A terahertz biosensor detection device and a biodetection method

Abstract

The present invention relates to the field of terahertz biosensor technology, and in particular to a terahertz biosensor detection device and a biodetection method. For detecting biological samples, the device comprises: a transmissive terahertz time-domain spectroscopy system for providing a transmissive terahertz optical path; a metasurface sensor located on the transmissive terahertz optical path, the metasurface sensor comprising a first gold film and a second gold film of a plurality of asymmetric structures in a periodic array, which excites a transmission spectrum resonance peak under the normal incidence of a terahertz wave in the polarization direction to characterize the biological sample; capture DNA modified on the surface of the metasurface sensor for capturing the biological sample; and a gold nanoprobe for specifically binding to the biological sample after capturing the biological sample to enhance the sensing sensitivity of the biological sample. The present invention adopts a combined detection method of a metasurface sensor and a gold nanoprobe, which can achieve high-sensitivity detection of trace small molecule biological samples.

Description

Technical Field

[0001] The present invention relates to the field of terahertz biosensor technology, and in particular to a terahertz biosensor detection device and a biodetection method. Background Art

[0002] Let-7a is a typical miRNA that can inhibit tumor growth and spread and holds promise for cancer diagnosis and treatment. Its concentration is significantly correlated with survival in cancer patients after surgery. With only 20–24 nucleotides, Let-7a has a relatively low molecular weight and is detected in trace amounts in living organisms. Therefore, developing a non-destructive and accurate Let-7 detection technology is crucial for cancer detection, prevention, and treatment.

[0003] Terahertz waves, due to their non-ionizing properties and minimal damage to biological tissues, are considered a powerful tool in biosensing research. Their frequencies match the skeletal vibration and rotation modes of many biomolecules and have been used to analyze numerous biochemical substances, including proteins, viruses, antibiotics, cells, and nucleic acids (DNA / RNA). However, terahertz biosensing faces severe challenges in the detection of trace substances. The mismatch between terahertz wavelength and the size of biomolecules makes it very difficult to achieve high-sensitivity sensing, especially in the detection of trace amounts in clinical samples. This is partly because metasurface-based biosensing essentially utilizes the response of the sensor device to changes in the dielectric environment of the surrounding space, and can only macroscopically reflect the refractive index changes caused by the substance to be tested. The sensor itself cannot specifically identify the target biomolecule. On the other hand, the presence of multiple different components in clinical samples can easily interfere with the detection results of the metasurface sensor, reducing its sensitivity to trace targets.

[0004] Traditional methods for identifying Let-7a, such as fluorescence detection, are complex, costly, and time-consuming, and fluorescent staining is prone to additional influencing factors. Furthermore, current terahertz biosensor metasurfaces struggle to achieve specific sensing of trace amounts of the small biomolecule Let-7a, particularly in serum samples, which are more realistic for testing. Furthermore, existing terahertz biosensors are generally complex in design and have limited detection capabilities, making it difficult to achieve multiplexing for multifunctional detection. Summary of the Invention

[0005] The present invention aims to at least improve one of the technical problems existing in the prior art. To this end, the present invention proposes a terahertz biosensor detection device and a biodetection method.

[0006] The technical solutions of the present invention are as follows:

[0007] A terahertz biosensor detection device for detecting biological samples, comprising:

[0008] A transmission terahertz time-domain spectroscopy system, used to provide a transmission terahertz optical path;

[0009] A sample holder is arranged on the transmission terahertz optical path of the transmission terahertz time-domain spectroscopy system;

[0010] A metasurface sensor is disposed on the sample holder, wherein the metasurface sensor includes a first gold film and a second gold film having a periodic array of asymmetric structures, and when a terahertz wave in a polarization direction is incident normally, a transmission spectrum resonance peak is excited to characterize the biological sample;

[0011] Capturing DNA, modified on the surface of the metasurface sensor, for capturing biological samples;

[0012] Gold nanoprobes are used to specifically bind to biological samples after capturing biological samples to enhance the sensing sensitivity of biological samples.

[0013] In a possible technical solution, further, the transmission-type terahertz time-domain spectroscopy system includes a terahertz lens emitting end, a first terahertz lens, a second terahertz lens, a third terahertz lens, a fourth terahertz lens and a terahertz detector arranged in sequence along the optical path, and the sample holder is arranged between the second terahertz lens and the third terahertz lens.

[0014] In a possible technical solution, further, the lengths of the first gold film and the second gold film located in the Y direction on the surface of the metasurface sensor are equal; the lengths of the first gold film and the second gold film located adjacent to each other in the X direction on the surface of the metasurface sensor are unequal to form an asymmetric structure, which is used to break the symmetry in the X direction. When the polarization direction of the incident terahertz wave is X, a Q-BIC resonance mode can be excited, and a Q-BIC resonance peak (transmission spectrum resonance peak) can be observed near 0.7 terahertz. The frequency shift of the resonance peak can be used to characterize the concentration of the biological sample.

[0015] It should be noted that the metasurface sensor has a glass substrate with a refractive index of 2 and a thickness of 300 μm, which is used to ensure that the transmittance of terahertz waves is greater than 90%.

[0016] In a possible technical solution, further, the thickness of the first gold film and the second gold film are equal, both 200 nm, which can enhance the optical performance while ensuring structural stability and biochemical compatibility, wherein the optical performance is mainly reflected in enhancing the plasma effect and suppressing background noise.

[0017] In a possible technical solution, further, the particle diameter of the gold nanoprobe is 60nm to 90nm, so that it has a stronger optical response, a higher molecular loading capacity and excellent field enhancement capability.

[0018] In a possible technical solution, further, the length and width of the first gold film are the same, both of which are 62 μm to 66 μm;

[0019] The second gold film has a length of 70 μm to 110 μm and a width of 62 μm to 66 μm.

[0020] It should be noted that the gold nanoprobe was modified with phenylboronic acid to enhance the electromagnetic response between the supersurface sensor and Let-7a.

[0021] The terahertz biosensor detection device of the present invention adopts a combined detection of a metasurface sensor and a gold nanoprobe, which can achieve high-sensitivity detection of trace small molecule biological samples (especially Let-7a), and the sensing sensitivity can reach 7.4GHz / nM.

[0022] A biological detection method, wherein the terahertz biosensor detection device as described above is used to perform biological detection, comprises the following steps:

[0023] Prepare biological sample solutions with different concentration gradients, solutions containing gold nanoprobes, and modification solutions containing thiolated single-stranded DNA (HS-ssDNA) and 6-mercapto-1-hexanol (MCH);

[0024] Cleaning the metasurface sensor, and immersing the cleaned metasurface sensor in the modification solution for modification to obtain captured DNA;

[0025] The supersurface sensor containing the captured DNA is placed in a biological sample solution and fully immersed therein, and then taken out and immersed in a solution containing gold nanoprobes for reaction to obtain a processed biological sample;

[0026] Take out the metasurface sensor, dry it, and place it on the sample holder;

[0027] The transmission terahertz time-domain spectroscopy system is turned on. After the incident terahertz wave is collimated and focused, it passes through the processed biological sample and metasurface sensor and is collimated and focused again. The outgoing terahertz wave is detected by the terahertz detector to obtain the terahertz transmission spectrum of the processed biological sample, thereby obtaining the transmission spectrum resonance peak of each biological sample with different concentrations;

[0028] The concentration of the biological sample is characterized according to the frequency shift of the resonance peak of the transmission spectrum of the biological sample with different concentrations.

[0029] In a possible technical solution, further, the modification solution is a KH2PO4 solution of thiolated single-stranded DNA (HS-ssDNA) and 6-mercapto-1-hexanol (MCH) in a 1:1 ratio, ensuring that there is sufficient spacing between HS-ssDNA molecules to promote hybridization reaction.

[0030] The bioassay method of the present invention utilizes a combination of different biomodifications to achieve multifunctional multiplexed biomolecule detection. It can not only specifically detect Let-7a, but also support the identification of RNA and DNA with the same sequence. The detection limit for complementary sequences is one order of magnitude higher than that for non-complementary sequences, and the response to RNA is more than three times that of DNA.

[0031] The terahertz biosensor detection device and biodetection method according to the present invention are simpler than the prior art, the device is easy to process and can be reused, and the manufacturing cost is low.

[0032] An electronic device includes a memory and a processor, wherein the memory is coupled to the processor; wherein the memory is used to store program data, and the processor is used to execute the program data to implement the above-mentioned biological detection method based on the terahertz biosensor detection device.

[0033] A computer storage medium, wherein instructions are stored in the computer storage medium, and when the instructions are executed on a computer, the computer is caused to execute the above-mentioned biological detection method based on the terahertz biological sensing detection device.

[0034] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. 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 any creative work.

[0036] Figure 1 is a schematic diagram of a transmission terahertz time-domain spectroscopy system of a terahertz biosensing detection device according to an embodiment of the present invention;

[0037] Figure 2 is a schematic diagram of a metasurface sensor of a terahertz biosensing detection device according to an embodiment of the present invention;

[0038] Figure 3 is a working schematic diagram of a terahertz biosensing detection device according to an embodiment of the present invention;

[0039] Figure 4 is a working schematic diagram of a terahertz biosensing detection device according to an embodiment of the present invention;

[0040] Figure 5 1 is a transmission spectrum diagram of the terahertz biosensing detection device for different concentrations of Let-7a according to an embodiment of the present invention;

[0041] Figure 6 1 is a diagram showing the detection results of Let-7a, DNA with the same sequence as Let-7a, and three types of random sequence DNA using the terahertz biosensor detection device according to an embodiment of the present invention.

[0042] Reference numerals:

[0043] Transmission terahertz time-domain spectroscopy system 1, terahertz lens transmitting end 100, first terahertz lens 110, second terahertz lens 120, third terahertz lens 130, fourth terahertz lens 140, terahertz detector 150;

[0044] Sample holder 2;

[0045] Metasurface sensor 3, first gold film 31, second gold film 32;

[0046] Capture DNA4;

[0047] Gold nanoprobe 5. DETAILED DESCRIPTION

[0048] The embodiments of the present invention are described in detail below. The embodiments described with reference to the accompanying drawings are exemplary. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0049] It should be noted that when an element is referred to as being “fixed to” another element, it may be directly on the other element or there may be an intermediate element. When an element is referred to as being “connected to” another element, it may be directly connected to the other element or there may be an intermediate element.

[0050] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention pertains. The terms used in this specification of the present invention are for the purpose of describing specific embodiments only and are not intended to limit the present invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0051] In the specification, claims, and accompanying drawings of this application, the terms "first," "second," "third," and the like are used to distinguish different objects and are not used to describe a particular order. Furthermore, the terms "including," "comprising," "having," and any variations thereof are intended to cover non-exclusive inclusions. For example, a list of steps or elements may be included, or alternatively, steps or elements not listed may be included, or other steps or elements may be included that are inherent to the process, method, product, or apparatus.

[0052] Only portions relevant to the present application are shown in the accompanying drawings, not all of them. Before discussing the exemplary embodiments in more detail, it should be noted that some exemplary embodiments are described as processes or methods depicted as flow charts. Although the flow charts describe the various operations (or steps) as sequential processes, many of the operations can be performed in parallel, concurrently, or simultaneously. In addition, the order of the various operations can be rearranged. The process can be terminated when its operations are completed, but can also have additional steps not included in the accompanying drawings. The process can correspond to a method, function, procedure, subroutine, subprogram, etc.

[0053] As used in this specification, the terms "component," "module," "system," "unit," and the like are used to refer to computer-related entities, hardware, firmware, a combination of hardware and software, software, or software in execution. For example, a unit can be, but is not limited to, a process running on a processor, a processor, an object, an executable file, an execution thread, a program, and / or distributed between two or more computers. In addition, these units can be executed from various computer-readable media having various data structures stored thereon. Units can communicate, for example, via local and / or remote processes based on signals having one or more data packets (e.g., data from a second unit interacting with another unit in a local system, a distributed system, and / or a network, such as the Internet, which interacts with other systems via signals).

[0054] Example 1

[0055] This embodiment provides a terahertz biosensor detection device for detecting biological samples, including:

[0056] A transmissive terahertz time-domain spectroscopy system 1 is configured to provide a transmissive terahertz optical path. The transmissive terahertz time-domain spectroscopy system 1 comprises a terahertz lens emitting end 100, a first terahertz lens 110, a second terahertz lens 120, a third terahertz lens 130, a fourth terahertz lens 140, and a terahertz detector 150, which are sequentially arranged along the optical path. The sample holder 2 is disposed between the second terahertz lens 120 and the third terahertz lens 130.

[0057] The sample holder 2 is arranged on the transmission terahertz optical path of the transmission terahertz time-domain spectroscopy system 1;

[0058] A metasurface sensor 3 is provided on the sample holder 2. The metasurface sensor 3 includes a first gold film 31 and a second gold film 32 in a periodic array of asymmetric structures. When a terahertz wave with a polarization direction is incident normally, the metasurface sensor 3 excites a transmission spectrum resonance peak to characterize the biological sample.

[0059] Capture DNA 4, modified on the surface of the metasurface sensor 3, for capturing biological samples;

[0060] The gold nanoprobe 5 is used to specifically bind to the biological sample after capturing the biological sample to enhance the sensing sensitivity of the biological sample.

[0061] It should be noted that the lengths of the first gold film and the second gold film located in the Y direction of the surface of the metasurface sensor are equal; the lengths of the first gold film and the second gold film located adjacent to each other in the X direction of the surface of the metasurface sensor are unequal to form an asymmetric structure, which is used to break the symmetry in the X direction. When the polarization direction of the incident terahertz wave is X, the Q-BIC resonance mode can be excited, and the Q-BIC resonance peak can be observed near 0.7 terahertz. The frequency shift of the resonance peak can be used to characterize the concentration of the biological sample.

[0062] It should be noted that the metasurface sensor 3 has a glass substrate with a refractive index of 2 and a thickness of 300 μm to 500 μm. In this embodiment, the thickness of the glass substrate is preferably 300 μm to ensure that the transmittance of the terahertz wave is greater than 90%.

[0063] It should be noted that the thickness of the first gold film 31 and the second gold film 32 are equal, both 200 nm, which can enhance the optical performance while ensuring structural stability and biochemical compatibility. The optical performance is mainly reflected in enhancing the plasma effect and suppressing background noise.

[0064] It should be noted that the particle diameter of the gold nanoprobe 5 ranges from 60 nm to 90 nm. In this embodiment, the particle diameter of the gold nanoprobe 5 is preferably 90 nm, so that it has a stronger optical response, a higher molecular loading capacity and excellent field enhancement capability.

[0065] It should be noted that the length and width of the first gold film 31 are identical, both ranging from 62μm to 66μm; the length of the second gold film 32 is 70μm to 110μm, the width is 62μm to 66μm, the period along the X-axis is 256μm to 260μm, and the period along the Y-axis is 256μm to 300μm. In this embodiment, the length and width of the first gold film 31 are identical, both ranging from 64μm; the length of the second gold film 32 is 110μm, the width is 64μm, the period along the X-axis is 256μm, and the period along the Y-axis is 256μm.

[0066] It should be noted that the gold nanoprobe 5 is modified with phenylboronic acid to enhance the electromagnetic response between the metasurface sensor and Let-7a.

[0067] It should be noted that, in this embodiment, the devices in the transmission terahertz time-domain spectroscopy system have no special requirements on the ambient temperature and can operate at room temperature. The operating frequency range of the devices is 0.1 THz to 3 THz.

[0068] The working principle of this embodiment is as follows: the transmission terahertz time-domain spectroscopy system can detect the transmission spectrum of the metasurface sensor, and use the transmission spectrum resonance peak generated by the Q-BIC metasurface structure of the metasurface sensor to characterize the biological sample to be tested. Figure 1 As shown in the transmission terahertz time-domain spectroscopy system, the terahertz wave is emitted from the terahertz lens emitting end 100. The incident linear polarization wave is collimated by the first terahertz lens 110 and focused by the second terahertz lens 120, and then is incident on the sample holder 2. The transmitted wave is collimated by the third terahertz lens 130 and focused by the fourth terahertz lens 140, and then converges to the terahertz detector 150. The outgoing terahertz wave can be detected, and then the complete terahertz transmission spectrum can be restored through theoretical calculation. In addition, the sensing sensitivity and sensing accuracy of the device are defined as:

[0069]

[0070]

[0071] in represents the frequency shift of resonance, Indicates the concentration change of the sample. =1GHz, refers to the frequency resolution of the terahertz time-domain spectroscopy system. The designed metasurface sensor is composed of an asymmetric metal wire array. The lengths of all metal wires in the Y direction are consistent, while the lengths of adjacent metal wires in the X direction are inconsistent. Therefore, it can be considered that the metasurface sensor breaks the symmetry in the X direction. When the terahertz wave with a polarization direction of x is incident normally, the metasurface sensor structure itself can excite the Q-BIC resonance mode and produce a very strong local electric field enhancement. When it is used to detect RNA / DNA molecules, the interaction between the terahertz wave and the molecules of the biological sample to be tested will be significantly enhanced, thereby achieving the purpose of trace sensing of RNA / DNA. In the present invention, the Q-BIC resonance peak can be observed near 0.7 terahertz, and the frequency shift of the resonance peak can be used to characterize the concentration of the biological sample. And as Figure 3 As shown, the present invention utilizes capture DNA to modify a metasurface sensor. The capture DNA can undergo nucleic acid complementary pairing with the sequence of the biological sample being tested (using Let-7a as an example in this example), capturing Let-7a through base pairing and preventing pairing with non-complementary sequences, enabling the metasurface to specifically sense Let-7a. Furthermore, the present invention utilizes phenylboronic acid-modified gold nanoprobes to achieve RNA / DNA discrimination and selectively amplify the Let-7a detection signal. After the metasurface sensor captures Let-7a and DNA of the same sequence, a gold nanoprobe solution is passed over the metasurface. Phenylboronic acid molecules specifically bind to vicinal diols. RNA is composed of ribonucleotides, whose pentose sugars contain two adjacent hydroxyl groups, which interact with phenylboronic acid molecules. In contrast, DNA is composed of deoxyribonucleotides, whose pentose sugars contain only one hydroxyl group and are therefore not specifically recognized by phenylboronic acid molecules. Because the phenylboronic acid-modified gold nanoprobe can only bind to Let-7a and not to DNA of the same sequence, the electromagnetic response between the metasurface and Let-7a is further enhanced, significantly improving sensing sensitivity and detection accuracy, thereby enabling RNA / DNA differentiation. Furthermore, the gold nanoprobe exhibits a surface ionization enhancement effect in the terahertz band, significantly improving the sensor's sensitivity and accuracy for Let-7a. In this embodiment of the present invention, the frequency shift of the Q-BIC resonance peak enables sample concentration sensing and nucleic acid identification.

[0072] The terahertz biosensing detection device according to the present invention adopts a combined detection of a metasurface sensor and a gold nanoprobe, which can achieve high-sensitivity detection of trace small molecule biological samples (Let-7a), and the sensing sensitivity can reach 7.4GHz / nM.

[0073] Example 2

[0074] This embodiment provides a biological detection method based on the above embodiment, wherein the biological detection is performed using the above-mentioned terahertz biosensor detection device, including the following steps:

[0075] Prepare biological sample solutions with different concentration gradients, solutions containing gold nanoprobes, and modification solutions containing thiolated single-stranded DNA (HS-ssDNA) and 6-mercapto-1-hexanol (MCH), where the modification solution is a KH2PO4 solution containing 1 pM HS-ssDNA and 1 pM MCH;

[0076] The metasurface sensor is cleaned and immersed in the modification solution to modify it and obtain captured DNA. In this example, the metasurface sensor is immersed in the modification solution for 2 hours. During this process, the HS-ssDNA acts as the captured DNA and is able to complementarily pair with the target Let-7a. Both HS-ssDNA and MCH can be chemically bound to the gold film surface via S-Au bonds. By using a 1:1 ratio of HS-ssDNA to MCH, sufficient spacing between HS-ssDNA molecules is ensured to promote hybridization reactions.

[0077] The supersurface sensor containing the captured DNA is placed in a prepared biological sample solution and fully immersed therein, and then taken out and immersed in a solution containing gold nanoprobes for reaction to obtain a processed biological sample;

[0078] Take out the metasurface sensor, dry it, and place it on the sample holder;

[0079] The transmission terahertz time-domain spectroscopy system is turned on. After the incident terahertz wave is collimated and focused, it passes through the processed biological sample and metasurface sensor and is collimated and focused again. The outgoing terahertz wave is detected by the terahertz detector to obtain the terahertz transmission spectrum of the processed biological sample, thereby obtaining the transmission spectrum resonance peak of each biological sample with different concentrations;

[0080] The concentration of the biological sample is characterized according to the frequency shift of the resonance peak of the transmission spectrum of the biological sample with different concentrations.

[0081] It should be noted that in the control experiment and serum sample detection, the metasurface sensor was immersed in a DNA solution with the same concentration as the Let-7a sequence and a human serum sample containing Let-7a, respectively. After sufficient soaking time, the metasurface was taken out and immersed in a solution containing gold nanoprobes. After a sufficient time, the metasurface sensor was taken out and dried, and also placed in Figure 1 The samples were characterized on the shown holders.

[0082] Experiments were conducted based on the above detection device and detection method, and the results are as follows:

[0083] like Figure 4 The transmission spectra of Let-7a at concentrations of 0.01 nM to 10 nM are shown in FIG. Figure 4 (a) in the figure is the experimental result without adding gold nanoprobes. When the concentration of Let-7a increases from 0.01 nM to 10 nM, the corresponding Q-BIC resonance peak also moves significantly. When the sample concentration is 10 nM, the Q-BIC resonance peak position is red-shifted by about 24 GHz compared with the initial value. At the same time, it can be seen that when the concentration of Let-7a is very low, it can still cause a slight change in the refractive index, but the displacement of the Q-BIC metasurface resonance peak is very small (about 3 GHz). At this time, it is necessary to use gold nanoprobes 11 to amplify the response. Figure 4 As shown in (b), the frequency shift response of the Q-BIC resonance peak was significantly improved by using gold nanoprobes. Within the same sample concentration range, the maximum frequency shift increased to 74 GHz. It is worth noting that the metasurface sensor exhibited a more pronounced frequency shift response to low-concentration samples, indicating that the gold nanoprobes improved the sensitivity of the metasurface sensor. According to calculations, the gold nanoprobes increased the sensitivity of the metasurface sensor from 2.4 GHz / nM to 7.4 GHz / nM, improving the biosensing performance by more than 3 times. Therefore, by utilizing the high sensitivity of the metasurface sensor to refractive index changes and the signal amplification from the gold nanoprobes, nanometer-scale sensing of Let-7a was achieved.

[0084] like Figure 5 Shown are the test results for Let-7a, DNA with the same sequence as Let-7a (CP DNA), and three random sequence DNAs. Figure 5 As can be seen in (a), the metasurface sensor exhibits significantly greater frequency shift responses to Let7a and CP DNA (both with complementary base sequences) compared to non-complementary random DNA. It is worth noting that for non-complementary sequence random DNA with a concentration as low as 0.01 nM, only an almost negligible frequency shift can be observed. Even with increasing concentration, a smaller frequency shift response can be gradually observed, but it is always lower than the frequency shift response of Let-7a and CP DNA at the same concentration. This shows that the detection limit of the sensor for complementary sequences is one order of magnitude higher than that for non-complementary sequences, demonstrating the sensor's ability to specifically recognize specific sequences. In addition, as Figure 5 As shown in (b), when the concentration of all samples is 10 nM, the resonance peak frequency shift caused by CP DNA (complementary sequence) is about 22 GHz, which is much lower than the 74 GHz caused by Let-7a. This is due to the high binding constant between the gold nanoprobe and RNA (containing cis-diol groups) compared with its weak interaction with DNA (lacking adjacent diol parts).

[0085] like Figure 6 Shown are the test results of human serum samples spiked with different concentrations (0.01 nM to 10 nM) of Let-7a. Figure 6 In (a), it can be observed that the sensor can still accurately detect four concentrations of Let-7a and maintain a discernible frequency shift response even for Let-7a concentrations as low as 0.01 nM, which demonstrates the potential application of the present invention in clinical sample analysis. The frequency shift amplitudes between purified samples and serum samples with different Let-7a concentrations were then compared, and linear regression analysis of these concentration-dependent responses was performed, as shown in Figure 2. Figure 6 As shown in (b). Visual quantitative proof (R 2 =0.970) matrix-induced signal attenuation in the serum environment reduced the detection sensitivity by 39.2% compared to the pure sample control. Crucially, the linear correlation between the logarithmic concentration of Let-7a and the frequency shift in the serum matrix validates the clinical applicability of the present invention.

[0086] The bioassay method of the present invention utilizes a combination of different biomodifications to achieve multifunctional multiplexed biomolecule detection. It can not only specifically detect Let-7a, but also support the identification of RNA and DNA with the same sequence. The detection limit for complementary sequences is one order of magnitude higher than that for non-complementary sequences, and the response to RNA is more than three times that of DNA.

[0087] The terahertz biosensor detection device and biodetection method according to the present invention are simpler than the prior art, the device is easy to process and can be reused, and the manufacturing cost is low.

[0088] Example 3

[0089] This embodiment provides an electronic device, which can be either a mobile electronic device or a non-mobile electronic device. For example, the mobile electronic device can be a mobile phone, tablet computer, laptop computer, PDA, ultra-mobile personal computer (UMPC), netbook, or personal digital assistant (PDA), while the non-mobile electronic device can be a server, network attached storage (NAS), personal computer (PC), etc., although this embodiment of the present application does not specifically limit this. The electronic device includes a memory and a processor, wherein the memory is coupled to the processor; the memory is used to store program data, and the processor is used to execute the program data to implement the aforementioned biodetection method based on a terahertz biosensor detection device. When the program or instructions are executed by the processor, each process of the aforementioned biodetection method embodiment can be implemented, and the same technical effects can be achieved. To avoid repetition, the details are not repeated here.

[0090] An embodiment of the present application also provides a computer storage medium, wherein the computer storage medium stores instructions. When the instructions are executed on a computer, the computer executes the various processes of the above-mentioned embodiment of the biological detection method based on the terahertz biosensor detection device, and can achieve the same technical effect. To avoid repetition, they are not repeated here.

[0091] The processor is the processor in the electronic device in the above embodiment. The readable storage medium includes a computer-readable storage medium, such as a computer read-only memory (ROM), random access memory (RAM), a magnetic disk, or an optical disk.

[0092] Throughout this specification, reference to terms such as "one embodiment," "some embodiments," "illustrative embodiments," "example," "specific example," or "some examples" means that a specific feature, structure, material, or characteristic described in conjunction with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, illustrative expressions of the above terms do not necessarily refer to the same embodiment or example.

[0093] Obviously, the described embodiments are only some of the embodiments of the present application, rather than all of the embodiments. Mentioning "embodiment" in this article means that the specific features, structures or characteristics described in conjunction with the embodiment may be included in at least one embodiment of the present embodiment application. The appearance of this phrase in various positions in the specification does not necessarily mean that they are all the same embodiments, nor are they independent or alternative embodiments that are mutually exclusive with other embodiments. It can be understood explicitly and implicitly by those skilled in the art that the embodiments described herein can be combined with other embodiments. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of this application.

[0094] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to the embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the claims and their equivalents.

Claims

1. A terahertz biosensor detection device, characterized in that: For testing biological samples, including: A transmission terahertz time-domain spectroscopy system (1) is used to provide a transmission terahertz optical path; A sample holder (2) is arranged on the transmission terahertz optical path of the transmission terahertz time-domain spectroscopy system (1); A metasurface sensor (3) is provided on the sample holder (2), wherein the metasurface sensor (3) comprises a first gold film (31) and a second gold film (32) of a plurality of asymmetric structures in a periodic array, and when a terahertz wave in a polarization direction is incident normally, a transmission spectrum resonance peak is excited to characterize the biological sample, wherein the lengths of the first gold film (31) and the second gold film (32) located in the Y direction on the surface of the metasurface sensor (3) are equal; and the lengths of the first gold film (31) and the second gold film (32) adjacent to each other in the X direction on the surface of the metasurface sensor (3) are unequal to form an asymmetric structure; Capturing DNA (4), modified on the surface of the supersurface sensor (3), for capturing biological samples; The gold nanoprobe (5) is used for specifically binding to the biological sample after capturing the biological sample to enhance the sensing sensitivity of the biological sample, wherein the particle diameter of the gold nanoprobe (5) is 60nm to 90nm.

2. The terahertz biosensor detection device according to claim 1, characterized in that: The transmission-type terahertz time-domain spectroscopy system (1) comprises a terahertz lens emitting end (100), a first terahertz lens (110), a second terahertz lens (120), a third terahertz lens (130), a fourth terahertz lens (140), and a terahertz detector (150) sequentially arranged along an optical path, wherein the sample holder (2) is arranged between the second terahertz lens (120) and the third terahertz lens (130).

3. The terahertz biosensor detection device according to claim 1, characterized in that: The thickness of the first gold film (31) and the second gold film (32) are equal, both being 200 nm.

4. The terahertz biosensor detection device according to claim 3, characterized in that: The length and width of the first gold film (31) are the same, both being 62 μm to 66 μm; The second gold film (32) has a length dimension of 70 μm to 110 μm and a width dimension of 62 μm to 66 μm.

5. A biological detection method, characterized in that: The terahertz biosensor detection device according to any one of claims 1 to 4 is used to perform biodetection, comprising the following steps: preparing biological sample solutions with different concentration gradients, a solution containing gold nanoprobes, and a modification solution containing thiolated single-stranded DNA and 6-mercapto-1-hexanol; Cleaning the metasurface sensor, and immersing the cleaned metasurface sensor in the modification solution for modification to obtain captured DNA; The supersurface sensor containing the captured DNA is placed in a biological sample solution and fully immersed therein, and then taken out and immersed in a solution containing gold nanoprobes for reaction to obtain a processed biological sample; Take out the metasurface sensor, dry it, and place it on the sample holder; The transmission terahertz time-domain spectroscopy system is turned on. After the incident terahertz wave is collimated and focused, it passes through the processed biological sample and metasurface sensor and is collimated and focused again. The outgoing terahertz wave is detected by the terahertz detector to obtain the terahertz transmission spectrum of the processed biological sample, thereby obtaining the transmission spectrum resonance peak of each biological sample with different concentrations; The concentration of the biological sample is characterized according to the frequency shift of the resonance peak of the transmission spectrum of the biological sample with different concentrations.

6. The biological detection method according to claim 5, characterized in that The modification solution is a KH2PO4 solution of thiolated single-stranded DNA and 6-mercapto-1-hexanol in a ratio of 1:

1.

7. An electronic device comprising a memory and a processor, characterized in that: The memory is coupled to the processor; The memory is used to store program data, and the processor is used to execute the program data to implement the biological detection method according to claim 5.

8. A computer storage medium, characterized in that The computer storage medium stores instructions, which, when executed on a computer, enable the computer to execute the biological detection method according to claim 5.

Images