding to the time when the integrated area of ​​the first spectrum curve reaches the maximum value at the acquisition temperature; is the preset hyperparameter, Represents the absolute value symbol.

[0037] It should be noted that Indicates The time difference when the molecular adsorption is stable in the Raman spectrum at each collection temperature. The larger the time difference is, the worse the adsorption stability of the sample molecule is. is a preset hyperparameter to ensure that the calculation results of molecular adsorption are valid. In this embodiment, , can be adjusted according to actual conditions, and this embodiment does not make any specific limitation.

[0038] Step 2.2, construct the characteristic significance of the adsorption equilibrium spectrum curve according to the performance of the Raman peak in the adsorption equilibrium spectrum curve at different collection temperatures.

[0039] It should be noted that when using surface enhanced Raman spectroscopy technology, metal nanostructures need to be selected as the substrate, and the electronic properties of metals are greatly affected by temperature. When the temperature rises, the metal nanostructures may deform, change the interaction with tumor marker molecules, and cause the Raman peak to move. At the same time, temperature changes will also affect the distribution of SERS active "hot spots", resulting in uneven distribution of hot spots. The hot spot distribution may cause the characteristic peaks in the spectrum to overlap, which will reduce the resolution of the spectrum. Therefore, it is necessary to determine the characteristic significance of the current spectrum in the molecule based on the above characteristics at different temperatures.

[0040] Firstly, the adsorption equilibrium spectral curves at several acquisition temperatures are determined, and the variation degree of the adsorption equilibrium spectral curve is calculated according to the position and peak width of the Raman peak in the adsorption equilibrium spectral curve.

[0041] It should be noted that the degree of change of the spectral curve is manifested as the movement of the Raman peak and the change of the peak width in the spectral curve at different acquisition temperatures. The degree of change of the curve is obtained by calculating the change of the peak value in the curve and the change of the peak width where the peak value is located.

[0042] As an embodiment, the specific calculation method of the variation degree of the adsorption equilibrium spectrum curve is: in, Indicates The degree of change of the adsorption equilibrium spectrum curve at each collection temperature; Indicates At the collection temperature, the adsorption equilibrium spectrum curve The Raman frequency shift corresponding to the peak; Indicates that at several acquisition temperatures, the adsorption equilibrium spectrum curve The mean of the Raman frequency shifts corresponding to the peaks; Indicates At the collection temperature, the adsorption equilibrium spectrum curve The peak width corresponding to the peak; Indicates that at several acquisition temperatures, the adsorption equilibrium spectrum curve The mean of the peak widths corresponding to the peaks; Represents the number of Raman peaks in the adsorption equilibrium spectrum curve; Represents the absolute value symbol.

[0043] It should be noted that Indicates The Raman frequency shift corresponding to the peak value in the adsorption equilibrium spectrum curve at each collection temperature; Indicates The change in peak width corresponding to the peak value in the adsorption equilibrium spectrum curve at each collection temperature. The more obvious the Raman frequency shift of the peak value, the greater the change in peak width, which indicates that the greater the change in the spectrum curve at the current collection temperature.

[0044] Then, for the adsorption equilibrium spectrum curves at several acquisition temperatures, the overlap factor of the overlapping peaks is constructed using the differences in the Raman frequency shifts corresponding to several peaks in the overlapping peaks and the peak widths of the overlapping peaks; The overlapping parameters of the Raman peaks in the adsorption equilibrium spectrum curve are determined according to the number of overlapping peaks and the overlapping factors of the overlapping peaks in the adsorption equilibrium spectrum curve.

[0045] It should be noted that when different molecules or molecular structures have similar vibration frequencies or Raman activities, their Raman peaks may overlap. In the same acquisition curve, the more overlapping peaks there are and the greater the degree of overlap between the Raman peaks, the more uneven the distribution of the current molecules is.

[0046] The absolute value of the difference in Raman frequency shifts corresponding to the peak values ​​of adjacent Raman peaks in the same overlapping peak is calculated, and the difference between the peak width of the overlapping peak and the absolute value of the difference in Raman frequency shifts corresponding to the peak values ​​of the adjacent Raman peaks is recorded as the overlapping factor of the overlapping peak.

[0047] As an embodiment, the specific calculation method of the overlapping factor of overlapping peaks is: in, Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, Overlap factor of overlapping peaks; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, The peak width of overlapping peaks; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, Overlapping peaks The Raman frequency shift of the Raman peak; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, Overlapping peaks The Raman frequency shift of the Raman peak; Indicates the number of Raman peaks contained in the overlapping peaks; Represents the absolute value symbol.

[0048] As an embodiment, the specific calculation method of the overlap parameter of the Raman peak in the adsorption equilibrium spectrum curve is: in, Indicates Overlap parameters of Raman peaks in the adsorption equilibrium spectrum curve at each acquisition temperature; Indicates The number of overlapping peaks in the adsorption equilibrium spectrum curve at each acquisition temperature; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, The peak width of overlapping peaks; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, Overlap factor of overlapping peaks.

[0049] It should be noted that It indicates the proportion of the overlapping degree of overlapping peaks to the total peak width value. The larger the proportion, the more serious the overlapping situation is; the more overlapping peaks there are, the more serious the overlapping situation is.

[0050] Finally, the characteristic significance of the spectral curve is constructed according to the variation degree of the adsorption equilibrium spectral curve and the overlap parameter of the Raman peak; the variation degree of the adsorption equilibrium spectral curve and the overlap parameter of the Raman peak are both negatively correlated with the characteristic significance of the adsorption equilibrium spectral curve.

[0051] As an embodiment, the method for constructing the characteristic significance of the adsorption equilibrium spectrum curve is: in, Indicates The characteristic significance of the adsorption equilibrium spectrum curve at each collection temperature is Indicates The degree of change of the adsorption equilibrium spectrum curve at each collection temperature is Indicates The overlap parameter of the Raman peaks in the adsorption equilibrium spectrum curve at different acquisition temperatures.

[0052] It should be noted that the more severe the overlap of the spectral curves at the current temperature and the larger the amplitude of the movement, the worse the characteristic significance of the spectral curves, which means that the molecular adsorption at the current temperature is more volatile and the current temperature is not suitable for tumor marker detection.

[0053] As an optional embodiment, according to the performance of Raman peaks in the adsorption equilibrium spectrum curve at different acquisition temperatures, the characteristic significance of the adsorption equilibrium spectrum curve is constructed, including the following method: Get the The number of overlapping peaks in the adsorption equilibrium spectrum curve at each acquisition temperature ,according to Get the The characteristic significance of the adsorption equilibrium spectrum curve at different collection temperatures ,in and Negatively correlated.

[0054] Step 2.3, using the characteristic significance of the sample molecular adsorption parameters and the adsorption equilibrium spectral curve, construct a stability index for tumor marker detection.

[0055] It should be noted that the change in temperature may cause deformation of the metal nanostructure, affect the stability of molecular adsorption, and the resolution of different molecules, and thus change the characteristic significance of the sample molecular adsorption parameters and the adsorption equilibrium spectral curve. Therefore, the stability index for tumor marker detection is constructed using the characteristic significance of the sample molecular adsorption parameters and the adsorption equilibrium spectral curve. At the current acquisition temperature, the more stable the metal nanostructure is, the better the stability of molecular adsorption is, so the better the stability is during tumor marker detection; the higher the resolution of different molecules at the current acquisition temperature, the smaller the overlap of the spectral curve, and the smaller the amplitude of movement, the greater the characteristic significance of the spectral curve, so the better the stability is during tumor marker detection.

[0056] The characteristic significance of the sample molecular adsorption parameters and the adsorption equilibrium spectral curve are both positively correlated with the stability index during tumor marker detection.

[0057] As an example, the specific calculation method of the stability index of tumor marker detection at any temperature is: in, Indicates The stability index of tumor marker detection at each collection temperature. Indicates The characteristic significance of the adsorption equilibrium spectrum curve at each collection temperature is Indicates The sample molecular adsorption parameters at the acquisition temperature are: Represents the normalization function.

[0058] At this point, the stability index for tumor marker detection is obtained through the above method.

[0059] Step S003: The optimal temperature for detecting tumor markers is obtained by comparing the stability indexes of tumor markers detected at several collection temperatures, and tumor marker detection is performed.

[0060] It should be noted that the intensity of the Raman signal is affected by temperature. The vibration state of some molecules may change at different temperatures. In addition, the adsorption properties of tumor markers on the SERS active surface will be affected by temperature changes. Therefore, it is necessary to select a temperature that makes the adsorption state of the marker molecules on the surface relatively stable. The optimal Raman spectroscopy detection temperature should be able to maintain the signal intensity and background stability to the greatest extent.

[0061] Specifically, the stability index of tumor marker detection at several collection temperatures within the collection temperature range is obtained, and the collection temperature when the stability index value is the highest is recorded as the optimal temperature for detecting tumor markers using surface enhanced Raman spectroscopy technology.

[0062] Molecular dynamics simulation was used to analyze the equilibrium adsorption spectrum curve at the optimal temperature to obtain the concentration of tumor markers in blood samples.

[0063] At this point, the detection of tumor markers is completed through the above method.

[0064] At this point, this embodiment is completed.

[0065] Compared with the traditional tumor marker detection method, the embodiment of the present invention takes into account the influence of temperature on surface enhanced Raman spectroscopy technology, and therefore determines the sample molecule adsorption parameters to more accurately reflect the adsorption degree of the sample molecules at different collection temperatures; and determines the characteristic significance of the adsorption equilibrium spectrum curve according to the performance of the Raman peak in the adsorption equilibrium spectrum curve at different collection temperatures, which more accurately reflects the resolution of the sample molecules by the Raman spectrum at different collection temperatures, so as to further construct a stability index during tumor marker detection, clearly reflect the stability of tumor marker detection at different collection temperatures, thereby obtaining the optimal temperature for detecting tumor markers, performing tumor marker detection, thereby reducing the influence of temperature on the metal nanostructure and the diffusion rate of molecules, and improving the accuracy of the detection results.

[0066] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for detecting tumor markers based on surface enhanced Raman spectroscopy, characterized in that: The method comprises the following steps: Obtaining Raman spectra of tumor markers at different temperatures, wherein the Raman spectra have Raman frequency shift as the horizontal axis and Raman scattering intensity as the vertical axis; According to the changing trend of the spectral curve in the Raman spectrum, the adsorption parameters of the sample molecules at different collection temperatures are determined; according to the performance of the Raman peak in the adsorption equilibrium spectral curve at different collection temperatures, the characteristic significance of the adsorption equilibrium spectral curve is constructed; using the sample molecule adsorption parameters and the characteristic significance of the adsorption equilibrium spectral curve, the stability index for tumor marker detection is constructed; By comparing the stability indicators of tumor markers detected at several collection temperatures, the optimal temperature for detecting tumor markers is obtained, and tumor marker detection is performed.

2. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 1, characterized in that: The specific method of obtaining the Raman spectra of tumor markers at different temperatures includes: A confocal micro-Raman spectrometer is used to detect the blood sample, and several groups of Raman spectral data are obtained at any collection temperature with a preset collection frequency and time interval; An image of several spectral curves formed by several groups of Raman spectral data at the same collection temperature is recorded as a Raman spectrum graph, and several Raman spectrum graphs at different collection temperatures are obtained within a set temperature range with a set change step.

3. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 1, characterized in that: The specific method of determining the sample molecule adsorption parameters at different collection temperatures according to the change trend of the spectral curve in the Raman spectrum is as follows: For several spectral curves in the Raman spectrum at any collection temperature, the Raman scattering intensity changes of the sample molecules are analyzed to obtain the adsorption equilibrium spectral curve; and then the adsorption parameters of the sample molecules at different collection temperatures are determined in combination with the adsorption time; in, Indicates The sample molecular adsorption parameters at the acquisition temperature; Indicates The integral area of ​​the adsorption equilibrium spectrum curve at each collection temperature; Indicates The collection frequency of the adsorption equilibrium spectrum curve corresponds to the time at each collection temperature; Indicates The acquisition frequency corresponding to the time when the integrated area of ​​the first spectrum curve reaches the maximum value at the acquisition temperature; is the preset hyperparameter, Represents the absolute value symbol.

4. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 1, characterized in that: The specific method of constructing the characteristic significance of the adsorption equilibrium spectrum curve according to the performance of the Raman peak in the adsorption equilibrium spectrum curve at different collection temperatures includes: The degree of change of the adsorption equilibrium spectrum curve is calculated, and the overlap parameters of the Raman peaks in the adsorption equilibrium spectrum curve are determined. The characteristic significance of the adsorption equilibrium spectrum curve is constructed through the degree of change of the adsorption equilibrium spectrum curve and the overlap parameters of the Raman peaks.

5. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 4, characterized in that: The specific method for calculating the degree of change of the adsorption equilibrium spectrum curve includes: Determine the adsorption equilibrium spectral curves at several acquisition temperatures, and calculate the degree of change of the adsorption equilibrium spectral curve according to the position and peak width of the Raman peak in the adsorption equilibrium spectral curve; in, Indicates The degree of change of the adsorption equilibrium spectrum curve at each collection temperature; Indicates At the collection temperature, the adsorption equilibrium spectrum curve The Raman frequency shift corresponding to the peak; Indicates that at several acquisition temperatures, the adsorption equilibrium spectrum curve The mean of the Raman frequency shifts corresponding to the peaks; Indicates At the collection temperature, the adsorption equilibrium spectrum curve The peak width corresponding to the peak; Indicates that at several acquisition temperatures, the adsorption equilibrium spectrum curve The mean of the peak widths corresponding to the peaks; Represents the number of Raman peaks in the adsorption equilibrium spectrum curve; Represents the absolute value symbol.

6. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 4, characterized in that: The specific method for determining the overlap parameter of the Raman peaks in the adsorption equilibrium spectrum curve includes: For the adsorption equilibrium spectral curves at several acquisition temperatures, the overlap factor of the overlapping peaks is constructed by using the differences in the Raman frequency shifts corresponding to several peaks in the overlapping peaks and the peak widths of the overlapping peaks; Determining the overlap parameter of the Raman peaks in the adsorption equilibrium spectrum curve according to the number of overlapping peaks and the overlapping factors of the overlapping peaks in the adsorption equilibrium spectrum curve; in, Indicates Overlap parameters of Raman peaks in the adsorption equilibrium spectrum curve at each acquisition temperature; Indicates The number of overlapping peaks in the adsorption equilibrium spectrum curve at each acquisition temperature; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, The peak width of overlapping peaks; Indicates In the adsorption equilibrium spectrum curve at the acquisition temperature, Overlap factor of overlapping peaks.

7. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 6, characterized in that: The method of constructing the overlapping factor of the overlapping peaks by using the difference in Raman frequency shifts corresponding to several peaks in the overlapping peaks and the peak width of the overlapping peaks includes the following specific methods: The absolute value of the difference in Raman frequency shifts corresponding to the peak values ​​of all adjacent Raman peaks in the same overlapping peak is calculated, and the difference between the peak width of the overlapping peak and the absolute value of the difference in Raman frequency shifts corresponding to the peak values ​​of the adjacent Raman peaks is recorded as the overlapping factor of the overlapping peak.

8. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 4, characterized in that: The characteristic significance degree of the adsorption equilibrium spectrum curve is constructed by the variation degree of the adsorption equilibrium spectrum curve and the overlap parameter of the Raman peak, wherein the variation degree of the adsorption equilibrium spectrum curve and the overlap parameter of the Raman peak are negatively correlated with the characteristic significance degree of the adsorption equilibrium spectrum curve.

9. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 1, characterized in that: The method of constructing a stability index for tumor marker detection by using the characteristic significance of the sample molecule adsorption parameters and the adsorption equilibrium spectrum curve includes the following specific methods: in, Indicates The stability index of tumor marker detection at each collection temperature. Indicates The characteristic significance of the adsorption equilibrium spectrum curve at each collection temperature is Indicates The sample molecular adsorption parameters at the acquisition temperature are: Represents the normalization function.

10. The method for detecting tumor markers based on surface enhanced Raman spectroscopy according to claim 1, characterized in that: The method of obtaining the optimal temperature for detecting tumor markers by comparing the stability indexes of tumor markers detected at several collection temperatures and performing tumor marker detection includes the following specific methods: Obtaining stability indexes of tumor markers detected at several collection temperatures within the collection temperature range, and recording the collection temperature at which the stability index value is the highest as the optimal temperature for detecting tumor markers using surface enhanced Raman spectroscopy technology; Molecular dynamics simulation was used to analyze the equilibrium adsorption spectrum curve at the optimal temperature to obtain the concentration of tumor markers in blood samples.

Images