Optical fiber biosensing probe for detecting biomarker in biliary tract

Abstract

The invention relates to the technical field of medical equipment and biomarker detection, and provides an optical fiber biosensing probe for detecting a biomarker in a biliary tract, the optical fiber biosensing probe comprises a computer, a spectrometer and a light source, the computer is electrically connected with the spectrometer, the spectrometer is electrically connected with the light source, the output end of the light source is connected with a probe, and the surface of the probe is of a spiral structure; the probe extends into a biliary tract system, the light source emits light with specific wavelength and intensity, so that the light can reach the probe along the incoming optical fiber, and a biomarker in the biliary tract is specifically combined with the surface of the probe, so that an optical signal is changed and output through the outgoing optical fiber; the spectrograph can be used for carrying out detailed spectral analysis on an optical signal output from the outgoing optical fiber, and the spectral analysis comprises the characteristics of wavelength, intensity, absorption peak and the like, so that in-vivo, rapid, accurate and stable detection on bile protein in the biliary tract is realized; during bile detection, collection, storage and in-vitro detection are not needed, the pollution deterioration possibility is small, and the in-vivo detection result is more real.

Description

Technical Field

[0001] The invention relates to the technical field of medical equipment and biomarker detection, in particular to an optical fiber biosensor probe for detecting biomarkers in the bile duct. Background Art

[0002] Bile is a digestive fluid that is secreted by liver cells, passes through the bile ducts at all levels, and is stored and concentrated in the gallbladder. When food, especially fat-rich food, enters the duodenum, it stimulates the gallbladder to contract and discharge the concentrated bile into the duodenum. It has physiological functions such as emulsifying fat, promoting the absorption of fat-soluble vitamins, and neutralizing gastric acid. Bile is mainly composed of bile pigments, bile salts, cholesterol, phospholipids, water, inorganic salts, various metabolites, and a very small amount of protein. Since bile is not only directly secreted by the human body's "metabolic factory" - the liver, but also directly contacts the bile ducts and gallbladder at all levels, The changes in bile composition are closely related to the occurrence and development of hepatobiliary and pancreatic diseases such as cholelithiasis, cholangitis, cholecystitis, cirrhosis, liver cancer, biliary tumors, biliary fibrosis, and pancreatitis. That is to say, when the above diseases occur, the expression levels of certain substances in the bile will undergo corresponding characteristic changes. Therefore, it is of great clinical value to accurately, conveniently and economically detect the changes in the composition of substances in bile for the diagnosis and treatment of various hepatobiliary and pancreatic diseases, especially the expression level of cluster protein CLU in bile that has been reported to be closely related to the diagnosis of cholangiocarcinoma.

[0003] So far, due to the difficulty in collecting bile samples, easy deterioration, and expensive testing in clinical practice, there are no instruments or products on the market specifically for in vivo testing of bile components. At present, the commonly used method for testing bile components is to collect bile first and then test it in vitro. There are generally three methods for collecting bile. One is to obtain bile through percutaneous gallbladder puncture and drainage under ultrasound guidance; the second is to obtain bile through biliary catheterization and drainage; the third is to directly extract bile during ERCP. The three methods of collecting bile increase the bile testing procedures, increase time consumption, increase additional storage and transportation costs, and increase the risk of contamination, which ultimately reduces the accuracy of the test results. Therefore, it is best to skip the bile collection procedure from the body and perform direct testing in the body to obtain the most accurate results.

[0004] Fiber optic biosensor probe technology has been applied to the detection of various body fluids, such as blood and amniotic fluid. However, due to limitations of factors such as manufacturing process, the front end of the antibody-coated probe has always adopted a "nose cone" design. Although this design reduces the difficulty of the manufacturing process, it also leads to a limited surface area of ​​the front end of the probe that is in contact with the body fluid to be tested. The probe cannot fully contact the body fluid to be tested, resulting in reduced detection accuracy of the probe and the generation of false negative results.

[0005] Therefore, we proposed a fiber optic biosensing spiral probe that can directly detect bile biomarkers in the bile duct in vivo to solve the above-mentioned problems. Summary of the invention

[0006] The present invention provides an optical fiber biosensor probe for detecting biomarkers in the bile duct. The probe is inserted into the bile duct system, so that bile does not need to be collected, stored, or tested in vitro when being detected. The possibility of contamination and deterioration is small, and the in vivo detection results are more realistic, so as to solve the problems mentioned in the background technology that the bile detection procedures in the current market are complicated, costly, slow, and have low accuracy.

[0007] The technical solution of the present invention is as follows:

[0008] A fiber optic biosensor probe for detecting biomarkers in the bile duct comprises: a computer, a spectrometer and a light source, wherein the computer is electrically connected to the spectrometer, the spectrometer is electrically connected to the light source, the output end of the light source is connected to a probe, and the probe surface is a spiral structure.

[0009] Furthermore, the probe includes an incoming optical fiber and an outgoing optical fiber, the incoming optical fiber and the outgoing optical fiber are entangled with each other, the input end of the incoming optical fiber is electrically connected to the light source, the output end of the outgoing optical fiber is electrically connected to the spectrometer, and the output end of the incoming optical fiber and the input end of the outgoing optical fiber are both fixedly connected to a probe; the length of the probe is 300 cm.

[0010] Furthermore, the incoming optical fiber and the outgoing optical fiber are multimode optical fibers, and the integrated diameter of the incoming optical fiber and the outgoing optical fiber is between 0.86-0.96 mm.

[0011] Furthermore, the main sensing area of ​​the probe is the coreless optical fiber part.

[0012] Furthermore, the probe surface is coated with a layer of gold film by magnetron sputtering, and the gold film surface is chemically cross-linked with specific biomarker protein antibodies. The probe is based on surface plasmon resonance (SPR for short); the specific biomarker protein antibodies are cross-linked with the gold film surface by chemical cross-linking.

[0013] Furthermore, the light source is polarized light; it can reduce the interference of tissue scattering on the light signal to a certain extent, so that the light signal can be transmitted and detected more clearly and accurately; and produce more specific interactions with specific biomarkers or sensing mechanisms, thereby improving the targetedness and sensitivity of detection; and help optimize the transmission characteristics of light in the bile duct, thereby improving the overall signal quality and stability.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] 1. The present invention extends a probe into the biliary system, and the light source emits light of a specific wavelength and intensity, so that the light can reach the probe along the incoming optical fiber. The biomarkers in the bile duct specifically bind to the probe surface, causing the light signal to change and be output through the outgoing optical fiber. The spectrometer can perform detailed spectral analysis of the light signal output from the outgoing optical fiber, including wavelength, intensity and other characteristics, thereby achieving in vivo, rapid, accurate and stable detection of bile protein in the bile duct, and analyzing the results through computer software; the present invention does not need to collect, store or test bile in vitro when testing bile, so the possibility of contamination and deterioration is small, and the in vivo test results are more realistic.

[0016] 2. The spiral structure of the probe of the present invention can increase the contact area of ​​the liquid to be tested, improve the detection accuracy, improve the operational convenience, and enhance the mechanical stability, biocompatibility, adaptability and versatility. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 It is a working principle diagram of the present invention;

[0018] Figure 2 is a stereogram of the probe of the present invention;

[0019] Figure 3 It is a cross-sectional view of the probe of the present invention.

[0020] In the figure:

[0021] 1. Probe; 11. Incoming optical fiber; 12. Outgoing optical fiber; 13. Probe. DETAILED DESCRIPTION

[0022] The following embodiments of the present invention are described in further detail in conjunction with the accompanying drawings and examples. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0023] like Figure 1-3 As shown, the present invention provides a fiber optic biosensor probe for detecting biomarkers in the bile duct, including: a computer, a spectrometer and a light source, the computer is electrically connected to the spectrometer, the spectrometer is electrically connected to the light source, the output end of the light source is connected to a probe 1, and the surface of the probe 1 is a spiral structure.

[0024] As a technical solution of the present invention, the probe 1 includes an incoming optical fiber 11 and an outgoing optical fiber 12, the incoming optical fiber 11 and the outgoing optical fiber 12 are intertwined, the input end of the incoming optical fiber 11 is electrically connected to the light source, the output end of the outgoing optical fiber 12 is electrically connected to the spectrometer, the output end of the incoming optical fiber 11 and the input end of the outgoing optical fiber 12 are both fixedly connected with a probe 13, and the length of the probe 1 is 300 cm.

[0025] As a technical solution of the present invention, the incoming optical fiber 11 and the outgoing optical fiber 12 are multimode optical fibers, and the integrated diameter of the incoming optical fiber 11 and the outgoing optical fiber 12 is between 0.86-0.96 mm. This size can ensure that they can pass through and be arranged in the bile duct relatively smoothly while ensuring a certain mechanical strength and stability.

[0026] As a technical solution of the present invention, the main sensing area of ​​the probe 13 is the coreless optical fiber part.

[0027] As a technical solution of the present invention, the surface of the probe 13 is coated with a layer of gold film by a magnetron sputtering instrument, and the gold film surface is chemically cross-linked with a specific biomarker protein antibody. The probe 13 is based on the SPR surface plasmon resonance. When conducting an experiment, a biological molecule (target molecule) is first fixed on the surface of the gold film, and then the molecule that interacts with it is dissolved in a solution (or a mixed solution) and flows over the surface of the gold film. In the process of binding and dissociation between the protein on the gold film and the molecule in the flow path, the resonance angle (i.e., the SPR angle) will change accordingly. The spectrometer detects this change. According to the change curve, the SPR angle changes with the refractive index of the metal surface, and the change in the refractive index is proportional to the molecular mass bound to the metal surface; therefore, the specific signal of the interaction between biological molecules and the concentration of the molecule to be tested can be obtained by the dynamic change of the SPR angle during the biological reaction process; the specific biomarker protein antibody is cross-linked with the gold film surface by chemical cross-linking, and the specific steps are as follows:

[0028] 1. Immerse the membrane in a solution containing an activation reagent, which can react with the functional groups on the membrane to form active intermediates; common activation reagents include EDC / NHS combinations, which can react with the carboxyl groups on the membrane to form active ester groups;

[0029] 2. Antibody dilution: Dissolve the antibody in a suitable buffer according to the antibody concentration and expected coating density;

[0030] 3. Binding of antibodies to membranes: The activated membrane is kept in contact with the antibody solution for a certain period of time, allowing the antibodies to bind to the active groups on the membrane through covalent bonds; the binding time and conditions depend on the specific activation reagents and experimental requirements;

[0031] 4. Block unreacted sites: To reduce nonspecific binding, treat the membrane with a blocking solution (such as a buffer containing BSA) to block the active groups that are not bound to the antibody;

[0032] 5. Washing: Wash the membrane thoroughly with buffer to remove unbound antibodies and blocking agents;

[0033] 6. Drying: Allow the film to dry naturally or by a suitable method, such as vacuum drying.

[0034] As a technical solution of the present invention, the light source is polarized light; it can reduce the interference of tissue scattering on the light signal to a certain extent, so that the light signal can be transmitted and detected more clearly and accurately; and produce more specific interactions with specific biomarkers or sensing mechanisms, thereby improving the targetedness and sensitivity of detection; and help optimize the transmission characteristics of light in the bile duct, thereby improving the overall signal quality and stability.

[0035] Working principle:

[0036] like Figure 1-2 As shown, the staff needs to use a fiber optic fusion splicer to fuse the incoming fiber 11 and the outgoing fiber 12 to the fiber optic probe respectively:

[0037] S1. Clean, flatten and perpendicular the fiber end face to the fiber axis. This usually involves stripping off the protective layer on the outside of the fiber and cutting the fiber end face with a special tool.

[0038] S2. The fusion splicer has a built-in precise alignment mechanism that can automatically detect and adjust the positions of the two optical fibers so that their central axes coincide as much as possible. Alignment can be achieved in a variety of ways, including but not limited to:

[0039] Outer diameter alignment: Alignment is based on the outer diameter of the optical fiber;

[0040] Core alignment: more precise alignment by observing the core of the optical fiber (i.e. the light-conducting part);

[0041] S3. When the two optical fibers are aligned, the fusion splicer generates a high-temperature arc discharge through the electrodes. The heat generated by the arc is enough to melt the optical fibers locally, so that the two optical fibers are fused together at the contact point. This process requires precise control of the energy, duration and position of the arc to ensure the quality of the fusion splicing.

[0042] S4. After welding is completed, a certain cooling time is required to allow the molten area to recrystallize and form a stable connection;

[0043] S5. The welding machine will automatically or manually check the welding effect, usually by observing the appearance of the welding point and measuring the welding loss to evaluate the quality of the connection.

[0044] like Figure 1-3 As shown, when in use, medical staff insert the probe 1 into the bile duct system through the duodenoscope, and the light source emits light of a specific wavelength and intensity, so that the light can reach the probe along the incoming optical fiber 11, and the biomarkers in the bile duct specifically bind to the surface of the probe 13, causing the light signal to change and be output through the outgoing optical fiber 12, and the spectrometer can perform detailed spectral analysis of the light signal output from the outgoing optical fiber 12, including wavelength, intensity and other characteristics, thereby realizing in vivo, rapid, accurate and stable detection of bile protein in the bile duct, and analyzing the results through computer software; the present invention does not require collection, storage, and in vitro detection when detecting bile, and the possibility of contamination and deterioration is small, and the in vivo detection results are more realistic; the conventional in vitro detection Elisa kit takes 3-5 hours to detect, and is expensive, and has high experimental operation requirements for the detection personnel. The present invention only takes about 10 seconds to detect, is price-friendly, and has low cost, and can be completed only in conjunction with ERCP (Encoscopic Retrograde Cho l angio-Pancreatography).

[0045] It should be noted that: the spiral structure of the probe 1 increases the coating area and coating quantity of the antibody, and the detection stability is higher and the precision is more accurate; and the integrated diameter of the incoming optical fiber 11 and the outgoing optical fiber 12 is between 0.86-0.96 mm, which can ensure that it can pass through and be arranged in the bile duct relatively smoothly, while ensuring a certain mechanical strength and stability; the length of the probe 1 is 300 cm, which can adapt to different detection needs and the anatomical structure of the bile duct, and can go deep into the appropriate position for the detection of biomarkers, providing a larger operating range and flexibility, so that the probe 1 can reach different depths and positions in the bile duct to obtain more comprehensive and accurate information. Such a design helps to improve the accuracy and reliability of the detection, and also provides convenience for clinical application;

[0046] And the spiral structure of the probe 13 has the following benefits:

[0047] 1. Improve detection accuracy

[0048] Increased contact area: The helical structure can provide a large surface area in a relatively small space, which means that more active sites of the sensor can contact bile, thereby improving detection sensitivity and response speed;

[0049] Optimizing fluid dynamics: The spiral design can improve the flow characteristics of the fluid around the probe 1, which helps to form a more uniform flow field, so that the substances to be tested in the bile can contact the sensor surface more evenly, thereby improving the consistency and reliability of the test results.

[0050] 2. Improve operational convenience

[0051] Easy insertion and positioning: The spiral structure makes it easier to insert the probe 1 into the bile duct and also facilitates positioning, especially when the bile duct is narrow or tortuous; the spiral shape can provide better grip and help to stably place the probe 1 in the bile duct;

[0052] Reduced risk of injury: Compared with the linear design, the spiral design can reduce friction on the bile duct wall during insertion and withdrawal, thereby reducing the risk of injury to the bile duct mucosa.

[0053] 3. Enhance mechanical stability

[0054] Improved mechanical strength: The spiral structure can enhance the overall mechanical strength of the probe 1, making it less likely to bend or deform in the complex environment of the bile duct, thereby ensuring the functional stability of the sensor;

[0055] 4. Biocompatibility

[0056] Reducing nonspecific adsorption: Reasonable spiral design can reduce the adsorption of nonspecific substances in bile on the surface of probe 1, thereby reducing background interference and improving the authenticity and accuracy of the detection signal.

[0057] 5. Adaptability

[0058] Adapt to different biliary environments: The spiral design can be flexibly adjusted according to the specific conditions of the bile duct, so that it can adapt to bile ducts of different sizes and shapes, increasing the flexibility of use.

[0059] 6. Versatility

[0060] Multi-point detection capability: If multiple detection points are arranged at different positions of the spiral structure, continuous monitoring of different parts of the bile duct can be achieved, thereby improving the value of diagnostic information.

[0061] The embodiments of the present invention are provided for the purpose of illustration and description. Although the embodiments of the present invention have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations of the present invention. Ordinary technicians in this field can change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. An optical fiber biosensor probe for detecting biomarkers in the bile duct, comprising: A computer, a spectrometer and a light source, characterized in that: the computer is electrically connected to the spectrometer, the spectrometer is electrically connected to the light source, the output end of the light source is connected to a probe (1), and the surface of the probe (1) is a spiral structure.

2. An optical fiber biosensor probe for detecting biomarkers in the bile duct as claimed in claim 1, characterized in that: The probe (1) comprises an incoming optical fiber (11) and an outgoing optical fiber (12), wherein the incoming optical fiber (11) and the outgoing optical fiber (12) are intertwined with each other, the input end of the incoming optical fiber (11) is electrically connected to the light source, the output end of the outgoing optical fiber (12) is electrically connected to the spectrometer, and the output end of the incoming optical fiber (11) and the input end of the outgoing optical fiber (12) are both fixedly connected to a probe (13).

3. An optical fiber biosensor probe for detecting biomarkers in the bile duct as claimed in claim 2, characterized in that: The incoming optical fiber (11) and the outgoing optical fiber (12) are multimode optical fibers.

4. The optical fiber biosensor probe for detecting biomarkers in the bile duct as claimed in claim 2, characterized in that: The main sensing area of ​​the probe (13) is the coreless optical fiber part.

5. An optical fiber biosensor probe for detecting biomarkers in the bile duct as claimed in claim 2, characterized in that: The surface of the probe (13) is coated with a layer of gold film by a magnetron sputtering apparatus, and the surface of the gold film is chemically cross-linked with a specific biomarker protein antibody.

6. An optical fiber biosensor probe for detecting biomarkers in the bile duct as claimed in claim 1, characterized in that: The light source is polarized light.

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