Digital MIRA instant diagnosis chip and analysis device

Abstract

The utility model belongs to the technical field of micro-fluidic chips, and particularly provides a digital MIRA real-time diagnosis chip and an analysis device.The digital MIRA real-time diagnosis chip comprises a top glass slide, a micro-pattern layer and a bottom glass slide which are sequentially arranged from top to bottom; a water-soluble film is embedded in a position where the micro-pattern layer is matched with the water filling nozzle; the sample injection port is suitable for sequentially injecting a reaction reagent and sealing oil. Through the design of a chip structure without an outlet, when reaction reagents are introduced from a sample injection port, all the reaction reagents simultaneously, equivalently and uniformly enter each micro-chamber, and the reaction reagents are separated into each micro-chamber by adopting an oil seal mode, so that the chip has the advantage of 100% sample discretization in the digital nucleic acid detection process, and no sample or reagent is lost. Meanwhile, each part of reaction reagent is accurately separated into each micro-chamber and cannot be pushed out of the chip, so that each molecule enters the corresponding micro-chamber, and the detection requirement of a low-concentration template is met.

Description

Technical Field

[0001] The utility model belongs to the technical field of microfluidic chips, and in particular relates to a digital MIRA instant diagnosis chip and an analysis device. Background Art

[0002] Point-of-Care Testing (POCT) refers to a testing method performed at the sampling site using portable analytical instruments and supporting reagents to rapidly obtain test results. Key criteria for POCT are that it does not require a fixed testing location and can produce results in a simple, step-by-step process. In recent years, rapid technological advancements and a fast-paced, efficient work style have made POCT, with its miniaturized instruments, simplified procedures, and immediate results, increasingly popular in various medical settings.

[0003] POCT testing requires the use of a digital MIRA instant diagnostic chip, which is a microfluidic chip that integrates sample pretreatment and multiple digital recombinase polymerase amplification detection. The design of this chip allows the entire process from sample distribution to nucleic acid amplification and detection to be completed on a single platform.

[0004] In conventional digital MIRA instant diagnostic chips, when a reagent is introduced, only a portion enters the microchamber for amplification. The portion remaining in the microchannel is pushed out of the chip, resulting in reagent loss. If the reagent contains only two molecules, there's a chance that none of them will enter the microchamber. Therefore, conventional digital nucleic acid detection chips are inadequate for low-concentration template detection.

[0005] How to avoid the waste of samples in digital MIRA instant diagnostic chips to meet the detection needs of low-concentration templates is a technical problem that needs to be solved urgently.

[0006] It should be noted that the above information disclosed in this background technology section is only used to understand the background technology of the present application concept, and therefore, the above description is not considered to constitute information of the prior art. Utility Model Content

[0007] The embodiments of the present disclosure at least provide a digital MIRA instant diagnostic chip and an analysis device.

[0008] In a first aspect, an embodiment of the present disclosure provides a digital MIRA instant diagnostic chip, which is provided with a top glass slide, a micro-pattern layer, and a bottom glass slide in order from top to bottom;

[0009] The micro-pattern layer is in communication with the sample injection port and the water injection port of the top glass slide;

[0010] A water-soluble film is embedded in the matching position between the micro-pattern layer and the water injection port;

[0011] The injection port is suitable for injecting reaction reagents and sealing oil in sequence.

[0012] In an optional embodiment, the micro-pattern layer includes a flow channel module, a micro-chamber array module, and a vacuum storage battery module;

[0013] The microchamber array module and the vacuum storage battery module are arranged side by side;

[0014] One end of the flow channel module is connected to the sample injection port of the top slide, and the other end is connected to the microchamber array module;

[0015] The water-soluble film is disposed above the vacuum storage battery module.

[0016] In an optional embodiment, the microchamber array module includes a plurality of microchambers with fractal symmetric structures.

[0017] In an optional embodiment, the flow channel module is a T-shaped branch channel network, and the end of each branch channel is connected to the corresponding microchamber;

[0018] Furthermore, the length of each branch channel from the sample injection port to the microchannel of the microchamber is the same.

[0019] In an optional embodiment, the vacuum storage battery module includes: multiple stages of batteries connected in series;

[0020] The adjacent batteries are independent and not connected, and a thin wall is provided between the adjacent batteries, and is suitable for discharging the gas in the space where the batteries are located from the thin wall into the flow channel module and out of the chip, so as to form a vacuum in the space where the batteries are located;

[0021] The water-soluble film is disposed above the battery adjacent to the microchamber array module.

[0022] In an optional embodiment, the sample injection port and the water injection port of the top glass slide are connected to a liquid storage tank and a water storage tank respectively.

[0023] In an optional embodiment, the top glass slide and the bottom glass slide are both made of an incompressible, airtight, and transparent material with good thermal conductivity.

[0024] In an optional embodiment, the micro-pattern layer is made of a compressible transparent material with a loose and porous interior.

[0025] In a second aspect, an embodiment of the present disclosure further provides an analysis device, comprising:

[0026] The device itself, the smart terminal, and the digital MIRA instant diagnostic chip as described above;

[0027] A chip fixing frame and a coaxial light source are provided inside the device body;

[0028] The top of the device body is provided with a fixing groove suitable for fixing the smart terminal;

[0029] The digital MIRA instant diagnostic chip is placed on the chip holder;

[0030] The smart terminal is fixed on the fixing slot and is arranged directly above the coaxial light source;

[0031] The beneficial effect of this utility model is that the digital MIRA instant diagnostic chip and analysis device utilizes a chip structure with no outlet. When reagents are introduced through the sample injection port, all reagents enter each microchamber simultaneously, in equal amounts, and evenly. Oil seals are used to separate the reagents within each microchamber. This provides the advantage of 100% sample discretization during digital nucleic acid testing, eliminating any sample or reagent loss. Furthermore, because each reagent is precisely separated into its own microchamber, it is prevented from being pushed out of the chip, ensuring that each molecule enters its corresponding microchamber, thus meeting the detection requirements of low-concentration templates.

[0032] Other features and advantages of the present invention will be described in the following description, and in part will become apparent from the description, or understood by practicing the present invention. The objectives and other advantages of the present invention are realized and obtained by the structures particularly pointed out in the description, claims and drawings.

[0033] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, preferred embodiments are specifically cited herein and described in detail with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the specific implementation methods of the utility model or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the utility model. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0035] Figure 1 This is a structural diagram of the digital MIRA instant diagnostic chip provided in an embodiment of the present disclosure.

[0036] Figure 2 A schematic structural diagram of a micro-pattern layer provided in an embodiment of the present disclosure.

[0037] Figure 3 A schematic diagram of a partial structure of an analysis device provided in an embodiment of the present disclosure.

[0038] Figure 4 Schematic diagram of the injection of reaction reagents and sealing oil into the microchamber provided in an embodiment of the present disclosure.

[0039] Figure 5 Schematic diagram of the oil seal provided in an embodiment of the present disclosure.

[0040] Figure 6 This is a schematic diagram of the principle of implementing the anti-evaporation of deionized water provided in an embodiment of the present disclosure.

[0041] In the figure: 110, bottom glass slide; 120, micro-pattern layer; 121, flow channel module; 122, microchamber array module; 123, vacuum storage battery module; 130, water-soluble film; 140, top glass slide; 141, sample injection port; 142, water injection port; 151, liquid storage tank; 152, water storage tank; 2, device body; 21, chip fixing frame; 22, coaxial light source; 23, fixing groove. DETAILED DESCRIPTION

[0042] To make the purpose, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0043] Glossary:

[0044] Multienzyme Isothermal Rapid Amplification (MIRA) technology is a constant-temperature rapid nucleic acid amplification technology that relies on the synergistic action of multiple functional proteins (helicase, recombinase, single-strand binding protein, DNA polymerase, etc.) to achieve rapid nucleic acid amplification at room temperature.

[0045] The following embodiments of the present invention are described in detail with reference to the accompanying drawings. In the absence of conflict, the following embodiments and features in the embodiments may be combined with each other.

[0046] See also Figure 1 , Figure 1The structure of the digital MIRA instant diagnostic chip is shown. The chip comprises, from top to bottom, a top glass slide 140, a micropatterned layer 120, and a bottom glass slide 110. The micropatterned layer 120 communicates with the sample injection port 141 and the water injection port 142 of the top glass slide 140. A water-soluble film 130 is embedded between the micropatterned layer 120 and the water injection port 142. The sample injection port 141 is suitable for sequentially injecting reagents and sealing oil. Thanks to the chip's outlet-free design, when reagents are introduced through the sample injection port 141, all reagents enter each microchamber simultaneously, in equal amounts, and evenly. Oil seals are used to separate the reagents within each microchamber, resulting in 100% sample discretization during digital nucleic acid testing, eliminating sample and reagent loss. Furthermore, because each reagent is precisely separated into its own microchamber, it cannot be pushed out of the chip, ensuring that each molecule enters its corresponding microchamber, thus meeting the detection requirements of low-concentration templates.

[0047] The micro-pattern layer 120 and the bottom glass slide 110 are sealed by using a heat-resistant transparent tape, heat bonding, heat-resistant adhesive bonding, hot pressing sealing, or air plasma treatment bonding.

[0048] Please continue reading Figure 1 In some embodiments, the sample injection port 141 and the water injection port 142 of the top glass slide 140 are respectively connected to a liquid storage tank 151 and a water storage tank 152. The liquid storage tank 151 and the water storage tank 152 are sealed to the body of the top glass slide 140 by using heat-resistant transparent tape, heat bonding, heat-resistant adhesive bonding, hot pressing sealing, or air plasma bonding.

[0049] The top glass slide 140 and the bottom glass slide 110 are both made of an incompressible, airtight, and thermally conductive transparent material, such as high-temperature-resistant glass. When heating the digital MIRA point-of-care diagnostic chip, the top and bottom glass slides 140 and 110 conduct heat to the reagents within the microchamber array module 122, enabling controlled heating and cooling. The micropatterned layer 120 is made of a compressible, porous, transparent material, such as polydimethylsiloxane.

[0050] See also Figure 2The structure of the micropatterned layer 120 is described below. In some embodiments, the micropatterned layer 120 includes a flow channel module 121, a microchamber array module 122, and a vacuum storage battery module 123. The microchamber array module 122 and the vacuum storage battery module 123 are arranged side by side. One end of the flow channel module 121 is connected to the injection port 141 of the top glass slide 140, and the other end is connected to the sample inlet of the microchamber array module 122. The water-soluble film 130 is disposed above the vacuum storage battery module 123. Reaction reagents flow through the injection port 141 of the top glass slide 140, through the flow channel module 121, and then into the microchamber array module 122 for sample dispersion. The vacuum storage battery module 123 provides driving force for the reaction reagents.

[0051] Specifically, the microchamber array module 122 comprises multiple microchambers with fractal symmetry. The flow channel module 121 comprises a T-shaped branch channel network, with each branch channel end connected to a corresponding microchamber; and each branch channel has the same length from the injection port 141 to the microchamber. The fractal structure of the T-shaped branch channel network facilitates chip expansion. Furthermore, each branch channel has the same length from the injection port 141 to the microchamber, ensuring uniform distribution of reaction reagents and template molecules.

[0052] For example, when the number of branch channels is 10, the number of microchambers can reach 1024. When the number of branch channels reaches 20, the total number of microchambers can reach 1,048,576. Compared with the million-pixel digital nucleic acid detection chip, the digital MIRA instant diagnostic chip provided in this embodiment is easier to apply to millions of single-molecule amplifications without any complex control system.

[0053] It should be noted that the detection range can be further adjusted for different pathogens by designing different micropore sizes. The total detection time can be shortened to 25 minutes (5 minutes for sample separation + 20 minutes for isothermal amplification), which is lower in cost and faster in readout than digital nucleic acid detection instruments in related technologies.

[0054] Please continue reading Figure 2In some embodiments, the vacuum storage battery module 123 includes: multiple stages of batteries connected in series; adjacent batteries are independent and not connected, and thin walls are provided between adjacent batteries, which are suitable for discharging gas in the battery space through the thin walls into the flow channel module 121 and out of the chip, thereby forming a vacuum in the battery space; the water-soluble film 130 is disposed above the battery adjacent to the microchamber array module 122. After deionized water is injected into the water inlet 142, the sacrificial valve of the water-soluble film 130 dissolves upon contact with the water, becoming permanently open. Under the action of the vacuum in the battery space, the deionized water fills the space where the battery is connected to the water-soluble film 130. This prevents the reaction reagent from being heated and converted into gas during the amplification process, and then entering the vacuum battery through the thin walls surrounding the microchamber, effectively preventing the reaction reagent from evaporating and ensuring the normal progress of the amplification reaction.

[0055] like Figure 3 As shown, at least one embodiment further provides an analysis device, comprising: a device body 2, an intelligent terminal, and a digital MIRA instant diagnostic chip as described above; a chip holder 21 and a coaxial light source 22 are provided inside the device body 2; a fixing groove 23 suitable for fixing the intelligent terminal is provided on the top of the device body 2; the digital MIRA instant diagnostic chip is placed on the chip holder 21; the intelligent terminal is fixed on the fixing groove 23 and is arranged directly above the coaxial light source 22; the chip holder 21 is arranged directly below the coaxial light source 22. The coaxial light source 22 ensures the uniformity and consistency of the excitation light during the image acquisition process. When performing fluorescence imaging, the intelligent terminal is vertically fixed directly above the coaxial light source 22, and the digital MIRA instant diagnostic chip is placed directly below the coaxial light source 22.

[0056] It should be noted that the analysis device can operate for a long time through a 30V power supply provided by an external adapter.

[0057] The working process of the analysis device is as follows:

[0058] S110: Place the digital MIRA instant diagnostic chip in a vacuum environment and degas at a preset negative pressure for 2-6 hours.

[0059] Specifically, during the degassing process, air from the series-connected cells enters the fluid channels through the porous thin walls inside and is discharged from the chip, thereby creating a vacuum within the battery. The preset negative pressure is -80kPa.

[0060] S120: Open the vacuum packaging and take out the digital MIRA instant diagnostic chip.

[0061] S130: Add reaction reagents and sealing oil to the liquid storage tank 151 in sequence. Under the action of vacuum and atmospheric pressure, the reaction reagents enter the microchannel quickly in the order of reaction reagents in front and sealing oil in the back. After passing through N fractal structures, the reagents will fill each microchamber equally and evenly, and the reaction reagents will be separated into each microchamber by the sealing oil.

[0062] Specifically, the schematic diagram of the reaction reagents and sealing oil entering the microchamber is as follows: Figure 4 The principle diagram of the oil seal is shown in Figure 5 shown.

[0063] S140 : Deionized water is injected into the water storage tank 152 to dissolve the water-soluble film 130 , so that the deionized water fills part of the batteries in the vacuum storage battery module 123 .

[0064] like Figure 6 Specifically, after deionized water is injected into the water inlet 142, the sacrificial valve of the water-soluble film 130 dissolves upon contact with the water, becoming permanently open. Under the influence of the vacuum in the cell space, the deionized water fills the space where the cell is located, which is connected to the water-soluble film 130. This prevents the reaction reagents from being heated and converted into gas during the amplification process, which then enters the vacuum cell through the thin walls surrounding the microchamber, effectively preventing the reaction reagents from evaporating and ensuring the normal progress of the amplification reaction.

[0065] S150: placing the digital MIRA instant diagnostic chip on the heating pack to perform a digital amplification reaction.

[0066] S160: The amplified digital MIRA instant diagnostic chip is placed on the chip holder 21 of the analysis device for analysis.

[0067] In summary, the present invention provides a digital MIRA instant diagnostic chip and analysis device. The digital MIRA instant diagnostic chip comprises, from top to bottom, a top glass slide 140, a micropatterned layer 120, and a bottom glass slide 110. The micropatterned layer 120 communicates with the sample injection port 141 and water injection port 142 of the top glass slide 140. A water-soluble film 130 is embedded at the interface between the micropatterned layer 120 and the water injection port 142. The sample injection port 141 is suitable for sequentially injecting reaction reagents and sealing oil. Through the chip's outlet-free structural design, when reaction reagents are introduced through the sample injection port 141, all reaction reagents enter each microchamber simultaneously, in equal amounts, and evenly. Oil seals are then used to separate the reaction reagents into the microchambers. This provides the advantage of 100% sample discretization during digital nucleic acid testing, preventing any sample or reagent loss. At the same time, since each reaction reagent is precisely separated into each microchamber, it will not be pushed out of the chip, so that each molecule enters the corresponding microchamber, thus meeting the detection needs of low-concentration templates.

[0068] Based on the above-mentioned ideal embodiment of the present invention, and in accordance with the above description, relevant personnel can make various changes and modifications without departing from the technical scope of the present invention. The technical scope of the present invention is not limited to the content of the specification, but must be determined according to the scope of the claims.

Claims

1. A digital MIRA instant diagnostic chip, characterized in that: From top to bottom, a top glass slide, a micro-pattern layer, and a bottom glass slide are sequentially arranged; The micro-pattern layer is in communication with the sample injection port and the water injection port of the top glass slide; A water-soluble film is embedded in the matching position between the micro-pattern layer and the water injection port; The injection port is suitable for injecting reaction reagents and sealing oil in sequence.

2. The digital MIRA instant diagnostic chip according to claim 1, wherein: The micro-pattern layer includes a flow channel module, a micro-chamber array module and a vacuum storage battery module; The microchamber array module and the vacuum storage battery module are arranged side by side; One end of the flow channel module is connected to the sample injection port of the top slide, and the other end is connected to the sample inlet of the microchamber array module; The water-soluble film is disposed above the vacuum storage battery module.

3. The digital MIRA instant diagnostic chip according to claim 2, wherein: The microchamber array module includes a plurality of microchambers with fractal symmetric structures.

4. The digital MIRA instant diagnostic chip according to claim 3, wherein: The flow channel module is a T-shaped branch channel network, and the end of each branch channel is connected to the corresponding microchamber; Furthermore, the length of each branch channel from the sample injection port to the microchannel of the microchamber is the same.

5. The digital MIRA instant diagnostic chip according to claim 2, wherein: The vacuum storage battery module includes: multiple stages of batteries connected in series; The adjacent batteries are independent and not connected, and a thin wall is provided between the adjacent batteries, and is suitable for discharging the gas in the space where the batteries are located from the thin wall into the flow channel module and out of the chip, so as to form a vacuum in the space where the batteries are located; The water-soluble film is disposed above the battery adjacent to the microchamber array module.

6. The digital MIRA instant diagnostic chip according to claim 1, wherein: The sample injection port and the water injection port of the top glass slide are respectively connected to a liquid storage tank and a water storage tank.

7. The digital MIRA instant diagnostic chip according to claim 1, wherein: The top glass slide and the bottom glass slide are both made of an incompressible, airtight, and transparent material with good thermal conductivity.

8. The digital MIRA instant diagnostic chip according to claim 1, wherein: The micro-pattern layer is made of a compressible transparent material with loose and porous interior.

9. An analysis device, characterized in that include: Device body, intelligent terminal and digital MIRA instant diagnostic chip according to any one of claims 1 to 8; A chip fixing frame and a coaxial light source are provided inside the device body; The top of the device body is provided with a fixing groove suitable for fixing the smart terminal; The digital MIRA instant diagnostic chip is placed on the chip holder; The smart terminal is fixed on the fixing slot and is arranged directly above the coaxial light source; The chip fixing frame is arranged directly below the coaxial light source.

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