Intelligent nucleic acid detection device and pathogenic bacteria dead or alive and free nucleic acid distinguishing method

Abstract

The application relates to an intelligent nucleic acid detection device and a pathogenic bacteria death and life and free nucleic acid distinguishing method. The device comprises a self-service swab sampling system and an intelligent sample processing system. Compared with a traditional swab sampling nucleic acid analysis method, the application can realize self-sampling and automatic nucleic acid amplification detection analysis, does not require help or assistance from others, and does not have site environment requirements, can realize standardized operation of swab sampling nucleic acid analysis of on-site, home, community, medical units at all levels, health and epidemic prevention departments and the like, and can avoid influence of human factors.

Description

Technical Field

[0001] This invention relates to an intelligent nucleic acid detection device and a method for distinguishing between live and dead nucleic acids of pathogens, and relates to the biomedical field. Background Technology

[0002] Nucleic acid testing is the gold standard for COVID-19 prevention and control, as well as for analyzing gynecological pathogen infections. Current nucleic acid analysis technologies using multiple instruments in series are time-consuming (results available within 24 hours), involve complex manual operations, are labor-intensive, require specialized molecular biology laboratories, and cannot distinguish between live and dead pathogens or those contaminated with nucleic acid. For example, in COVID-19 nucleic acid test-positive cases, only live bacteria present in the infected person are infectious; asymptomatic individuals with dead bacteria or residual nucleic acid contamination are not infectious. Current nucleic acid testing instruments cannot accurately differentiate between asymptomatic infected individuals, which is a current blind spot in COVID-19 nucleic acid testing for prevention and control.

[0003] In clinical practice, accurately distinguishing between live and dead pathogens and residual nucleic acids in infected subjects is equally crucial. This not only helps predict disease progression, assess drug treatment effectiveness, and guide precise medication use, but also prevents drug resistance due to overuse and effectively reduces testing and treatment costs. Therefore, the development of intelligent sample collection and nucleic acid analysis systems, as well as methods for differentiating between live and dead pathogens and free nucleic acids, is urgently needed and is equally important for precision medicine diagnosis and guiding correct drug treatment in clinical pathogen infections. Summary of the Invention

[0004] To address the aforementioned problems, the purpose of this invention is to provide an intelligent nucleic acid detection device and a method for distinguishing between live and dead pathogens and free nucleic acids, which can significantly reduce manual operation and effectively avoid cross-contamination and infection.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] In a first aspect, the present invention provides an intelligent sample processing system, the device comprising a lead screw drive motor, a pulley drive motor and / or a rotary motor, an injection tube structure module, a heating and oscillating clamp and a reagent storage device, wherein the injection tube structure module comprises a piston with a central opening;

[0007] The pulley-driven motor is used to drive the overall movement of the injection tube structure module;

[0008] The lead screw drive motor is used to drive the piston to move up and down. When the piston rises to a set position, it is connected to the rotary motor to rotate, thereby realizing the elution of biological samples.

[0009] The heated oscillating clamp is used to lyse biological samples in the injection tube structure module to separate nucleic acids.

[0010] The reagent storage device is used to store various reaction reagents and to recycle waste liquid.

[0011] Furthermore, the injection tube structure module includes an injection tube barrel, a piston holder, a filter membrane, a needle, and a protective sleeve;

[0012] The piston holder is provided at the top of the injection tube, the needle is provided at the bottom of the injection tube, the filter membrane is provided at the connection between the needle and the injection tube, and the protective sleeve is provided on the outside of the needle. The injection tube includes an upper injection tube and a lower injection tube connected by threads. The lower injection tube is pre-stored with swab-preserved elution lysis solution and surface-modified magnetic beads or glass beads or other nanomaterials or polymer materials.

[0013] Furthermore, the piston holder includes one or more sealing rings and a rotary holder. The sealing rings are fixed to the outside of the bottom end of the piston. The rotary holder is inserted from the lower end of the central opening of the piston and can be rotated and slid up and down in the piston. The rotary holder is driven to rotate at high speed by the rotary motor.

[0014] Furthermore, the heated oscillating clamp includes a centrally perforated clamping sleeve, an oscillating motor or piezoelectric ceramic, one or more heating devices and / or a magnetically controllable adsorber; the heating device is installed in the centrally perforated clamping sleeve, the oscillating motor or piezoelectric ceramic is connected to the centrally perforated clamping sleeve from the side via an eccentric wheel, and the magnetically controllable adsorber is installed on the upper part of the centrally perforated clamping sleeve near the central opening, thereby achieving the lysis of the biological sample in the injection tube and releasing nucleic acid proteins through oscillating lysis, high-temperature lysis and / or chemical lysis.

[0015] Furthermore, the intelligent sample processing system also includes a photolysis processor, which is used to perform photolysis processing on the biological samples in the injection tube structure module.

[0016] Furthermore, the photolysis processor includes a frame or sleeve and one or more blue LEDs, all of which are uniformly and symmetrically distributed around the frame or sleeve, wherein the photolysis reagent is pre-mixed in the lysis reagent of the injection tube.

[0017] Furthermore, the intelligent sample processing system also includes a sample amplification system, an optical detection system, a temperature control system, a motion control system, and / or an intelligent analysis and identity entry early warning direct reporting system;

[0018] The sample amplification system is configured to distribute the processed biological samples to each detection unit to achieve multi-channel parallel nucleic acid amplification;

[0019] The optical detection system is configured to measure the results of nucleic acid amplification reactions;

[0020] The temperature control system is configured to heat and control the temperature during the biological sample processing and detection process;

[0021] The motion control system is configured to perform motion control on the biological sample processing and detection process;

[0022] The intelligent analysis and identity entry early warning system is configured to control the biological sample processing and detection process, and to store, analyze, display and / or report early warnings on the biological sample detection results.

[0023] Secondly, the present invention also provides an intelligent nucleic acid detection device, including the aforementioned intelligent sample processing system and self-service swab sampling system;

[0024] The self-service swab sampling system includes a substrate;

[0025] A swab sampler is used to hold and / or release a sampling swab.

[0026] A compound gear set for revolution and rotation is disposed in the base body. One end of the compound gear set for revolution and rotation is connected to the screw drive motor via a lead screw, and the other end of the compound gear set for revolution and rotation is connected to the swab holder.

[0027] A controller, located within the base, controls the movement of the lead screw drive motor.

[0028] Furthermore, the end of the substrate is provided with a flaring tongue depressor, which is used to flare the tube / cavity wall of the sampling object. It adopts two separate plate-like structures, cylindrical tubes, or elliptical tube structures. The end of the substrate near the flaring tongue depressor is provided with a position and / or force feedback device. A protective sleeve is also provided on the outside of the substrate.

[0029] Furthermore, the composite gear set for revolution and rotation includes a small revolution gear, a large revolution gear, a small rotation gear, a large rotation gear ring, and a universal adapter. The small revolution gear is connected to the lead screw of the lead screw drive motor and moves with the lead screw drive motor, driving the large revolution gear to rotate when it meshes with it. The small rotation gear is fixed on the large revolution gear and revolves around it in a circle. The circumference of the revolution is concentric with the large rotation gear ring. The small rotation gear meshes with the large rotation gear ring, and the small rotation gear rotates simultaneously with the large revolution gear. One end of the universal adapter is connected to the small rotation gear, and the other end is connected to the swab holder.

[0030] Furthermore, the substrate is also provided with a switch button, which is used to control the clamping, movement and release of the sampling swab. Different switch controls send three position IO commands (clamping, forward and backward movement, and release) through the controller, so that the lead screw drive motor drives the planetary and rotational compound gear set to reach three different position states: top, middle and bottom.

[0031] Thirdly, the present invention provides a method for distinguishing between live and dead pathogens and free nucleic acids, comprising:

[0032] The collected biological samples were divided into three equal parts, including sample 1, sample 2 and sample 3. Sample 1 and sample 2 were added to the photolysis reagent.

[0033] The sample No. 1 will be irradiated with blue LED light for a set time using the intelligent sample processing system described in claim 6, so that the photolysis reagent can bind to the free nucleic acid and the nucleic acid in the dead bacteria in sample No. 1.

[0034] Samples No. 2 and No. 3 are processed using the intelligent sample processing system described in any one of claims 1-5;

[0035] Samples No. 1 and No. 2 were subjected to sample concentration, lysis, nucleic acid extraction, purification, mixing with amplification reagents, amplification reaction, and real-time amplification signal detection, respectively. Sample No. 3 was directly subjected to nucleic acid extraction, purification, mixing with amplification reagents, amplification reaction, and real-time amplification signal detection.

[0036] By analyzing the differences in real-time amplification signals generated from three numbered samples, it is possible to distinguish between live and dead nucleic acids of pathogens using variable-temperature or isothermal nucleic acid amplification methods.

[0037] Furthermore, by analyzing the differences in real-time amplification signals generated by three numbered samples, it was achieved to distinguish between live and dead nucleic acids of pathogens using temperature-controlled or isothermal nucleic acid amplification methods, including:

[0038] The real-time amplification signal generated by sample No. 1 corresponds to the nucleic acid content of live bacteria; the real-time amplification signal generated by sample No. 2 corresponds to the total nucleic acid content of live bacteria, dead bacteria, and free nucleic acid; and the real-time amplification signal generated by sample No. 3 corresponds to the content of free nucleic acid. The nucleic acid content corresponding to dead bacteria is obtained by subtracting the nucleic acid content corresponding to the real-time amplification signals generated by samples No. 1 and No. 3 from the total nucleic acid content corresponding to the real-time amplification signal generated by biological sample No. 2.

[0039] Because the present invention adopts the above technical solution, it has the following characteristics:

[0040] 1. Compared with traditional swab sampling nucleic acid analysis methods, the intelligent nucleic acid detection device of the present invention allows users to sample themselves and automatically perform nucleic acid amplification and detection analysis without the help or assistance of others, and there are no site environment requirements. It can be used in a standardized manner in the field, at home, in the community, in medical units at all levels, and in health and epidemic prevention departments, avoiding the influence of human factors.

[0041] 2. Compared with traditional swab elution methods, this invention is compatible with two methods: adding the sampling swab eluent or directly adding the sampling swab to the sealed elution separation sample. It efficiently transfers the eluent in situ, and the tube is not opened again after sampling. The whole process is sealed, which effectively avoids serious safety and reliability problems such as infection of subsequent operators and cross-contamination throughout the process.

[0042] 3. The intelligent sample processing system of the present invention is equipped with a rotary clamp, a heating and piston squeezing unit. By using a combination of high-speed rotational shearing mechanical crushing and / or oscillatory lysis and / or high-temperature lysis and / or chemical lysis with the rotary clamp, the sample adhering to the swab can be more fully eluted and separated, nucleic acid components can be efficiently lysed, and the recovery efficiency of the sample adhering to the swab can be improved.

[0043] 4. The intelligent sample processing system of the present invention adopts strategies such as photolysis of biological samples, filter membranes or filter paper and / or magnetic beads or anion exchange adsorption for nucleic acid concentration and purification. It can not only distinguish between dead and free nucleic acids of pathogens, but is also suitable for nucleic acid and protein detection and analysis of different biological samples such as tissue cells, exosomes, and exovesicles. It can also effectively reduce the amount of elution buffer used and increase the initial concentration of recovered adsorbed samples or lysed nucleic acids for subsequent use by nearly 100 times.

[0044] 5. Compared with conventional test tubes, well plates, and other carriers (which typically require 25 μL / index) for detection, the sample amplification unit of this invention adopts a microchannel and microcavity structure, which not only significantly reduces the consumption of sample reagents (<1 μL / index), but also allows for the simultaneous detection of dozens or even hundreds of specific gene protein analysis indicators in a single injection. This greatly reduces manual operation steps and labor intensity, while avoiding the problem of human operation errors, making swab sampling and detection safer, more robust, and more efficient.

[0045] 6. This invention is compatible with wired / wireless communication transmission devices and can be directly displayed on computers, mobile phones or other terminals. It can also automatically send abnormal detection results to cloud servers or national infectious disease health and epidemic prevention monitoring network platforms, etc., so as to assess or predict and warn of the incidence risk in the whole country or various regions, and meet the practical application needs of human health monitoring and direct reporting of epidemics in the whole country or various regions.

[0046] 7. The self-service swab sampling system of the present invention adopts a single motor to control the combined motion of the swab's revolution and rotation, realizing the automatic control and positioning of the swab head on the pharyngeal wall and its periodic movement along the pharyngeal wall. It is not only simple in structure and convenient and quick to operate, but also ensures that the swab can move along the oropharyngeal wall to closely rub and scrape to collect sufficient samples. The synchronous rotation of the swab attaches the sample to the circumferential surface of the swab head, ensuring the effectiveness, accuracy and reliability of the sampling.

[0047] 8. The method for distinguishing between inactivated and free nucleic acids of pathogens according to the present invention can distinguish between inactivated and free nucleic acids of pathogens through temperature-switching or temperature-switching nucleic acid amplification methods.

[0048] In summary, this invention addresses the need for advanced diagnostic technologies and equipment in convenient smart healthcare systems, enabling the advancement of public health and precision medicine from large hospitals and third-party testing centers to townships, communities, and homes. This effectively alleviates the pressure on large hospitals and achieves intelligent network coverage of healthcare and epidemic prevention in townships, communities, homes, large hospitals, third-party testing centers, and public health departments. It effectively improves the level of advanced science and technology in precision medicine and public health and epidemic prevention for the entire population. This invention can be widely applied to swab sampling and testing in field settings, homes, communities, township clinics, clinical medical units at all levels, public health departments, biomedical research, or other fields. Attached Figure Description

[0049] Various other advantages and benefits will become apparent to those skilled in the art upon reading the following detailed description of preferred embodiments. The accompanying drawings are for illustrative purposes only and are not intended to limit the invention. Throughout the drawings, the same reference numerals denote the same parts. In the drawings:

[0050] Figure 1 This is a schematic diagram of the structure of the intelligent nucleic acid detection device according to an embodiment of the present invention;

[0051] Figure 2 This is a schematic diagram of the self-service swab sampling system according to an embodiment of the present invention;

[0052] Figure 3 This is a schematic diagram of the structure of the composite gear set of revolution and rotation according to an embodiment of the present invention;

[0053] Figure 4 (a) and (b) are schematic diagrams of the slide bar structure according to an embodiment of the present invention;

[0054] Figure 5 (a) to (c) are schematic diagrams of the swab holder structure and usage status according to embodiments of the present invention.

[0055] Figure 6 This is a schematic diagram of the intelligent sample collection and nucleic acid analysis system according to an embodiment of the present invention;

[0056] Figure 7 This is a schematic diagram of the injection tube structure module according to an embodiment of the present invention;

[0057] Figure 8 (a) is a schematic diagram of a piston clamp structure according to one embodiment of the present invention;

[0058] Figure 8 (b) is a schematic diagram of another piston clamp structure according to an embodiment of the present invention;

[0059] Figure 9 This is a schematic diagram of the structure of the heating and oscillating clamp according to an embodiment of the present invention;

[0060] Figure 10 This is a schematic diagram of the optical resolution processor structure according to an embodiment of the present invention;

[0061] Figure 11 (a) is a schematic diagram of a reagent storage device according to one embodiment of the present invention;

[0062] Figure 11 (b) is a schematic diagram of another reagent storage device structure according to an embodiment of the present invention;

[0063] Figure 12 (a) is a schematic diagram of the base structure of a sample amplification system according to one embodiment of the present invention;

[0064] Figure 12 (b) is a schematic diagram of the cover plate structure of a sample amplification system according to one embodiment of the present invention;

[0065] Figure 12 (c) is a schematic diagram of the cover plate structure of another sample amplification system according to an embodiment of the present invention;

[0066] Figure 12 (d) is a schematic diagram of the base structure of another sample amplification system according to an embodiment of the present invention;

[0067] Figure 13 (a) and (b) are schematic diagrams of the base structure of the third sample amplification system according to an embodiment of the present invention. Detailed Implementation

[0068] It should be understood that the terminology used herein is for the purpose of describing particular exemplary embodiments only and is not intended to be limiting. Unless the context clearly indicates otherwise, the singular forms “a,” “an,” and “described” as used herein may also include the plural forms. The terms “comprising,” “including,” “containing,” and “having” are inclusive and therefore indicate the presence of the stated features, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, steps, operations, elements, components, and / or combinations thereof. The method steps, processes, and operations described herein are not construed as requiring them to be performed in a particular order described or illustrated unless the order of performance is explicitly indicated. It should also be understood that additional or alternative steps may be used.

[0069] Although terms such as first, second, third, etc., may be used in this document to describe multiple elements, components, regions, layers, and / or segments, these elements, components, regions, layers, and / or segments should not be limited by these terms. These terms may be used only to distinguish one element, component, region, layer, or segment from another. Unless the context clearly indicates otherwise, terms such as "first," "second," and other numerical terms used herein do not imply order or sequence. Therefore, the first element, component, region, layer, or segment discussed below may be referred to as the second element, component, region, layer, or segment without departing from the teachings of the exemplary embodiments.

[0070] For ease of description, spatial relative terms may be used in the text to describe the relationship of one element or feature relative to another element or feature as shown in the figure. These relative terms include, for example, "inside," "outside," "middle," "outer," "below," "above," etc. Such spatial relative terms are intended to include different orientations of the device in use or operation, other than those depicted in the figure.

[0071] Compared with traditional swab sampling nucleic acid analysis methods, the intelligent nucleic acid detection device and the method for distinguishing between inactive and free nucleic acids of pathogens provided by this invention allow users to sample themselves and automatically perform nucleic acid amplification and detection analysis without the need for assistance from others or any site requirements. It can achieve standardized operation and use in fields, homes, etc., avoiding the influence of human factors. Moreover, this invention can distinguish between inactive and free nucleic acids of pathogens and can achieve the distinction between inactive and free nucleic acids of pathogens through variable temperature or isothermal nucleic acid amplification methods.

[0072] Exemplary embodiments of the invention will now be described in more detail with reference to the accompanying drawings. While exemplary embodiments of the invention are shown in the drawings, it should be understood that the invention can be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided to enable a more thorough understanding of the invention and to fully convey the scope of the invention to those skilled in the art.

[0073] Example 1: As Figure 1 As shown, the intelligent nucleic acid testing device provided in this embodiment includes a self-service swab sampling system 100 and an intelligent sample processing system 200.

[0074] The self-service swab sampling system 100 can use a single motor to drive the swab to move forward or backward quickly, and after reaching the set position, it can simultaneously drive the swab to rotate and periodically revolve according to a certain trajectory to complete standardized automatic sampling.

[0075] In this embodiment, as Figure 2 As shown, the self-service swab sampling system 100 provided in this embodiment includes a base screw-driven motor 101, a revolution and rotation compound gear set 102, a swab holder 103, and a controller 109. The revolution and rotation compound gear set 102 is disposed within the base. One end of the revolution and rotation compound gear set 102 is connected to the screw-driven motor 101 via a screw, and the other end is connected to the swab holder 103. The revolution and rotation compound gear set 102 is controlled by the screw-driven motor 101, enabling the swab holder 103 to move back and forth and / or rotate in both revolution and rotation directions. A controller 109 is disposed at the bottom of the base for controlling the movement of the screw-driven motor 101.

[0076] In a preferred embodiment, a tongue depressor 104 is detachably connected to the end of the base corresponding to the swab holder 103. The base can be detachably connected to the tongue depressor 104 by means of snaps or threads, etc., which is not limited here. The tongue depressor 104 is for one-time replacement to avoid cross-contamination.

[0077] Furthermore, the tongue depressor 104 is used to flare the tube / cavity wall of the sampling object. It can be a two-piece plate structure with the swab holder 103 placed between the two plate structures. Alternatively, the tongue depressor 104 can be a simple cylindrical or elliptical cylinder structure with the swab holder 103 placed at the center of the cylindrical or elliptical cylinder structure. As long as it can flare the tube / cavity wall of the sampling object, it is not limited here.

[0078] Furthermore, the tube / cavity wall of the sampled object can be the wall of the human oral cavity, pharynx, nasal cavity, vagina, uterus, ear canal, human epidermis, or the tube / cavity wall and surface of the sampled object; biological samples collected by swabs include, but are not limited to, pathogens, tissue cells, exosomes, and exovesicles.

[0079] In a preferred embodiment, a position and / or force feedback device 105 is provided near the center of the base tip near the diaphragm tongue depressor 104. The position and / or force feedback device 105 is connected to a controller 109. The controller 109 controls the movement of the lead screw drive motor 101 by acquiring signals from the position and / or force feedback device 105. The position feedback device can be an infrared or ultrasonic rangefinder, used to test the pharyngeal depth and provide the maximum forward distance for the swab. The force feedback device can be a pressure sensor, used to monitor the pressure of the swab upon contact with the pharyngeal wall, ensuring that the swab stops moving forward after contact to avoid over-impact damage to the pharyngeal wall.

[0080] In a preferred embodiment, a power supply 106 and an indicator light 108 are also provided in the substrate. The power supply 106 is used to supply power to various electrical devices and can be flexibly powered by batteries, chargers or direct connection to mains power. The indicator light 108 is used to indicate whether the power supply 106 has power.

[0081] In a preferred embodiment, a cross-contamination protection sleeve 110 is also provided on the outside of the substrate to wrap and protect other components except for the sampling swab and the tongue depressor 104. The cross-contamination protection sleeve 110 is provided with a small window that can transmit infrared laser or ultrasound at the position and / or force feedback device 105. It can be replaced after sampling is completed, and is used only once to ensure that there is no cross-contamination.

[0082] In a preferred embodiment, such as Figure 3As shown, the compound gear set 102 for revolution and rotation includes a small revolution gear 1021, a large revolution gear 1022, a small rotation gear 1023, a large rotation gear ring 1024, and a universal joint 1025. The small revolution gear 1021 is fixedly connected to the top of the lead screw of the lead screw drive motor 101 and moves together with the lead screw drive motor 101. When the small revolution gear 1021 moves and meshes with the large revolution gear 1022, it drives the large revolution gear 1022 to rotate, and the rotation... The small gear 1023 is fixed to the large revolving gear 1022 by a bearing. It revolves in a circle with the large revolving gear 1022. The revolving circle is concentric with the rotating large gear ring 1024. The rotating small gear 1023 meshes with the rotating large gear ring 1024. The large revolving gear 1022 drives the rotating small gear 1023 to rotate simultaneously. One end of the universal conversion head 1025 is connected to the rotating small gear 1023, and the other end is connected to the swab holder 103. Furthermore, the rotating large gear ring 1024 can also adopt a gear structure, which is concentrically fixed on the central shaft of the revolution large gear 1022 and meshes with the rotating small gear 1023. The revolution of the revolution large gear 1022 simultaneously drives the rotation of the rotating small gear 1023. Even further, the revolution-rotation compound gear set 102 can also adopt a motor-driven slip ring structure. The motor connects to the rotating small gear 1023 to drive the swab holder 103 to rotate. The motor provides power and control signals through the slip ring structure, ensuring that the power supply wires and control signal wires do not become tangled due to the revolution and rotation. It should be noted that the entire revolution-rotation compound gear set 102 can be fixed by the connecting bracket 1026, the details of which will not be elaborated further.

[0083] In a preferred embodiment, such as Figure 4 As shown in (a), a slide bar structure 113 is also provided in the base to improve the smoothness of movement. The slide bar structure 113 includes a guide shaft 113-1 and a bushing 113-2. The bushing 113-2 is provided on the guide shaft 113-1 and is fixed on the connecting bracket 1026 of the compound gear set 102 of revolution and rotation. The lead screw drive motor 101 includes a lead screw motor 101-1, a lead screw 101-2, a nut 101-3, and a coupling 101-4. It is fixed on the connecting bracket 1026 of the compound gear set 102 of revolution and rotation by the nut 101-3. The motor 101-1 is connected to one end of the lead screw 101-2 through the coupling 101-4, and the small revolution gear 1021 is fixed to the other end of the lead screw 101-2. Figure 4 As shown in (b), when the compound gear set 102 of revolution and rotation moves forward to the position of the small gear 1021, the small gear 1021 meshes with the large gear 1022, causing the large gear 1022 to rotate. When the compound gear set 102 of revolution and rotation moves in the opposite direction and disengages from the position of the small gear 1021, the large gear 1022 stops rotating.

[0084] In a preferred embodiment, photoelectric switches are also provided at the front and rear ends of the substrate, including a front photoelectric switch 111-1 and a rear photoelectric switch 111-2, for feedback of the front and rear limit positions of the lead screw drive motor 101, as a second layer of protection to prevent overshoot of the sampling swab's front and rear movements and to provide a correction indication of the coordinates of the starting position of the movement.

[0085] Furthermore, two baffles 114 are also provided below the connecting bracket 1026 of the revolution and rotation compound gear set 102, such as... Figure 2 As shown, the baffle 114 is fixedly connected to the connecting bracket 1026 of the compound gear set 102 of revolution and rotation. As a whole, it can move back and forth along the lead screw structure. When the front end of the baffle 114 reaches the photoelectric switch 111-1, the lead screw moves to the top limit position. When the rear end of the baffle 114 reaches the photoelectric switch 111-2, the lead screw moves to the bottom limit position.

[0086] In a preferred embodiment, such as Figure 5 As shown, the swab holder 103 includes a telescopic sleeve 103-1, a holder shaft 103-2, a spring 103-3, and a sliding bushing 103-4. The holder shaft 103-2 is installed in the telescopic sleeve 103-1 and can slide back and forth. Figure 5 As shown in (a); one end of the gripper shaft 103-2 is connected to the universal adapter 1025, and the other end of the gripper shaft 103-2 can be used to attach and detach the swab. When the gripper shaft 103-2 moves forward to the top of the telescopic sleeve 103-1, the blocking sliding sleeve 103-4 compresses the spring 103-3, and the forked head of the gripper shaft 103-2 disengages from the fastening of the sliding sleeve 103-4 and is in a relaxed state, as shown in (a). Figure 5 As shown in (c), this facilitates the loading and unloading of the swab; and when the clamping shaft 103-2 moves in the opposite direction away from the top of the retractable sleeve 103-1, the spring 103-3 pushes the sliding bushing 103-4 forward to tighten the forked head of the clamping shaft 103-2, thus clamping the swab. Figure 5 As shown in (b), this facilitates the periodic revolution and rotation of the swab, meeting the requirements for standardized swab sampling.

[0087] In a preferred embodiment, a switch button 107 is provided on the side wall of the substrate, which controls the clamping, movement and release of the swab holder 103. There are three switch buttons, which are used to control the clamping, movement, and release of the sampling swab. Different switch controls can be sent to the controller 109 via three position IO commands: clamping, forward / backward movement, and release. This causes the lead screw drive motor 101 to drive the combined rotation and revolution gear set 102 to reach three different position states: top, middle, and bottom. The top position corresponds to the released state of the swab holder 103, where the swab holder 103 is fully exposed to the tongue depressor 104 for easy loading and unloading of the swab. The middle position corresponds to the clamping state of the swab holder 103, which allows the lead screw drive motor 101 to move the swab forward / backward. The bottom position corresponds to the clamping state of the swab holder 103, where the swab is hidden in the tongue depressor 104, making it easy for the subject to open their mouth and put the swab in. Alternatively, the functions of the three switch buttons can be combined into one switch button to create a cyclical control function, simplifying the structure of the switch button 107. This can be configured as needed.

[0088] It should be noted that each structure in this embodiment can be supported and fixed using a corresponding structure. The specific fixing method is not limited, as long as it can meet the corresponding function of the present invention. In order to facilitate use and reduce weight, the base of this embodiment can be implemented using a hollow cylinder. This is just an example and is not limited to it. Any corresponding structure can be used to implement it.

[0089] The present invention also provides a method for using the self-service swab sampling system 100, the process of which includes: first, controlling the swab holder 103 to be in the top position so that the swab holder 103 is in a relaxed state, and inserting the sampling swab 112 into the swab holder 103; then, controlling the swab holder 103 to be in the bottom position, clamping and retracting the sampling swab into the diaphragm tongue depressor 104 for easy placement into the mouth; next, quickly extending the sampling swab 112 forward, and after the forward extension reaches the pharyngeal cavity depth position detected by the infrared or ultrasonic rangefinder and / or the pressure sensor senses a large force of the swab head hitting the pharyngeal cavity wall, driving the sampling swab 112 to rotate and synchronously move along a certain tube / cavity. The cross-sectional trajectory of the wall revolves periodically, keeping the rotating swab periodically rubbing and scraping around the tube / cavity wall of the sampled object to achieve standardized sampling, including but not limited to the requirements of the COVID-19 specimen collection standards stipulated in the COVID-19 Prevention and Control Plan (9th Edition) and above, and / or other clinical medical and health epidemic prevention specimen collection standards. These can all be specifically set in the sampling control program. After sampling is completed, the sampling swab is controlled to retract into the flared tongue depressor 104 and taken out of the mouth; finally, the sampling swab 112 is quickly extended forward to the top, so that the swab holder 103 is released and the sampling swab is exposed, which facilitates the loading and unloading of the sampling swab 112.

[0090] In a preferred embodiment, the intelligent sample processing system 200 is used to encapsulate and conceal the sampling swab head, photolyze biological samples, lyse biological samples, concentrate and purify the lysed nucleic acids, and uniformly mix the concentrated and purified nucleic acids with amplification reaction reagents to form a reaction mixture.

[0091] In this embodiment, as Figure 6 As shown, the intelligent sample processing system 200 includes a lead screw drive motor 201, a pulley drive motor 202 and / or a rotary motor 203, an injection tube structure module 204, a heating and oscillating clamp 205 and / or a photolysis processor 206, and a reagent storage device 207.

[0092] The pulley-driven motor 202 is used to drive the injection tube structure module 204 to move upward as a whole. After reaching the position of the photolysis processor 206, the photolysis processor 206 performs photolysis processing on the biological sample in the injection tube structure module 204.

[0093] The lead screw drive motor 201 drives the piston in the injection tube structure module 204 (e.g., Figure 5 As shown, the piston moves up and down. After rising to a certain position, it connects to the rotary motor 203, which then drives the swab to rotate, thus eluting the biological sample. The biological sample can be separated from the eluted nucleic acid by the shearing force generated by the high-speed rotation, which can be used to abrade, break down, and lyse the sample.

[0094] The heated and oscillating clamp 205 is used to perform high-temperature and oscillating lysis and separation of nucleic acids from biological samples in the injection tube structure module 204;

[0095] The reagent storage container 207 is used to store various reaction reagents and to recover waste liquid.

[0096] In this embodiment, as Figure 7 As shown, the injection tube structure module 204 includes an injection tube 2041, a piston holder 2042, a filter membrane or filter paper 2043, a needle 2044, and a protective sleeve 2045 connected by upper and lower threads. The piston holder 2042 is provided at the top of the injection tube 2041, and the needle 2044 is provided at the bottom of the injection sleeve 2041. The filter membrane or filter paper 2043 is provided at the connection between the needle 2044 and the injection tube 2041, and the protective sleeve 2045 is provided on the outside of the needle 2044. The injection tube 2041 includes two parts, injection tube 2041-1 and 2041-2, connected by threads.

[0097] Furthermore, the injection tube 2041 is pre-stored with swab preservation elution and lysis buffer 2046 and surface-modified magnetic beads 2047, glass beads, or other nanomaterials or polymers. The surface-modified magnetic beads 2047, glass beads, or other nanomaterials include, but are not limited to, anion exchange-type broad-spectrum modifications that result in negative charges in acidic environments, and specific group modifications that target targets of interest based on nucleic acid hybridization or protein immune binding principles. These modifications adsorb negatively charged nucleic acids or other biological samples in acidic environments and release negatively charged nucleic acids or other biological samples in alkaline environments. Alternatively, the modifications may involve coating with poly(N-isopropylacrylamide) macromolecule (PNIPAAm), which adsorbs nucleic acids or other biological samples at low temperatures below 32°C and collapses to release nucleic acids or other biological samples at high temperatures of 60°C or above.

[0098] In this embodiment, as Figure 8 As shown, the piston holder 2042 includes a piston 2042-1 with a central opening, one or more sealing rings 2042-2, and a rotary holder 2042-3. The sealing rings 2042-2 are fixed to the outside of the bottom end of the piston 2042-1, and the rotary holder 2042-3 is inserted from the lower end of the central opening of the piston and can be rotated and slidably placed in the piston 2042-1. Wherein, as... Figure 8 As shown in (a), the rotary gripper 2042-3 can be a hollow cylinder at the bottom, suitable for gripping and concealing the sampling swab head; as Figure 8As shown in (b), the rotary clamp 2042-3 can also be a hollow cylinder divided into two or more symmetrical or asymmetrical blade-shaped, conical, or cylindrical structures. The rotary motor 203 drives the rotary clamp 2042-3 to rotate at high speed, generating mechanical shearing force to grind and crush the biological sample in the injection tube 2041 and release nucleic acid proteins.

[0099] In this embodiment, as Figure 9 As shown, the heated oscillating clamp 205 includes a centrally perforated clamping sleeve 2051, an oscillating motor or piezoelectric ceramic 2052, one or more heating rods or heating resistance wires or heating films 2053, and / or a magnetically controllable adsorbent 2054. The heating rods or heating resistance wires or heating films 2053 are installed in the centrally perforated clamping sleeve 2051. The oscillating motor or piezoelectric ceramic 2052 is connected to the centrally perforated clamping sleeve 2051 from the side via an eccentric wheel. The magnetically controllable adsorbent 2054 is installed on the upper part of the centrally perforated clamping sleeve 2051 near the central opening. It achieves the lysis of biological samples in the injection tube 341 and releases nucleic acid proteins through oscillating lysis and / or high-temperature lysis. The magnetically controllable adsorbent 2054 can be an electromagnet or a pair or more permanent magnets, and the magnetic field can be controlled by adjusting the relative positions.

[0100] In this embodiment, as Figure 10 As shown, the photolysis processor 206 includes a sleeve or casing 2061 and one or more blue LEDs 2062. The blue LEDs 2062 are uniformly and symmetrically distributed around the sleeve or casing 2061. The photolysis reagent is pre-mixed in the lysis reagent in the injection tube 2041. After irradiation with the blue LEDs 2062 for, for example, 30 minutes, the photolysis reagent binds to free nucleic acids or nucleic acids in dead bacteria, inhibiting these nucleic acids from undergoing subsequent nucleic acid amplification reactions. The photolysis reagent includes, but is not limited to, PMA. TM PMAxx TM EMA or other similar azide dyes, etc.

[0101] In this embodiment, as Figure 11 As shown, the reagent reservoir 207 includes three or more cavities 2071-2073 and sealing rubber caps or gaskets 2070, with a rubber cap or gasket 2070 on the top of each cavity. Cavity 2071 is used for waste liquid recovery. Cavities 2072 and 2073 are pre-filled with different reagents as needed, such as lysis reagents, buffer reagents, elution reagents, nucleic acid amplification reagents, etc., and can be arranged in a ring, such as... Figure 11 As shown in (a), a linear arrangement of stacked layers or staggered front and back layers can also be used, such as... Figure 11 As shown in (b), the lysis reagent, elution reagent, and nucleic acid amplification reagent can be liquid or lyophilized solid, and the nucleic acid amplification reagent can also be stored in the sample amplification system.

[0102] In this embodiment, the intelligent sample processing system 200 may further include a sample amplification system 300, an optical detection system 400, a temperature control system 500, a motion control system 600, and an intelligent analysis and identity entry early warning direct reporting system 700.

[0103] The self-service swab sampling system 100 is configured to complete standardized automatic sampling by moving the swab.

[0104] The intelligent sample processing system 200 is configured to elute, lyse, separate, purify, and concentrate nucleic acids from biological samples collected by swabs, and to uniformly mix the nucleic acids with amplification reaction reagents to form a reaction mixture; the biological samples include pathogens, tissue cells, exosomes, exovesicles, etc.

[0105] The sample amplification system 300 is configured to distribute the reaction mixture to each detection unit to achieve multi-channel parallel nucleic acid amplification.

[0106] The optical detection system 400 is configured to measure fluorescence, turbidity, phase contrast, color, etc., of the nucleic acid amplification reaction results of each detection unit. Alternatively, a silicon-based silica microchip can be fixed at the bottom of the detection cavity first, and then primer probes can be covalently linked and fixed to the surface of the silica layer on the microchip. The optical detection system can then perform super-resolution measurement of the thickness of the white light interference hyperspectral signal.

[0107] The temperature control system 500 is configured to heat and control the temperature during biological sample processing and detection.

[0108] The motion control system 600 is configured to perform motion control on biological sample processing and detection processes.

[0109] The Intelligent Analysis and Identity Entry Early Warning and Direct Reporting System 700 is configured to perform human-computer interactive control of the biological sample processing and testing process, and to store, analyze, display and / or report early warnings for the biological sample testing results.

[0110] In a preferred embodiment, the sample amplification system 300, together with the temperature control system 500 and the motion control system 600, achieves temperature control and motion control, enabling uniform distribution of multi-channel samples and variable-temperature or isothermal nucleic acid amplification reactions.

[0111] In this embodiment, as Figure 12 As shown, the sample amplification system 300 includes a base and a cover plate. The cover plate is set on the top of the base and is sealed by adhesive film, thermoforming, or photocuring. The base is designed with microchannels to connect linear, circular, fan-shaped, or matrix-shaped microcavity structures. Samples are introduced by positive / negative pressure or centrifugation. Different microcavities are isolated from each other by air, oil, paraffin, thermoforming, or mechanical pressure to prevent cross-contamination, thus realizing multi-channel parallel detection with one sample addition.

[0112] In this embodiment, the base includes three or more detection chambers C1 to C10, distribution chambers H1 to H10, a sample inlet I11 or I21, an exhaust port I12 or I22, group connection ports L11 and L12, a redundant sample collection port F1, and multiple connecting microchannels Ch1 to Ch2. Figure 12 (a) The detection cavities can be arranged in a linear array C1 to C10; such as Figure 12 As shown in (d), the detection chambers can also be arranged in a planar array structure C1 to C16. Nucleic acid amplification reagents are pre-ly lyophilized and stored in the dispensing chambers H1 to H10. At the bottom of each detection chamber C1 to C10, primers and probes for detecting different nucleic acid analytical indicators can be directly embedded and fixed using low-melting-point paraffin or low-melting-point agarose gel. These are released by heating through a temperature control system 500 to induce a nucleic acid amplification reaction, and the optical detection system 400 measures fluorescence, turbidity, phase contrast, and color. Alternatively, a silicon-based silica microchip can be first fixed to the bottom of the detection chambers C1 to C10, and then primers and probes can be covalently linked and fixed to the surface of the silica layer on the microchip. The optical detection system 400 then performs super-resolution measurement of the thickness of the white light interference hyperspectral signal.

[0113] The cover plate has a small number of connecting microchannels T1 to T3, such as Figure 12 (b) or Figure 12 As shown in (c), different samples can be added to the detection chambers C1 to C10 for group detection, as in one implementation using... Figure 12 (b) The cover plate, through connecting microchannels T1 to T2, respectively connects the group connection hole L11 to the vent hole I12 and the group connection hole L12 to the vent hole I22, meeting the requirement of adding 2 samples and analyzing 5 nucleic acid detection indicators for each sample; another implementation uses... Figure 12 (c) Cover plate, which connects the group connection hole L11 and the connection hole L12 through the micro-channel T3 to meet the needs of adding 1 sample and analyzing 10 nucleic acid detection indicators for each sample.

[0114] Furthermore, such as Figure 13As shown, the base of the sample amplification system 300 can also arrange the dispensing chambers FP1-FP19 and the detection chambers C1-C19 into a circular, annular, or fan-shaped structure, including the detection chambers C1-C19, dispensing chambers FP1-FP19, buffer chambers M1-M19, sample inlet In, exhaust outlet Out, mixing chamber MC, excess sample collection port FY, and connecting microchannels, etc. Nucleic acid amplification reagents can be pre-frozen and stored in the mixing chamber MC. A rotary motor is connected at the positioning center WZ of the base to drive the rotation and generate centrifugal force. Through the action of the siphon valve HX, the sample is uniformly transferred from the mixing chamber MC to the dispensing chambers FP1-FP19, and then the detection chambers C1-C19 are filled. Excess sample is stored in the buffer chambers M1-M19.

[0115] In a preferred embodiment, the optical detection system 400 is used to detect the sample amplification system 300 using a single imaging method with a CCD or other area array detector; or to detect the sample amplification system 300 by region using a scanning method; or to detect it using white light interference, phase contrast, turbidity or other label-free methods, etc.

[0116] In this embodiment, the optical detection system 400 adopts existing technology, including an objective lens, an illumination source, an excitation filter, an imaging lens, an emission filter, and a detector. It can detect fluorescence signals by switching the filter, or perform super-resolution measurement of white light interference hyperspectral signal thickness decoding by placing the filter in an empty position. It can also perform turbidity, phase contrast, color, or other spectrophotometric measurements using a mobile phone or human eye, which will not be elaborated here.

[0117] In a preferred embodiment, the temperature control system 500 can use existing technology to heat and control the intelligent sample processing system 200 and the sample amplification system 300, and the specific structure will not be described in detail. The motion control system 600 realizes motion control of the intelligent sample processing system 200, the sample amplification system 300 and the optical detection system 400 through motion control of each motor. The specific control is programmed and set as needed, and will not be described in detail here.

[0118] In a preferred embodiment, the intelligent analysis and identity entry early warning direct reporting system 700 includes a signal storage, processing and analysis unit, a wired or wireless communication interface, a display terminal and a cloud big data server.

[0119] The signal storage, processing, and analysis unit is equipped with a big data prior knowledge base for the detection of specific gene proteins for precision medical molecular diagnosis of various viruses, pathogens, tumors, and cancers. Based on the big data prior knowledge base for the detection of specific gene proteins, the unit performs artificial intelligence and / or big data analysis on the nucleic acid and protein molecular diagnostic detection results of swab samples to obtain specific gene protein detection and identification results of various biomedical indicators.

[0120] The display terminal is used to display the detection and identification results of specific gene proteins for various biomedical indicators.

[0121] Wired or wireless communication interfaces are used to transmit the results of specific gene protein detection and identification to the mobile phones of relevant personnel, or directly report them to the cloud big data server or the national health and epidemic prevention and health risk monitoring network platform for infectious diseases, major chronic diseases, malignant tumors and cancers. This enables smart medical networking between individuals, families, communities, township clinics and medical units and health and epidemic prevention departments at all levels, and provides prediction and early warning for highly prevalent infectious diseases, major chronic diseases, malignant tumors and cancers.

[0122] The intelligent sample processing system 200 provided in this embodiment, in use, works in conjunction with the temperature control system 500 and the motion control system 600 to achieve heating temperature control and motion control during operation. First, the upper half 2041-1 of the injection tube is unscrewed. Then, a swab or swab eluent is placed in the lower half 2041-2 of the injection tube after sampling, or the swab is manually used to elute the sample directly in the lower half 2041-2 of the injection tube. The two halves of the injection tube, 2041-1 and 2041-2, are then closed and tightened. Next, the injection tube structure module 204 is driven upwards by the pulley-driven motor 202 until it reaches the position of the photolysis processor 206. The photolysis processor 206 then performs photolysis on the biological sample in the injection tube 2041. The process involves inhibiting the subsequent nucleic acid amplification reaction of free nucleic acids or nucleic acids in dead bacteria. Then, a screw-driven motor 201 operates the piston 2042 in the injection tube module 204, causing it to move up and down. When the piston 2042 descends to a certain position, it clamps the swab. Then, when the piston rises to a certain position, a rotary motor 203 operates the rotary gripper 2042-3 at the center of the piston 2042 in the injection tube module 204, causing it to rotate and elute the biological sample from the swab. Finally, the piston 2042 descends near the bottom, encasing and concealing the swab within it, and emptying most of the swab from the injection tube 2041. The eluent is then transferred to the first chamber 2071 of the reagent storage tank 207. In the process, the biological sample eluted by the swab is blocked by the filter membrane 2043 and left in the injection tube 2041. When the injection tube 2041 reaches the position of the heated oscillating clamp 205, the heated oscillating clamp 205 lyses the biological sample in the injection tube 2041 to separate nucleic acids. Simultaneously, the surface-modified magnetic beads 2047 pre-installed in the injection tube 2041 adsorb the lysed and separated nucleic acids. Finally, the magnetic field generated by the magnetic field-controlled adsorber 2054 holds the magnetic beads. Simultaneously, with the combined movement of the lead screw drive motor 201 and the pulley drive motor 202, the liquid in the injection tube 2041 is discharged into the first cavity 2071 of the reagent storage 207, and from the second cavity 2072 of the reagent storage 207. The cleaning reagent is drawn in, purifying the biological sample in the injection tube 2041 and discharging the cleaning reagent into the first chamber 2071. Then, a small volume of elution and amplification reagent is drawn in from the third chamber 2073 of the reagent reservoir 207 to elute the nucleic acid from the magnetic beads 2047, thus concentrating the nucleic acid separated from the injection tube 2041 and mixing it uniformly with the amplification reaction reagent. If the injection tube is pre-filled with surface-modified glass beads or other nanomaterials, the filter membrane 2043 in the injection tube structure module 204 will block them, keeping the glass beads or other nanomaterials inside the injection tube 2041, thus achieving the elution, purification, and concentration of nucleic acid captured on the glass beads or other nanomaterials.

[0123] The usage process of the intelligent sample collection and nucleic acid analysis device in this embodiment includes:

[0124] First, the subject or another person inserts the swab into the self-service swab sampling system 100. Then, the self-service swab sampling system 100 is brought close to the area to be sampled. The self-service swab sampling system 100 completes the rotation of the swab and periodically revolves around the tube / cavity wall of the sampled object at the area to be sampled, thus achieving standardized sampling of the sampling site.

[0125] Then, the sampled swabs are placed in the intelligent sample processing system 200. Through the combined operation of the intelligent sample processing system 200, temperature control system 500, and motion control system 600, the biological samples attached to the swabs are automatically eluted, concentrated, lysed, and nucleic acid extracted, purified, and mixed with amplification reagents to form a reaction mixture. The reaction mixture is then added to the sample amplification system 300. The sample amplification system 300, temperature control system 500, and motion control system 600 then work together to automatically and evenly distribute the reaction mixture into the detection chambers for various nucleic acid analysis indicators. Subsequently, these detection chambers are automatically subjected to variable-temperature or isothermal nucleic acid amplification reactions.

[0126] Finally, the optical detection system 400 and the motion control system 600 work together to perform real-time signal detection of multiple nucleic acid analysis indicators on these detection chambers, and the detection results are stored, analyzed, processed, displayed, and reported as early warnings by the intelligent analysis and identity entry early warning system 700.

[0127] Example 2: The method for distinguishing between live and dead pathogens and free nucleic acids provided in this example includes the following steps:

[0128] S1. Divide the biological samples collected by swabs or other means into three parts: No. 1, No. 2, and No. 3. Add sample No. 1 and sample No. 2 to the photolysis reagent.

[0129] S2. Select sample No. 1 and irradiate it with blue LED light for ≤60 minutes to allow the photolysis reagent to bind with the free nucleic acid and the nucleic acid in the dead bacteria in sample No. 1, thereby inhibiting their nucleic acid amplification reaction; samples No. 2 and No. 3 are not treated with blue LED light, so the photolysis reagent will not bind with the free nucleic acid or the nucleic acid in the dead bacteria in sample No. 2.

[0130] S3. Samples No. 1 and No. 2 are subjected to sample concentration, lysis, nucleic acid extraction, purification, mixing with amplification reagents, amplification reaction, and real-time amplification signal detection, respectively. Sample No. 3 is subjected to nucleic acid extraction, purification, mixing with amplification reagents, amplification reaction, and real-time amplification signal detection directly. In this way, only the nucleic acid produced by live bacteria in sample No. 1 undergoes nucleic acid amplification reaction, all the nucleic acids produced by live and dead bacteria and free nucleic acids in sample No. 2 undergo nucleic acid amplification reaction, and sample No. 3, because it has not undergone lysis treatment, only free nucleic acid undergoes nucleic acid amplification reaction.

[0131] S4. By analyzing and comparing the differences in real-time amplification signals generated by these three samples, it is possible to distinguish between live and dead nucleic acids of pathogens using variable-temperature or isothermal nucleic acid amplification methods.

[0132] Specifically, the real-time amplification signal generated by sample No. 1 mainly corresponds to the nucleic acid content of live bacteria, the real-time amplification signal generated by sample No. 2 corresponds to the total nucleic acid content of live bacteria, nucleic acid released from dead bacteria lysis, and free nucleic acid, and the real-time amplification signal generated by sample No. 3 mainly corresponds to the content of free nucleic acid. By subtracting the nucleic acid content corresponding to the real-time amplification signals generated by samples No. 1 and No. 3 from the total nucleic acid content corresponding to the real-time amplification signal generated by sample No. 2, the nucleic acid content corresponding to the nucleic acid released from dead bacteria lysis can be obtained.

[0133] Furthermore, other commercial separation instruments and equipment can be used to perform manual step-by-step operations according to the above method, so as to distinguish between dead and free nucleic acids of pathogens through temperature-controlled or isothermal nucleic acid amplification methods.

[0134] In summary, the intelligent sample collection and nucleic acid analysis device provided in this embodiment allows users to operate a self-service swab sampling system for standardized swab sampling. The sampled swabs are then directly placed into the injection tube of the intelligent sample processing system, enabling simultaneous parallel detection of specific genes for two or more pathogenic bacteria (bacteria, fungi, viruses, microorganisms, model organisms, etc.) infection indicators or tumor and cancer analysis indicators. The sample reagent consumption for each detection indicator can be reduced from tens of microliters to less than 1 microliter as needed. Furthermore, the method for distinguishing between inactivated and cell-free nucleic acids of pathogens provided by this invention can achieve this distinction through temperature-switched or isothermal nucleic acid amplification methods.

[0135] The various embodiments in this specification are described in a progressive manner. Similar or identical parts between embodiments can be referred to interchangeably. Each embodiment focuses on describing the differences from other embodiments. In the description of this specification, references to terms such as "an embodiment," "furthermore," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the embodiments in this specification. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any suitable manner in one or more embodiments or examples. Furthermore, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0136] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An intelligent nucleic acid detection device, characterized in that, This includes intelligent sample processing systems and self-service swab sampling systems; The self-service swab sampling system includes a substrate; A swab sampler is used to hold or release a sampling swab. A compound gear set for revolution and rotation is disposed within the base. One end of the compound gear set is connected to a screw-driven motor via a lead screw, and the other end is connected to the swab sampler. The compound gear set includes a small revolution gear, a large revolution gear, a small rotation gear, a large rotation gear ring, and a universal joint. The small revolution gear is connected to the lead screw of the screw-driven motor and moves with the motor, meshing with the large revolution gear to rotate it. The small rotation gear is fixed to the large revolution gear and revolves in a circular path with the large revolution gear. The circumference of the revolution is concentric with the large rotation gear ring, and the small rotation gear meshes with the large rotation gear ring, driving the small rotation gear to rotate simultaneously with the revolution of the large revolution gear. One end of the universal joint is connected to the small rotation gear, and the other end is connected to the swab sampler. A controller, located within the base, controls the movement of the lead screw drive motor.

2. The intelligent nucleic acid detection device according to claim 1, characterized in that, Corresponding to the location of the swab sampler, a flaring tongue depressor is provided at the end of the base. The flaring tongue depressor is used to flare the tube / cavity wall of the sampled object. It adopts two separate plate-like structures, cylindrical tubes, or elliptical tube structures. A position feedback device and / or force feedback device are provided at the end of the base near the flaring tongue depressor. A protective sleeve is also provided on the outside of the base.

3. The intelligent nucleic acid detection device according to claim 1, characterized in that, The substrate is also equipped with a switch button, which is used to control the clamping, movement and release of the sampling swab. Different switch controls send three position IO commands (clamping, forward and backward movement, and release) through the controller, so that the lead screw drive motor drives the planetary and rotational compound gear set to reach three different position states: top, middle and bottom.

4. The intelligent nucleic acid detection device according to claim 1, characterized in that, The intelligent sample processing system includes a lead screw drive motor, a pulley drive motor and / or a rotary motor, an injection tube structure module, a heating and oscillating clamp, and a reagent storage device. The injection tube structure module includes a piston with a central opening. The pulley-driven motor is used to drive the overall movement of the injection tube structure module; The lead screw drive motor is used to drive the piston to move up and down. When the piston rises to a set position, it is connected to the rotary motor to rotate, thereby realizing the elution of biological samples. The heated oscillating clamp is used to lyse biological samples in the injection tube structure module to separate nucleic acids. The reagent storage device is used to store various reaction reagents and to recycle waste liquid.

5. The intelligent nucleic acid detection device according to claim 4, characterized in that, The injection tube structure module includes an injection tube barrel, a piston holder, a filter membrane, a needle, and a protective sleeve; The piston holder is provided at the top of the injection tube, the needle is provided at the bottom of the injection tube, the filter membrane is provided at the connection between the needle and the injection tube, and the protective sleeve is provided on the outside of the needle. The injection tube includes an upper injection tube and a lower injection tube connected by threads. The lower injection tube is pre-stored with elution lysis solution and surface-modified magnetic beads or glass beads.

6. The intelligent nucleic acid detection device according to claim 5, characterized in that, The piston holder includes one or more sealing rings and a rotary holder. The sealing rings are fixed to the outside of the bottom end of the piston. The rotary holder is inserted into the piston from the lower end of the central opening and can be rotated and slid up and down in the piston. The rotary holder is driven to rotate at high speed by the rotary motor.

7. The intelligent nucleic acid detection device according to claim 5, characterized in that, The heated oscillating clamp includes a centrally perforated clamping sleeve, an oscillating motor or piezoelectric ceramic, one or more heating devices and / or a magnetically controlled adsorbent; the heating device is installed in the centrally perforated clamping sleeve, the oscillating motor or piezoelectric ceramic is connected to the centrally perforated clamping sleeve from the side via an eccentric wheel, and the magnetically controlled adsorbent is installed on the upper part of the centrally perforated clamping sleeve near the central opening, thereby achieving the lysis of biological samples in the injection tube and releasing nucleic acid proteins through oscillating lysis, high-temperature lysis and / or chemical lysis.

8. The intelligent nucleic acid detection device according to claim 5, characterized in that, The intelligent sample processing system also includes a photolysis processor, which is used to perform photolysis processing on the biological samples in the injection tube structure module.

9. The intelligent nucleic acid detection device according to claim 8, characterized in that, The photolysis processor includes a frame or sleeve and one or more blue LEDs, all of which are uniformly and symmetrically distributed around the frame or sleeve. The photolysis reagent is pre-mixed in the lysis reagent of the injection tube.

10. The intelligent nucleic acid detection device according to claim 4, characterized in that, The intelligent sample processing system also includes a sample amplification system, an optical detection system, a temperature control system, a motion control system, and / or an intelligent analysis and identity entry early warning reporting system; The sample amplification system is configured to distribute the processed biological samples to each detection unit to achieve multi-channel parallel nucleic acid amplification; The optical detection system is configured to measure the results of nucleic acid amplification reactions; The temperature control system is configured to heat and control the temperature during the biological sample processing and detection process; The motion control system is configured to perform motion control on the biological sample processing and detection process; The intelligent analysis and identity entry early warning system is configured to control the biological sample processing and detection process, and to store, analyze, display, and / or report early warnings on the biological sample detection results.

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