A chip-level biomolecule detection system and a detection method

Abstract

The application provides a chip-level biomolecule detection system, which comprises a biomolecule detection chip and an integrated module; the integrated module comprises a temperature control chip and an information acquisition assembly; the temperature control chip is in heat-conducting contact with the biomolecule detection chip; the information acquisition assembly comprises an optical waveguide chip and an image sensing chip; the optical waveguide chip is covered on the image sensing chip, and a filter layer is arranged between the optical waveguide chip and the image sensing chip. Nucleic acid extraction, purification, enrichment and amplification can be realized through the biomolecule chip; low power consumption and high power density can be realized by using the temperature control chip; the optical waveguide chip does not need to be installed with geometric optical components, and the optical path calibration and maintenance in the later stage are avoided, which is beneficial to multi-channel large-scale parallel detection and can be manufactured on a large scale; the structure is simple, and the application requirements of equipment miniaturization and portability are met through chip-level integration. The application also provides a detection method, which is simple in steps, accurate in temperature control, high in detection efficiency and reliable in results.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biomolecule detection, and particularly relates to a chip-level biomolecule detection system and a corresponding biomolecule detection method. BACKGROUND

[0002] Biomolecule detection is the most direct, reliable and sensitive method for early, rapid and specific detection of pathogens. Taking nucleic acid detection as an example, it can quickly detect pathogen nucleic acids in the detection sample, providing a scientific test basis for the diagnosis of infectious cases. The premise of nucleic acid analysis is nucleic acid amplification, which amplifies the nucleic acid sequence to be detected by the action of an enzyme, i.e. polymerase chain reaction (PCR), referred to as PCR detection. PCR technology is most widely used because of its good specificity, low cost and other advantages. Taking variable temperature PCR as an example, its general process includes three basic reaction steps of denaturation, annealing and extension. For PCR detection that needs to cycle the temperature up and down, the existing PCR reaction container has low heat conduction efficiency, resulting in slow temperature rising and falling speed, and thus a long process time. There are also constant temperature PCR in the prior art that does not need to cycle the temperature up and down, such as LAMP loop-mediated isothermal amplification, RPA recombinase polymerase amplification (also known as RAA technology), RCA rolling circle amplification, SDA enzyme-mediated DNA in vitro isothermal amplification method, CRP cross primer isothermal amplification technology, TMA transcription-mediated nucleic acid amplification, NASBA nucleic acid sequence-based amplification, SAT real-time fluorescent isothermal amplification detection, etc. Although these constant temperature PCR do not need to cycle the temperature up and down, the efficiency is better than that of variable temperature PCR, but in constant temperature PCR, complex and expensive detection equipment is still needed, and the temperature provided needs to be kept at a precise value. In addition, in the existing biomolecule detection method, the extraction, purification, enrichment and amplification of nucleic acids are implemented by different independent devices, and the detection cycle is long, the steps are numerous, and the detection place, environment and equipment configuration of the detection system are all very high, the detection system is complex and needs to be built before detection. The existing detection system has great limitations for outdoor or specific places and mobile detection work. In the detection process of biomolecules, photoelectric detection equipment is generally used for signal conversion, and the photoelectric detection equipment in the existing detection system is generally a spatial geometric optical design, which has complex structure, complex manufacturing and installation of geometric optical components, high cost, and needs to be calibrated after a period of time, which has a great impact on the continuity and timeliness of the detection process. It can be seen that the existing biomolecule detection system does not meet the development trend of device miniaturization, and the existing biomolecule detection system has great limitations in further improving the detection efficiency and effectively implementing the detection work in application.

[0003] Therefore, it is very necessary to study a chip-level biomolecule detection system and a corresponding detection method, which can be applied to biomolecule detection work using variable temperature or constant temperature PCR, has low cost, high integration, flexible application and is conducive to further optimizing detection efficiency, and has good reliability of detection results, so as to further promote the in-depth development and wide application of biomolecule detection technology. SUMMARY

[0004] The present application is to solve all or part of the problems of the prior art, one aspect provides a chip-level biomolecule detection system, which can be applied to in vitro nucleic acid detection, but not limited to. Another aspect of the present application provides a detection method, which uses the chip-level biomolecule detection system of the present application for detection.

[0005] One aspect of the present application provides a chip-level biomolecule detection system, which comprises a biomolecule detection chip and an integrated module; the integrated module comprises a temperature control chip and an information acquisition assembly, a cavity is formed between the temperature control chip and the information acquisition assembly, for accommodating the biomolecule detection chip; the temperature control chip is in thermal contact with the biomolecule detection chip; the information acquisition assembly comprises an optical waveguide chip and an image sensor chip, the optical waveguide chip is located on the side close to the cavity; the optical waveguide chip is covered on the image sensor chip, and a filter layer is arranged between the two.

[0006] The biomolecule detection chip can be integrated with high precision by microfluidic technology, and traditional nucleic acid extraction, purification, enrichment and amplification can be completed in the micro-nano scale channel and microcavity in the biomolecule detection chip. The integrated module integrates the temperature control chip, the optical waveguide chip and the image sensor chip, and the temperature control, signal excitation and signal acquisition in the biomolecule detection process can be implemented by chip-level components. The temperature control by the temperature control chip realizes low power consumption and high power density, and can accurately control the temperature, meet the control of temperature rising and falling cycle in variable temperature PCR application, and control to keep at a constant temperature in constant temperature PCR application; the optical waveguide chip transfers the optical path to the chip, and the optical waveguide can be manufactured on the chip, which can be manufactured on a large scale, without installing geometric optical components and without later optical path calibration and maintenance, so the cost can be reduced, and when large-scale parallel detection is needed, large-scale parallel detection fluorescence excitation channels can be designed on the chip, filling the gap that cannot be realized by spatial optical structure; the chip-level structure reduces the necessary structure design space of the biomolecule detection system, improves the portability of the biomolecule detection system, and further enhances the flexibility, efficiency and portability of the biomolecule detection system application.

[0007] The filter layer is deposited on the image sensor chip; the optical waveguide chip is a multilayer thin film structure, which is deposited on the filter layer.

[0008] In one specific implementation, the filter layer is deposited on the image sensor chip by an ultra-low temperature thin film deposition method, and then the multi-layer thin film structure of the optical waveguide chip is also deposited by the low temperature deposition method. The optical waveguide chip is generated by ICPCVD / PECVD, photolithography, etching, CMP, etc. The image sensor chip is selected according to different fluorescence detection intensity and detection range; the filter layer is generated on the image sensor chip by a low temperature thin film deposition process, the multi-layer thin film structure is deposited on the filter layer by a low temperature thin film deposition process, and then the optical waveguide structure is generated to obtain the optical waveguide chip. The multi-layer thin film structure includes but is not limited to silica, silicon nitride, etc.

[0009] The optical waveguide chip is provided with a plurality of fluorescence excitation channels; the fluorescence excitation channel includes a plurality of optical waveguide units; the optical waveguide unit includes an incident grating structure, a multimode interference coupling structure, an exit grating structure arranged in sequence, and a waveguide arranged between the incident grating structure and the multimode interference coupling structure, and between the multimode interference coupling structure and the exit grating structure. The waveguide can be a multimode or single-mode waveguide. The optical waveguide structure can be used to control light on the optical waveguide chip, including but not limited to light combination and light splitting, and the exit grating structure can be used to project light into space.

[0010] The refractive index of the optical waveguide chip ranges from 1.79 to 2.1. The optical waveguide chip with the refractive index range can accurately project light to a specific position on the biomolecule detection chip inserted into the cavity.

[0011] The biomolecule detection chip is one of a silicon-based chip, a plastic chip, or a hybrid integrated chip of plastic and silicon-based chip, and is provided with a microfluidic structure to achieve nucleic acid extraction, purification, enrichment, and amplification. The biomolecule detection chip has high heat conduction efficiency and can quickly complete variable temperature or constant temperature PCR detection; it can be mass-produced and designed as an integrated microfluidic chip to achieve nucleic acid extraction, purification, enrichment, and amplification. The use of plastic and other heat-conducting materials has low cost and is more suitable for mass production, which is conducive to the manufacture of the biomolecule detection chip and the popularization and application of the chip-level biomolecule detection system.

[0012] The biomolecule detection chip is also provided with a sealing mechanism and a sample addition area; the sample addition area is provided with a sample inlet and an air outlet; and the sealing mechanism is used to seal the sample inlet and the air outlet. In biomolecule detection, biological safety is the first priority, and it is very important to prevent liquid leakage and aerosol generation during the detection process. The sealed biomolecule detection chip can achieve a fully enclosed PCR detection environment.

[0013] The temperature control chip is provided with a limiting structure, and the corresponding biomolecule detection chip is provided with a positioning structure. The limiting structure cooperates with the positioning structure to limit the contact position of the temperature control chip and the biomolecule detection chip. The limiting structure and the positioning structure make the contact position of the temperature control chip and the biomolecule detection chip not easy to relatively displace in the process of heat conduction, keep the action area of heat conduction unchanged, and are beneficial to the stability of heat conduction and the consistency of temperature change of the action position.

[0014] The biomolecule detection system is also provided with a structure for keeping the temperature control chip in heat conduction contact with the biomolecule detection chip. The structure can adopt a buckle structure, a magnet structure, or a pressing structure formed by the upper and lower surfaces of the cavity (an upper cover is additionally provided on the temperature control chip, and the downward pressure structure of the upper cover cooperates with the bearing surface of the cavity to press the temperature control chip and the biomolecule detection chip tightly).

[0015] The surface of the temperature control chip in contact with the biomolecule detection chip is provided with a layer of heat-conducting material. By arranging the layer of heat-conducting material between the temperature control chip and the biomolecule detection chip, the close heat-conduction contact of the temperature control chip and the biomolecule detection chip is further ensured, and the influence of the contact gap caused by slight errors in the structure design on the heat conduction effect is effectively avoided. In some implementation cases, the layer of heat-conducting material can also be omitted, and the temperature control chip and the biomolecule detection chip can be directly heated by being pressed tightly, which is not limited.

[0016] The temperature control range of the temperature control chip is 20℃ to 110℃, the temperature control precision is 0.1℃, and the temperature control rate range is 0.1℃ / second to 30℃ / second.

[0017] The side of the optical waveguide chip facing the cavity is provided with a light-transmitting layer, and the light-transmitting layer material includes but is not limited to one of inorganic glass or organic glass. Preferably, the light-transmitting layer material adopts colorless and transparent organic glass or inorganic glass. The light-transmitting layer is arranged between the biomolecule detection chip and the optical waveguide chip, plays a role in protecting the optical waveguide chip, avoids the direct contact of the biomolecule detection chip inserted into the cavity with the optical waveguide chip, makes the optical waveguide chip not easy to be damaged in multiple uses, and is beneficial to keeping the stability of the chip-level biomolecule detection system in multiple uses and prolonging the service life of the chip-level biomolecule detection system.

[0018] The chip-level biomolecule detection system further comprises a light source assembly and an information analysis assembly; the light source assembly is optically connected with the optical waveguide chip and is used to provide a working light beam; the information analysis assembly is electrically connected with the image sensing chip and is used to perform information processing to obtain a detection result.

[0019] According to different fluorescent substances, the working light beam has multiple wavelengths, including but not limited to 470 nm, 530 nm, 580 nm and 630 nm, and different fluorescent substances are excited correspondingly. The light source of the light source assembly can be a laser or an LED, and the working light beam provided by the light source has multiple selectable wavelengths, which can meet the excitation of multiple fluorescent substances in a parallel detection fluorescent excitation channel, including but not limited to 470 nm for fluorescent substance FAM, 530 nm for HEX, 580 nm for ROX and 630 nm for CY5.

[0020] The information analysis assembly comprises a touch display screen and a memory, and is used to display and record the detection result.

[0021] Another aspect of the present application provides a detection method, which is performed by using the chip-level biomolecule detection system, and comprises: PCR detection to obtain a fluorescent signal; collection of fluorescent light excited by biomolecules and imaging, and extraction of the fluorescent signal for detection and judgment; the PCR detection comprises a control target of a preset temperature, and the reaction temperature is controlled through a temperature control program.

[0022] The PCR detection further comprises: obtaining a biomolecule sample, placing the biomolecule sample in a biomolecule detection chip, and performing nucleic acid extraction, purification, enrichment and amplification.

[0023] Before the PCR detection, the biomolecule detection chip is inserted into the cavity, and the temperature control chip and the light source assembly are started. During the entire PCR detection process, fluorescent excitation and fluorescent collection are performed synchronously, the fluorescent signal can be extracted synchronously for detection and judgment, and the entire detection result is more comprehensive and complete.

[0024] The PCR detection is semi-quantitative PCR or quantitative PCR.

[0025] The temperature control program is based on a PID algorithm; the control target of the preset temperature includes but is not limited to: the number of temperature rising and falling cycles, the denaturation temperature value and duration of the PCR detection, the annealing temperature value and duration, and the extension temperature value and duration. The PID algorithm is a control algorithm that combines proportional, integral and differential links. Based on the PID algorithm, accurate control can be achieved, which can effectively prevent temperature overshoot and ensure that the temperature platform period temperature is stable and the jitter is less than 0.1℃.

[0026] The extraction of the fluorescent signal for detection and judgment is executed by running a preset algorithm, including judging the category of biological molecules and whether viruses and bacteria exist through the intensity change of fluorescence; the algorithm includes an image processing algorithm based on threshold analysis and / or a clustering analysis algorithm based on machine learning.

[0027] Compared with the prior art, the main beneficial effects of the present application are:

[0028] 1、The chip-level biological molecule detection system of the present application adopts an on-chip integrated structure to realize biological molecule detection, has simple structure design, low material cost, meets the application requirements of device miniaturization and portability, is easy to mass-produce, and further widens the applicable scenarios of the biological molecule detection system, providing a positive and effective solution for nucleic acid detection, daily work and research and development of biological medicine. The biological molecule detection chip can integrally realize the functions of nucleic acid extraction, purification, enrichment and amplification, and has low material cost; the temperature control chip is arranged in heat conduction contact with the biological molecule detection chip, accurate temperature control can be realized, and low power consumption and high power density can be realized by using the temperature control chip; the optical waveguide chip realizes the transfer of spatial geometric optics to on-chip optics, generates an optical waveguide structure on the chip, is suitable for large-scale manufacturing combined with semiconductor manufacturing processes, is not dependent on the structure limitation of spatial optics, does not need to install geometric optical components, and does not need to be maintained by later optical path calibration, greatly reducing the equipment cost, improving the detection efficiency, and optimizing the continuity of the detection process, and when large-scale parallel detection is performed, a large number of parallel detection fluorescence excitation channels can be arranged on the optical waveguide chip. The chip-level biological molecule detection system of the present application is applicable to not only variable temperature PCR, but also constant temperature PCR such as LAMP, further meeting the breadth and depth of the application of the biological molecule detection system.

[0029] 2、The detection method of the present application has corresponding advantages when the chip-level biological molecule detection system of the present application is used, each link of the detection is integrally implemented by using the chip-level detection component, and the detection result is extracted combined with an intelligent algorithm, so that the overall steps are simple, the temperature control is accurate, the detection efficiency is high, and the detection result has high reliability. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 Figure 1 is a schematic diagram of a chip-level biomolecule detection system according to an embodiment of the present application.

[0031] Figure 2 Figure 2 is a schematic diagram of a light waveguide unit structure according to an embodiment of the present application.

[0032] Figure 3 Figure 3 is a schematic diagram of a chip-level biomolecule detection system according to another embodiment of the present application.

[0033] Figure 4 Figure 4 is a schematic diagram of a detection method according to a further embodiment of the present application.

[0034] Legend of reference signs:

[0035] Biomolecule detection chip 1, cavity 10, integrated module 2, information acquisition component 20, temperature control chip 21, light waveguide chip 22, image sensor chip 23, light filter layer 24, light transmission layer 25, incident grating structure 221, multimode interference coupling structure 222, outgoing grating structure 223, waveguide 224, light source component 3, information analysis component 4. DETAILED DESCRIPTION

[0036] It should be noted that the flowcharts and block diagrams in the drawings illustrate the operation processes that can be implemented according to the embodiments of the present application. It should also be noted that in some alternative implementations, the functions labeled in the blocks can also occur in a different order from that labeled in the drawings. For example, two blocks that are represented successively can actually be executed substantially in parallel, and they can also be executed in an interleaved manner, depending on the purpose to be achieved by the steps involved.

[0037] Embodiment 1

[0038] In the embodiment 1 of the present application, as shown in Figure 1, a chip-level biomolecule detection system comprises a biomolecule detection chip 1, a light source component 3, an information acquisition component 20, and an information analysis component 4. Figure 1As shown, the chip-level biomolecule detection system comprises a biomolecule detection chip 1 and an integrated module 2. The example biomolecule detection chip 1 is a plastic and silicon-based hybrid integrated chip, which is used to integrally complete nucleic acid extraction, purification, enrichment and amplification. The example biomolecule detection chip 1 is provided with a microfluidic structure (not shown), a sealing mechanism (not shown) and a sample loading area (not shown). The sample loading area is provided with a sample inlet and a gas outlet, and the sealing mechanism is used to seal the sample inlet and the gas outlet. The example sealing mechanism can adopt a PCR sealing film, a sealing plug, an elastic adhesive tape and the like. In the example case, the sample inlet and the gas outlet are sealed by the sealing plug to achieve a fully closed PCR detection environment. The biomolecule detection chip 1 can also adopt a silicon-based chip, a plastic chip or a chip made of other heat-conducting materials. The heat-conducting material is not limited here and can be selected according to the actual application and the manufacturing cost. The biomolecule detection chip 1 of the embodiment can realize rapid temperature rising and falling in the variable-temperature PCR application, thereby greatly shortening the PCR reaction time and can shorten the PCR reaction process, which usually takes more than one hour, to five minutes at the fastest.

[0039] The example integrated module 2 comprises an information acquisition assembly 20 and a temperature control chip 21, and a cavity 10 accommodating the biomolecule detection chip 1 is formed between the temperature control chip 21 and the information acquisition assembly 20. The temperature control chip 21 is in heat-conducting contact with the biomolecule detection chip 1 inserted into the cavity 10. As shown in the figure, Figure 1 As shown, the information acquisition assembly 20 comprises a light waveguide chip 22 and an image sensing chip 23. The light waveguide chip 22 covers the image sensing chip 23, and a filter layer 24 is arranged between the two. In the embodiment, the filter layer 24 is deposited on the image sensing chip 23 by a low-temperature thin film deposition process. The example image sensing chip 23 is a commercial CMOS chip, and other types of photoelectric conversion devices can also be used to realize image sensing in some specific applications, which are not limited. The filter layer 24 is deposited on the CMOS chip by an ultra-low-temperature thin film deposition method. The example process of the ultra-low-temperature thin film deposition method comprises: depositing a layer of silicon nitride thin film on a substrate material by inductively coupled chemical vapor deposition (ICPCVD), and the deposition temperature is 25-150°C, which is a low-temperature process. The silicon nitride is realized by adjusting the flow ratio of the silicon source and the nitrogen source in the reaction gas. The silicon source can be silane, and the nitrogen source can be ammonia or nitrogen. In the embodiment, after the filter layer 24 is deposited on the image sensing chip 23, the low-temperature deposition method is also used to deposit the multi-layer thin film structure of the light waveguide chip 22. The medium of the adjacent two layers in the general multi-layer film structure is silicon dioxide or silicon nitride. The entire preparation process adopts processes such as ICPCVD or PECVD, photolithography, etching and CMP.

[0040] In the embodiment, the light waveguide chip 22 is provided with a plurality of fluorescence excitation channels. As shown in the figure, Figure 2As shown, each fluorescence excitation channel can include an independent light waveguide unit, or multiple light waveguide units can be set according to actual application, and is not limited. An example of a light waveguide unit includes, in sequence, an incident grating structure 221, a multimode interference coupling structure 222, an exit grating structure 223, and a waveguide 224 arranged between the incident grating structure 221 and the multimode interference coupling structure 222 and between the multimode interference coupling structure 222 and the exit grating structure 223. The waveguide 224 can be a single-mode waveguide or a multimode waveguide, and is not limited.

[0041] As Figure 1As shown, the temperature control chip 21 in the example adopts a TEC chip. The TEC chip, i.e. TEC (Thermoelectric Cooler), is widely used in temperature control. The temperature control chip 21 in the example can adopt a conventional cast TEC chip. In some embodiments, a TEC chip based on semiconductor thin film technology can also be adopted, without limitation. The temperature control range of the temperature control chip 21 in the example is 20-110°C, and the temperature control accuracy under PID temperature control program control can reach 0.1°C, and the temperature control rate range is 0.1°C / s-30°C / s. In the application example of the temperature cycler, the temperature control chip 21 can accurately control the temperature in the cyclic temperature rising and falling process. In some embodiments, when applied to constant temperature PCR such as LAMP, the temperature control chip 21 can control to maintain a precise constant temperature. The temperature control chip 21 is combined with the corresponding settings of the specific application, which is not limited here. In the present embodiment, the temperature control chip 21 is provided with a limiting structure, and the biological molecule detection chip 1 is provided with a positioning structure. The contact position of the temperature control chip 21 and the biological molecule detection chip 1 is fixed by the cooperation of the limiting structure and the positioning structure during the working process and does not move. In the example, a limiting groove is provided on the surface of the temperature control chip 21 facing the cavity 10, and a corresponding elastic lug positioning structure is provided on the outer surface of the biological molecule detection chip 1, so that the contact position of the biological molecule detection chip 1 and the temperature control chip 21 does not change during the working process after the biological molecule detection chip 1 is inserted into the cavity 10. The limiting structure and the positioning structure of the biological molecule detection chip 1 and the temperature control chip 21 adopt a quick release structure, and the biological molecule detection chip 1 can also be taken out of the cavity 10 after the detection process is completely finished according to actual needs. In the present embodiment, the structure is designed to maintain the close heat-conducting contact between the temperature control chip 21 and the biological molecule detection chip 1, so as to avoid the separation of the two due to external factors such as vibration during the detection application process. The example structure can be to provide a clamping hole on the temperature control chip 21, and a corresponding clamping hook on the biological molecule detection chip 1, so that the contact position of the two cannot be separated during the working process through the clamping structure. The distance between the upper and lower sides of the cavity 10 can also be designed, and an upper cover plus downward pressure structure is provided on the side of the temperature control chip 21 away from the cavity 10. After the biological molecule detection chip 1 is inserted into the cavity 10, the extrusion structure formed by the upper and lower surfaces of the cavity 10 maintains the heat-conducting contact between the biological molecule detection chip 1 and the temperature control chip 21. In a better implementation of the present embodiment, a magnet is embedded in the biological molecule detection chip 1, and a corresponding magnet is also embedded in the temperature control chip 21, so as to maintain the two by magnetic attraction. In the present embodiment, in order to make the contact more close, a layer of heat-conducting material can also be filled between the biological molecule detection chip 1 and the temperature control chip 21. In the specific implementation, a layer of heat-conducting material is provided on the surface of the temperature control chip 21 in contact with the biological molecule detection chip 1, which further avoids the existence of gaps between the biological molecule detection chip 1 and the temperature control chip 21 to affect the heat-conducting effect.

[0042] Example 2

[0043] like Figure 3 As shown, the main difference between Embodiment 2 and Embodiment 1 is that a light-transmitting layer 25 is also provided on the side of the information acquisition component 20 facing the cavity 10. The light-transmitting layer 25 in this example is an optical glass layer, made of colorless and transparent organic glass. The surface of the optical glass layer facing the cavity 10 forms a sidewall of the cavity 10. When the biomolecular detection chip 1 is inserted into the cavity 10, the side of it facing away from the temperature control chip 21 contacts the surface of the optical glass layer.

[0044] In this embodiment, the chip-level biomolecule detection system further includes a light source component 3 and an information analysis component 4. The light source component 3 in this example is a laser, and its working beam output is optically connected to the optical waveguide chip 22 via an optical fiber and an optical fiber coupler to provide the working beam. The light source can also be an LED light source or other types of light sources; this is not limited here. The information analysis component 4 is electrically connected to the image sensing chip 23 to process information and obtain the detection result. In the example implementation, such as... Figure 3 As shown, the integrated module 2 and the information analysis component 4 are integrated through a housing, which has an optical window for introducing the working beam. The example can use more than one laser. The wavelength of the working beam provided by the light source component 3 can satisfy the excitation of various fluorescent substances in the parallel fluorescence excitation channels, including but not limited to 470nm for FAM, 530nm for HEX, 580nm for ROX, and 630nm for CY5. The width and height of the waveguide structure of the corresponding optical waveguide unit also change accordingly for different wavelengths of the working beam.

[0045] The example information analysis component 4 may also include a touch screen and memory to display and record the detection results.

[0046] Example 3

[0047] Example 3 illustrates the detection method provided by the present invention. Figure 4The detection method includes: obtaining a biomolecule sample, placing it in the biomolecule detection chip 1, and performing nucleic acid extraction, purification, enrichment, and amplification; PCR detection to obtain a fluorescence signal; collecting the fluorescence of the biomolecules excited and imaging, and extracting the fluorescence signal for detection and judgment. Through PCR detection, a fluorescence signal is obtained. During PCR detection, the fluorescence of the biomolecules excited is collected and imaged synchronously, and the fluorescence signal is extracted. There are preset temperature control targets during PCR detection, which include, in this embodiment, the number of temperature rising and falling cycles, the denaturation temperature value and duration of PCR detection, the annealing temperature value and duration, and the extension temperature value and duration. For example, 60°C, 95°C, and 60°C are one cycle, and the entire PCR process is 40 cycles. In this embodiment, the specific temperature control is precisely controlled by a PID algorithm to prevent temperature overshoot and ensure that the temperature platform period is stable with a jitter of less than 0.1°C. It should be noted that in other specific implementation cases, such as when applied to constant-temperature PCR, the preset temperature control target can also include a preset temperature value to control the heat conduction process and maintain a constant specific temperature value. Here, it is not limited.

[0048] By reference Figure 1 , Figure 2 and Figure 3 , in this embodiment, the cavity 10 as shown in Embodiment One or Two is prepared before PCR detection. The image sensor chip 23 is selected according to different fluorescence detection intensities and detection ranges. The dynamic range, dark noise, readout noise, quantum efficiency, pixel size, chip size, full-frame speed, and other technical parameters of the image sensor chip 23 are considered according to different fluorescence detection intensities and detection ranges, and the image sensor chip 23 that meets the detection requirements is selected. The nucleic acid extraction, purification, enrichment, and amplification are integrated in the biomolecule detection chip 1. In this embodiment, the biomolecule detection chip 1 is inserted into the cavity 10 to start the light source assembly 3 and the temperature control chip 21. During the entire PCR detection process, fluorescence excitation is performed synchronously, detection judgment can be performed synchronously, and the entire detection result is more comprehensive and complete. The example PCR detection can use semi-quantitative PCR or quantitative PCR, and is not limited.

[0049] In this embodiment, the example light waveguide unit can form a matrix detection structure. The exit grating structure 223 is located below the microfluidic channel of the biomolecule detection chip 1 to guide light upward into the microfluidic channel. The refractive index of the example light waveguide chip 22 ranges from 1.79 to 2.1. Different exit directions of the exit grating structure 223 can be designed according to actual application to facilitate increased detection flexibility.

[0050] In the present embodiment, the extraction of the fluorescent signal for detection and judgment is performed by running a preset algorithm, including judging the category of the biomolecule and whether there is a virus, bacteria, etc. by the intensity change of the fluorescence. The example algorithm includes an image processing algorithm based on threshold analysis and a clustering analysis algorithm based on machine learning. Only one of the algorithms or other algorithms can also be selected according to the actual application needs, and is not limited. The fluorescent signal can be converted by an analog / digital converter ADC and operated by an FPGA / ASIC, and then sent to the processor of the computer to run the algorithm to perform image and data processing to obtain the detection result.

[0051] The above description of the embodiments is only used to help understand the method and core idea of the present application. For those skilled in the art, some improvements and modifications can be made to the present application without departing from the principles of the present application, and these improvements and modifications also fall within the scope of protection of the claims of the present application.

Claims

1. A chip-scale biomolecule detection system, characterized by: The chip comprises a biomolecule detection chip and an integrated module. The integrated module comprises an information acquisition component and a temperature control chip, and a cavity is formed between the information acquisition component and the temperature control chip to accommodate the biomolecule detection chip. The biomolecule detection chip is one of a silicon-based chip, a plastic chip, or a plastic and silicon-based hybrid integrated chip, and is provided with a microfluid channel structure to achieve nucleic acid extraction, purification, enrichment, and amplification. The temperature control chip is in thermal contact with the biomolecule detection chip. The information acquisition component comprises an optical waveguide chip and an image sensing chip, the optical waveguide chip is located on the side close to the cavity, and the optical waveguide chip is covered on the image sensing chip, and a filter layer is arranged between the optical waveguide chip and the image sensing chip.

2. The chip-scale biomolecule detection system of claim 1, wherein: The filter layer is deposited on the image sensing chip, and the optical waveguide chip is a multi-layer thin film structure deposited on the filter layer.

3. The chip-scale biomolecule detection system of claim 1, wherein: The optical waveguide chip is provided with a plurality of fluorescence excitation channels, each fluorescence excitation channel comprises a plurality of optical waveguide units, each optical waveguide unit comprises an incident grating structure, a multimode interference coupling structure, an exit grating structure arranged in sequence, and a waveguide arranged between the incident grating structure and the multimode interference coupling structure and between the multimode interference coupling structure and the exit grating structure.

4. The chip-scale biomolecule detection system of claim 1, wherein: The refractive index of the optical waveguide chip ranges from 1.79 to 2.

1.

5. The chip-scale biomolecule detection system of claim 1, wherein: The biomolecule detection chip is further provided with a sealing mechanism and a sample adding area, the sample adding area is provided with a sample inlet and an air outlet, and the sealing mechanism is used to seal the sample inlet and the air outlet.

6. The chip-scale biomolecule detection system of claim 1, wherein: The temperature control chip is provided with a limiting structure, and the biomolecule detection chip is correspondingly provided with a positioning structure, the limiting structure and the positioning structure cooperate to limit the contact position of the temperature control chip and the biomolecule detection chip.

7. The chip-scale biomolecule detection system of claim 1, wherein: A structure for maintaining the thermal contact between the temperature control chip and the biomolecule detection chip is further provided.

8. The chip-scale biomolecule detection system of claim 1, wherein: The surface of the temperature control chip in contact with the biomolecule detection chip is provided with a layer of thermal conductive material.

9. The chip-scale biomolecule detection system of claim 1, wherein: The temperature control range of the temperature control chip is 20-110℃, the temperature control accuracy is 0.1℃, and the temperature control rate ranges from 0.1℃ / s to 30℃ / s.

10. The chip-scale biomolecule detection system of claim 1, wherein: The side of the optical waveguide chip facing the cavity is provided with a light transmission layer, and the material of the light transmission layer includes but is not limited to one of inorganic glass or organic glass.

11. The chip-scale biomolecule detection system of any of claims 1-10, wherein: The chip further comprises a light source component and an information analysis component, the light source component is optically connected with the optical waveguide chip to provide a working light beam, and the information analysis component is electrically connected with the image sensing chip to process information and obtain a detection result.

12. The chip-scale biomolecule detection system of claim 11, wherein: According to different fluorescent substances, the wavelength of the working light beam has multiple values, including but not limited to 470nm, 530nm, 580nm, and 630nm, which correspond to different fluorescent substances.

13. A method of detection, characterized by: The chip-level biomolecule detection system is used to perform the following operations: PCR detection to obtain a fluorescent signal; Collecting the fluorescence of the biomolecule excited and imaging, and extracting the fluorescent signal for detection and judgment; The PCR detection process includes a control target of a preset temperature, and the reaction temperature is controlled by a temperature control program.

14. The detection method of claim 13, wherein: Before the PCR detection, the biomolecule detection chip is inserted into the cavity, and a temperature control chip and a light source assembly are started. The light source assembly is optically connected with the optical waveguide chip and is used to provide a working light beam.

15. The method of claim 13, wherein: The PCR detection is semi-quantitative PCR or quantitative PCR.

16. The method of claim 13, wherein: The temperature control program is based on a PID algorithm. The control target of the preset temperature includes but is not limited to the number of temperature rising and falling cycles, the denaturation temperature value and duration of the PCR detection, the annealing temperature value and duration, and the extension temperature value and duration.

17. The method of claim 13, wherein: The extraction of the fluorescence signal for detection and judgment is performed by running a preset algorithm, including judging the category of the biomolecule and whether there is a virus or bacteria through the intensity change of the fluorescence. The algorithm includes an image processing algorithm based on threshold analysis and / or a clustering analysis algorithm based on machine learning.

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