A nucleic acid detection kit based on a negative pressure system

Abstract

The application discloses a nucleic acid detection kit based on a negative pressure system, which comprises a sample injection liquid cavity, a nucleic acid amplification cavity and a liquid-stopping ventilation cavity which are sequentially connected, the sample injection liquid cavity is internally provided with a nucleic acid lysis and extraction liquid, and the nucleic acid amplification cavity is internally provided with a nucleic acid amplification reagent, wherein the nucleic acid amplification reagent is a solid reagent; the kit further comprises a negative pressure cavity which is in communication with the liquid-stopping ventilation cavity, a negative pressure opening device is arranged at the communication position of the negative pressure cavity and the liquid-stopping ventilation cavity, a sample injection switch is arranged at the connecting position of the sample injection liquid cavity and the nucleic acid amplification cavity, and the sample injection liquid cavity, the nucleic acid amplification cavity, the liquid-stopping ventilation cavity and the negative pressure cavity form a closed system, so that the whole reaction system can be formed into a negative pressure, the sample injection liquid can be rapidly injected, the freeze-dried reagent can be dissolved in a negative pressure mode, the reaction speed is improved, the reaction time is shortened, and the detection efficiency is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of in vitro diagnostic devices, in particular to a nucleic acid detection kit based on a negative pressure system. BACKGROUND

[0002] In vitro diagnostic (IVD) technology refers to obtaining relevant clinical diagnostic information by detecting samples such as blood, body fluid and tissue outside the human body, so as to help judge diseases or body functions. The IVD field involves a very wide range, including blood testing, clinical biochemistry, cell diagnosis, microbiological diagnosis, coagulation diagnosis, immunochemistry, molecular diagnosis, point-of-care testing (POCT) and the like. The IVD with the greatest development potential and the fastest growth at present refers to molecular diagnosis and point-of-care testing (POCT) technology based on nucleic acid detection.

[0003] In recent years, with the progress of science and technology, advanced nucleic acid detection integrated, fully automated POCT reagent kits and supporting equipment have gradually appeared in the industry, such as Xpert products of Cypheid, filmarray products of Biofire, and Vivalytic products of Bosch. These products all use external power to operate the components in the reagent kit to control the flow of various liquids in the reagent kit, so as to realize the automation of the whole process of nucleic acid sample lysis, extraction, amplification and detection. However, the existing nucleic acid detection reagent kit is automatically flowed into the amplification chamber for detection by the action of liquid gravity after lysis and extraction of the sample. The liquid after lysis and extraction of the sample dissolves the lyophilized reagent after entering the amplification chamber, and then realizes the amplification reaction. However, relying on gravity to fall into the amplification chamber for dissolution, the lyophilized ball dissolves slowly, which increases the amplification time and affects the detection efficiency. SUMMARY

[0004] In view of the above problems, the present application provides a nucleic acid detection kit based on a negative pressure system, which has the advantages of improving liquid flow rate, accelerating reaction time and improving detection efficiency.

[0005] The technical scheme of the present application is as follows:

[0006] A nucleic acid detection kit based on a negative pressure system comprises a sample injection liquid cavity, a nucleic acid amplification cavity and a liquid-stopping ventilation cavity connected in sequence, the sample injection liquid cavity is internally provided with a nucleic acid lysis extraction liquid, the nucleic acid amplification cavity is internally provided with a nucleic acid amplification reagent, the nucleic acid amplification reagent is a solid reagent, the kit further comprises a negative pressure cavity, the negative pressure cavity is communicated with the liquid-stopping ventilation cavity, a negative pressure opening device is arranged at the communication position of the negative pressure cavity and the liquid-stopping ventilation cavity, a sample injection switch is arranged at the connecting position of the sample injection liquid cavity and the nucleic acid amplification cavity, and the sample injection liquid cavity, the nucleic acid amplification cavity, the liquid-stopping ventilation cavity and the negative pressure cavity form a closed system.

[0007] The kit further comprises a box body and a reagent card capable of relatively sliding, the sample injection liquid cavity and the negative pressure cavity are arranged in the box body, the nucleic acid amplification cavity and the liquid-stopping ventilation cavity are arranged on the reagent card, and the negative pressure opening device and the sample injection switch are on the reagent card.

[0008] A first puncture membrane is arranged at the outlet of the sample injection liquid cavity, a second puncture membrane is arranged at the outlet of the negative pressure cavity, the sample injection switch is a first puncture needle corresponding to the first puncture membrane, and the negative pressure opening device is a second puncture needle corresponding to the second puncture membrane.

[0009] The top height of the first puncture needle is lower than that of the second puncture needle.

[0010] An adding port is arranged at the end of the sample injection liquid cavity away from the nucleic acid amplification cavity, and an easy-to-rip membrane is arranged at the adding port.

[0011] The nucleic acid amplification cavity is internally provided with a nucleic acid amplification reagent, and the nucleic acid amplification reagent is a solid reagent.

[0012] The liquid-stopping ventilation cavity is internally provided with a liquid-stopping ventilation device, and the liquid-stopping ventilation device is a PE sintered filter element.

[0013] The kit has the following beneficial effects:

[0014] 1. The kit is provided with a negative pressure cavity, and the whole reaction system can be formed into a negative pressure through a negative pressure opening device, so that the sample injection liquid can be rapidly injected, the freeze-dried reagent can be dissolved through the negative pressure, the reaction speed is improved, the reaction time is shortened, and the detection efficiency is improved.

[0015] 2. The kit comprises a reagent card and a box body capable of relatively sliding, the sample injection liquid cavity and the negative pressure cavity can be opened through the sliding mode, the controllability of the detection process can be improved, and the safety and reliability of use can be improved.

[0016] 3. The puncture membranes are arranged at the outlets of the sample injection liquid cavity and the negative pressure cavity, the sample injection switch and the negative pressure opening device are both puncture needles, and the communication is realized through the puncture mode, so that the reliability of the communication is improved.

[0017] 4、The first puncture needle and the second puncture needle have a height difference, which can ensure that negative pressure is formed in the reagent card first during use, and the solution after lysis and extraction can quickly enter the amplification cavity, and the safety of use can be improved;

[0018] 5、The sample liquid cavity, the amplification cavity, the gas-liquid separation cavity and the negative pressure cavity form a closed system as a whole, which can prevent aerosol from being generated, can achieve closed and pollution-free throughout, and can improve the safety of use.

[0019] 6、The liquid-stopping ventilation device is a PE sintered filter element, which has a simple structure and can effectively prevent solution overflow, thereby improving the safety and reliability of use. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 is the overall structure schematic diagram of the nucleic acid detection kit based on the negative pressure system according to the embodiment of the present application;

[0021] Figure 2 is the structure schematic diagram of the reagent card of the nucleic acid detection kit based on the negative pressure system according to the embodiment of the present application;

[0022] Figure 3 is the exploded structure schematic diagram of the nucleic acid detection kit based on the negative pressure system according to the embodiment of the present application;

[0023] Figure 4 is the perspective view of the nucleic acid detection kit based on the negative pressure system according to the embodiment of the present application;

[0024] Figure 5 is the front view of the nucleic acid detection kit based on the negative pressure system according to the embodiment of the present application;

[0025] Figure 6 is the rear view of the nucleic acid detection kit based on the negative pressure system according to the embodiment of the present application.

[0026] BRIEF DESCRIPTION OF DRAWINGS

[0027] 1 is a sample liquid cavity, 2 is a nucleic acid amplification cavity, 3 is a liquid-stopping ventilation cavity, 4 is a negative pressure cavity, 5 is a box body, 6 is a reagent card, 7 is a first puncture membrane, 8 is a second puncture membrane, 11 is a nucleic acid lysis and extraction liquid, 12 is a sample injection switch, 13 is a sample injection port, 21 is a nucleic acid amplification reagent, 31 is a liquid-stopping ventilation device, 41 is a negative pressure opening device, 121 is a first puncture needle, and 411 is a second puncture needle. DETAILED DESCRIPTION

[0028] The embodiments of the present application will be further described below with reference to the drawings.

[0029] EMBODIMENT:

[0030] AsFigures 1-6 As shown, a nucleic acid detection kit based on a negative pressure system comprises a sample injection liquid chamber 1, a nucleic acid amplification chamber 2 and a liquid-stopping ventilation chamber 3 connected in sequence, the sample injection liquid chamber 1 is internally provided with a nucleic acid lysis extraction liquid 11, the nucleic acid amplification chamber 2 is internally provided with a nucleic acid amplification reagent 21, the nucleic acid amplification reagent 21 is a solid reagent, the kit further comprises a negative pressure chamber 4, the negative pressure chamber 4 is in communication with the liquid-stopping ventilation chamber 3, the negative pressure chamber 4 is provided with a negative pressure opening device 41 at the communication position with the liquid-stopping ventilation chamber 3, the sample injection liquid chamber 1 is provided with a sample injection switch 12 at the connection position with the nucleic acid amplification chamber 2, and the sample injection liquid chamber 1, the nucleic acid amplification chamber 2, the liquid-stopping ventilation chamber 3 and the negative pressure chamber 4 connected in sequence form a closed system.

[0031] The working principle of the above technical solution is as follows:

[0032] The sample injection liquid chamber 1, the nucleic acid amplification chamber 2, the liquid-stopping ventilation chamber 3 and the negative pressure chamber 4 connected in sequence form a closed system, the sample injection liquid chamber 1 is provided with the sample injection switch 12 and internally provided with the nucleic acid lysis extraction liquid 11, the nucleic acid amplification chamber 2 is internally provided with the solid nucleic acid amplification reagent 21, the negative pressure chamber 4 is provided with the negative pressure opening device 41 between the negative pressure chamber 4 and the liquid-stopping ventilation chamber 3, by opening the negative pressure opening device 41, a negative pressure is formed in the closed system, then the sample injection switch 12 is opened, the sample injection into the nucleic acid amplification chamber 2 can be quickly realized, the dissolution of the nucleic acid amplification reagent 21 can be accelerated, the reaction speed is improved, the reaction time is shortened, and the detection efficiency is improved.

[0033] The kit further comprises a box body 5 and a reagent card 6 capable of relatively sliding, the sample injection liquid chamber 1 and the negative pressure chamber 4 are arranged in the box body 5, the nucleic acid amplification chamber 2 and the liquid-stopping ventilation chamber 3 are arranged on the reagent card 6, and the negative pressure opening device 41 and the sample injection switch 12 are arranged on the reagent card 6.

[0034] The reagent card 6 is internally provided with a microfluidic structure, the reagent card 6 and the box body 5 can relatively slide, the sample injection liquid chamber 1 and the negative pressure chamber 4 are arranged in the box body 5, the negative pressure opening device 41 and the sample injection switch 12 are arranged on the reagent card 6, the sample injection liquid chamber 1 and the negative pressure chamber 4 can be opened through the sliding mode, the controllability of the detection process can be improved, and the safety and reliability of use can be improved.

[0035] The first puncture membrane 7 is arranged at the outlet of the sample injection liquid chamber 1, the second puncture membrane 8 is arranged at the outlet of the negative pressure chamber 4, the sample injection switch 12 is a first puncture needle 121 corresponding to the first puncture membrane 7, and the negative pressure opening device 41 is a second puncture needle 411 corresponding to the second puncture membrane 8.

[0036] The puncture membranes are arranged at the outlets of the sample injection liquid chamber 1 and the negative pressure chamber 4, the sample injection switch 12 and the negative pressure opening device 41 are both puncture needles, the cavity communication can be realized through the puncture mode, and the reliability of the communication can be improved.

[0037] The top height of the first puncture needle 121 is lower than the top height of the second puncture needle 411.

[0038] The first puncture needle 121 and the second puncture needle 141 have a height difference, which can ensure that a negative pressure is formed in the reagent card 6 during use, and the solution after lysis and extraction can quickly enter the nucleic acid amplification cavity 2, thereby improving the reliability of use.

[0039] The end of the sample liquid cavity 1 away from the nucleic acid amplification cavity 2 is provided with a sample port 13, and the sample port 13 is provided with a peel-off film.

[0040] The peel-off film can seal the sample port 13, prevent the safety and reliability of the nucleic acid lysis solution extraction liquid 11 inside the sample liquid cavity 1, and facilitate the safety and reliability of the equipment during transportation.

[0041] The liquid-stopping and air-passing cavity 3 is internally provided with a liquid-stopping and air-passing device 31, and the liquid-stopping and air-passing device 31 is a PE sintered filter element.

[0042] The sintered filter element has a simple structure, can effectively prevent solution overflow, and can improve the safety and reliability of use.

[0043] The solid reagent for nucleic acid amplification includes dry powder reagent or reagent freeze-dried ball or quality control internal standard reagent; the volume of the sample liquid cavity 1 is 100 mu L-3000 mu L, and the volume of the nucleic acid lysis and extraction liquid is 100 mu L-3000 mu L.

[0044] The sample port 13 communicates with the sample liquid cavity 1, and the sample port 13 is sealed by the peel-off film in a state that the reagent box is not used. The peel-off film can be an easy-to-peel aluminum film. When the reagent box is used, the peel-off film needs to be peeled off, a swab with a sample is used to stir in the sample liquid, and the box body 5 is further provided with an upper cover. A recess is arranged on the upper cover, and a protrusion is arranged in the recess. The protrusion is a screw cap. After the swab is stirred in the sample liquid for a period of time, the sample port 13 is sealed by the screw cap. Since the screw cap is pressed into the sample port 13, a certain space is left between the sample port 13 and the liquid reagent, so that the volume of the sample liquid cavity 1 is reduced, the air pressure in the sample liquid cavity 1 is increased, that is, the sample liquid is in a high-pressure environment, and the liquid can also flow into the amplification cavity quickly.

[0045] The above-described embodiments only express the specific implementation of the present application, and the description is more specific and detailed, but it cannot be understood as a limitation on the scope of the patent of the present application. It should be noted that for ordinary skilled persons in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which belong to the protection scope of the present application.

Claims

1. A nucleic acid detection kit based on a negative pressure system, comprising a sample injection liquid chamber, a nucleic acid amplification chamber, and a liquid-stopping and venting chamber connected in sequence, wherein the sample injection liquid chamber contains nucleic acid lysis extraction solution, the nucleic acid amplification chamber contains nucleic acid amplification reagent, and the nucleic acid amplification reagent is a solid reagent, characterized in that, The kit also includes a negative pressure chamber, which is connected to a liquid-stopping and venting chamber. A negative pressure opening device is provided at the connection between the negative pressure chamber and the liquid-stopping and venting chamber. An injection switch is provided at the connection between the sample injection liquid chamber and the nucleic acid amplification chamber. The sequentially connected sample injection liquid chamber, nucleic acid amplification chamber, liquid-stopping and venting chamber and negative pressure chamber form a closed system. The kit also includes a relatively sliding box and a reagent card. The sample injection chamber and the negative pressure chamber are located in the box, the nucleic acid amplification chamber and the liquid-stopping and venting chamber are located on the reagent card, and the negative pressure opening device and the sample injection switch are located on the reagent card. A first puncture membrane is provided at the outlet of the sample injection chamber, a second puncture membrane is provided at the outlet of the negative pressure chamber, the sample injection switch is a first puncture needle corresponding to the first puncture membrane, and the negative pressure opening device is a second puncture needle corresponding to the second puncture membrane. The height of the tip of the first puncture needle is lower than the height of the tip of the second puncture needle.

2. The nucleic acid detection kit based on a negative pressure system according to claim 1, characterized in that, The sample injection chamber is provided with a sample dispensing port at the end away from the nucleic acid amplification chamber, and an easy-to-remove membrane is provided at the sample dispensing port.

3. The nucleic acid detection kit based on a negative pressure system according to claim 1, characterized in that, The liquid-stopping and venting cavity has a built-in liquid-stopping and venting device, which is a PE sintered filter element.

Images