TW202100971A - Portable nucleic acid molecule analysis device quickly and simply executes detection of constant temperature circular amplification method at low energy consumption - Google Patents

Abstract

A portable nucleic acid molecule analysis device suitable for analyzing a sample is disclosed. The device includes a heating unit, a control unit for controlling the heating unit, a flashlight for providing an ultraviolet light source that excits the sample, a mask surrounding and defining a placement space suitable for placing the sample and made of an opaque material, and a handheld device removably placed on the mask. The handheld device includes a camera head for taking pictures of the sample placed in the placement space and a program interface informationally connected to the camera head and executing analysis. The heating unit having a small size is controlled by a simple circuit, the sample can be executed with required heating. The sample can be photographed by a handheld device, such as a mobile phone, and analysis of the constant temperature circular amplification method is exactly executed.

Description

Portable nucleic acid molecule analysis device

The invention relates to a biological molecule analysis device, in particular to a portable nucleic acid molecule analysis device.

At present, in the diagnosis and identification of animal and plant quarantine, in addition to traditional morphological identification and disease diagnosis, biochemical, physiological and other related detection techniques are also widely used, especially the direct analysis of nucleic acid molecules of specific organisms, which can be the most direct To achieve the purpose of diagnosis and identification. For example: Enzyme-Linked Immunosorbent Assay (ELISA) that uses the specific binding of antigen and antibody, Polymerase Chain Reaction (PCR) that uses specific types of polymerases with high sensitivity, and even At the same time, it has the advantages of sensitivity and short detection time. However, because the quarantine requirements in various fields are becoming more and more extensive, there are special requirements in terms of sensitivity, detection time, and even cost reduction. There is no single detection method that can meet all needs, so in related fields In terms of testing, further improvement and research and development are still needed.

In recent years, a new nucleic acid molecule detection technology-Loop-mediated isothermal Amplification (LAMP) has emerged. It mainly uses three sets of primers, namely outer primers and inner primers. (inner primers), and loop primers. Wherein, the primer pair must be designed for the specific region of the target sequence, thereby generating the specificity of detecting the target, so that very high accuracy and sensitivity can be achieved. In addition to the primers, polymerase purified and produced by specific types of bacteria needs to be added to the reaction to polymerize with the primer pair at a specific optimum temperature to amplify self-complementary DNA fragments to form a stem The dumbbell-shaped structure of the stem-loop allows continuous and repeated reactions to produce a large number of DNA fragments. During the reaction, accompanied by the release of pyrophosphate, the pyrophosphate will react with the magnesium ions in the reaction solution to form a white salt precipitate of magnesium pyrophosphate. As the reaction time increases, it can be done with the naked eye. A large amount of white precipitate is observed, which means that a specific specific reaction has occurred, which means that the target is detected, and the existence or concentration of the target can be determined based on this. In addition, in addition to direct visual observation, in order to ensure the accuracy of the result verification, nucleic acid dyes can be added, and the dye molecules are excited by ultraviolet light (UV) irradiation to observe the reaction results. The dye molecules are excited. The extra light emitted achieves better accuracy than direct observation of the white deposit with the naked eye.

However, the constant temperature loop amplification method usually needs to be completed in a laboratory with a bulky and expensive machine, especially when the image analysis technology is used to determine the test result, a machine with an external power supply is required. Visualization, calculation, analysis. Taking into account the needs of field experiments, real-time testing, etc. outside the laboratory, and even to overcome the risk of contamination that may occur in the process of transporting samples to the laboratory, if a portable device with a smaller volume can be developed, it can meet the requirements of the experiment. The same detection effect of the desktop machine in the room is bound to be able to more thoroughly utilize the detection advantages of the constant temperature ring amplification method, and to reduce the implementation threshold of the constant temperature ring amplification method and increase the constant temperature ring amplification The scope of application of the method.

Therefore, the object of the present invention is to provide a portable nucleic acid molecule analysis device that can reliably perform the constant temperature circular amplification method and is small in size and low in energy consumption.

Therefore, the portable nucleic acid molecule analysis device of the present invention is suitable for analyzing a sample, and includes a heating unit, a control unit connected to the heating unit with information, a flashlight for providing an ultraviolet light source for exciting the sample, and The surrounding defines a mask made of an opaque material suitable for placing the sample, and a handheld device that is removably placed on the mask.

The heating unit includes a heat-conducting block defining at least one detection groove for placing the sample, and a heating rod penetrating the heat-conducting block for heating the sample placed in the detection groove.

The handheld device includes a camera for photographing the sample placed in the placement space, and a program interface for information connection to the camera and for performing analysis based on the shooting results of the camera.

The effect of the present invention is that the configuration of the heating unit is simple and can cooperate with the control of the control unit to achieve the required heating effect with less electric energy. At the same time, the flashlight is used to provide an ultraviolet light source to the sample, for example, The camera of the handheld device of the mobile phone captures the development effect, and it is easy to directly use the installed program interface to perform analysis. It is indeed possible to quickly and easily perform constant temperature ring expansion with low energy consumption when the device is simple and easy to carry. Increased detection.

Referring to FIG. 1, an embodiment of the portable nucleic acid molecule analysis device of the present invention is suitable for analyzing a sample 800 and includes a heating unit 1, a control unit 2 connected to the heating unit 1, and a control unit 2 for providing excitation The sample 800 includes a flashlight 3 with an ultraviolet light source, a shield 4 made of opaque material, and a handheld device 9 that is removably placed on the shield 4.

Referring to FIGS. 1 and 2 at the same time, it should be noted that the sample 800 is usually contained in a test tube 81 to facilitate the detection of the constant temperature circular amplification method. The heating unit 1 includes a heat-conducting block 11 that defines two detection tanks 110 for placing the test tube 81, and a heat-conducting block 11 for heating the sample 800 placed in the detection tank 110. The heating rod 12, a temperature sensor 13 passing through the heat conducting block 11 and spaced adjacent to the detection grooves 110, and a heat insulating sleeve 14 covering the heat conducting block 11 and used to maintain temperature. Among them, the thermally conductive block 11 is preferably made of aluminum with good thermal conductivity, but it is not limited to this. The heat-conducting block 11 can be formed by die casting, machining, etc., to facilitate the formation of the detection grooves 110. The number of the detection grooves 110 can be changed as required, as long as the number and position distribution are designed to ensure that the heat energy provided by the heating rod 12 can be reliably transferred to the detection groove 110 via the heat conducting block 11 The test tube 81 can be used to generate the required heating effect on the sample 800. In addition, the thermal insulation sleeve 14 can use styrofoam with excellent thermal insulation properties, thereby heating the sample 800 at “constant temperature” as much as possible.

Referring to FIG. 3 and in conjunction with FIG. 1, the control unit 2 includes a circuit board 21, a proportional-integral-derivative controller 22 (Propotional-Integral and Derivatice control, PID) configured on the circuit board 21 and connected to the heating rod 12. ), a temperature measuring device 23 electrically connected to the circuit board 21 and suitable for directly detecting the temperature of the sample 800, a display panel 24 arranged on the circuit board 21 and information connected to the temperature sensor 13, An operating interface 25 arranged on the circuit board 21 and used to operate the proportional integral derivative controller 22, and a single chip 26 arranged on the circuit board 21 and capable of programming directly. In order to reduce the volume of the overall device, the circuit board 21 can be a printed circuit board (PCB) that is easy to arrange pre-designed circuits, so that the proportional integral derivative controller 22, the display panel 24, and the operation interface 25 can be easily It is arranged on the circuit board 21 to cooperate with the pre-designed circuit, thereby reducing the weight of the whole device as much as possible. Through the operation interface 25, the temperature information sensed by the temperature sensor 13 displayed on the display panel 24 can be used to input an operation command to the proportional integral derivative controller 22, and then the proportional integral derivative controller 22 is sufficient. The heating rod 12 is controlled to heat in a specific heating mode.

In addition, considering that the proportional-integral-derivative controller 22, the display panel 24, and the control program of the operation interface 25 may be adjusted according to different needs, the single chip 26 that is easy to modify the program in real time is used for programming. That is beneficial to immediately modify the code to meet different needs.

It is worth noting that the main design of this embodiment is "easy to carry", "low energy consumption", and "simple execution". Therefore, in the application of electric energy, it will naturally be designed on the premise that batteries can be used. In order to get rid of the restriction that the high-energy power supply must be used in the laboratory, the present embodiment can achieve the purpose of being carried out of the laboratory for use. Therefore, the electric energy of the heating unit 1 and the control unit 2 is of course also mainly batteries. In this embodiment, it is preferable to cooperate with a small-sized lithium battery to form an appropriate driving voltage in series with each other. Taking three 18650 lithium batteries with a reference voltage of 3.6V~4.2V in series with each other as an example, the heating rod 12 with an operating power of 14W can be driven, and the heating time of 90 minutes is sufficient to heat the sample 800 to 65℃~70℃ demand.

Referring to FIG. 3 and FIG. 4 in conjunction with FIG. 1, the mask 4 is preferably made of black acrylic material, but it is not limited to use a material that can shield external light. The shield 4 includes a body 41 that defines a space 410 for placing the test tube 81 containing the sample 800, a receptacle 42 recessed in the body 41 for the handheld device 9 to be placed, and A through hole 43 penetrates the main body 41 and communicates with the placement space 410 and the receiving groove 42.

It should be particularly noted that the power source of the flashlight 3 is also based on easy-to-obtain, small-sized, and replaceable batteries, and is preferably fixed on other components in a movable and irradiating direction to facilitate long-term irradiation of the placement space 410 The sample 800 in. Among them, in order to enable the flashlight 3 to emit ultraviolet light required for stimulating fluorescent light, the installed lamp preferably adopts an ultraviolet LED bulb.

The handheld device 9 includes a camera 91 for photographing the sample 800 placed in the placement space 410, and a program interface (not shown) that is connected to the camera 91 with information and used to perform analysis based on the shooting results of the camera 91 Label). Taking the handheld device 9 as a smart phone as an example, the camera 91 equipped with the program interface can be configured to continuously shoot for a set period of time, or to shoot at intervals of several seconds within a fixed period of time, so as to observe the sample 800 The change. And the program interface can be developed with the operating system of the smartphone, using the JAVA language that can be executed in the operating system, to meet the analysis needs of the constant temperature ring amplification method, and is operated by the smartphone’s own processor. Perform analysis on the captured images. Wherein, when the handheld device 9 is placed in the receiving slot 42, the position of the perforation 43 of the mask 4 corresponds to the camera 91 of the handheld device 9, ensuring that the camera 91 can pass through the perforation 43 to take pictures in the placement space The sample 800 in 410.

Referring to FIG. 5 in conjunction with FIG. 3, the sample 800 is a donkey DNA sample, and the constant temperature circular amplification method of this embodiment is used as an example for illustration. The sample 800 is contained in a test tube 81, and the required fluorescent dye and other matching component reagents are added, then the test tube 81 can be put into the detection tank 110 of the heat conducting block 11, and pass through the control unit 2 to control the heating mode of the heating rod 12, and to match the heat preservation effect of the heat insulation sleeve 14, to heat the sample 800 in the test tube 81 at a specific temperature. At this time, matching the temperature displayed on the display panel 24 of the control unit 2 and directly measuring the temperature of the sample 800 using the temperature measuring device 23 can ensure that the sample 800 is maintained at the correct temperature. After the heated sample 800 is contained in another test tube 81, the test tube 81 can be placed in the placement space 410 defined by the body 41 of the shield 4, and the ultraviolet light emitted by the flashlight 3 can be used to irradiate The sample 800 is excited to emit specific fluorescence. At the same time, the camera 91 of the handheld device 9 placed in the receptacle 42 of the shield 4 will continue to photograph the sample 800 through the perforation 43. Using the program interface pre-installed in the handheld device 9, the image captured by the camera 91 can be used to perform image analysis that analyzes the component ratios of the three primary lights of red light, green light, and blue light. The ratio of blue light can determine the analysis result of the sample 800 after the constant temperature circular amplification method.

It should be particularly emphasized that the above is only one of the ways to perform the constant temperature circular amplification method using this embodiment. In actual implementation, the user can use the control unit 2 to perform necessary control, adjust the heating temperature, heating time and other parameters of the heating unit 1, and change according to the characteristics of the analyzed sample, the reagents used, and other variables. Whether the flashlight 3 is irradiated or not, the irradiation time, even the shooting mode of the camera 91 of the handheld device 9, and the analysis settings of the program interface, etc., to perform the analysis of the constant temperature ring amplification method according to specific needs, and It is not limited to the above-mentioned embodiment.

In summary, the use of the portable nucleic acid molecule analysis device of the present invention can indeed effectively reduce the overall volume of the device, and when performing a constant temperature circular amplification method, the control unit 2 and the heating unit 1 can be used to compare The low energy consumption ensures sufficient heating effect, and the handheld device 9 can be used directly to perform shooting and analysis, which effectively reduces the threshold for the implementation of the constant temperature circular amplification method, and can greatly expand the application scope of the constant temperature circular amplification method. It can indeed achieve the purpose of the invention.

However, the above are only examples of the present invention. When the scope of implementation of the present invention cannot be limited by this, all simple equivalent changes and modifications made in accordance with the scope of the patent application of the present invention and the content of the patent specification still belong to This invention patent covers the scope.

1: heating unit 11: Thermal block 110: detection slot 12: heating rod 13: Temperature sensor 14: Insulation sleeve 2: control unit 21: circuit board 22: Proportional integral derivative controller 23: Temperature measuring device 24: display panel 25: Operation interface 26: Single chip 3: flashlight 4: Mask 41: body 410: place space 42: Tolerance 43: Piercing 800: sample 81: test tube 9: Handheld device 91: camera

Other features and effects of the present invention will be clearly presented in the embodiments with reference to the drawings, in which: Figure 1 is a block diagram illustrating an embodiment of the portable nucleic acid molecule analysis device of the present invention; Figure 2 is a perspective exploded view illustrating a heating unit of this embodiment; Figure 3 is a top view illustrating a control unit of this embodiment; Figure 4 is a schematic diagram illustrating a mask and a handheld device of the embodiment; and FIG. 5 is a flowchart illustrating the process of performing the constant temperature circular amplification method using this embodiment.

1: heating unit

2: control unit

3: flashlight

4: Mask

800: sample

9: Handheld device

Claims (9)

A portable nucleic acid molecule analysis device, suitable for analyzing a sample, and includes: A heating unit, including a heat conduction block defining at least one detection groove for placing the sample, and a heating rod penetrating the heat conduction block for heating the sample placed in the detection groove; A control unit with information connected to the heating unit and used to control the heating condition of the heating rod; A flashlight for providing an ultraviolet light source for exciting the sample; A mask that surrounds and defines a space suitable for placing the sample, and is made of opaque material; and A handheld device, which is removably placed on the mask, and includes a camera for photographing the sample placed in the placement space, and an information connected to the camera and used for the shooting result of the camera Program interface for performing analysis. The portable nucleic acid molecule analysis device according to claim 1, wherein the heating unit further includes a temperature sensor penetrating the heat conducting block and adjacent to the at least one detecting groove. The portable nucleic acid molecule analysis device according to claim 2, wherein the control unit includes a circuit board and a proportional integral derivative controller arranged on the circuit board and connected to the heating rod. The portable nucleic acid molecule analysis device according to claim 3, wherein the control unit further includes a temperature measuring device electrically connected to the circuit board and suitable for directly detecting the temperature of the sample. The portable nucleic acid molecule analysis device according to claim 3 or 4, wherein the control unit further includes a display panel disposed on the circuit board and connected to the temperature sensor for information. The portable nucleic acid molecule analysis device according to claim 5, wherein the control unit further includes an operating interface arranged on the circuit board and used for operating the proportional integral derivative controller. The portable nucleic acid molecule analysis device according to claim 3, wherein the control unit further includes a single chip which is arranged on the circuit board and can directly burn programs. The portable nucleic acid molecule analysis device according to claim 1 or 2, wherein the heating unit further includes a heat insulating sleeve covering the heat conducting block and used for maintaining temperature. The portable nucleic acid molecule analysis device according to claim 1, wherein the mask includes a body defining the placement space, a container recessed in the body and used for placing the handheld device, and a It penetrates the main body and is connected between the placing space and the container, and the position corresponds to the hole of the camera of the handheld device.

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