Multi-primer response PCR (Polymerase Chain Reaction) sealed detection kit

Abstract

The invention discloses a PCR (Polymerase Chain Reaction) sealed detection kit with multi-primer response. The PCR sealed detection kit comprises a clamping plate, a chip and a sealing element, the clamping plate comprises a first [-shaped plate and a second [-shaped plate which form a rectangular structure; the chip inserted into the rectangular structure comprises a cover plate and a substrate which are overlapped and glued; flow channel mechanisms are arranged on the substrate, each flow channel mechanism is communicated with at least two reaction cavities, and a specific primer and a reaction enzyme are arranged in each reaction cavity; the sealing pieces are located on the side, away from the base, of the cover piece and comprise the first sealing piece and the second sealing piece, each of the first sealing piece and the second sealing piece comprises a shell, a silica gel piece and a partition plate, the silica gel pieces are bonded to the sides, away from the lower bottom face, of the four side faces of the shell, and the partition plates are arranged between the silica gel pieces and the lower bottom face of the shell; a sealing cavity is formed among part of the silica gel sheets, part of the side faces and the two adjacent partition plates, and water stop glue is bonded in the sealing cavity. The problems that multi-primer response cannot be achieved, sample injection is slow, clamping is inconvenient, and in addition, self-sealing cannot be achieved after sample injection are solved.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and more particularly to a multi-primer responsive PCR sealed detection box. Background Art

[0002] Chinese patent application document 1 (application number: 2024105587025, application date: 2024.05.08) discloses a PCR sealed detection box, including a PCR microfluidic chip body and a chip support plate. The PCR microfluidic chip body includes a base and a cover plate that are bonded to each other. At least one reaction chamber 3' is formed on the base, and injection holes 4' are respectively formed on the cover plate at the end position relative to each reaction chamber 3'; the second surface of the chip support plate is provided with multiple chip protrusions 5' and elastic limit cards 6'. The multiple chip protrusions 5' are used to limit and fix the PCR microfluidic chip body. The elastic limit card 6' is arranged on a side of the PCR microfluidic chip body that is different from the multiple chip protrusions 5'. In the above scheme, the sealant is immersed in the four injection holes 4' at one time after the samples are injected respectively. At least four injections result in a too long injection time, and aerosol contamination is likely to occur between different samples. In addition, the small area of ​​the chip makes it difficult for the chip to resist its own bending stress and the chip support plate, requiring strong buckling. At the same time, the buckling process is more likely to cause the silicon substrate and the glass cover to break. At the same time, only one reaction chamber 3' is reacted at one time, resulting in serious sample waste.

[0003] Chinese patent application document 2 (application number: 2019100709326, application date: 2019.01.25) discloses a digital PCR chip and its preparation method, preparation device, and use method. The digital PCR chip includes: a substrate and a structural layer arranged above the substrate, the structural layer has at least one sample channel, each sample channel includes a microporous structure with multiple micropores and an overflow cavity for holding excess sample, and the overflow cavity is located on at least one side of the microporous structure. The above solution can only perform one sample detection on a film-coated chip. At the same time, the equipment is large in size and cumbersome to use. During use, the sample needs to be rolled and squeezed immediately so that the sample is filled into the microporous structure. The assembly time is too long, which does not significantly shorten the time required for the sample PCR reaction cycle, and the detection efficiency cannot be significantly improved.

[0004] Chinese patent application document 3 (application number: 2017108784701, application date: 2017.09.26) discloses a multi-flux microfluidic nucleic acid detection chip with active control flow path and its use method. The detection chip includes a chip body, on which a sample loading chamber, a reaction chamber, and a microfluidic flow channel are provided; the reaction chamber includes several reaction chambers; the microfluidic flow channel includes a main sample outlet channel and several sample distribution channels; each sample distribution channel is separately provided, including a sample outlet channel for the sample distribution chamber and a sample inlet channel for the reaction chamber; the sample outlet channel for the sample distribution chamber and the sample inlet channel for the reaction chamber can be connected by a sample distribution connection channel formed by heat-sealing a thin film on the chip body, and the thin film portion of the sample distribution connection channel can be heat-sealed to form a sample distribution blocking portion to intercept the sample distribution channel. The above scheme also has the problem of only being able to perform single sample detection.

[0005] Therefore, it is a technical problem that urgently needs to be solved in this field to provide a PCR sealed detection box that not only responds to multiple primers but also has rapid sampling, secure snap-in fixation, and can self-seal after sampling to reduce aerosol contamination. Summary of the Invention

[0006] In view of this, the present invention provides a multi-primer response PCR sealed detection box to solve the problems of no multi-primer response, slow sampling, inconvenient snap-in fixation, and failure to self-seal after sampling, which easily leads to aerosol contamination.

[0007] The present application provides a multi-primer response PCR sealed detection box, comprising a card board, a rectangular chip and a strip-shaped sealing member, wherein:

[0008] The card plate includes a first N-shaped plate and a second N-shaped plate, the N-shaped opening of the first N-shaped plate faces the N-shaped opening of the second N-shaped plate, and the N-shaped opening of the second N-shaped plate faces the first N-shaped plate. The first N-shaped plate and the second N-shaped plate are spliced ​​together to form a rectangular structure. The rectangular structure has a hollow portion that penetrates the rectangular structure along the thickness direction of the chip, and the shape of the hollow portion is rectangular;

[0009] Along the peripheral direction of the hollow portion, a first ⌚-shaped groove is formed on a side of the first ⌚-shaped plate close to the hollow portion and is recessed in a direction away from the hollow portion, and a second ⌚-shaped groove is formed on a side of the second ⌚-shaped plate close to the hollow portion and is recessed in a direction away from the hollow portion; the first ⌚-shaped groove and the second ⌚-shaped groove are connected and penetrated to form a rectangular groove;

[0010] A chip, the edges of which are inserted into a rectangular groove, comprising a cover sheet and a base that are overlapped and glued to each other along the thickness direction of the chip, the cover sheet being located above the base, and the orthographic projection of the hollow portion overlaps within the orthographic projections of the cover sheet and the base along the thickness direction of the chip; at least two flow channel mechanisms are provided on a side of the base close to the cover sheet, and the at least two flow channel mechanisms extend along the length direction of the chip and are arranged along the width direction of the chip; each flow channel mechanism is a groove opening toward the cover sheet, and each flow channel mechanism is connected to at least two reaction chambers, and at least two reaction chambers are arranged along the length direction of the chip, and the reaction chambers are fan-shaped grooves, and specific primers and reaction enzymes are provided in each reaction chamber; the cover sheet is provided with an injection hole and an outlet hole at both ends of each flow channel mechanism along the length direction of the chip, and the injection hole and the outlet hole respectively pass through the cover sheet along the thickness direction of the chip, and both the injection hole and the outlet hole are circular holes;

[0011] The sealing member is located on the side of the cover away from the base, and includes a first sealing member corresponding to the injection hole and a second sealing member corresponding to the output hole. The first sealing member and the second sealing member each include a rectangular column shell, a rectangular silicone sheet and a partition. The shell includes an upper bottom surface, a lower bottom surface and a side surface. The upper bottom surface and the lower bottom surface are two parallel and congruent rectangles. The side surface is composed of four rectangles. The long side of each side surface is parallel to the corresponding side of the upper bottom surface and the lower bottom surface, and the short side of each side surface is perpendicular to the upper bottom surface. The upper bottom surface, the lower bottom surface and the four side surfaces are parallel to each other. The same rectangular column is formed, and a strip opening is opened on the upper bottom surface. The strip opening passes through the upper bottom surface along the thickness direction of the chip and extends along the width direction of the chip. The periphery of the silicone sheet is bonded to the four side surfaces away from the lower bottom surface respectively. A partition is provided between the silicone sheet and the lower bottom surface. The partition is made of plastic, and the number of partitions is twice the number of flow channel mechanisms. A sealed cavity is formed between part of the silicone sheet, part of the side surface, and two adjacent partitions. The sealed cavity in the first seal is connected to the sample inlet, and the sealed cavity in the second seal is connected to the sample outlet.

[0012] The silicone sheet is provided with a positioning hole that penetrates the silicone sheet along the thickness direction of the chip, and the positioning hole is connected to the sealing cavity; a water-stop glue is bonded to the side of the partition close to the sealing cavity. The material of the water-stop glue is polyurethane sealant. Along the thickness direction of the chip, the water-stop glue and the central axis of the positioning hole do not coincide, and the orthographic projection of the positioning hole is located within the orthographic projection of the sealing cavity.

[0013] Compared with the prior art, the multi-primer response PCR sealed detection kit provided by the present invention achieves at least the following beneficial effects:

[0014] First, the multi-primer responsive PCR sealed detection box provided by the present invention can be self-sealed during the sampling process to prevent aerosol contamination between each other; first, the present invention provides a seal with an independent sealing cavity to seal the two ends of the flow channel mechanism to prevent different flow channels from sharing a sealing cavity and being unable to seal individually. Furthermore, the present invention provides a water-swelling water-stopping glue in the sealing cavity. After the sample liquid is filled, the water-stopping glue can serve as a mark of the completion of the filling of the sample liquid; on the other hand, the water-stopping glue in the first sealing member and the second sealing member at both ends of the flow channel mechanism can be automatically fixed and sealed by ultraviolet light to prevent the sample liquid in the flow channel from diffusing and contaminating, and also play a better anti-overflow effect.

[0015] Second, through the multi-primer responsive PCR sealed detection box provided by the present invention, the chip is simply and safely snapped in and fixed, and is convenient and quick to use; first, the card plate provided by the present invention is a two-piece splicing structure, which avoids the need to perform a certain bending operation of the chip with low resistance to silicon material bending stress when a one-piece card plate is snapped into the chip, thereby improving the safety of chip use; further, the card plate includes a first ⌚-shaped plate with a first ⌚-shaped groove and a second ⌚-shaped plate with a second ⌚-shaped groove, and the first ⌚-shaped plate and the second ⌚-shaped plate are docked with each other, ensuring the safety of chip use while snapping in and installing quickly.

[0016] Third, the multi-primer responsive PCR sealed detection box provided by the present invention takes into account multi-channel and multi-primer responses at the same time, and the injection process is faster, which effectively reduces the time for PCR amplification and detection of a large number of samples; first, the water-stop glue in the first seal and the second seal provided by the present invention self-cures and seals the flow channel mechanism, and no additional sealing operation is required after the injection operation, which reduces the time for PCR amplification of a large number of samples from the aspect of simplifying the operating procedures; at the same time, the needle withdrawal operation and UV curing of the injection needle of the present invention can be carried out simultaneously, further reducing the time required for injection; further, after the injection time and operation of a flow channel mechanism are simplified, the injection time and operation of the entire chip are simplified exponentially by the number of flow channel mechanisms, which greatly saves a large number of samples for PCR amplification and detection as a whole; at the same time, diffusion contamination between different reaction chambers is avoided, that is, amplification and detection have been completed.

[0017] Of course, any product implementing the present invention does not necessarily need to achieve all of the technical effects described above at the same time.

[0018] Further features and advantages of the present invention will become apparent from the following detailed description of exemplary embodiments of the present invention with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and, together with the description, serve to explain the principles of the invention.

[0020] Figure 1 This is a schematic diagram of the structure of a PCR sealed detection box provided by Chinese patent application document 1;

[0021] Figure 2 This is a schematic structural diagram of a multi-primer response PCR sealed detection kit provided by the present invention;

[0022] Figure 3 1 is a schematic structural diagram of a first profile plate provided by the present invention;

[0023] Figure 4 2 is a schematic structural diagram of a second molded plate provided by the present invention;

[0024] Figure 5 This is a schematic diagram of the assembly of the chip and the seal provided by the present invention;

[0025] Figure 6 yes Figure 5 Enlarged view of point A in the middle;

[0026] Figure 7 is a BB' cross-sectional view of the sealing member provided by the present invention;

[0027] Figure 8 yes Figure 7 Enlarged view of point C in the middle;

[0028] Figure 9 Schematic diagram of the splicing structure of the first shaped plate, the second shaped plate and the metal strip provided by the present invention;

[0029] Figure 10 This is a schematic structural diagram of another multi-primer response PCR sealed detection box provided by the present invention. DETAILED DESCRIPTION

[0030] Various exemplary embodiments of the present invention will now be described in detail with reference to the accompanying drawings. It should be noted that unless otherwise specifically stated, the relative arrangement of components and steps, numerical expressions and numerical values ​​set forth in these embodiments do not limit the scope of the present invention.

[0031] The following description of at least one exemplary embodiment is merely illustrative in nature and is in no way intended to limit the invention, its application, or uses.

[0032] Technologies, methods, and equipment known to ordinary technicians in the relevant art may not be discussed in detail, but where appropriate, the technologies, methods, and equipment should be considered part of the specification.

[0033] In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not limiting. Therefore, other examples of the exemplary embodiments may have different values.

[0034] It should be noted that like reference numerals and letters refer to like items in the following figures, and therefore, once an item is defined in one figure, it need not be further discussed in subsequent figures.

[0035] Example 1

[0036] Reference Figure 2-Figure 8 As shown, Figure 2 This is a schematic structural diagram of a multi-primer response PCR sealed detection kit provided by the present invention; Figure 3 1 is a schematic structural diagram of a first profile plate provided by the present invention; Figure 4 2 is a schematic structural diagram of a second molded plate provided by the present invention; Figure 5 This is a schematic diagram of the assembly of the chip and the seal provided by the present invention; Figure 6 yes Figure 5 Enlarged view of point A in the middle; Figure 7 is a BB' cross-sectional view of the sealing member provided by the present invention; Figure 8 yes Figure 7 The enlarged view of the middle C; This embodiment provides a multi-primer response PCR sealed detection box, comprising a card board 1, a rectangular chip 2 and a strip-shaped seal 3, wherein,

[0037] The card board 1 includes a first ⌚-shaped plate 11 and a second ⌚-shaped plate 12. The ⌚-shaped opening of the first ⌚-shaped plate 11 faces the ⌚-shaped opening of the second ⌚-shaped plate 12, and the ⌚-shaped opening of the second ⌚-shaped plate 12 faces the first ⌚-shaped plate 11. The first ⌚-shaped plate 11 and the second ⌚-shaped plate 12 are spliced ​​together to form a rectangular structure. The rectangular structure has a hollow portion 13 that penetrates the rectangular structure along the thickness direction Z of the chip 2. The shape of the hollow portion 13 is rectangular;

[0038] Along the circumference of the hollow portion 13, a first ⌚-shaped groove 111 is formed on a side of the first ⌚-shaped plate 11 close to the hollow portion 13 and is recessed away from the hollow portion 13. A second ⌚-shaped groove 121 is formed on a side of the second ⌚-shaped plate 12 close to the hollow portion 13 and is recessed away from the hollow portion 13. The first ⌚-shaped groove 111 and the second ⌚-shaped groove 121 are interconnected and penetrated to form a rectangular groove.

[0039] The chip 2 has its edges inserted into the rectangular groove, and includes a cover sheet 21 and a base 22 that are overlapped and glued together along the thickness direction of the chip 2. The cover sheet 21 is located above the base 22. Along the thickness direction Z of the chip 2, the orthographic projection of the hollow portion 13 overlaps within the orthographic projections of the cover sheet 21 and the base 22; at least two flow channel mechanisms 212 are provided on one side of the base 22 close to the cover sheet 21. The at least two flow channel mechanisms 212 extend along the length direction Y of the chip 2 and are arranged along the width direction X of the chip 2; each flow channel mechanism 212 is open toward the cover sheet. 21, each of the flow channel mechanisms 212 is connected to at least two reaction chambers 214, and the at least two reaction chambers 214 are arranged along the length direction Y of the chip 2. The reaction chambers 214 are fan-shaped grooves, and each reaction chamber 214 is provided with a specific primer and a reaction enzyme; the cover plate 21 is provided with an injection hole 211 and an outlet hole 213 at both ends along the length direction Y of the chip 2 corresponding to each flow channel mechanism 212. The injection hole 211 and the outlet hole 213 respectively penetrate the cover plate 21 along the thickness direction Z of the chip 2, and the injection hole 211 and the outlet hole 213 are both circular holes;

[0040] The seal 3 is located on the side of the cover away from the base, and includes a first seal 3F corresponding to the injection hole 211 and a second seal 3S corresponding to the output hole 213. The first seal 3F and the second seal 3S each include a rectangular column shell 31, a rectangular silicone sheet 32 ​​and a partition 33. The shell 31 includes an upper bottom surface 311, a lower bottom surface 312 and a side surface 313. The upper bottom surface 311 and the lower bottom surface 312 are two parallel and congruent rectangles. The side surface 313 is composed of four rectangles. The long side of each side surface 313 is parallel to the corresponding side of the upper bottom surface 311 and the lower bottom surface 312, and the short side of each side surface 313 is perpendicular to the upper bottom surface 311. The upper bottom surface 311, the lower bottom surface 312 and the four side surfaces 313 together constitute a rectangular column. A strip opening is provided on the upper bottom surface 311, which passes through the upper bottom surface 311 along the thickness direction Z of the chip 2 and extends along the width direction X of the chip 2. The periphery of the silicone sheet 32 ​​is bonded to the four side surfaces 313 away from the lower bottom surface 312. A partition 33 is provided between the silicone sheet 32 ​​and the lower bottom surface 312. The material of the partition 33 is plastic. The number of flow channel mechanisms 212 is n, n is greater than or equal to 2 and is a positive integer, and the number of partitions 33 is twice the number of flow channel mechanisms 212; a sealed cavity 34 is formed between part of the silicone sheet 32, part of the side surface 313 and two adjacent partitions 33. The sealed cavity 34 in the first sealing member 3F is connected to the sampling hole 211, and the sealed cavity in the second sealing member 3S is connected to the sampling hole 213;

[0041] The silicone sheet 32 ​​is provided with a positioning hole 321 that passes through the silicone sheet 32 ​​along the thickness direction Z of the chip 2, and the positioning hole 321 is connected to the sealing cavity 34; a water-stop glue 35 is bonded to the side of the partition 33 close to the sealing cavity 34. The material of the water-stop glue 35 is polyurethane sealant. Along the thickness direction Z of the chip 2, the water-stop glue 35 does not coincide with the central axis of the positioning hole 321, and the orthographic projection of the positioning hole 321 is located within the orthographic projection of the sealing cavity 34.

[0042] Specifically, continue to refer to Figure 2 and Figure 3 As shown, the card board 1 includes a first shaped plate 11 and a second shaped plate 12 of the same size. The first shaped plate 11 and the second shaped plate 12 are both rectangular strips bent in a shaped manner. The materials of the first shaped plate 11 and the second shaped plate 12 can be plastic or rubber. Plastic and rubber have the advantages of being light, economical and easy to manufacture. Along the width direction X of the chip 2, the two ends of the first shaped plate 11 and the second shaped plate 12 are not in the same horizontal plane, that is, the lengths of the two ends of the first shaped plate 11 and the second shaped plate 12 after being bent in a shaped manner are different. The t-shaped opening of the first t-shaped plate 11 faces the t-shaped opening of the second t-shaped plate 12, and the t-shaped opening of the second t-shaped plate 12 faces the first t-shaped plate 11; the short end of the first t-shaped plate 11 is engaged with the long end of the second t-shaped plate 12, and the long end of the first t-shaped plate 11 is engaged with the short end of the second t-shaped plate 12. The first t-shaped plate 11 and the second t-shaped plate 12 are engaged with each other to form a hollow rectangular structure. The width direction of the rectangular structure is parallel to the width direction X of the chip 2, and the length direction of the rectangular structure is parallel to the length direction Y of the chip 2. The central axis of the rectangular structure coincides with the central axis of the chip 2; the thickness direction Z of the chip 2, the length direction Y of the chip 2, and the width direction X of the chip 2 are perpendicular to each other; the rectangular structure has a hollow portion 13 that passes through the rectangular structure along the thickness direction Z of the chip 2, and the hollow portion 13 is a rectangular shape that is similar to the rectangular structure in mathematical definition. The rectangular hollow portion 13 is used to accommodate the chip 2 . Along the thickness direction Z of the chip 2 , the orthographic projection of the hollow portion 13 coincides with the orthographic projection of the chip 2 .

[0043] Along the circumference of the hollow portion 13, a first ⌚-shaped groove 111 is formed on the side of the first ⌚-shaped plate 11 close to the hollow portion 13, and a second ⌚-shaped groove 121 is formed on the side of the second ⌚-shaped plate 12 close to the hollow portion 13, and is recessed away from the hollow portion 13. The first ⌚-shaped groove 111 and the second ⌚-shaped groove 121 are both ⌚-shaped groove bodies. When the first ⌚-shaped plate 11 and the second ⌚-shaped plate 12 are mutually engaged to form a rectangular structure, the first ⌚-shaped groove 111 and the second ⌚-shaped groove 121 are interconnected and connected to form a rectangular groove that opens toward the central axis of the rectangular structure. The first n-shaped groove 111 and the second n-shaped groove 121 respectively clamp the chip 2 along the periphery of the chip 2; along the thickness direction Z of the chip 2, the orthographic projections of the hollow portion 13, the first n-shaped groove 111 and the second n-shaped groove 121 coincide with the orthographic projection of the chip 2, and the chip 2 can be tightly clamped in the rectangular structure of the first n-shaped plate 11 and the second n-shaped plate 12 through the hollow portion 13, the first n-shaped groove 111 and the second n-shaped groove 121.

[0044] Continue to refer to Figure 2-Figure 6 As shown, the chip 2 includes a cover sheet 21 and a base 22 that overlap and are glued together along its thickness direction. The cover sheet 21 and the base 22 are both rectangular in shape, and the sum of the thicknesses of the cover sheet 21 and the base 22 along the thickness direction Z of the chip 2 is equal to the slot width of the first ∘-shaped groove 111 or the second ∘-shaped groove 121 along the thickness direction Z of the chip 2. The cover sheet 21 can be a transparent glass slide, and the base 22 can be made of silicon. Both silicon and transparent glass slides are conventional techniques in the art and will not be further explained in this embodiment. Two strip-shaped sealing members 3 are fixedly glued to the side of the cover sheet 21 away from the chip 2.

[0045] A flow channel mechanism 212 extending along the length direction is provided on the side of the base 22 close to the cover plate 21. The flow channel mechanism 212 is a linear groove extending along the length direction Y of the chip 2, and the notch of the flow channel mechanism 212 faces the cover plate 21. At least two flow channel mechanisms 212 are provided along the width direction X of the chip 2. At least two flow channel mechanisms 212 are parallel to each other, and their orthographic projections along the width direction X of the chip 2 coincide with each other. This embodiment uses three flow channel mechanisms 212 as a specific example, and does not serve as a limitation on the specific number of flow channel mechanisms 212. Each flow channel mechanism 212 is provided with at least two fan-shaped reaction chambers 214 along its length direction; this embodiment uses three reaction chambers 214 on each flow channel mechanism 212 as a specific embodiment, and does not serve as a specific limitation on this device; the reaction chambers 214 are connected and connected to the flow channel mechanism 212, and different reaction chambers 214 are independent of each other. Different specific primers and enzymes that catalyze the corresponding specific primers are fixedly disposed on one side of each reaction chamber 214 away from the flow channel mechanism 212; both the specific primers and the enzymes are freeze-dried reagents. The fan-shaped reaction chamber 214 facilitates the construction of a buffer region between the specific primers and the flow channel mechanism 212 after the sample liquid enters the reaction chamber 214 from the flow channel mechanism 212; this allows the sample liquid to flow unidirectionally and simultaneously into each reaction chamber 214; once the reaction chamber 214 is full, no further liquid is introduced. The fan-shaped roots of the reaction chamber 214 and the flow channel mechanism 212 can increase the necessary diffusion distance required to prevent mutual contamination between the different reaction chambers 214, thereby increasing the necessary diffusion time, thereby reducing the uneven and unclear detection color development caused by mutual contamination.

[0046] Each flow channel mechanism 212 has a corresponding inlet hole 211 and outlet hole 213 formed on the cover sheet 21 at both ends along the length direction Y of the chip 2. The inlet hole 211 and outlet hole 213 are circular holes extending through the cover sheet 21 along the thickness direction Z of the chip 2. The n inlet holes 211 and outlet holes 213 are arranged in a direction parallel to the width direction X of the chip 2, and the n inlet holes 211 and outlet holes 213 are arranged in a direction parallel to each other. In this embodiment, three flow channel mechanisms 212 are taken as an example. The cover sheet 21 is provided with three injection holes 211 arranged along the width direction X of the chip 2 at one end along the length direction Y of the chip 2. The cover sheet 21 is provided with three outlet holes 213 arranged along the width direction X of the chip 2 at one end away from the three injection holes 211. The three injection holes 211 and the three outlet holes 213 correspond to the two ends of the three flow channel mechanisms 212, so that the two ends of the three flow channel mechanisms 212 are connected to the outside at the side of the cover sheet 21 away from the base 22.

[0047] Continue to refer to Figure 2-Figure 8As shown, this embodiment is provided with three flow channel mechanisms 212, corresponding to which are provided three injection holes 211 and three outlet holes 213. A first sealing member 3F is glued together and extends through at least three injection holes 211 along the width direction X of the chip 2. A second sealing member 3S is glued together and extends through at least three outlet holes 213 along the width direction X of the chip 2. The sealing member 3 comprises a hollow cylindrical housing 31, a rectangular silicone sheet 32, and a partition 33 disposed within the housing 31.

[0048] The hollow rectangular cylindrical cavity passes through the shell 31 along the thickness direction Z of the chip 2. The shell 31 includes an upper bottom surface 311, a lower bottom surface 312 and a side surface 313. The upper bottom surface 311 and the lower bottom surface 312 are two parallel and congruent rectangles, and the length direction of the upper bottom surface 311 and the lower bottom surface 312 is the width direction X of the chip 2. The side surface 313 is composed of four rectangles, and the long side of each side surface 313 is parallel to the corresponding side of the upper bottom surface 311 and the lower bottom surface 312, and the short side of each side surface 313 is perpendicular to the upper bottom surface 311. The upper bottom surface 311, the lower bottom surface 312 and the four side surfaces 313 together form a rectangular column extending along the width direction X of the chip 2. A strip-shaped opening extends through the upper bottom surface 311 along the thickness direction Z of the chip 2 and along the width direction X of the chip 2. Along the thickness direction Z of the chip 2, the orthographic projection of the sample inlet 211 coincides with the orthographic projection of the strip-shaped opening of the first sealing member 3F; the orthographic projection of the sample outlet 213 coincides with the orthographic projection of the strip-shaped opening of the second sealing member 3S. The lower bottom surface 312 of the housing 31 is fixedly bonded to the side of the cover 21 facing away from the base 22. A silicone sheet 32 ​​is provided along the upper bottom surface 311 of the housing 31, enclosing the strip-shaped opening. The silicone sheet 32 ​​is a rectangular sheet with a Shore hardness of no more than 60. Along the thickness direction Z of the chip 2, the orthographic projection of the silicone sheet 32 ​​coincides with the orthographic projection of the hollow cavity of the housing 31. A partition 33 is bonded perpendicularly to the side of the silicone sheet 32 ​​near the base 22; the partition 33 can be made of plastic. The number of partitions 33 is equal to twice the number of flow channel mechanisms 212, that is, 2n; this embodiment provides three flow channel mechanisms 212, and six partitions 33 are correspondingly provided; part of the silicone sheet 32, six partitions 33 and part of the side surface 313 respectively constitute three quadrangular prism-shaped sealed cavities 34 in the inner cavity of the shell 31, which are respectively connected to the three flow channel mechanisms 212; the three sealed cavities 34 are independent of each other, and the three quadrangular prism-shaped sealed cavities 34 are respectively connected to the three injection holes 211.

[0049] Furthermore, the silicone sheet 32 ​​is provided with positioning holes 321 extending through the sheet along the thickness direction Z of the chip 2. The number of positioning holes 321 is the same as the number of flow channel mechanisms 212, i.e., n. In this embodiment, there are three flow channel mechanisms 212, and the first seal 3F and the second seal 3S each have three positioning holes 321. The three positioning holes 321 of the first seal 3F correspond to the three injection holes 211 of the three flow channel mechanisms 212, while the three positioning holes 321 of the second seal 3S correspond to the three outlet holes 213 of the three flow channel mechanisms 212. The positioning holes 321 are elastic, contractible circular holes, which are formed by the elastic stress of the silicone sheet 32. In their natural state, the positioning holes 321 are closed before use. The user uses the positioning holes 321 to pass the sample needle through the silicone sheet 32 ​​and the sealing cavity 34, and then into the injection hole 211 for sample injection.

[0050] The side of the silicone sheet 32 ​​closest to the cover 21, along with the two partitions 33 closest to the positioning hole 321, forms a sealed cavity 34. The sealed cavity 34 of the first sealing member 3F is connected to the sample inlet 211 on the cover, while the sealed cavity 34 of the second sealing member 3S is connected to the sample outlet 213 on the cover. Within the sealed cavity 34, a waterstop 35 is adhered to the side 313 of the housing 31 or the partitions 33. This waterstop 35 is a single-component, solvent-free polyurethane sealant that exhibits both water-expanding properties and elastic sealing properties. The water-stop glue 35 is glued to the partition 33, and the central axis of the water-stop glue 35 and the positioning hole 321 do not coincide with each other, so as to prevent the trace sample liquid at the tip of the injection needle from coming into contact with the water-stop glue 35 during the injection process, and to prevent the tip of the injection needle from sticking to the water-stop glue 35 and carrying the water-stop glue 35 into the injection hole 211, further causing blockage problems in the injection hole 211 or the flow channel mechanism 212.

[0051] In the specific implementation process, continue to refer to Figure 2-Figure 4 As shown, this embodiment can be used for specific amplification of multiple specific genes and their sealing detection, and can be specifically applied to gene cloning, gene expression analysis, detection of disease-related genes and DNA fingerprint analysis, etc. The specific operations are as follows:

[0052] The first step is to load the already integrally bonded chip 2 and the seal 3 into the card board 1: First, slide the chip 2 along the length direction Y of the chip 2 and snap it into the first C-shaped groove 111 of the first C-shaped plate 11; then hold the outer side of the first C-shaped plate 11 away from the chip 2, orient the C-shaped opening of the second C-shaped plate 12 towards the first C-shaped plate 11, align the second C-shaped groove 121 of the second C-shaped plate 12 with the side of the chip 2 away from the first C-shaped plate 11, and slide the second C-shaped plate 12 along the length direction Y of the chip 2 so that the short end of the first C-shaped plate 11 is engaged with the long end of the second C-shaped plate 12, and the long end of the first C-shaped plate 11 is engaged with the short end of the second C-shaped plate 12.

[0053] The second step is to magnetically snap the card board 1 into the PCR amplifier. Magnetic snap fitting is an existing technology in the art, and this step will not be elaborated too much here.

[0054] The third step is to pretreat the sample containing the target gene amplification template and dissolve it in a reaction solution containing dNTP (deoxyribonucleoside triphosphate), magnesium ions and buffer to prepare a sample solution to be initiated for amplification; the preparation of the sample solution is common knowledge technology in the art, and this embodiment will not explain this operation too much.

[0055] The fourth step is to automatically inject the sample into the chip 2 by an injection needle: This embodiment is provided with 3 flow channel mechanisms 212. Along the width direction X of the chip 2, they are the first flow channel mechanism, the second flow channel mechanism and the third flow channel mechanism in sequence. First, inject the sample into the first flow channel mechanism. Specifically, the injection needle passes through the positioning holes 321 on the silica gel sheet 32 of the first seal 3F along the thickness direction Z of the chip 2, the sealing cavity 34 corresponding to the first flow channel mechanism and the injection hole 211 of the cover sheet 21 corresponding to the first flow channel mechanism in sequence to inject the sample solution into the first flow channel mechanism; the sample solution flows from one injection hole 211 corresponding to the first flow channel mechanism to the outlet hole 213 at the other end of the first flow channel mechanism; during the flowing process, the sample solution fills the 3 reaction cavities 214 and the first flow channel mechanism in sequence; after the reaction cavities 214 and the first flow channel mechanism are filled, the sealing cavities 34 corresponding to the first flow channel mechanism at both ends are finally filled by the first seal 3F and the second seal 3S. The water in the sample solution contacts the water-stop glue 35 in the first seal 3F and the second seal 3S, and the water-stop glue 35 expands in the first seal 3F and the second seal 3S corresponding to the first flow channel mechanism. After the injection needle retracts, the PCR amplifier emits ultraviolet light to seal and cure the water-stop glue 35 in the sealing cavity 34 corresponding to the first flow channel mechanism, completing the injection and sealing of the first flow channel mechanism. Then, the injection and sealing of the second flow channel mechanism and the third flow channel mechanism are carried out in sequence. The injection and sealing operations of the second flow channel mechanism and the third flow channel mechanism are the same as those of the first flow channel mechanism, and this embodiment will not elaborate too much on this.

[0056] Step 5: Controlled amplification and automatic detection by PCR amplification instrument.

[0057] Compared with the prior art, the multi-primer response sealed PCR detection kit provided in this embodiment achieves at least the following beneficial effects:

[0058] First, the multi-primer responsive PCR sealed detection box provided by this embodiment can be self-sealed during the sampling process to prevent aerosol contamination between each other; first, this embodiment provides a seal 3 with an independent sealing cavity 34 to seal the two ends of the flow channel mechanism 212, to prevent the shared sealing cavity 34 between different flow channels from being unable to be individually sealed. Furthermore, this embodiment provides a water-swelling water-stopping glue 35 in the sealing cavity 34. After the sample liquid is filled, the water-stopping glue 35 can serve as a mark of the completion of the filling of the sample liquid; on the other hand, the water-stopping glue 35 in the first seal 3F and the second seal 3S at both ends of the flow channel mechanism 212 can be automatically fixed and sealed by ultraviolet light, to prevent the sample liquid in the flow channel from diffusing and contaminating, and also to achieve a better anti-overflow effect.

[0059] Second, through the multi-primer responsive PCR sealed detection box provided in this embodiment, the snap-fitting and fixing of the chip 2 is simple and safe, and it is convenient and quick to use; first, the card plate 1 provided in this embodiment is a two-piece splicing structure, which avoids the need to perform a certain bending operation of the chip 2 with a low resistance to silicon material bending stress when the one-piece card plate 1 is snapped onto the chip 2, thereby improving the safety of the chip 2; further, the card plate 1 includes a first ∘-shaped plate 11 with a first ∘-shaped groove 111 and a second ∘-shaped plate 12 with a second ∘-shaped groove 121, and the first ∘-shaped plate 11 and the second ∘-shaped plate 12 are docked with each other, ensuring the safety of the chip 2 in use while snapping and installing quickly.

[0060] Third, the multi-primer responsive PCR sealed detection box provided by this embodiment takes into account multi-channel and multi-primer responses, and the injection process is faster, which effectively reduces the time for PCR amplification and detection of a large number of samples; first, the water-stop glue 35 in the first seal 3F and the second seal 3S provided in this embodiment self-cures and seals the flow channel mechanism 212, and no additional sealing operation is required after the injection operation, thereby reducing the time for PCR amplification of a large number of samples from the aspect of simplifying the operating procedures; at the same time, the needle withdrawal operation and UV curing of the injection needle of this embodiment can be performed simultaneously, further reducing the time required for injection; further, after the injection time and operation of a flow channel mechanism 212 are simplified, the injection time and operation of the entire chip 2 are simplified exponentially by the number of flow channel mechanisms 212, which greatly saves a large number of samples for PCR amplification and detection as a whole; at the same time, diffusion contamination between different reaction chambers 214 is avoided, that is, amplification and detection have been completed.

[0061] In an alternative embodiment, continue to refer to Figure 2 、 Figure 3 and Figure 4 as shown; the first C-shaped plate 11 and the second C-shaped plate 12 both include a C-shaped short segment 01S, a C-shaped middle segment 01I, and a C-shaped long segment 01L that are sequentially and perpendicularly connected. The C-shaped short segment 01S and the C-shaped long segment 01L are parallel to each other; along the length direction Y of the chip 2, the length of the C-shaped short segment 01S is less than the length of the C-shaped long segment 01L; the C-shaped short segment 01S in the first C-shaped plate 11 is clamped with the C-shaped long segment 01L in the second C-shaped plate 12, and the C-shaped long segment 01L in the first C-shaped plate 11 is clamped with the C-shaped short segment 01S in the second C-shaped plate 12. The C-shaped short segment 01S in the first C-shaped plate 11, the C-shaped middle segment 01I in the first C-shaped plate 11, the C-shaped long segment 01L in the first C-shaped plate 11, the C-shaped short segment 01S in the second C-shaped plate 12, the C-shaped middle segment 01I in the second C-shaped plate 12, and the C-shaped long segment 01L in the second C-shaped plate 12 form a rectangular structure.

[0062] Specifically, the first C-shaped plate 11 and the second C-shaped plate 12 both include a C-shaped short segment 01S, a C-shaped middle segment 01I, and a C-shaped long segment 01L that are integrally connected in sequence; the C-shaped short segment 01S, the C-shaped middle segment 01I, and the C-shaped long segment 01L are all cuboids with the same thickness. The C-shaped short segment 01S and the C-shaped long segment 01L are parallel to each other, and the length extension direction is along the length direction Y of the chip 2; along the length direction Y of the chip 2, the length of the C-shaped short segment 01S is less than the length of the C-shaped long segment 01L. One end of the C-shaped short segment 01S and the C-shaped long segment 01L along their length directions are respectively perpendicularly connected to both ends of the C-shaped middle segment 01I. The length extension direction of the C-shaped middle segment 01I is along the width direction X of the chip 2. <00,00160>Specifically, one end of the C-shaped short segment 01S in the first C-shaped plate 11 that is far from its C-shaped middle segment 01I is movably clamped with one end of the C-shaped long segment 01L in the second C-shaped plate 12 that is far from its C-shaped middle segment 01I; one end of the C-shaped long segment 01L in the first C-shaped plate 11 that is far from its C-shaped middle segment 01I is movably clamped with one end of the C-shaped short segment 01S in the second C-shaped plate 12 that is far from its C-shaped middle segment 01I; the C-shaped short segment 01S in the first C-shaped plate 11, the C-shaped middle segment 01I in the first C-shaped plate 11, the C-shaped long segment '01L in the first C-shaped plate 11, the C-shaped short segment 01S in the second C-shaped plate 12, the C-shaped middle segment 01I in the second C-shaped plate 12, and the C-shaped long segment 01L in the second C-shaped plate 12 form a rectangular structure for clamping the chip.

[0064] With the above solution, the U-shaped long segments 01L and U-shaped short segments 01S of different lengths are such that the two splicing joints of the first U-shaped plate 11 and the second U-shaped plate 12 are not on the same parallel line along the width direction X of the chip 2, so that the clamping plate 1 can resist torsional shear stress to a certain extent on the clamping surface of the chip 2, enhancing the connection stability between the first U-shaped plate 11 and the second U-shaped plate 12.

[0065] In an alternative embodiment, continue to refer to Figure 2 , Figure 3 , Figure 4 and Figure 9 as shown, Figure 9 is a schematic diagram of the splicing structure of the first U-shaped plate, the second U-shaped plate and the metal strip provided by the present invention; at one end of the U-shaped short segment 01S in the first U-shaped plate 11 away from the U-shaped middle segment 01I in the first U-shaped plate 11, a rectangular first tab 011 is integrally provided, and rectangular first protrusions 0111 are respectively provided on both sides of the first tab 011 along the thickness direction Z of the chip 2; at one end of the U-shaped long segment 01L in the first U-shaped plate 11 away from the U-shaped middle segment 01I in the first U-shaped plate 11, a first fixing groove 012 is opened, the first fixing groove 012 is recessed towards the U-shaped middle segment 01I in the first U-shaped plate 11 along the length direction Y of the chip 2, and a first inner groove 0121 is opened corresponding to the inner surface of the U-shaped long segment 01 in the first U-shaped plate 11 along the thickness direction Z of the chip 2;

[0066] At one end of the U-shaped short segment 01S in the second U-shaped plate 12 away from the U-shaped middle segment 01I in the second U-shaped plate 12, a second tab 013 matching the first fixing groove 012 is integrally provided, and second protrusions 0131 matching the first inner groove 0121 are provided on both sides of the second tab 013 along the thickness direction Z of the chip 2, and the second tab 013 and the second protrusions 0131 are respectively clamped into the first fixing groove 012 and the first inner groove 0121;

[0067] At one end of the U-shaped long segment 01L in the second U-shaped plate 12 away from the U-shaped middle segment 01I in the second U-shaped plate 12, a second fixing groove 014 matching the first tab 011 is opened, and a second inner groove 0141 matching the first tab 011 is opened corresponding to the inner surface of the U-shaped long segment 01 in the second U-shaped plate 12 along the thickness direction Z of the chip 2, and the first tab 011 and the first protrusions 0111 are respectively clamped into the second fixing groove 014 and the second inner groove 0141.

[0068] Specifically, a first tab 011 is integrally provided at one end of the short ⌚-shaped section 01S in the first ⌚-shaped plate 11, distal from the middle ⌚-shaped section 01I. The first tab 011 is a rectangular body; the thickness of the first tab 011 along the thickness direction Z of the chip 2 is less than the thickness of the short ⌚-shaped section 01S in the first ⌚-shaped plate 11 along the thickness direction Z of the chip 2; the length direction of the first tab 011 is the length direction Y of the chip 2. Rectangular first protrusions 0111 are provided on both sides of the first tab 011 along the thickness direction Z of the chip 2. The first protrusion 0111 is a rectangular block. The combined thickness of the first tab 011 and the first protrusion 0111 along the thickness direction Z of the chip 2 is less than the thickness of the short ⌚-shaped section 01S in the first ⌚-shaped plate 11 along the thickness direction Z of the chip 2. A first fixing groove 012 is integrally formed at one end of the long ⌚-shaped section 01L of the first ⌚-shaped plate 11, distal from the middle ⌚-shaped section 01I, to mate with the first protrusion 011. The first fixing groove 012 is recessed toward the middle ⌚-shaped section 01I of the first ⌚-shaped plate 11 along the length direction Y of the chip 2. The length, width, and thickness of the first fixing groove 012 are identical to those of the first protrusion 011. The thickness of the first fixing groove 012 along the thickness direction Z of the chip 2 is less than the thickness of the long ⌚-shaped section 01L along the thickness direction Z of the chip 2. The length direction of the first fixing groove 012 is the length direction Y of the chip 2. First inner grooves 0121 are formed on both sides of the first fixing groove 012 along the thickness direction Z of the chip 2, mate with the first protrusion 0111. The combined thickness of the first inner groove 0121 and the first fixing groove 012 in the thickness direction Z of the chip 2 is smaller than the thickness of the N-shaped short segment 01S in the first N-shaped plate 11 along the thickness direction Z of the chip 2 .

[0069] A second tab 013 is integrally provided at one end of the short tab section 01S in the second ∘-shaped plate 12, distal from the middle ∘-shaped plate 01I. The second tab 013 is a rectangular body; its thickness along the thickness direction Z of the chip 2 is less than the thickness of the short tab section 01S in the second ∘-shaped plate 12 along the thickness direction Z of the chip 2; the length of the second tab 013 is along the length direction Y of the chip 2. Rectangular second protrusions 0131 are provided on either side of the second tab 013 along the thickness direction Z of the chip 2. The second protrusion 0131 is a rectangular block. The combined thickness of the second tab 013 and the second protrusion 0131 along the thickness direction Z of the chip 2 is less than the thickness of the short tab section 01S in the second ∘-shaped plate 12 along the thickness direction Z of the chip 2.

[0070] At one end of the middle C-shaped long section 01L of the second C-shaped plate 12 away from its C-shaped middle section 01I, a second fixing groove 014 that is engaged with the second tab 013 in a matching manner is integrally provided. The second fixing groove 014 is recessed from the C-shaped middle section 01I in the second C-shaped plate 12 along the length direction Y of the chip 2. The length, width, and thickness of the second fixing groove 014 are correspondingly the same as the length, width, and thickness of the second tab 013; the thickness of the second fixing groove 014 along the thickness direction Z of the chip 2 is smaller than the thickness of the C-shaped long section 01L along the thickness direction Z of the chip 2; the length direction of the second fixing groove 014 is the length direction Y of the chip 2. Second inner grooves 0141 that are engaged with the second protrusions 0131 are provided on both sides of the second fixing groove 014 along the thickness direction Z of the chip 2. The combined thickness of the second inner grooves 0141 and the second fixing groove 014 in the thickness direction Z of the chip 2 is smaller than the thickness of the C-shaped short section 01S in the second C-shaped plate 12 along the thickness direction Z of the chip 2.

[0071] Through the above solution, the C-shaped long section 01L of the first C-shaped plate 11 and the C-shaped short section 01S of the second C-shaped plate 12 are fixedly clamped through the second tab 013 and the first fixing groove 012, and the C-shaped short section 01S of the first C-shaped plate 11 and the C-shaped long section 01L of the second C-shaped plate 12 are fixedly clamped through the first tab 011 and the second fixing groove 014, which can further enhance the connection stability between the first C-shaped plate 11 and the second C-shaped plate 12.

[0072] In an optional embodiment, continue to refer to Figure 2 、 Figure 3 and Figure 9 as shown, Figure 9 is a schematic diagram of the splicing structure of the first C-shaped plate, the second C-shaped plate, and the metal strip provided by the present invention; this embodiment further includes a rectangular metal strip 14, and the material of the metal strip 14 is a ferromagnetic metal;

[0073] Third C-shaped grooves 112 are respectively provided on the sides of the C-shaped short section 01S in the first C-shaped plate 11 and the C-shaped long section 01L in the second C-shaped plate 12 away from the hollow part 13, and the third C-shaped grooves 112 are recessed toward the side close to the hollow part 13; the side of the first C-shaped groove 111 away from the hollow part 13 and the side of the third C-shaped groove 112 close to the hollow part 13 are not connected; fourth C-shaped grooves 122 are respectively provided on the sides of the C-shaped long section 01L in the first C-shaped plate 11 and the C-shaped short section 01S in the second C-shaped plate 12 away from the hollow part 13, and the fourth C-shaped grooves 122 are recessed toward the side close to the hollow part 13; the side of the second C-shaped groove 121 away from the hollow part 13 and the side of the fourth C-shaped groove 122 close to the hollow part are not connected;

[0074] There are two metal strips 14. After the short ∘-shaped section 01S in the first ∘-shaped plate 11 and the long ∘-shaped section 01L in the second ∘-shaped plate 12, and the short ∘-shaped section 01S in the second ∘-shaped plate 12 and the long ∘-shaped section 01L in the first ∘-shaped plate 11 are clamped together, one metal strip 14 is clamped to the third ∘-shaped groove 112, and the other metal strip 14 is clamped to the fourth ∘-shaped groove 122.

[0075] Specifically, a third n-shaped groove 112 is respectively provided on the side of the n-shaped short section 01S in the first n-shaped plate 11 and the side of the n-shaped long section 01L in the second n-shaped plate 12 away from the hollow portion 13, and the third n-shaped groove 112 is recessed toward the side close to the hollow portion 13; a fourth n-shaped groove 122 is respectively provided on the side of the n-shaped long section 01L in the first n-shaped plate 11 and the side of the n-shaped short section 01S in the second n-shaped plate 12 away from the hollow portion 13, and the fourth n-shaped groove 122 is recessed toward the side close to the hollow portion 13; the recessed directions of the third n-shaped groove 112 and the fourth n-shaped groove 122 are opposite. The lengths of the third n-shaped groove 112 and the fourth n-shaped groove 122 are equal; and the length extension directions of the third n-shaped groove 112 and the fourth n-shaped groove 122 are both in the length direction Y of the chip 2. Perpendicular to the length direction thereof, the cross-sectional shape of the third N-shaped groove 112 and the fourth N-shaped groove 122 is N-shaped.

[0076] Along the thickness direction Z of the chip 2, the orthographic projection of the third n-shaped groove 112 and the orthographic projection of the fourth n-shaped groove 122 are parallel to each other; the orthographic projections of the third n-shaped groove 112 and the fourth n-shaped groove 122 do not overlap with the orthographic projections of the first n-shaped groove 111 or the second n-shaped groove 121. When the first n-shaped plate 11 and the second n-shaped plate 12 are interlocked to form a rectangular structure, the third n-shaped groove 112 and the fourth n-shaped groove 122 can each engage a metal strip 14 on one side along the length direction Y of the chip 2.

[0077] The metal strip 14 is a rectangular ferromagnetic metal. Along the thickness direction Z of the chip 2, the width of the metal strip 14 is equal to the width of the third N-shaped groove 112 or the fourth N-shaped groove 122. The length of the metal strip 14 along the length direction Y of the chip 2 is equal to the length of the third N-shaped groove 112 or the fourth N-shaped groove 122 along the length direction Y of the chip 2.

[0078] Along the thickness direction Z of the chip 2, the cross-sectional shape of the short N-shaped segments 01S and the long N-shaped segments 01L in the first N-shaped plate 11 and the second N-shaped plate 12 is an I-shaped segment. Along the length direction Y of the chip 2, the cross-sectional shape of the middle N-shaped segments 01I in the first N-shaped plate 11 and the second N-shaped plate 12 is an I-shaped segment. The I-shaped short N-shaped segments 01S and the long N-shaped segments 01L facilitate secure engagement with both sides of the chip 2 along its thickness direction, while also facilitating engagement with the metal strip 14 on the side away from the chip 2.

[0079] Through the above solution, an invisible magnetic attraction structure is provided between the first C-shaped plate 11 and the second C-shaped plate 12. By engaging two metal strips 14 with the third C-shaped groove 112 and the fourth C-shaped groove 122, the first C-shaped plate 11 and the second C-shaped plate 12 are magnetically fixed in the PCR amplifier; the surface where the metal strips 14 are arranged is concealed and not easy to fall off; moreover, the rectangular metal strips 14 enable the clamping plate 1 to have a larger effective magnetic attraction area, and the adsorption is more stable; in addition, the metal strips 14 on both sides form a "connection bridge" at the clamping joint of the first C-shaped plate 11 and the second C-shaped plate 12, enhancing the clamping stability of the first C-shaped plate 11 and the second C-shaped plate 12.

[0080] In an alternative embodiment, continue to refer to Figure 2-Figure 8 As shown, on the side of the silica gel sheet 32 away from the water stop glue 35, a rectangular water-absorbing cotton 36 is adhesively bonded. The water-absorbing cotton 36 is provided with water-absorbing holes 361 corresponding to and matching the positioning holes 321 along the thickness direction Z of the chip 2. The shape of the water-absorbing holes 361 is circular, and along the thickness direction Z of the chip 2, the water-absorbing holes 361 communicate with the positioning holes 321.

[0081] Specifically, on the side of the silica gel sheet 32 away from the water stop glue 35, a rectangular water-absorbing cotton 36 is adhesively bonded; the material of the water-absorbing cotton 36 is pure cotton. The side of the water-absorbing cotton 36 away from the silica gel sheet 32 and the side of the housing 31 away from the base 22 are in the same plane. The length and width of the water-absorbing cotton 36 are equal to the length and width of the silica gel sheet 32. Along the thickness direction Z of the chip 2, the orthographic projection of the water-absorbing cotton 36 coincides with the orthographic projection of the silica gel sheet 32.

[0082] The water-absorbing cotton 36 is provided with n water-absorbing holes 361 corresponding to and matching n positioning holes 321 along the thickness direction Z of the chip 2. In this embodiment, 3 positioning holes 321 are provided, and 3 corresponding water-absorbing holes 361 are provided. The shape of the water-absorbing holes 361 is circular, and the radius of the water-absorbing holes 361 is not less than the radius of the positioning holes 321. The sampling needle sequentially passes through the water-absorbing holes 361 and the positioning holes 321 and pierces into the sampling hole 211.

[0083] Through the above solution, during the sampling process of the sampling needle, due to the too fast sampling speed, the water stop glue 35 may not have time to cure, and a large amount of sample liquid may reverse and leak out from the sealing cavity 34 of the first seal 3F and the second seal 3S through the silica gel sheet 32; the porous structure of the water-absorbing cotton 36 enables it to quickly absorb the reversely leaking sample liquid, effectively preventing excessive sample liquid from overflowing and causing dirt inside the PCR amplifier. And the water-absorbing cotton 36 made of pure cotton can absorb sample liquid several times its own volume. [[ID=IS]]

[0084] In an alternative embodiment, continue to refer to Figure 2-Figure 8 As shown, the water-absorbing cotton 36 is a water-color-changing water-absorbing cotton, and a water-color-changing agent is adhesively bonded to the side away from the upper bottom surface 311.

[0085] Specifically, when the absorbent cotton 36 is hydrochromic, the sample liquid that seeps back through the silicone sheet 32 ​​first flows out through the water absorption hole 361. Then, along the water absorption hole 361 as the center, the hydrochromic agent on the absorbent cotton 36 absorbs the water in the sample liquid, causing an instant color change. The hydrochromic agent is specifically bonded to the side of the absorbent cotton 36 away from its housing 31. The hydrochromic agent includes a color-changing cobalt complex salt, a sensitizer, and a color enhancer. The color-changing cobalt complex salt can be cobalt sulfate, which is light pink when dry and changes to a striking dark blue when exposed to water. The sensitizer can be hexamethylenetetramine, which enhances the color-changing effect of the color-changing cobalt complex salt. The color enhancer can be an azo dye, phthalocyanine dye, or the like. When mixed with the color-changing cobalt complex salt, the azo dye, phthalocyanine dye, and the color can be greatly enhanced.

[0086] Through the above scheme, in cases of excessive sampling and too fast sampling speed, the sample liquid reversely seeping out from the sealed cavity 34 can be immediately subjected to color detection; it is convenient to detect the sampling process, on the one hand to avoid contamination of the inside of the PCR amplifier, and on the other hand to avoid waste of sample liquid.

[0087] Example 2

[0088] Continue to refer to Figure 2 、 Figure 3 、 Figure 9 and Figure 10 As shown, Figure 10 This is a schematic structural diagram of another multi-primer response PCR sealed detection box provided by the present invention;

[0089] In this embodiment, along the length direction Y of the chip 2, the sum of the lengths of the short N-shaped segment 01S in the first N-shaped plate 11 and the long N-shaped segment 01L in the second N-shaped plate 12, and the sum of the lengths of the short N-shaped segment 01S in the second N-shaped plate 12 and the long N-shaped segment 01L in the first N-shaped plate 11 are both a, and the length of the N-shaped middle segment 01I along the width direction Y of the chip 2 is b, where a=2b;

[0090] A fifth ∘-shaped groove is respectively formed on the side of the ∘-shaped middle section 01I of the first ∘-shaped plate 11 away from the hollow portion 13 in each card plate 1, and the fifth ∘-shaped groove is recessed toward the side close to the hollow portion 13; the side of the first ∘-shaped groove 111 away from the hollow portion 13 and the side of the fifth ∘-shaped groove close to the hollow portion 13 are not connected; a sixth ∘-shaped groove is respectively formed on the side of the ∘-shaped middle section 01I of the second ∘-shaped plate 12 away from the hollow portion 13, and the sixth ∘-shaped groove is recessed toward the side close to the hollow portion 13; the side of the second ∘-shaped groove 121 away from the hollow portion 13 and the side of the sixth ∘-shaped groove close to the hollow portion are not connected;

[0091] When the number of the card boards 1 is two, the two card boards 1 are connected side by side along the length direction Y of the chip 2, the first metal strip 14 is engaged into the fifth U-shaped groove along the width direction X of the chip 2, and the second metal strip 14 is engaged into the sixth U-shaped groove along the width direction X of the chip 2.

[0092] Specifically, the length direction of the card board 1 is the length direction Y of the chip 2, the width direction of the card board 1 is the width direction X of the chip 2, the length of the card board 1 is the sum a of the length of the short U-shaped segment 01S in the first U-shaped plate 11 and the length of the long U-shaped segment 01L in the second U-shaped plate 12, and the width of the card board 1 is the length b of the middle U-shaped segment 01I; the length of the card board 1 is twice its width, a = 2b. After the two groups of card boards 1 are connected side by side along the length direction Y of the chip 2, the first metal strip 14 can be connected in series into the fifth U-shaped grooves on one side of the two groups of card boards 1 along the width direction X of the chip 2 at the same time along the width direction X of the chip 2, that is, one metal strip 14 is connected in series with the two groups of card boards 1 along the width direction of the card board 1 at the same time, and the combined width of the two groups of card boards 1 is equal to the length of the metal strip 14; the second metal strip 14 is connected in series into the sixth U-shaped grooves on the other side of the two groups of card boards 1 along the width direction X of the chip 2 at the same time along the width direction X of the chip 2, that is, the second metal strip 14 is connected in series with the two groups of card boards 1 on the side far from the previous metal strip 14 along the width direction of the card board 1 at the same time, and the combined width of the two groups of card boards 1 is equal to the length of the metal strip 14.

[0093] Through the above scheme, after the two groups of card boards 1 are connected side by side along their length directions, a square double-board can be formed, so that the two metal strips 14 for fixing a single card board 1 can magnetically fix the two groups of card boards 1 along their width extension directions in the PCR amplifier at the same time, enhancing the expandability of the number of detections of the card board 1 at the same time, and thus enhancing the practicability of this embodiment in the simultaneous detection of multiple samples; and the operation is convenient and fast, which is beneficial to reducing the operation time in the fixing process.

[0094] There is a relationship of mutual cooperation and functional support among the technical features of the present invention, that is, the technical solution is not a "simple superposition" of the technical features between multiple comparative documents, so the present invention does not fall into the situation of "simple superposition" in Chapter 4, Part II of the "Patent Examination Guidelines".

[0095] Although some specific embodiments of the present invention have been described in detail through examples, those skilled in the art should understand that the above examples are only for the purpose of illustration and not for the purpose of limiting the scope of the present invention. Those skilled in the art should understand that the above embodiments can be modified without departing from the scope and spirit of the present invention. The scope of the present invention is defined by the appended claims.

Claims

1. A multi-primer response PCR sealed detection kit, characterized in that: It includes a card board, a rectangular chip and a strip-shaped sealing member, wherein: The card plate includes a first N-shaped plate and a second N-shaped plate, wherein the N-shaped opening of the first N-shaped plate faces the N-shaped opening of the second N-shaped plate, and the N-shaped opening of the second N-shaped plate faces the first N-shaped plate, and the first N-shaped plate and the second N-shaped plate are spliced ​​together to form a rectangular structure, and the rectangular structure has a hollow portion that passes through the rectangular structure along the thickness direction of the chip, and the shape of the hollow portion is rectangular; Along the peripheral direction of the hollow portion, a first N-shaped groove is formed on a side of the first N-shaped plate close to the hollow portion and is recessed away from the hollow portion, and a second N-shaped groove is formed on a side of the second N-shaped plate close to the hollow portion and is recessed away from the hollow portion; the first N-shaped groove and the second N-shaped groove are connected and penetrated to form a rectangular groove; The chip, with its four edges inserted in the rectangular groove, comprises a cover sheet and a base that are overlapped and glued to each other along the thickness direction of the chip, the cover sheet is located above the base, and along the thickness direction of the chip, the orthographic projection of the hollow part overlaps with the orthographic projection of the cover sheet and the base; the base is provided with at least two flow channel mechanisms on one side close to the cover sheet, and at least two of the flow channel mechanisms extend along the length direction of the chip and are arranged along the width direction of the chip; each of the flow channel mechanisms is a groove opening toward the cover sheet, and each of the flow channel mechanisms is connected to at least two reaction chambers, at least two of the reaction chambers are arranged along the length direction of the chip, the reaction chambers are fan-shaped grooves, and each of the reaction chambers is provided with a specific primer and a reaction enzyme; the cover sheet is provided with an injection hole and an outlet hole at both ends of each flow channel mechanism along the length direction of the chip, and the injection hole and the outlet hole respectively pass through the cover sheet along the thickness direction of the chip, and the injection hole and the outlet hole are both circular holes; The seal is located on the side of the cover sheet away from the substrate, and includes a first seal corresponding to the sample inlet hole and a second seal corresponding to the sample outlet hole. Both the first seal and the second seal include a rectangular prism-shaped housing, a rectangular silica gel sheet, and a partition. The housing includes an upper bottom surface, a lower bottom surface, and side surfaces. The upper bottom surface and the lower bottom surface are two parallel and congruent rectangles. The side surfaces are composed of four rectangles. The long sides of each side surface are respectively parallel to the corresponding sides of the upper bottom surface and the lower bottom surface, and the short sides of each side surface are perpendicular to the upper bottom surface. The upper bottom surface, the lower bottom surface, and the four side surfaces together form a rectangular prism. A strip-shaped opening is provided on the upper bottom surface, and the strip-shaped opening penetrates the upper bottom surface along the thickness direction of the chip and extends along the width direction of the chip. The periphery of the silica gel sheet is adhesively bonded to one side of the four side surfaces away from the lower bottom surface. A partition is provided between the silica gel sheet and the lower bottom surface. The material of the partition is plastic, and the number of partitions is twice the number of the flow channel mechanisms. Part of the silica gel sheet, part of the side surface, and the adjacent two partitions form a sealing cavity. The sealing cavity in the first seal is communicated with the sample inlet hole, and the sealing cavity in the second seal is communicated with the sample outlet hole; The silica gel sheet is provided with a positioning hole that penetrates the silica gel sheet along the thickness direction of the chip, and the positioning hole is communicated with the sealing cavity. A water-stop glue is adhesively bonded to one side of the partition close to the sealing cavity. The material of the water-stop glue is polyurethane-type sealant. Along the thickness direction of the chip, the water-stop glue does not coincide with the central axis of the positioning hole, and the orthographic projection of the positioning hole is located within the orthographic projection of the sealing cavity.

2. The multi-primer response PCR sealed detection kit according to claim 1, characterized in that: Both the first U-shaped plate and the second U-shaped plate include a U-shaped short section, a U-shaped middle section, and a U-shaped long section that are sequentially and perpendicularly connected. The U-shaped short section and the U-shaped long section are parallel to each other. Along the length direction of the chip, the length of the U-shaped short section is less than the length of the U-shaped long section. The U-shaped short section in the first U-shaped plate is clamped with the U-shaped long section in the second U-shaped plate, and the U-shaped long section in the first U-shaped plate is clamped with the U-shaped short section in the second U-shaped plate. The U-shaped short section in the first U-shaped plate, the U-shaped middle section in the first U-shaped plate, the U-shaped long section in the first U-shaped plate, the U-shaped short section in the second U-shaped plate, the U-shaped middle section in the second U-shaped plate, and the U-shaped long section in the second U-shaped plate form the rectangular structure.

3. A multi-primer response PCR sealed detection kit according to claim 2, characterized in that: One end of the short C-shaped section of the first C-shaped plate, which is away from the middle C-shaped section of the first C-shaped plate, is integrally provided with a rectangular first tab. On both sides of the first tab along the thickness direction of the chip, rectangular first protrusions are respectively provided; one end of the long C-shaped section of the first C-shaped plate, which is away from the middle C-shaped section of the first C-shaped plate, is provided with a first fixing groove. The first fixing groove is recessed towards the middle C-shaped section of the first C-shaped plate along the length direction of the chip. Along the thickness direction of the chip, a first inner groove is provided corresponding to the inner surface of the long C-shaped section of the first C-shaped plate in the first fixing groove; One end of the short C-shaped section of the second C-shaped plate, which is away from the middle C-shaped section of the second C-shaped plate, is integrally provided with a second tab that matches the first fixing groove. On both sides of the second tab along the thickness direction of the chip, second protrusions that match the first inner groove are provided. The second tab and the second protrusions are respectively snapped into the first fixing groove and the first inner groove; One end of the long C-shaped section of the second C-shaped plate, which is away from the middle C-shaped section of the second C-shaped plate, is provided with a second fixing groove that matches the first tab. Along the thickness direction of the chip, a second inner groove that matches the first tab is provided corresponding to the inner surface of the long C-shaped section of the second C-shaped plate in the second fixing groove. The first tab and the first protrusions are respectively snapped into the second fixing groove and the second inner groove.

4. The multi-primer response PCR sealed detection kit according to claim 2, characterized in that: It further includes a rectangular metal strip, and the material of the metal strip is ferromagnetic metal; On one side of the short C-shaped section of the first C-shaped plate and the long C-shaped section of the second C-shaped plate that are away from the hollow part, third C-shaped grooves are respectively provided, and the third C-shaped grooves are recessed towards the side close to the hollow part; the side of the first C-shaped groove away from the hollow part and the side of the third C-shaped groove close to the hollow part are not connected; on one side of the long C-shaped section of the first C-shaped plate and the short C-shaped section of the second C-shaped plate that are away from the hollow part, fourth C-shaped grooves are respectively provided, and the fourth C-shaped grooves are recessed towards the side close to the hollow part; the side of the second C-shaped away from the hollow part and the side of the fourth C-shaped groove close to the hollow part are not connected; The number of the metal strips is two. After the short C-shaped section of the first C-shaped plate and the long C-shaped section of the second C-shaped plate, and the short C-shaped section of the second C-shaped plate and the long C-shaped section of the first C-shaped plate are mutually snapped, one metal strip is snapped into the third C-shaped groove, and the other metal strip is snapped into the fourth C-shaped groove.

5. The multi-primer response PCR sealed detection kit according to claim 2, characterized in that: It further includes a rectangular metal strip, and the material of the metal strip is ferromagnetic metal; Along the length direction of the chip, the sum of the lengths of the short C-shaped section of the first C-shaped plate and the long C-shaped section of the second C-shaped plate, and the sum of the lengths of the short C-shaped section of the second C-shaped plate and the long C-shaped section of the first C-shaped plate are both a. The length of the middle C-shaped section along the width direction of the chip is b, and a = 2b; In each of the card boards, a fifth U-shaped groove is respectively formed on one side of the U-shaped middle section of the first U-shaped plate背离 the hollow part, and the fifth U-shaped groove is recessed toward the side close to the hollow part; one side of the first U-shaped groove away from the hollow part and one side of the fifth U-shaped groove close to the hollow part are not connected; a sixth U-shaped groove is respectively formed on one side of the U-shaped middle section of the second U-shaped plate背离 the hollow part, and the sixth U-shaped groove is recessed toward the side close to the hollow part; one side of the second U-shaped groove away from the hollow part and one side of the sixth U-shaped groove close to the hollow part are not connected; The two card boards are connected side by side along the length direction of the chip, the first metal strip is clamped into the fifth U-shaped groove along the width direction of the chip, and the second metal strip is clamped into the sixth U-shaped groove along the width direction of the chip.

6. The multi-primer response PCR sealed detection kit according to claim 1, characterized in that: A rectangular water-absorbing cotton is adhesively bonded to the side of the silica gel sheet away from the water-stop glue, a water-absorbing hole corresponding to and matching the positioning hole is formed in the water-absorbing cotton along the thickness direction of the chip, the shape of the water-absorbing hole is circular, and the water-absorbing hole is communicated with the positioning hole along the thickness direction of the chip.

7. The multi-primer response PCR sealed detection kit according to claim 6, characterized in that: The water-absorbing cotton is water-induced color-changing water-absorbing cotton, and a water-induced color-changing agent is adhesively bonded to the side of the water-absorbing cotton away from the upper bottom surface.

8. The multi-primer response PCR sealed detection kit according to claim 6, characterized in that: The material of the water-absorbing cotton is pure cotton.

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