Carcino-embryonic antigen detection method based on ultraviolet resonance Raman

Abstract

The invention discloses a carcino-embryonic antigen detection method based on ultraviolet resonance Raman, which adopts an adjustable ultraviolet laser source, adopts an immune enrichment method, captures CEA (Carcino Embryonic Antigen) by using a specific antibody, places a treated sample in an optical pool, adjusts the laser wavelength, and excites the sample to generate a Raman scattering signal through laser. Carrying out preliminary signal processing, analyzing the obtained Raman spectrum by adopting a chemometrics method, and extracting a CEA characteristic peak; and establishing a standard curve, and carrying out calibration through a CEA sample with known concentration so as to realize quantitative detection of an unknown sample. According to the present invention, the CEA can be accurately detected at the low concentration, the common fluorescence interference problem in the traditional Raman spectrum is avoided, the Raman signal intensity can be significantly enhanced through the resonance effect with the target molecule by the ultraviolet resonance Raman, the signal-to-noise ratio is improved, and the carcino-embryonic antigen can be clearly identified and quantified.

Description

Technical Field

[0001] The invention relates to a method for detecting carcinoembryonic antigen, in particular to a method for detecting carcinoembryonic antigen based on ultraviolet resonance Raman. Background Art

[0002] Carcinoembryonic antigen (CEA) is a glycoprotein that is mainly produced in tumor cells and is often used to monitor the progression and treatment effects of various cancers, especially in types of cancer such as colorectal cancer, breast cancer and lung cancer. Early detection of changes in CEA concentration is crucial for timely diagnosis and treatment. Traditional detection methods such as immunological methods such as enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) mostly rely on biochemical analysis, but these methods have limitations in sensitivity, specificity and operational complexity. Raman spectroscopy produces characteristic scattering spectra through molecular vibrations, provides molecular structure information, and is suitable for qualitative and quantitative analysis of biological molecules. UV resonance Raman enhances the Raman signal of the target molecule through ultraviolet light, significantly improving the detection sensitivity, and is suitable for the detection of biological molecules.

[0003] Technical solution of prior art 1:

[0004] Carcinoembryonic antigen detection method based on enzyme-linked immunosorbent assay (ELISA). This method uses specific antibodies to bind to CEA to form an antigen-antibody complex, and generates a measurable signal through an enzyme-catalyzed reaction for quantitative analysis. The whole process usually includes steps such as sample dilution, sample addition, incubation, washing and substrate reaction, and finally the results are read by colorimetry.

[0005] Disadvantages of prior art 1:

[0006] At present, ELISA is not only technically complex but also expensive. Because enzyme-labeled antigens or antibodies are required, and not all antigens or antibodies are suitable for labeling, the cost of enzyme-linked labeling is relatively high. If two antibodies are used for analysis, the antigen needs to have more than two antibody binding sites, and cross-reactions will also affect its specific detection. ELISA performs poorly in low-concentration CEA detection and is difficult to meet the needs of early screening. There are many steps and it is easily affected by the skills of the operator, which increases the possibility of experimental errors.

[0007] Technical solution of the second prior art:

[0008] Electrochemical immunoassay is a detection method that converts biological signals into electrochemical signals. By loading antibodies onto the surface of electrodes or substrate materials, the specific antigen and antibody bind to each other, and the electron transfer rate between the solution interface and the electrode surface changes, generating membrane potential, causing current response changes, and then the response signal is converted into recognizable electrical signals such as current, voltage, and resistance through the conversion system.

[0009] Disadvantages of the second prior art:

[0010] It is difficult to select suitable luminescent materials, the nanoprobes used to label CEA are easy to be lost, and the immunoactive substances modified on the electrode surface have poor conductivity.

[0011] In summary, as an important tumor marker, the detection of carcinoembryonic antigen (CEA) is of great significance in the early diagnosis of cancer and the monitoring of therapeutic efficacy. Existing detection methods have problems such as insufficient sensitivity, complex operation and high cost, which cannot meet the clinical needs for rapid and accurate detection.

[0012] In view of this, the present invention is proposed, which aims to provide a highly sensitive, rapid and low-cost CEA detection method based on ultraviolet resonance Raman spectroscopy (UVRR). Summary of the invention

[0013] The purpose of the present invention is to provide a method for detecting carcinoembryonic antigen based on ultraviolet resonance Raman to solve the above technical problems existing in the prior art.

[0014] The objective of the present invention is achieved through the following technical solutions:

[0015] The method for detecting carcinoembryonic antigen based on ultraviolet resonance Raman of the present invention comprises the following steps:

[0016] A. Collect serum samples and remove cellular components through centrifugation steps;

[0017] B. Place the processed sample in the optical cell;

[0018] C. Excite the sample with ultraviolet laser to generate Raman scattering signal and perform preliminary signal processing;

[0019] D. Analyze the obtained Raman spectrum using chemometric methods to extract the characteristic peak of carcinoembryonic antigen;

[0020] E. Establish a standard curve and calibrate it with samples of known concentration of carcinoembryonic antigen to achieve quantitative detection of unknown samples.

[0021] Compared with the prior art, the carcinoembryonic antigen detection method based on ultraviolet resonance Raman provided by the present invention can accurately detect CEA at low concentrations, avoiding the common fluorescence interference problem in traditional Raman spectroscopy. Ultraviolet resonance Raman can significantly enhance the Raman signal intensity through the resonance effect with the target molecule, improve the signal-to-noise ratio, and more clearly identify and quantify carcinoembryonic antigen. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 A schematic flow chart of a method for detecting carcinoembryonic antigen based on ultraviolet resonance Raman provided in an embodiment of the present invention.

[0023] Figure 2 Schematic diagram of the device structure of an embodiment of the present invention. DETAILED DESCRIPTION

[0024] The following is a clear and complete description of the technical solutions in the embodiments of the present invention in conjunction with the drawings in the embodiments of the present invention; it is obvious that the described embodiments are only part of the embodiments of the present invention, not all of the embodiments, which does not constitute a limitation of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the protection scope of the present invention.

[0025] First, the terms that may be used in this article are explained as follows:

[0026] The term “and / or” means that either or both of them can be realized at the same time. For example, X and / or Y means both “X” or “Y” and “X and Y”.

[0027] The terms "include", "comprises", "contains", "has" or other descriptions with similar semantics should be interpreted as non-exclusive inclusion. For example, including certain technical feature elements (such as raw materials, components, ingredients, carriers, dosage forms, materials, dimensions, parts, components, mechanisms, devices, steps, procedures, methods, reaction conditions, processing conditions, parameters, algorithms, signals, data, products or products, etc.) should be interpreted as including not only certain technical feature elements explicitly listed, but also other technical feature elements known in the art that are not explicitly listed.

[0028] The term "consisting of..." means excluding any technical feature elements not explicitly listed. If this term is used in a claim, it will make the claim closed, so that it does not contain technical feature elements other than the technical feature elements explicitly listed, except for the conventional impurities related to them. If this term only appears in a clause of a claim, it only limits the elements explicitly listed in the clause, and the elements recorded in other clauses are not excluded from the overall claim.

[0029] The contents not described in detail in the examples of the present invention belong to the prior art known to professionals in the field. If no specific conditions are specified in the examples of the present invention, the conditions are carried out according to the conventional conditions in the field or the conditions recommended by the manufacturer. If the manufacturer is not specified in the reagents or instruments used in the examples of the present invention, they are all conventional products that can be purchased commercially.

[0030] The method for detecting carcinoembryonic antigen based on ultraviolet resonance Raman of the present invention comprises the following steps:

[0031] A. Collect serum samples and remove cellular components through centrifugation steps;

[0032] B. Place the processed sample in the optical cell;

[0033] C. Excite the sample with ultraviolet laser to generate Raman scattering signal and perform preliminary signal processing;

[0034] D. Analyze the obtained Raman spectrum using chemometric methods to extract the characteristic peak of carcinoembryonic antigen;

[0035] E. Establish a standard curve and calibrate it with samples of known concentration of carcinoembryonic antigen to achieve quantitative detection of unknown samples.

[0036] In step A, an immune enrichment method is used to capture carcinoembryonic antigen using specific antibodies to reduce background noise and increase the concentration of carcinoembryonic antigen.

[0037] In step C, an adjustable ultraviolet laser source is used to optimize the resonance Raman scattering of the specific structure of the carcinoembryonic antigen to increase the signal intensity;

[0038] By adjusting the laser wavelength and selecting the most suitable wavelength, the Raman signal can be made strongest and fluorescence interference can be eliminated.

[0039] In summary, it can be seen that the carcinoembryonic antigen detection method based on ultraviolet resonance Raman of the embodiment of the present invention can accurately detect CEA at low concentrations through an adjustable ultraviolet light source, meeting the needs of early diagnosis. The detection process is short, and can be completed within 30 minutes from sample processing to result analysis, which is suitable for rapid clinical detection. The automated data analysis module makes the operation simpler and suitable for non-professionals, improving the feasibility of clinical application. The ultraviolet light source avoids the common fluorescence interference problem in traditional Raman spectroscopy and improves the accuracy of detection. Ultraviolet resonance Raman can significantly enhance the Raman signal intensity through the resonance effect with the target molecule, improve the signal-to-noise ratio, and more clearly identify and quantify carcinoembryonic antigen.

[0040] In order to more clearly demonstrate the technical solution and technical effects provided by the present invention, the embodiments of the present invention are described in detail with specific examples below.

[0041] Example 1

[0042] like Figure 1 , Figure 2 As shown:

[0043] An adjustable UV laser source is used to optimize the resonant Raman scattering of the specific structure of CEA and improve the signal intensity.

[0044] Serum samples were collected and cellular components were removed by a simple centrifugation step. In order to increase the concentration of CEA, an immunoenrichment method was used to capture CEA using specific antibodies and reduce background noise.

[0045] Place the processed sample in an optical cell, adjust the laser wavelength, and select the most suitable wavelength to make the Raman signal the strongest and eliminate fluorescence interference.

[0046] The Raman scattering signal is generated by laser excitation of the sample and preliminary signal processing is performed.

[0047] The obtained Raman spectra were analyzed by chemometric method to extract the characteristic peaks of CEA.

[0048] A standard curve was established and calibrated with CEA samples of known concentration to achieve quantitative detection of unknown samples.

[0049] The key technical points of the present invention are:

[0050] An ultraviolet laser source with adjustable wavelength is selected so that it can effectively excite the specific structure of carcinoembryonic antigen (CEA) and enhance the Raman scattering signal.

[0051] The immune enrichment method is used to enrich CEA in serum samples, reduce background interference, and improve detection accuracy.

[0052] The CEA detection method based on UV resonance Raman spectroscopy combines a specific UV laser source and immune enrichment technology to form a new detection idea.

[0053] Protect specific data processing algorithms used for Raman signal analysis, including the identification of characteristic peaks and the methods and implementation of quantitative analysis.

[0054] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by any technician familiar with the technical field within the technical scope disclosed in the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims. The information disclosed in the background technology section of this article is only intended to deepen the understanding of the overall background technology of the present invention, and should not be regarded as an admission or in any form that the information constitutes prior art known to those skilled in the art.

Claims

1. A method for detecting carcinoembryonic antigen based on ultraviolet resonance Raman, characterized in that: Includes steps: A. Collect serum samples and remove cellular components through centrifugation steps; B. Place the processed sample in the optical cell; C. Excite the sample with ultraviolet laser to generate Raman scattering signal and perform preliminary signal processing; D. Analyze the obtained Raman spectrum using chemometric methods to extract the characteristic peak of carcinoembryonic antigen; E. Establish a standard curve and calibrate it with samples of known concentration of carcinoembryonic antigen to achieve quantitative detection of unknown samples.

2. The method for detecting carcinoembryonic antigen based on ultraviolet resonance Raman according to claim 1, characterized in that: In step A, an immune enrichment method is used to capture carcinoembryonic antigen using specific antibodies to reduce background noise and increase the concentration of carcinoembryonic antigen.

3. The method for detecting carcinoembryonic antigen based on ultraviolet resonance Raman according to claim 2, characterized in that: In step C, an adjustable ultraviolet laser source is used to optimize the resonance Raman scattering of the specific structure of the carcinoembryonic antigen to increase the signal intensity; By adjusting the laser wavelength and selecting the most suitable wavelength, the Raman signal can be made strongest and fluorescence interference can be eliminated.

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