Integrated microfluidic pathogen nucleic acid detection chip and detection device

Abstract

The utility model discloses an integrated microfluidic pathogen nucleic acid detection chip and a detection device, relates to the technical field of biological detection, and aims to solve the problems of design charge and high manufacturing cost of pathogen nucleic acid detection equipment in the prior art. The detection chip comprises a feeding layer plate, a liquid storage layer plate, a mixing layer plate, a detection layer plate and two fixing sleeves, and the detection device comprises the detection chip, a detection device shell, a pushing mechanism, an upper limiting mechanism, a heating assembly, a mixing vibration assembly and a back cover plate. The rapid, efficient and sensitive detection on multiple respiratory tract infection pathogens is realized. The system has the advantages of one-time sample adding, reagent sequential mixing, sequential temperature control and full-automatic operation, and is particularly suitable for pathogen detection in a resource limited environment. Through combination of the multi-layer totally-closed sliding type microfluidic chip and the colloidal gold paper chip, high-sensitivity and high-specificity pathogen nucleic acid detection is realized.

Description

Technical Field

[0001] The utility model relates to the technical field of biological detection, and particularly relates to an integrated microfluidic pathogen nucleic acid detection chip and a detection device. Background Art

[0002] Recently, new and sudden infectious diseases caused by respiratory diseases have occurred frequently worldwide. These diseases pose a major threat to human health and bring severe challenges to social public safety. Traditional nucleic acid detection relies on professional laboratories and technicians, faces the risk of environmental pollution, and is time-consuming due to cumbersome sample processing and transportation, resulting in delayed result issuance. Technical sensitivity requires strict temperature control and complex amplification steps, increasing the operation difficulty. At the same time, high costs hinder the wide application of this technology in large-scale screening and economically backward regions.

[0003] Colloidal gold test strip detection is commonly used for rapid screening of various pathogens and is suitable for the preliminary diagnosis of pathogens. Although it has obvious advantages in rapid screening, it may lack sensitivity and specificity in the detection of viral diseases. In order to improve the detection accuracy, the combined use of nucleic acid amplification technology and colloidal gold test strips not only enhances the accuracy but also maintains simplicity in operation and cost-effectiveness.

[0004] However, currently, the high-throughput automated respiratory pathogen nucleic acid detection technologies and devices with colloidal gold test strips on the market are often complex in design and relatively high in production cost, which to a certain extent limits the wide promotion and application of these technologies. Therefore, it is very necessary to develop safe and efficient automated nucleic acid detection technologies and devices. Summary of the Utility Model

[0005] The utility model aims to solve the problems of complex design and high production cost existing in the prior art for pathogen nucleic acid detection equipment, and further provides an integrated microfluidic pathogen nucleic acid detection chip and a detection device;

[0006] An integrated microfluidic pathogen nucleic acid detection chip, the detection chip includes a dosing layer plate, a liquid storage layer plate, a mixing layer plate, a detection layer plate and two fixing sleeves. The dosing layer plate, the liquid storage layer plate, the mixing layer plate and the detection layer plate are stacked in sequence from top to bottom to form a stacked structure. The two fixing sleeves are sleeved on the stacked structure and limit the stacked structure in the width direction. A lubricating oil layer is provided between adjacent two layer plates in the stacked structure and sliding connection is achieved through the lubricating oil;

[0007] Furthermore, a plurality of dosing holes are sequentially processed on the top of the dosing layer plate along the length extension direction of the dosing layer plate, and a plurality of liquid storage holes are sequentially processed on the top of the liquid storage layer plate along the length extension direction of the liquid storage layer plate, and each liquid storage hole is correspondingly arranged with a dosing hole;

[0008] Further, mixing holes and slots for the first heating sheet are machined at the top of the mixing laminate, detection holes and slots for the second heating sheet are machined at the top of the detection laminate, a test strip slot is machined at the bottom of the detection laminate along the length extension direction of the detection laminate, and the test strip slot is communicated with the detection holes;

[0009] Further, the mixing laminate and the detection laminate have the same length, the feeding laminate and the liquid storage laminate have the same length, and the length of the mixing laminate and the detection laminate is greater than the length of the feeding laminate and the liquid storage laminate;

[0010] An integrated microfluidic pathogen nucleic acid detection device, the detection device includes a detection chip, a detection device housing, a pushing mechanism, an upper limit mechanism, a heating assembly, a mixing and vibrating assembly, and a back cover plate. The pushing mechanism, the upper limit mechanism, the heating assembly, and the mixing and vibrating assembly are all integrated in the detection device housing. The pushing mechanism is installed at the upper part of the detection device housing, and the execution end of the pushing mechanism passes through the top of the detection device housing and extends outside the detection device housing. The mixing and vibrating assembly is installed at the inner bottom of the detection device housing, and the mixing and vibrating assembly is located in the extension direction of the pushing mechanism. The detection chip is installed on the top of the mixing and vibrating assembly, and the detection chip is correspondingly arranged with the execution end of the pushing mechanism. The upper limit mechanism is arranged between the pushing mechanism and the mixing and vibrating assembly. The upper limit mechanism is installed at the inner bottom of the detection device housing, and the limiting end of the upper limit mechanism is correspondingly arranged with the execution end of the pushing mechanism. The heating assembly is installed at the inner bottom of the detection device housing, and the heating end of the heating assembly is correspondingly arranged with the detection chip. The heating end of the heating assembly can be inserted and removed relative to the detection chip. The back cover plate is arranged on the back side of the detection device housing, and the back cover plate is detachably connected to the detection device housing;

[0011] Further, the pushing mechanism includes a pushing cylinder, a connecting plate, and a push rod. The pushing cylinder is installed on the inner wall of the detection device housing through a mounting bracket, and the extending end of the pushing cylinder faces the mixing and vibrating assembly. The connecting plate is an L-shaped plate. The connecting plate is arranged on the extending end of the pushing cylinder, and the vertical part of the connecting plate is detachably connected to the extending end of the pushing cylinder. The push rod is arranged on the horizontal part of the connecting plate, and the bottom end of the push rod is fixedly connected to the connecting plate. The top end of the push rod passes through the top of the detection device housing and extends outside the detection device housing:

[0012] The push rod includes a push rod body, a limit rod section, a liquid storage layer push rod section, and a mixing layer push rod section. The push rod body is arranged vertically on the horizontal part of the connecting plate, and the bottom end of the push rod body is fixedly connected to the connecting plate. The limit rod section, the liquid storage layer push rod section, and the mixing layer push rod section are arranged in sequence from top to bottom on one side of the push rod body facing the mixing vibration assembly, and the limit rod section, the liquid storage layer push rod section, and the mixing layer push rod section are all fixedly connected to the push rod body. There is an arrangement gap between the limit rod section and the liquid storage layer push rod section. The limit rod section is correspondingly arranged with the limit end of the limit mechanism. The liquid storage layer push rod section is correspondingly arranged with the liquid storage layer plate in the detection chip. The mixing layer push rod section is correspondingly arranged with the mixing layer plate in the detection chip;

[0013] Further, the upper limit mechanism includes a longitudinal push rod and a limit frame. The longitudinal push rod is arranged between the pushing mechanism and the mixing vibration assembly. The longitudinal push rod is vertically erected on the inner bottom of the detection device housing, and the housing of the longitudinal push rod is detachably connected to the inner bottom of the detection device housing by bolts. The limit frame is erected at the end of the piston rod in the longitudinal push rod, and the bottom of the limit frame is detachably connected to the piston rod of the longitudinal push rod by bolts. The top of the limit frame is correspondingly arranged with the limit rod section;

[0014] Further, the heating assembly includes a pneumatic source, a pneumatic slide rail pair, an air inlet pipe, an air outlet pipe, and a heating sheet moving plate. The pneumatic source is installed on the inner bottom of the detection device housing. The pneumatic slide rail pair is arranged between the upper limit mechanism and the mixing vibration assembly, and the length extension direction of the pneumatic slide rail pair is the same as the width extension direction of the detection device housing. One end of the pneumatic guide rail in the pneumatic slide rail pair is detachably connected to the detection device housing by bolts. The air inlet pipe and the air outlet pipe are arranged between the pneumatic source and the pneumatic slide rail pair. One end of the air inlet pipe is communicated with the air outlet end of the pneumatic source, and the other end of the air inlet pipe is communicated with the air inlet end of the pneumatic guide rail in the pneumatic slide rail pair. One end of the air outlet pipe is communicated with the air suction end of the pneumatic source, and the other end of the air outlet pipe is communicated with the air outlet end of the pneumatic guide rail in the pneumatic slide rail pair. The heating sheet moving plate is detachably connected to the sliding block in the pneumatic slide rail pair by bolts. The pneumatic slide rail pair drives the heating sheet moving plate to reciprocate along the width extension direction of the detection device housing;

[0015] On one side of the heating sheet moving plate facing the inner side wall of the detection device housing, a first heating sheet, a second heating sheet, and a third heating sheet are fixedly installed from top to bottom. The first heating sheet is correspondingly arranged with the first heating sheet slot, the second heating sheet is correspondingly arranged with the second heating sheet slot, and the third heating sheet is correspondingly arranged with the third heating sheet slot in the mixing vibration assembly;

[0016] Further, the hybrid vibration assembly includes a vibration cylinder, a chip holder, an extended support plate, a pressing cylinder, an L-shaped pressing rod, a lower limit cylinder, and a lower limit block. The vibration cylinder is vertically installed at the inner bottom of the detection device housing, and the housing of the vibration cylinder is detachably connected to the detection device housing. The chip holder is arranged at the end of the piston rod of the vibration cylinder, and the chip holder is detachably connected to the piston rod of the vibration cylinder by bolts. The extended support plate is arranged at the lower part of the chip holder on the side facing the back plate, and the extended support plate is integrally formed with the chip holder. The pressing cylinder is vertically installed on the extended support plate, and the cylinder body of the pressing cylinder is detachably connected to the extended support plate by bolts. The L-shaped pressing rod is arranged at the end of the piston rod in the pressing cylinder, and the vertical part of the L-shaped pressing rod is fixedly connected to the piston rod in the pressing cylinder. The horizontal part of the L-shaped pressing rod does not extend to the top of the chip holder. The detection chip is placed in the chip holder and fixed by the L-shaped pressing rod. A groove is machined on one side of the chip holder away from the upper limit mechanism. The lower limit cylinder is vertically installed in the groove, and the cylinder body of the lower limit cylinder is fixed on the side wall of the groove by a locking clip. The lower limit block is fixed at the end of the piston rod in the lower limit cylinder;

[0017] Further, a plurality of insertion rods are provided on the side of the back cover plate facing the detection device housing. One end of each insertion rod is fixedly connected to the back cover plate. A plurality of insertion sleeves are provided on the inner side wall of the detection device housing. One end of each insertion sleeve is fixedly connected to the detection device housing. The back cover plate is detachably connected to the detection device housing through the plurality of insertion rods and the plurality of insertion sleeves;

[0018] Advantages of the present application over the prior art:

[0019] An integrated microfluidic pathogen nucleic acid detection chip and a detection device provided by the present application can realize the functions of one-time sample addition, reagent order mixing, sequential temperature control, and fully automated operation, simplify the operation process, reduce human intervention, and improve work efficiency.

[0020] An integrated microfluidic pathogen nucleic acid detection chip provided by the present application improves the sensitivity and specificity of detection and better avoids the problem of nucleic acid aerosol contamination by integrating a multi-layer passive drive fully enclosed sliding microfluidic chip and a colloidal gold paper chip.

[0021] An integrated microfluidic pathogen nucleic acid detection device provided by the present application improves the convenience of nucleic acid detection operation, the precise transfer of reagents, the uniformity of reactions, and the accuracy of detection by introducing a pushing mechanism, a limiting mechanism, a heating component, and a hybrid vibration component during design, directly improving the detection efficiency and accuracy.

[0022] An integrated microfluidic pathogen nucleic acid detection chip and a detection device provided by the present application have significant advantages in terms of cost-effectiveness, reduce the detection cost, and improve the popularity of detection. Description of the Drawings

[0023] Figure 1 Isometric schematic view of the detection chip described in this application;

[0024] Figure 2 Structural schematic view of the detection chip described in this application;

[0025] Figure 3 Bottom view schematic of the detection layer board in the detection chip described in this application;

[0026] Figure 4 Front view of the detection device described in this application;

[0027] Figure 5 Internal schematic view of the detection device described in this application;

[0028] Figure 6 Internal schematic view of the detection device described in this application;

[0029] Figure 7 Internal schematic view of the detection device described in this application;

[0030] Figure 8 Top view schematic of the detection device described in this application;

[0031] Figure 9 Structural schematic view of the pushing mechanism in the detection device described in this application;

[0032] Figure 10 Structural schematic view of the push rod in the detection device described in this application;

[0033] Figure 11 Front view schematic of the upper limit mechanism in the detection device described in this application;

[0034] Figure 12 Side view schematic of the upper limit mechanism in the detection device described in this application;

[0035] Figure 13 Structural schematic view of the heating component in the detection device described in this application;

[0036] Figure 14 Structural schematic view of the heating sheet moving plate in the detection device described in this application;

[0037] Figure 15 Structural schematic view of the vibration component in the detection device described in this application;

[0038] Figure 16 Structural schematic view of the L-shaped pressure rod in the detection device described in this application;

[0039] In the figure, there are detection chip 1, dispensing layer board 11, dispensing holes 111, liquid storage layer board 12, liquid storage holes 121, mixing layer board 13, mixing holes 131, first heating sheet slots 132, detection layer board 14, detection holes 141 machined at the top, second heating sheet slots 142, test strip slots 143, fixing sleeves 15, detection device housing 2, pushing mechanism 3, pushing cylinder 31, connecting plate 32, push rod 33, push rod main body 331, limiting rod section 332, liquid storage layer push rod section 333, mixing layer push rod section 334, upper limit mechanism 4, longitudinal push rod 41, limit frame 42, heating component 5, pneumatic source 51, pneumatic slide rail pair 52, intake pipe 53, exhaust pipe 54, heating sheet moving plate 55, first heating sheet 551, second heating sheet 552, third heating sheet 553, mixing and vibrating component 6, vibrating cylinder 61, chip seat 62, extending support plate 63, pressing cylinder 64, L-shaped pressing rod 65, lower limit cylinder 66, lower limit block 67 and back cover plate 7. Detailed implementation manners

[0040] Detailed implementation manner one: Combining Figures 1 to 16 To illustrate this implementation manner, in this implementation manner, an integrated microfluidic pathogen nucleic acid detection chip is provided. The detection chip 1 includes a dispensing layer board 11, a liquid storage layer board 12, a mixing layer board 13, a detection layer board 14 and two fixing sleeves 15. The dispensing layer board 11, the liquid storage layer board 12, the mixing layer board 13 and the detection layer board 14 are stacked in sequence from top to bottom to form a stacked structure. The two fixing sleeves 15 are sleeved on the stacked structure to limit the stacked structure in the width direction. A lubricating oil layer is provided between adjacent two layer boards in the stacked structure, and sliding connection is achieved through the lubricating oil.

[0041] Detailed implementation manner two: Combining Figures 1 to 16 To illustrate this implementation manner, the difference between this implementation manner and the first detailed implementation manner is that a plurality of dispensing holes 111 are sequentially machined at the top of the dispensing layer board 11 along the length extension direction of the dispensing layer board 11, and a plurality of liquid storage holes 121 are sequentially machined at the top of the liquid storage layer board 12 along the length extension direction of the liquid storage layer board 12, and each liquid storage hole 121 is correspondingly arranged with a dispensing hole 111. Other compositions and connection manners are the same as those in the first detailed implementation manner.

[0042] Detailed implementation manner three: Combining Figures 1 to 16 To illustrate this implementation manner, the difference between this implementation manner and the second detailed implementation manner is that a mixing hole 131 and a first heating sheet slot 132 are machined at the top of the mixing layer board 13, a detection hole 141 and a second heating sheet slot 142 are machined at the top of the detection layer board 14, and a test strip slot 143 is machined at the bottom of the detection layer board 14 along the length extension direction of the detection layer board 14, and the test strip slot 143 is communicated with the detection hole 141. Other compositions and connection manners are the same as those in the second detailed implementation manner.

[0043] Embodiment 4: In combination with Figures 1 to 16 This embodiment is described. The difference between this embodiment and Embodiment 3 is that the lengths of the mixing layer plate 13 and the detection layer plate 14 are the same, the lengths of the feeding layer plate 11 and the liquid storage layer plate 12 are the same, and the lengths of the mixing layer plate 13 and the detection layer plate 14 are greater than the lengths of the feeding layer plate 11 and the liquid storage layer plate 12. Other components and connection methods are the same as those in Embodiment 3.

[0044] Combined with Embodiments 1 to 4, the feeding layer plate 11, the liquid storage layer plate 12, the mixing layer plate 13, and the detection layer plate 14 in this application are all made of PMMA material, and are designed to implement key steps such as step-by-step injection, mixing, and heating of samples. The chip consists of four layers. The top layer is the feeding layer plate 11, which is configured with three feeding holes 111 for adding samples and different reagents respectively to provide the necessary starting materials for detection. The second layer is the liquid storage layer plate 12, which is configured with three liquid storage holes 121 for storing corresponding detection reagents to ensure the stable supply of reagents. The third layer is the mixing layer plate 13, which integrates a mixing hole 131 and a first heating sheet slot 132. The mixing pool is responsible for effectively mixing different reagents and samples, and the heating area provides a constant temperature environment to ensure the smooth progress of the nucleic acid amplification reaction. The fourth layer is the detection layer plate 14, which is provided with a test strip slot 143 and a detection hole 141. The purpose of the detection hole 141 is to introduce the mixed sample into the test strip slot 143 and react with the colloidal gold test strip for detection. The detection layer plate 14 is also provided with a second heating sheet slot 142, the purpose of which is to ensure a constant temperature environment for the reaction. The surface of each layer of the layer plate is treated with oil seal to ensure the lubrication and sealing of the chip layer plate. Alignment holes are designed on the side of each layer of the layer plate to facilitate the alignment and assembly of each layer of the chip.

[0045] Embodiment 5: In combination with Figures 1 to 16To describe this embodiment, this embodiment provides a detection device with an integrated microfluidic pathogen nucleic acid detection chip structure. The detection device includes a detection chip 1, a detection device housing 2, a pushing mechanism 3, an upper limit mechanism 4, a heating component 5, a mixing and vibrating component 6, and a back cover plate 7. The pushing mechanism 3, the upper limit mechanism 4, the heating component 5, and the mixing and vibrating component 6 are all integrated in the detection device housing 2. The pushing mechanism 3 is installed in the upper part of the detection device housing 2, and the execution end of the pushing mechanism 3 passes through the top of the detection device housing 2 and extends outside the detection device housing 2. The mixing and vibrating component 6 is installed at the inner bottom of the detection device housing 2, and the mixing and vibrating component 6 is located in the extending direction of the pushing mechanism 3. The detection chip 1 is installed on the top of the mixing and vibrating component 6, and the detection chip 1 is correspondingly arranged with the execution end of the pushing mechanism 3. The upper limit mechanism 4 is arranged between the pushing mechanism 3 and the mixing and vibrating component 6. The upper limit mechanism 4 is installed at the inner bottom of the detection device housing 2, and the limiting end of the upper limit mechanism 4 is correspondingly arranged with the execution end of the pushing mechanism 3. The heating component 5 is installed at the inner bottom of the detection device housing 2, and the heating end of the heating component 5 is correspondingly arranged with the detection chip 1. The heating end of the heating component 5 can be inserted and removed relative to the detection chip 1. The back cover plate 7 is arranged on the back side of the detection device housing 2, and the back cover plate 7 is detachably connected to the detection device housing 2. Other compositions and connection methods are the same as those in the fourth specific embodiment.

[0046] In this embodiment, a detection device is customized for the microfluidic chip, which integrates a motion control module (pushing mechanism 3) and a temperature control module (heating component 5) to achieve precise control of the detection process. The motion control module is responsible for precisely moving and positioning the microfluidic chip, while the temperature control module ensures that the temperature inside the chip remains constant throughout the detection process, providing ideal conditions for nucleic acid amplification.

[0047] Specific embodiment six: Combining Figures 1 to 16 To describe this embodiment, the difference between this embodiment and the fifth specific embodiment is that the pushing mechanism 3 includes a pushing cylinder 31, a connecting plate 32, and a push rod 33. The pushing cylinder 31 is installed on the inner wall of the detection device housing 2 through a mounting bracket, and the extending end of the pushing cylinder 31 faces the mixing and vibrating component 6. The connecting plate 32 is an L-shaped plate. The connecting plate 32 is arranged on the extending end of the pushing cylinder 31, and the vertical part of the connecting plate 32 is detachably connected to the extending end of the pushing cylinder 31. The push rod 33 is arranged on the horizontal part of the connecting plate 32, and the bottom end of the push rod 33 is fixedly connected to the connecting plate 32. The top end of the push rod 33 passes through the top of the detection device housing 2 and extends outside the detection device housing 2;

[0048] The push rod 33 includes a push rod body 331, a limit rod section 332, a liquid storage layer push rod section 333, and a mixing layer push rod section 334. The push rod body 331 is arranged vertically on the horizontal part of the connecting plate 32, and the bottom end of the push rod body 331 is fixedly connected to the connecting plate 32. The limit rod section 332, the liquid storage layer push rod section 333, and the mixing layer push rod section 334 are sequentially arranged from top to bottom on the side of the push rod body 331 facing the mixing vibration assembly 6, and the limit rod section 332, the liquid storage layer push rod section 333, and the mixing layer push rod section 334 are all fixedly connected to the push rod body 331. There is an arrangement gap between the limit rod section 332 and the liquid storage layer push rod section 333. The limit rod section 332 is correspondingly arranged with the limit end of the limit mechanism 4. The liquid storage layer push rod section 333 is correspondingly arranged with the liquid storage layer plate 12 in the detection chip 1. The mixing layer push rod section 334 is correspondingly arranged with the mixing layer plate 13 in the detection chip 1. Other compositions and connection methods are the same as those in the fifth specific embodiment.

[0049] In this embodiment, the purpose of the pushing mechanism 3 is a structure for controlling the relative movement between the layer plates in the detection chip 1. When the push rod 33 performs an action, it contacts the layer plates in the chip and pushes the corresponding layer plates to make the layer plates reach the accurate positions of each process, facilitating the sequential movement of the liquid, ensuring the precise movement and distribution of the sample and reagent in the microfluidic chip. The limit rod section 332 is used to contact the upper limit mechanism 4, and the upper limit mechanism 4 is used to define the accurate working position of the push rod 33. The liquid storage layer push rod section 333 and the mixing layer push rod section 334 are correspondingly arranged with the liquid storage layer plate 13 and the mixing layer plate 14 respectively, and respectively push the liquid storage layer plate 13 and the mixing layer plate 14 to move. It should be noted that the liquid storage layer push rod section 333 and the mixing layer push rod section 334 move synchronously with the push rod 33. According to the detection process, in order to prevent the pushing actions of the two layer plates from interfering with each other, the length of the liquid storage layer push rod section 333 is longer than that of the mixing layer push rod section 334.

[0050] Specific embodiment seven: Combined with Figures 1 to 16 This embodiment is described. The difference between this embodiment and the sixth specific embodiment is that the upper limit mechanism 4 includes a longitudinal push rod 41 and a limit frame 42. The longitudinal push rod 41 is arranged between the pushing mechanism 3 and the mixing vibration assembly 6. The longitudinal push rod 41 is vertically erected on the inner bottom of the detection device housing 2, and the housing of the longitudinal push rod 41 is detachably connected to the inner bottom of the detection device housing 2 by bolts. The limit frame 42 is erected at the end of the piston rod in the longitudinal push rod 41, and the bottom of the limit frame 42 is detachably connected to the piston rod of the longitudinal push rod 41 by bolts. The top of the limit frame 42 is correspondingly arranged with the limit rod section 332. Other compositions and connection methods are the same as those in the sixth specific embodiment.

[0051] In this practical implementation manner, the upper limit mechanism 4 is used to limit the push rod 33, which can control the movement range of the microfluidic chip pushing structural member and accurately control the displacement of the chip. The limit frame 42 is the contact component with the limit rod section 332. The longitudinal push rod 41 is used to drive the limit frame 42 to move longitudinally. When there is no need to limit the push rod 33, the limit frame 42 will move downward to between the limit rod section 332 and the liquid storage layer push rod section 333 to release the interference with the push rod 33 and expand the movement range of the push rod 33.

[0052] Specific implementation manner eight: In combination with Figures 1 to 16 This implementation manner is described. The difference between this implementation manner and the seventh specific implementation manner is that the heating component 5 includes a pneumatic source 51, a pneumatic slide rail pair 52, an air inlet pipe 53, an air outlet pipe 54, and a heating sheet moving plate 55. The pneumatic source 51 is installed at the inner bottom of the detection device housing 2. The pneumatic slide rail pair 52 is arranged between the upper limit mechanism 4 and the mixing and vibration component 6, and the length extension direction of the pneumatic slide rail pair 52 is the same as the width extension direction of the detection device housing 2. One end of the pneumatic guide rail in the pneumatic slide rail pair 52 is detachably connected to the detection device housing 2 by bolts. The air inlet pipe 53 and the air outlet pipe 54 are arranged between the pneumatic source 51 and the pneumatic slide rail pair 52. One end of the air inlet pipe 53 is communicated with the air outlet end of the pneumatic source 51, and the other end of the air inlet pipe 53 is communicated with the air inlet end of the pneumatic guide rail in the pneumatic slide rail pair 52. One end of the air outlet pipe 54 is communicated with the air suction end of the pneumatic source 51, and the other end of the air outlet pipe 54 is communicated with the air outlet end of the pneumatic guide rail in the pneumatic slide rail pair 52. The heating sheet moving plate 55 is detachably connected to the sliding block in the pneumatic slide rail pair 52 by bolts. The pneumatic slide rail pair 52 drives the heating sheet moving plate 55 to perform reciprocating motion along the width extension direction of the detection device housing 2

[0053] On the side of the heating sheet moving plate 55 facing the inner side wall of the detection device housing 2, a first heating sheet 551, a second heating sheet 552, and a third heating sheet 553 are fixedly arranged from top to bottom. The first heating sheet 551 is correspondingly arranged with the first heating sheet slot 132, the second heating sheet 552 is correspondingly arranged with the second heating sheet slot 142, and the third heating sheet 553 is correspondingly arranged with the third heating sheet slot in the mixing and vibration component 6. Other compositions and connection methods are the same as those in the seventh specific implementation manner.

[0054] In this embodiment, the heating component 5 is used to provide the required constant temperature environment during the nucleic acid detection process. It includes three heating elements, namely the first heating element 551, the second heating element 552, and the third heating element 553. They are important components of the temperature control module and can adjust the temperature within the range of 20 - 130 degrees, providing the required constant temperature environment for nucleic acid amplification. It has high control precision and can control the temperature error within a range not greater than 0.1 degree after stabilization, ensuring the efficiency and accuracy of the nucleic acid amplification reaction. At the same time, to ensure the normal working state of the detection chip, the heating element moving plate 55 in the heating component 5 can selectively enter the specified position as needed to ensure the flexibility of the work.

[0055] Specific Embodiment Nine: In combination with Figures 1 to 16 This embodiment is described. The difference between this embodiment and Specific Embodiment Eight is that the hybrid vibration component 6 includes a vibration cylinder 61, a chip holder 62, an extended support plate 63, a pressing cylinder 64, an L-shaped pressing rod 65, a lower limit cylinder 66, and a lower limit block 67. The vibration cylinder 61 is vertically installed on the inner bottom of the detection device housing 2, and the housing of the vibration cylinder 61 is detachably connected to the detection device housing 2. The chip holder 62 is arranged at the end of the piston rod of the vibration cylinder 61, and the chip holder 62 is detachably connected to the piston rod of the vibration cylinder 61 by bolts. The extended support plate 63 is arranged at the lower part of the chip holder 62 on the side facing the back plate 7, and the extended support plate 63 is integrally formed with the chip holder 62. The pressing cylinder 64 is vertically installed on the extended support plate 63, and the cylinder body of the pressing cylinder 64 is detachably connected to the extended support plate 63 by bolts. The L-shaped pressing rod 65 is arranged at the end of the piston rod in the pressing cylinder 64, and the vertical part of the L-shaped pressing rod 65 is fixedly connected to the piston rod in the pressing cylinder 64. The horizontal part of the L-shaped pressing rod 65 extends to the top of the chip holder 62. The detection chip 1 is placed in the chip holder 62 and fixed by the L-shaped pressing rod 65. A groove is machined on one side of the chip holder 62 away from the upper limit mechanism 4. The lower limit cylinder 66 is vertically installed in the groove, and the cylinder body of the lower limit cylinder 66 is fixed on the side wall of the groove by a locking clip. The lower limit block 67 is fixed at the end of the piston rod in the lower limit cylinder 66. The other components and connection methods are the same as those in Specific Embodiment Eight.

[0056] In this embodiment, the hybrid vibration assembly 6 is a component for mixing samples and also a component for supporting the detection chip 1. The mixing principle of this application is based on gravity-assisted reagent mixing technology and vibration mixing mechanism. By driving the chip to vibrate through an external device, uniform mixing of the sample and the reagent is achieved. It can adjust the vibration frequency and amplitude precisely according to the sample characteristics and mixing requirements. The vibration cylinder 61 is used for precise adjustment of the vibration frequency and amplitude, the chip seat 62 is used to carry the chip, and the chip fixing structure composed of the pressing cylinder 64 and the L-shaped pressing rod 65 can fix the chip on the chip seat 62. The end limit structure composed of the lower limit cylinder 66 and the lower limit block 67 stops the pushing structure member when the device moves to the specified position, preventing over-pushing or under-pushing, and ensuring the precise positioning of the chip component during the detection process.

[0057] Specific Embodiment Ten: In combination with Figures 1 to 16 To illustrate this embodiment, the difference between this embodiment and Specific Embodiment Nine is that there are a plurality of insertion rods on the side of the back cover plate 7 facing the detection device housing 2. One end of each insertion rod is fixedly connected to the back cover plate 7, and there are a plurality of insertion sleeves on the inner side wall of the detection device housing 2. One end of each insertion sleeve is fixedly connected to the detection device housing 2. The back cover plate 7 is detachably connected to the detection device housing 2 through a plurality of insertion rods and a plurality of insertion sleeves. Other compositions and connection methods are the same as those in Specific Embodiment Nine.

[0058] The present utility model has been disclosed above with preferred embodiments. However, it is not intended to limit the present utility model. Any person skilled in the art, without departing from the scope of the technical solution of the present utility model, can make some modifications or decorations using the disclosed structure and technical content to form equivalent embodiments of equivalent changes. However, any simple modification, equivalent change, and decoration made to the above embodiments based on the technical essence of the present utility model without departing from the content of the technical solution of the present utility model still fall within the scope of the technical solution of the present utility model.

[0059] Working Principle

[0060] The working process of an integrated microfluidic pathogen nucleic acid detection device provided by this application is as follows:

[0061] Step 1: Carefully inspect the contact surfaces of each layer of the microfluidic chip to ensure that there is no dust or other impurities.

[0062] Step 2: On the basis of confirming that the contact surfaces are clean, evenly apply an appropriate amount of silicone oil to the contact surfaces to form a sealing film.

[0063] Step 3: Assemble the aligned laminates in a predetermined order and direction to complete the overall assembly process of the microfluidic chip.

[0064] Step 4: After completing the chip assembly, through the operation of a precision mechanical device (manual push), when the dispensing holes 111 in the top dispensing layer plate 11 are completely aligned with the liquid storage holes 121 in the second-layer liquid storage layer plate 12, sample addition is carried out.

[0065] Step 5: After injecting the sample and reagent, restore the chip to its original position and ensure that there is no reagent leakage during subsequent operations.

[0066] Step 6: Perform necessary fixing operations on the detection chip 1 containing the reagent sample and install it on the chip holder 62 to ensure its stability during subsequent mechanical movements.

[0067] Step 7: Start the automated control system of the device and set the target temperature according to the experimental requirements. The system will automatically adjust and maintain the temperature of the reagent in the chip until the set target temperature is reached. Specifically, this process is to control the first heating sheet 551 to enter the corresponding hole position in the first heating sheet slot 132 to heat the liquid stored in the liquid storage layer plate 12. After the heating is completed, control the first heating sheet 551 to retract. It should be noted that since the three heating sheets are integrated on the heating sheet moving plate 55, the three heating sheets perform insertion and retraction actions, but only one heating sheet is controlled to perform the heating work as needed.

[0068] Step 8: After the heating is completed, through the precise control of the push rod 33, push the second-layer liquid storage layer plate 12 of the chip to move, and at the same time control the moving speed and force to ensure smooth sliding between the chip layers.

[0069] Step 9: When the limiting rod section 332 on the push rod 33 contacts the limiting frame 42, the structural member stops moving, ensuring that the chip is in the correct position and preparing for the next operation. At this time, the reagent samples in the second-layer liquid storage layer plate 12 of the chip all fall into the mixing holes 131 in the third-layer mixing layer plate 13 under the action of gravity.

[0070] Step 10: After completing the task, the push rod 33 retracts to the starting position. Subsequently, the microfluidic chip and the base perform several precisely controlled falling actions to fully mix the reagent under the action of gravity, ensuring the uniformity and efficiency of the reaction.

[0071] Step 11: At this time, control the heating sheet moving plate 55 to insert the second heating sheet 552 into the second heating hole 142 to heat the third-layer mixing layer plate 13. After the heating is completed, the heating sheet moving plate 55 resets, the lower limit block 67 moves down, and the push rod 33 continues to move to the right. Transfer all the reagents to the test paper slots 143 in the fourth-layer detection layer plate 14.

[0072] Step 12: At this time, control the heating sheet moving plate 55 to insert the third heating sheet 553 into the third heating hole located on the chip holder 62 to provide a constant temperature environment for the colloidal gold test strip. After a few minutes of constant temperature reaction, the colloidal gold test strip will have a color reaction. By observing the band changes on the colloidal gold test strip, the result of the nucleic acid test can be interpreted. This step provides a quick and intuitive conclusion for the entire detection process.

Claims

1. An integrated microfluidic pathogen nucleic acid detection chip, characterized in that: The detection chip (1) includes a dosing layer plate (11), a liquid storage layer plate (12), a mixing layer plate (13), a detection layer plate (14), and two fixing sleeves (15). The dosing layer plate (11), the liquid storage layer plate (12), the mixing layer plate (13), and the detection layer plate (14) are stacked in sequence from top to bottom to form a stacked structure. The two fixing sleeves (15) are sleeved on the stacked structure and limit the stacked structure in the width direction. A lubricating oil layer is provided between adjacent layer plates in the stacked structure, and sliding connection is achieved through the lubricating oil.

2. The integrated microfluidic pathogen nucleic acid detection chip according to claim 1, wherein: A plurality of dosing holes (111) are sequentially processed on the top of the dosing layer plate (11) along the length extension direction of the dosing layer plate (11). A plurality of liquid storage holes (121) are sequentially processed on the top of the liquid storage layer plate (12) along the length extension direction of the liquid storage layer plate (12), and each liquid storage hole (121) is correspondingly arranged with a dosing hole (111).

3. The integrated microfluidic pathogen nucleic acid detection chip according to claim 2, wherein: A mixing hole (131) and a first heating sheet slot (132) are processed on the top of the mixing layer plate (13). A detection hole (141) and a second heating sheet slot (142) are processed on the top of the detection layer plate (14). A test strip slot (143) is processed on the bottom of the detection layer plate (14) along the length extension direction of the detection layer plate (14), and the test strip slot (143) is communicated with the detection hole (141).

4. An integrated microfluidic pathogen nucleic acid detection chip according to claim 3, characterized in that: The mixing layer plate (13) and the detection layer plate (14) have the same length. The dosing layer plate (11) and the liquid storage layer plate (12) have the same length. Moreover, the length of the mixing layer plate (13) and the detection layer plate (14) is greater than the length of the dosing layer plate (11) and the liquid storage layer plate (12).

5. An integrated microfluidic pathogen nucleic acid detection device, characterized in that: The detection device includes the detection chip (1) according to any one of claims 1 to 4, and further includes a detection device housing (2), a pushing mechanism (3), an upper limit mechanism (4), a heating component (5), a mixing and vibrating component (6), and a back cover plate (7). The pushing mechanism (3), the upper limit mechanism (4), the heating component (5), and the mixing and vibrating component (6) are all integrated in the detection device housing (2). The pushing mechanism (3) is installed at the upper part of the detection device housing (2), and the execution end of the pushing mechanism (3) passes through the top of the detection device housing (2) and extends outside the detection device housing (2). The mixing and vibrating component (6) is installed at the inner bottom of the detection device housing (2), and the mixing and vibrating component (6) is located in the extending direction of the pushing mechanism (3). The detection chip (1) is installed on the top of the mixing and vibrating component (6), and the detection chip (1) is correspondingly arranged with the execution end of the pushing mechanism (3). The upper limit mechanism (4) is arranged between the pushing mechanism (3) and the mixing and vibrating component (6). The upper limit mechanism (4) is installed at the inner bottom of the detection device housing (2), and the limiting end of the upper limit mechanism (4) is correspondingly arranged with the execution end of the pushing mechanism (3). The heating component (5) is installed at the inner bottom of the detection device housing (2), and the heating end of the heating component (5) is correspondingly arranged with the detection chip (1). The heating end of the heating component (5) can perform a plugging and unplugging action relative to the detection chip (1). The back cover plate (7) is arranged on the back side of the detection device housing (2), and the back cover plate (7) is detachably connected to the detection device housing (2).

6. The integrated microfluidic pathogen nucleic acid detection device according to claim 5, characterized in that: The pushing mechanism (3) includes a pushing cylinder (31), a connecting plate (32), and a push rod (33). The pushing cylinder (31) is installed on the inner wall of the detection device housing (2) through a mounting bracket, and the extending end of the pushing cylinder (31) faces the mixing and vibrating component (6). The connecting plate (32) is an L-shaped plate. The connecting plate (32) is arranged on the extending end of the pushing cylinder (31), and the vertical part of the connecting plate (32) is detachably connected to the extending end of the pushing cylinder (31). The push rod (33) is arranged on the horizontal part of the connecting plate (32), and the bottom end of the push rod (33) is fixedly connected to the connecting plate (32). The top end of the push rod (33) passes through the top of the detection device housing (2) and extends outside the detection device housing (2): The push rod (33) includes a push rod body (331), a limit rod section (332), a liquid storage layer push rod section (333) and a mixing layer push rod section (334). The push rod body (331) is arranged vertically on the horizontal part of the connecting plate (32), and the bottom end of the push rod body (331) is fixedly connected to the connecting plate (32). The limit rod section (332), the liquid storage layer push rod section (333) and the mixing layer push rod section (334) are arranged in sequence from top to bottom on the side of the push rod body (331) facing the mixing vibration assembly (6), and the limit rod section (332), the liquid storage layer push rod section (333) and the mixing layer push rod section (334) are all fixedly connected to the push rod body (331). There is an arrangement gap between the limit rod section (332) and the liquid storage layer push rod section (333). The limit rod section (332) is correspondingly arranged with the limit end of the limit mechanism (4). The liquid storage layer push rod section (333) is correspondingly arranged with the liquid storage layer plate (12) in the detection chip (1). The mixing layer push rod section (334) is correspondingly arranged with the mixing layer plate (13) in the detection chip (1).

7. An integrated microfluidic pathogen nucleic acid detection device according to claim 6, wherein: The upper limit mechanism (4) includes a longitudinal push rod (41) and a limit frame (42). The longitudinal push rod (41) is arranged between the pushing mechanism (3) and the mixing vibration assembly (6). The longitudinal push rod (41) stands vertically on the inner bottom of the detection device housing (2) in the vertical direction, and the housing of the longitudinal push rod (41) is detachably connected to the inner bottom of the detection device housing (2) by bolts. The limit frame (42) stands at the end of the piston rod in the longitudinal push rod (41), and the bottom of the limit frame (42) is detachably connected to the piston rod of the longitudinal push rod (41) by bolts. The top of the limit frame (42) is correspondingly arranged with the limit rod section (332).

8. An integrated microfluidic pathogen nucleic acid detection device according to claim 7, characterized in that: The heating assembly (5) includes a pneumatic source (51), a pneumatic slide rail pair (52), an air inlet pipe (53), an air outlet pipe (54) and a heating sheet moving plate (55). The pneumatic source (51) is installed on the inner bottom of the detection device housing (2). The pneumatic slide rail pair (52) is arranged between the upper limit mechanism (4) and the mixing vibration assembly (6), and the length extension direction of the pneumatic slide rail pair (52) is the same as the width extension direction of the detection device housing (2). One end of the pneumatic guide rail in the pneumatic slide rail pair (52) is detachably connected to the detection device housing (2) by bolts. The air inlet pipe (53) and the air outlet pipe (54) are arranged between the pneumatic source (51) and the pneumatic slide rail pair (52). One end of the air inlet pipe (53) is communicated with the air outlet end of the pneumatic source (51), and the other end of the air inlet pipe (53) is communicated with the air inlet end of the pneumatic guide rail in the pneumatic slide rail pair (52). One end of the air outlet pipe (54) is communicated with the air suction end of the pneumatic source (51), and the other end of the air outlet pipe (54) is communicated with the air outlet end of the pneumatic guide rail in the pneumatic slide rail pair (52). The heating sheet moving plate (55) is detachably connected to the sliding block in the pneumatic slide rail pair (52) by bolts, and the pneumatic slide rail pair (52) drives the heating sheet moving plate (55) to move reciprocally along the width extension direction of the detection device housing (2); On one side of the heating sheet moving plate (55) facing the inner wall of the detection device housing (2), a first heating sheet (551), a second heating sheet (552), and a third heating sheet (553) are fixedly arranged from top to bottom. The first heating sheet (551) is correspondingly arranged with the first heating sheet slot (132), the second heating sheet (552) is correspondingly arranged with the second heating sheet slot (142), and the third heating sheet (553) is correspondingly arranged with the third heating sheet slot in the hybrid vibration assembly (6).

9. An integrated microfluidic pathogen nucleic acid detection device according to claim 8, wherein: The hybrid vibration assembly (6) includes a vibration cylinder (61), a chip holder (62), an extended support plate (63), a pressing cylinder (64), an L-shaped pressing rod (65), a lower limit cylinder (66), and a lower limit block (67). The vibration cylinder (61) is vertically arranged on the inner bottom of the detection device housing (2), and the housing of the vibration cylinder (61) is detachably connected to the detection device housing (2). The chip holder (62) is arranged at the end of the piston rod of the vibration cylinder (61), and the chip holder (62) is detachably connected to the piston rod of the vibration cylinder (61) by bolts. The extended support plate (63) is arranged at the lower part of the side of the chip holder (62) facing the back cover plate (7), and the extended support plate (63) is integrally formed with the chip holder (62). The pressing cylinder (64) is vertically arranged on the extended support plate (63), and the cylinder body of the pressing cylinder (64) is detachably connected to the extended support plate (63) by bolts. The L-shaped pressing rod (65) is arranged at the end of the piston rod in the pressing cylinder (64), and the vertical part of the L-shaped pressing rod (65) is fixedly connected to the piston rod in the pressing cylinder (64). The horizontal part of the L-shaped pressing rod (65) does not extend to the top of the chip holder (62). The detection chip (1) is placed in the chip holder (62) and fixed by the L-shaped pressing rod (65). An embedding groove is machined on one side of the chip holder (62) away from the upper limit mechanism (4). The lower limit cylinder (66) is vertically arranged in the embedding groove, and the cylinder body of the lower limit cylinder (66) is fixed on the side wall of the embedding groove by a locking clip. The lower limit block (67) is fixed at the end of the piston rod in the lower limit cylinder (66).

10. An integrated microfluidic pathogen nucleic acid detection device according to claim 9, characterized in that: On the side of the back cover plate (7) facing the detection device housing (2), a plurality of insertion rods are provided. One end of each insertion rod is fixedly connected to the back cover plate (7). On the inner side wall of the detection device housing (2), a plurality of insertion sleeves are provided. One end of each insertion sleeve is fixedly connected to the detection device housing (2). The back cover plate (7) is detachably connected to the detection device housing (2) through the plurality of insertion rods and the plurality of insertion sleeves.

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