CN207596860U - A kind of nucleic acid integration Multiple detection box body - Google Patents

Abstract

The utility model relates to a nucleic acid integrated multiple detection box body, which comprises a box body shell and a two-layer board inside the shell. The two-layer boards are stacked up and down in the shell, and the upper-layer board and the lower-layer board can slide relative to each other. The lower plate first communicates with the four hemispherical groove chambers of the upper plate communicating with the flow channel to form four spherical chambers, and makes the embedded chambers of the upper and lower plates communicate. Liquid can flow from the lower plate to the upper plate. As the upper plate slides, the lower plate can communicate with four relatively independent hemispherical groove chambers of the upper plate to form four spherical chambers. The nucleic acid liquid extracted from the lower plate flows into the four spherical chambers, and is mixed with the dry powder of the isothermal amplification detection reagent pre-packaged in the chamber, and the temperature control and real-time fluorescence detection are carried out outside to realize the integrated multiple detection of nucleic acid. The utility model combines high-temperature one-step nucleic acid extraction, isothermal amplification, and real-time fluorescence detection, which greatly shortens the time of traditional nucleic acid detection and reduces the cost requirements for equipment. The detection is completed in a closed environment, effectively avoiding the External contamination and nucleic acid cross-contamination.

Description

A nucleic acid integrated multiple detection cartridge

technical field

The utility model relates to a nucleic acid detection chip in the field of molecular biology, in particular to a nucleic acid integrated multiple detection box body and a detection method, belonging to the fields of biomedical engineering and molecular diagnosis.

Background technique

The current biomedical engineering research is going deep into the molecular level from the overall and cellular level. Nucleic acid is an important class of biomolecules in cells and participates in the regulation of most of the functions of cells. Understanding the content and distribution of nucleic acids in different cells is very important for in-depth research on their functions and the biological significance behind them. The detection, diagnosis and treatment of diseases and various infectious diseases are of great significance. Existing nucleic acid detection mainly adopts PCR technology, which has high sensitivity and good specificity, and is currently the most commonly used genetic diagnosis method. However, the PCR method is more complicated to operate, and the experiment takes a long time, which requires relatively high personnel and equipment, and is not suitable for rapid diagnosis at the grassroots level or on the spot.

Isothermal amplification technology is currently a popular amplification technology. The reaction process is always maintained at a constant temperature, and the purpose of rapid amplification is achieved by adding enzymes with different activities and their respective specific primers. It has the advantages of simple operation, short reaction time and high sensitivity, and is suitable for the field of rapid diagnosis. However, at the same time, the high efficiency of isothermal amplification can easily cause contamination of amplification products during downstream detection. If it can be detected, the upstream sample processing, that is, nucleic acid extraction, amplification, and downstream detection, are enclosed in a box for integration. , it can effectively avoid detection contamination. At the same time, in order to meet the needs of rapid detection at the grassroots level or on-site, the detection device needs to be miniaturized and portable.

In order to solve the above problems, the utility model proposes a nucleic acid integrated multiple detection box.

Contents of the invention

The purpose of the utility model is to provide a nucleic acid integrated multiple detection box for the existing basic level or on-site rapid detection requirements. contamination and cross-contamination between samples.

In order to realize the purpose and advantages of the utility model, the utility model is realized through the following technical solutions:

A nucleic acid integrated multiple detection box body, the box body includes a shell, and two layers of flow channel plates in the shell. The two-layer runner plates are stacked one above the other and can be put into the casing.

The shell is square, one side of the shell is open, the other side has a small hole, and the other sides are all closed. The two-layer flow channel plate is pushed into the casing through the open side to form a complete box body.

The two-layer flow channel plate is divided into an upper layer plate and a lower layer plate, and a plurality of liquid flow channels and chambers are engraved in each of the two layer plates.

Preferably, the upper layer has nine chambers, eight of which are hemispherical groove chambers, and the other is a chamber embedded in the upper layer; of the eight hemispherical groove chambers, four of them are One is connected to the flow channel, and at the same time communicates with one end of the inner cavity of the upper plate; the other four hemispherical groove chambers are not connected to the flow channel, and are relatively independent. The other end of the chamber embedded in the upper plate communicates with a short air pressure balance channel, and the outlet of the channel communicates with the small hole on one side of the shell.

Preferably, the lower plate is provided with five chambers, four of which are hemispherical groove chambers, and the other is a chamber embedded in the lower plate; the four hemispherical groove chambers are all connected to the flow channel, and at the same time communicate with one end of the embedded chamber of the lower plate through the flow channel; sample device.

The position of the hemispherical groove chamber of the upper plate and the position of the hemispherical groove chamber of the lower plate correspond to each other; the notch of the hemispherical groove chamber of the upper plate is downward, and the lower plate The notch of the hemispherical groove chamber is upward, and the diameters of the chamber grooves of the two-layer boards are the same, so that a complete spherical chamber can be formed by one-to-one correspondence.

The upper board and the lower board can slide relative to each other. The lower plate first communicates with the four hemispherical groove chambers of the upper plate communicating with the flow channel to form four spherical chambers, and makes the embedded chambers of the upper and lower plates communicate. As the upper plate slides, the lower plate can communicate with four relatively independent hemispherical groove chambers of the upper plate to form four spherical chambers.

The four relatively independent hemispherical groove chambers of the upper plate are pre-loaded with reagent solid dry powder for nucleic acid isothermal amplification detection, which can be different disease detection reagents of the same sample or the detection of different sites of the same target gene Reagents, in order to achieve the purpose of multiple detection.

The external sampling system first feeds the mineral oil into the embedded chamber of the lower board through the sampling flow channel of the lower board, and then drives the mineral oil in the chamber to the connected flow channel through the pressure system, and drains out the inside of the lower board. The mineral oil in the embedded chamber is discharged into the embedded chamber of the upper plate; then the sample to be tested and the extraction solution are passed into the embedded chamber of the lower plate through the sampling system, and the external heat is applied to the embedded chamber. Control, control the reaction temperature at 100°C, and react for 10 minutes to achieve high-temperature one-step extraction of nucleic acids. The upper plate is slid so that the lower plate communicates with four relatively independent hemispherical groove chambers of the upper plate to form four spherical chambers. Drive the reacted liquid in the second process into the four spherical chambers, and fill the chambers, dissolve the solid dry powder of reagents for isothermal amplification detection of nucleic acids pre-packaged in the hemispherical groove chambers of the upper plate, and mix the reagents with external assistance . The sampling system feeds mineral oil into the embedded chamber of the lower plate again, and drives it into the flow channel of the lower plate. The heating temperature of the four relatively independent hemispherical groove chambers on the upper plate was controlled, and the reaction temperature was controlled at 65° C. for 30 minutes. During the reaction process, a fluorescence detection device should be used for real-time detection of reactants. After the reaction is completed, the results of multiple detections can be judged by observing the fluorescence curves of each chamber displayed by the fluorescence detection device.

The beneficial effects of the utility model:

The utility model adopts a box layer structure, integrates the whole nucleic acid detection process in a closed environment, and combines high-temperature one-step nucleic acid extraction, isothermal amplification, and real-time fluorescence detection, which greatly shortens the traditional nucleic acid detection time. , and the cost requirements for equipment are greatly reduced, effectively avoiding external pollution and cross-contamination of nucleic acids; the utility model has less difficulty in processing the box body, simple and portable operation, and meets the needs of on-site testing.

Description of drawings

Fig. 1 is a schematic diagram of the structural appearance of the utility model.

Fig. 2 is a schematic perspective view of the structure of the utility model.

Fig. 3 is a schematic diagram of the structure of the upper plate of the utility model.

Fig. 4 is a schematic diagram of the structure of the lower plate of the utility model.

Fig. 5 The plan view (A) and the side view (B) of the initial relative positions of the upper and lower layers of the utility model.

Fig. 6 is a plan view (A) and a side view (B) of the relative positions of the nucleic acid amplification detection process of the middle and lower plate of the utility model.

Among them: 1-box body, 2-shell, 3-upper plate, 4-lower plate, 301-upper flow channel, 302-embedded chamber in the upper layer plate, 303-upper layer flow channel connected groove chamber, 304- Independent chamber, 305-balance flow channel, 401-lower layer flow channel, 402-chamber embedded in the lower layer plate, 403-lower layer flow channel connected to groove chamber, 404-injection flow channel.

Detailed ways

Below in conjunction with embodiment and accompanying drawing 1-6, the utility model is further explained, so that those skilled in the art can implement according to referring to the description. The following examples are only used to illustrate the utility model, but are not intended to limit the implementation scope of the utility model.

A nucleic acid integrated multiple detection box and detection method, as shown in Figures 1 and 2, the box 1 includes a shell 2, and two layers of flow channel plates (upper plate 3 and lower plate 4) in the shell 2. The two-layer runner plates are stacked one above the other and can be put into the casing. Described casing 2 is square, and one side is open, and the other side has a small hole, and other sides are all closed. The two-layer flow channel plate can be pushed into the casing through the open side to form a complete box body 1 . The upper plate 3 and the lower plate 4 are respectively engraved with a plurality of liquid channels and a plurality of chambers.

As shown in FIG. 3 , it is a schematic structural diagram of the upper plate 3 , which has nine chambers and several upper flow channels 301 . One of them is a chamber 302 embedded in the upper deck, and the other eight are hemispherical recessed chambers. Four of the eight hemispherical groove chambers are connected to the flow channel and the chamber 302 embedded in the upper plate, which is called the upper flow channel communication groove chamber 303, and the other four are not connected to the flow channel and are relatively independent. , called the independent chamber 304 . The other end of the cavity 302 embedded in the upper plate communicates with a section of air pressure balance channel 305 .

As shown in FIG. 4 , it is a schematic structural diagram of the lower plate 4 , which has five chambers and several lower flow channels 401 . One of them is a chamber 402 embedded in the lower plate, and the other four are hemispherical groove chambers, which are all connected to the flow channel and the chamber 402 embedded in the lower plate, called the lower flow channel communication groove chamber 403 . The other end of the chamber 402 embedded in the lower plate communicates with a sample injection flow channel 404 .

Relative sliding can be performed between the upper plate 3 and the lower plate 4, the flow channel of the upper plate 3 communicates with the groove chamber 303 and the notch of the independent chamber 304 downwards, and the flow channel of the lower plate 4 communicates with the groove cavity The notches of the chamber 403 are upward, and the notches of the two layers of boards have the same diameter, so that one-to-one correspondence can form a complete spherical chamber.

For the above nucleic acid integrated multiple detection cartridges, the following methods are used in the case:

Before the upper plate 3 and the lower plate 4 are stacked together and put into the housing 2, the independent chamber 304 of the upper plate 3 is pre-loaded with reagent solid dry powder for nucleic acid isothermal amplification detection, which can be different samples of the same sample. Disease detection reagents or detection reagents targeting different sites of the same gene, so as to achieve the purpose of multiple detection.

Before the nucleic acid integrated multiple detection experiment, the initial relative positions of the upper plate 3 and the lower plate 4 in the housing 2 are shown in Figure 5, the upper flow channel communicates with the groove chamber 303 and the lower flow channel communicates with the groove chamber 403 notches correspond to form four spherical chambers and communicate with each channel. At the beginning of the experiment, the external sampling system first passes mineral oil through the sampling channel 404 of the lower plate 4 to the embedded chamber 402 of the lower plate 4, and then drives the mineral oil in the embedded chamber 402 through the pressure system. into the connected flow channel 401, and empty the mineral oil in the embedded cavity of the lower plate, and drain it into the embedded cavity 302 of the upper plate 3 and the upper flow channel 301.

Then, the sample to be tested and the extraction solution are passed into the embedded chamber 402 of the lower plate 4 through the sampling system, and the heating temperature of the embedded chamber 402 is controlled by the outside world, and the reaction temperature is controlled at 100°C, and the reaction is performed for 10 minutes to realize nucleic acid extraction. high-temperature one-step extraction.

After the extraction is completed, slide the upper plate 3 so that the flow channel of the lower plate 4 communicates with the notches of the groove chamber 403 and the notches of the four independent chambers 304 of the upper plate 3 to form four spherical chambers. Drive the extracted nucleic acid liquid from the embedded chamber 402 of the lower plate 4 into the four spherical chambers, and fill the chambers to dissolve the nucleic acid isothermal amplification detection reagents pre-packaged in the hemispherical groove chamber of the upper plate Solid dry powder, external auxiliary mixing reagent.

The sampling system feeds mineral oil into the embedded chamber 402 of the lower plate 4 again, and drives it into the flow channel 401 of the lower plate to seal the spherical chamber and prevent reagents from evaporating at high temperature. The heating temperature of the four independent chambers 304 of the upper plate 3 is controlled, the reaction temperature is controlled at 65° C., and the reaction is performed for 30 minutes. During the reaction process, a fluorescence detection device should be used for real-time detection of reactants. After the reaction is completed, the results of multiple detections can be judged by observing the fluorescence curves of each chamber displayed by the fluorescence detection device.

The utility model combines high-temperature one-step nucleic acid extraction, isothermal amplification, and real-time fluorescence detection, which greatly shortens the time of traditional nucleic acid detection, and greatly reduces the cost requirements for equipment. The entire nucleic acid detection process is integrated in a closed It is completed in the environment, effectively avoiding external pollution and cross-contamination of nucleic acid, and the operation is simple and portable, which can meet the needs of on-site testing.

Claims (7)

1. a kind of nucleic acid integration Multiple detection box body, which is characterized in that including：

Two-ply in one shell and shell, two-ply mounted on top in the shell；

Top plate in the two-ply is equipped with nine chambers, wherein eight are hemispherical groove chambers, another is upper The chamber that laminate embeds；Eight hemispherical groove chambers, wherein there are four being connected with runner, while with the upper strata Plate embeds chamber and is connected by runner；Four additional hemispherical groove chamber does not connect with runner, respectively relatively independent；

Lower plywood in the two-ply is set there are five chamber, wherein four are hemispherical groove chambers, another is under The chamber that laminate embeds；Four hemispherical groove chambers are connected with runner, while embed chamber with the lower plywood One end is connected by runner, and the lower plywood embeds the chamber other end and connected with sample introduction runner one end in lower plywood.

2. a kind of nucleic acid integration Multiple detection box body according to claim 1, which is characterized in that the top plate Hemispherical groove chamber location and the hemispherical groove chamber location of the lower plywood correspond.

3. a kind of nucleic acid integration Multiple detection box body according to claim 1, which is characterized in that the top plate Hemispherical groove chamber notch is downward, and the hemispherical groove chamber notch of the lower plywood is upward, the chamber notch diameter of two-ply Size is identical, and one-to-one correspondence can form fully spherical shape chamber.

4. a kind of nucleic acid integration Multiple detection box body according to claim 1, which is characterized in that the top plate with Opposite slide can be carried out between the lower plywood.

5. a kind of nucleic acid integration Multiple detection box body according to claim 1, which is characterized in that the shell is side Shape, one side of shell are opened wide, and another side is provided with an aperture, other each faces are closed；Two layers of runner plate is by opening wide Side push-in shell in, form complete box body.

6. a kind of nucleic acid integration Multiple detection box body according to claim 1, which is characterized in that the top plate embeds The chamber other end connect a bit of air pressure balance runner, runner exit is communicated with shell one side aperture.

7. a kind of nucleic acid integration Multiple detection box body according to claim 1, which is characterized in that the top plate The reagent solid dry powder detected for nucleic acid isothermal amplification is preinstalled in four relatively independent hemispherical groove chambers.