Portable isothermal amplification nucleic acid detection device

Abstract

The utility model relates to a portable isothermal amplification nucleic acid detection device which comprises a shell, a reaction tube bracket, an excitation light source, a photoelectric sensor, a heating resistance wire and a control module, a reaction chamber with an opening in the side wall is arranged in the shell; the reaction tube bracket is inserted into the reaction chamber through the side wall opening and is used for fixing the reaction tube; the excitation light source is mounted in the reaction chamber; the photoelectric sensor is arranged at the top of the reaction chamber, and a condensing lens and an optical filter are sequentially arranged at the front end of the photoelectric sensor; the heating resistance wire is arranged in the reaction chamber and is used for controlling the temperature of the reaction tube; the control module is installed in the shell and connected with the excitation light source, the photoelectric sensor and the heating resistance wire. The device controls the temperature of the reaction tube through the heating resistance wire, excites the fluorophore in the reaction tube through laser to generate fluorescence, monitors the fluorescence intensity through the photoelectric sensor, and can be carried to the field for on-site detection, so that the detection instantaneity is improved.

Description

Technical Field

[0001] This utility model belongs to the technical field of agricultural field nucleic acid testing equipment, specifically relating to a portable isothermal amplification nucleic acid testing device. Background Technology

[0002] Nucleic acid testing is a DNA or RNA detection technology that can be applied to the detection of pathogenic microorganisms such as fungi, bacteria, viruses, and nematodes. Exposure of plants to plant pathogens can lead to the spread of these pathogens, causing outbreaks, spread, and epidemics of plant diseases in agriculture. Therefore, nucleic acid testing of plants, soil, and water during the planting process is of great significance for monitoring and early warning of plant diseases, and rapid on-site detection of agricultural pathogens is a crucial step in this process.

[0003] Nucleic acid testing technology based on polymerase chain reaction (PCR) requires large instruments, which are not portable, expensive, and difficult to move. In point-of-care testing (POCT) scenarios that only require a small number of samples, such as in the field, large instruments are not suitable for use in the field. Samples must be brought back to the testing room, resulting in poor immediacy and low work efficiency.

[0004] Therefore, there is an urgent need to develop a portable nucleic acid testing device that is suitable for field use to improve the immediacy of testing. Utility Model Content

[0005] To address at least one of the problems in the prior art, the purpose of this invention is to provide a portable isothermal amplification nucleic acid detection device that can be carried to the field for on-site testing, thereby improving the immediacy of the test.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A portable isothermal amplification nucleic acid detection device, comprising:

[0008] The shell contains a reaction chamber with openings in the side walls.

[0009] A reaction tube support is inserted into the reaction chamber through the opening in the side wall, and the reaction tube support is used to fix the reaction tube.

[0010] An excitation light source is installed in the reaction chamber, and the laser emitted by the excitation light source is used to excite the fluorescent groups in the reaction tube to produce fluorescence;

[0011] A photoelectric sensor is disposed at the top of the reaction chamber. A focusing lens and a filter are sequentially disposed at the front end of the photoelectric sensor. The fluorescence generated by the fluorescent group passes through the filter and the focusing lens and is received by the photoelectric sensor. The photoelectric sensor is used to detect the intensity of the fluorescence.

[0012] A heating resistance wire is placed in the reaction chamber to control the temperature of the reaction tube;

[0013] The control module is installed on one side of the housing and is connected to the excitation light source, the photoelectric sensor and the heating resistance wire, respectively.

[0014] Preferably, a temperature sensor is provided at the bottom of the reaction chamber, and the temperature sensor is connected to the control module.

[0015] Preferably, a vibration motor is provided at the bottom of the reaction chamber, and the vibration motor is connected to the control module to mix the liquid in the reaction tube.

[0016] Preferably, the condenser lens is mounted above the filter, and the focal length of the condenser lens is in the range of 6 mm to 12 mm.

[0017] Preferably, the transmission wavelength of the filter is 500 to 550 nm.

[0018] Preferably, the wavelength range of the excitation light source is 475 to 495 nm.

[0019] Preferably, the resistance value of the heating resistance wire is in the range of 6 to 20 ohms.

[0020] Preferably, the outer wall of the housing is provided with an LED indicator for displaying the test results, and the LED indicator is connected to the control module.

[0021] Preferably, the optical path from the excitation source to the reaction tube is the excitation optical path, and the optical path from the reaction tube to the photoelectric sensor is the fluorescence detection optical path, wherein the excitation optical path is perpendicular to the fluorescence detection optical path.

[0022] This utility model has the following advantages due to the adoption of the above technical solution:

[0023] 1. The portable isothermal amplification nucleic acid detection device provided by this utility model controls the temperature of the reaction tube by heating the resistance wire and generates fluorescence by exciting the fluorescent group in the reaction tube with a laser. The fluorescence intensity is monitored by a photoelectric sensor. The device can be powered by a USB power bank and can be carried to the field for on-site detection, which improves the immediacy of the detection.

[0024] 2. The portable isothermal amplification nucleic acid detection device provided by this utility model uses a vibration motor to make the entire device vibrate, thereby making the liquid in the reaction tube fully mixed. Attached Figure Description

[0025] Figure 1 This is a schematic diagram of the structure of a portable isothermal amplification nucleic acid detection device provided in one embodiment of the present invention.

[0026] Figure 2 This is a schematic diagram of the internal perspective structure of a portable isothermal amplification nucleic acid detection device provided in an embodiment of this utility model.

[0027] 1 is the housing, 2 is the condenser lens, 3 is the filter, 4 is the reaction tube support, 5 is the reaction chamber, 6 is the vibration motor, 7 is the heating resistance wire, 8 is the excitation light source, 9 is the temperature sensor, 10 is the control module, and 11 is the photoelectric sensor. Detailed Implementation

[0028] To make the objectives, technical solutions, and advantages of this utility model clearer, the technical solutions of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this utility model. All other embodiments obtained by those skilled in the art based on the embodiments of this utility model without creative effort are within the scope of protection of this utility model.

[0029] In the description of this utility model, it should be noted that the terms "upper", "lower", "front", "rear", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the system or component referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model.

[0030] In the description of this utility model, it should be noted that, unless otherwise explicitly specified and limited, the terms "assembly," "setup," and "connection" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model based on the specific circumstances.

[0031] This invention provides a portable isothermal amplification nucleic acid detection device. It generates fluorescence by exciting fluorescent groups in the reaction tube with a laser and monitors the fluorescence intensity with a photoelectric sensor. The device can be carried to the field for on-site detection, improving the immediacy of the detection.

[0032] The embodiments of this utility model will now be described in detail with reference to the accompanying drawings.

[0033] Example

[0034] Please refer to the reference. Figure 1 and Figure 2This embodiment provides a portable isothermal amplification nucleic acid detection device, which includes a shell 1, a reaction tube support 4, an excitation light source 8, a photoelectric sensor 11, a heating resistance wire 7, and a control module 10.

[0035] The shell 1 has a reaction chamber 5 with side wall openings;

[0036] The reaction tube support 4 is inserted into the reaction chamber 5 through the side wall opening. The reaction tube support 4 is used to fix the reaction tube.

[0037] The excitation light source 8 is installed inside the reaction chamber 1. The laser emitted by the excitation light source 8 is used to excite the fluorescent groups in the reaction tube to produce fluorescence.

[0038] A photoelectric sensor 11 is installed at the top of the reaction chamber 5. A condensing lens 2 and a filter 3 are sequentially arranged at the front end of the photoelectric sensor 11. The fluorescence generated by the fluorescent group is received by the photoelectric sensor 11 after passing through the filter 3 and the condensing lens 2. The photoelectric sensor 11 is used to detect the intensity of the fluorescence.

[0039] Heating resistance wire 7 is installed inside reaction chamber 5 to control the temperature of reaction tube;

[0040] The control module 10 is installed on one side of the housing 1 and is connected to the excitation light source 8, the photoelectric sensor 11 and the heating resistance wire 7 respectively.

[0041] In practical applications, the housing 1 is a rectangular box made of engineering plastic, which ensures strength while reducing weight, facilitating use in different locations and improving portability. The housing 1 contains a reaction chamber 5, with an opening on the side wall of the chamber 1. The reaction tube support 4 can be inserted into the reaction chamber 5 through this opening. The reaction tube support 4 has a reaction tube fixing hole into which the reaction tube is inserted. The reaction tube support 4 is suitable for 100 μL and 200 μL reaction tubes and can be adjusted according to different detection tube sizes. The inner surface of the reaction tube support 4 is coated with a low-reflection paint to reduce interference from excitation light reflection on fluorescence detection.

[0042] Optionally, both the housing 1 and the reaction tube support 4 are 3D printed as a single unit, made of black plastic, which has the advantages of being lightweight, inexpensive, and readily available.

[0043] The excitation light source 8 is installed on the side wall at the bottom of the reaction chamber 5. The optical path from the excitation light source 8 to the reaction tube is the excitation optical path, and the optical path from the reaction tube to the photoelectric sensor 11 is the fluorescence detection optical path. The excitation optical path and the fluorescence detection optical path are perpendicular. The light outlet of the excitation light source 8 is aligned with the reaction tube. The excitation light source 8 emits a laser, which irradiates the fluorescent group inside the reaction tube, causing it to fluoresce. The wavelength range of the excitation light source is 475 to 495 nm. Specifically, a solid-state laser with a center wavelength of 488 nm can be used.

[0044] The photoelectric sensor 11 is mounted on top of the reaction chamber 5. A condenser lens 2 and a filter 3 are mounted on the front end of the photoelectric sensor 11 via a bracket. Both the filter 3 and the condenser lens 2 are detachable and replaceable to accommodate the detection of different fluorescent groups and to adapt to a wider range of applications. When a fluorescent group is excited by a laser, the fluorescence is filtered out by the filter 3 to remove stray light, and then focused onto the photoelectric sensor 11 by the condenser lens 2. The photoelectric sensor 11 detects the fluorescence intensity and transmits the signal to the control module 10. The photoelectric sensor 11 is a digital photoelectric sensor using an IIC bus, saving I / O input / output points for the control module 10.

[0045] In this embodiment, the condenser lens 2 is mounted above the filter, and the focal length of the condenser lens ranges from 6 mm to 12 mm. The transmission wavelength of the filter 3 is 500 to 550 nm. The filter 3 can reduce the amount of excitation light transmitted, improve the contrast between different fluorescence intensities, and improve the detection sensitivity.

[0046] A heating resistance wire 7 is installed on the side wall at the bottom of the reaction chamber 5. The resistance value of the heating resistance wire 7 ranges from 6 to 20 ohms, and it is connected to the control module 10 via wires. The heating resistance wire 7 heats the reaction chamber 5, thereby controlling the temperature of the reaction tube. Heating the resistance wire 7 keeps the reaction chamber 5 at a constant temperature.

[0047] In this embodiment, a temperature sensor 9 is installed at the bottom of the reaction chamber 5. The temperature sensor 9 is connected to the control module 10. The temperature sensor 9 can be a digital temperature sensor using an IIC bus, which facilitates connection with the control module 10. The temperature sensor 9 can provide real-time feedback of the temperature of the reaction chamber 5 to the control module 10, thereby allowing the control module 10 to adjust the heating temperature of the heating resistance wire 7 in real time.

[0048] In this embodiment, a vibration motor 6 is provided at the bottom of the reaction chamber 5 to mix the liquid in the reaction tube.

[0049] Specifically, the vibration motor 6 is installed at the bottom of the reaction chamber 5. There can be one or more vibration motors 6. The vibration motor 6 causes the entire device to vibrate, thereby making the liquid in the reaction tube fully mixed. The diameter of the vibration motor 6 is 3 to 10 mm, and it is connected to the control module 10 through wires.

[0050] In this embodiment, the outer wall of the housing 1 is provided with an LED indicator for displaying the detection results, and the LED indicator is connected to the control module 10. The fluorescence detection results are displayed by the flashing of the LED indicator.

[0051] In this embodiment, the control module 10 includes a circuit board based on the ATmega328P, on which a power conversion module is provided. The power conversion module can convert a 3 to 20 volt input voltage to 5V for the device to use. The device can operate normally using a common USB power bank, making it convenient for use in the field. The control module 10 is connected to the excitation light source 8, photoelectric sensor 11, heating resistance wire 7, temperature sensor 9, vibration motor 6, and LED indicator light.

[0052] The working principle of the portable isothermal amplification nucleic acid detection device in this embodiment includes the following steps:

[0053] Step 1: Place the reaction tube containing the sample to be tested and the nucleic acid detection reagent into reaction tube holder 4;

[0054] Step 2: Insert the reaction tube holder 4 containing the reaction tube into the reaction chamber 5;

[0055] Step 3: Connect the power supply to the portable isothermal amplification nucleic acid detection device;

[0056] Step 4: The excitation light source 8 and photoelectric sensor 11 operate. The laser emitted by the excitation light source 8 excites the fluorescent group of the reaction tube. The photoelectric sensor 11 reads the fluorescence value and compares it with the fluorescence detection threshold preset in the control module 10. If the detected value is greater than the set fluorescence detection threshold, an error signal is output, that is, the LED indicator flashes rapidly, prompting the user to replace the detection tube; if the detected value is less than or equal to the set fluorescence detection threshold, the excitation light source 8 and photoelectric sensor 11 stop working and proceed to the next step.

[0057] Step 5: Start the temperature sensor 9 and heating resistance wire 7 to control the temperature of reaction chamber 5. After reaching the set temperature in control module 10, proceed to the next step.

[0058] Step 6: Start the vibration motor 6 and vibrate for 10 seconds. After 5 minutes, the light source 8 and photoelectric sensor 11 will start working. The photoelectric sensor 11 will read the fluorescence intensity value once and compare it with the set fluorescence detection threshold to determine whether the fluorescence detection threshold has been reached.

[0059] Step 7: If the detection result is less than or equal to the fluorescence detection threshold, repeat step 6, and repeat the number of times less than the number set in control module 10, specifically eight times; if the detection result is greater than the fluorescence detection threshold or the number of repetitions is greater than or equal to the set number of times, proceed to the next step.

[0060] Step 7: Flash different LED indicators according to the number of times Step 6 is repeated to display the fluorescence detection result. If the number of times Step 6 is repeated is greater than or equal to the set number, the detection result is determined to be greater than the fluorescence detection threshold. If the number of times Step 6 is repeated is less than the set number, the detection result is determined to be less than or equal to the fluorescence detection threshold.

[0061] The portable isothermal amplification nucleic acid detection device of this embodiment is used to detect the fluorescence intensity generated by nucleic acid amplification or CRISPR reaction in the reaction tube fixed in the reaction tube support 4. It can quickly perform on-site nucleic acid detection and analysis of pathogenic microorganisms in the field, improve the immediacy of detection, facilitate subsequent pathogen detection and field disease epidemic monitoring, and is of great significance.

[0062] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this utility model, and not to limit it. Although this utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this utility model.

Claims

1. A portable isothermal amplification nucleic acid detection device, characterized in that, include: The shell contains a reaction chamber with openings in the side walls. A reaction tube support is inserted into the reaction chamber through the opening in the side wall, and the reaction tube support is used to fix the reaction tube. An excitation light source is installed in the reaction chamber, and the laser emitted by the excitation light source is used to excite the fluorescent groups in the reaction tube to produce fluorescence; A photoelectric sensor is disposed at the top of the reaction chamber. A focusing lens and a filter are sequentially disposed at the front end of the photoelectric sensor. The fluorescence generated by the fluorescent group passes through the filter and the focusing lens and is received by the photoelectric sensor. The photoelectric sensor is used to detect the intensity of the fluorescence. A heating resistance wire is placed in the reaction chamber to control the temperature of the reaction tube; The control module is installed on one side of the housing and is connected to the excitation light source, the photoelectric sensor and the heating resistance wire, respectively.

2. The portable isothermal amplification nucleic acid detection device according to claim 1, characterized in that, A temperature sensor is installed at the bottom of the reaction chamber, and the temperature sensor is connected to the control module.

3. The portable isothermal amplification nucleic acid detection device according to claim 1, characterized in that, A vibration motor is installed at the bottom of the reaction chamber. The vibration motor is connected to the control module to mix the liquid in the reaction tube.

4. The portable isothermal amplification nucleic acid detection device according to claim 1, characterized in that, The condenser lens is mounted above the filter, and the focal length of the condenser lens ranges from 6 mm to 12 mm.

5. The portable isothermal amplification nucleic acid detection device according to claim 1, characterized in that, The transmission wavelength of the filter is 500 to 550 nm.

6. The portable isothermal amplification nucleic acid detection device according to claim 1, characterized in that, The wavelength range of the excitation light source is 475 to 495 nm.

7. The portable isothermal amplification nucleic acid detection device according to claim 1, characterized in that, The resistance value of the heating resistance wire ranges from 6 to 20 ohms.

8. The portable isothermal amplification nucleic acid detection device according to claim 1, characterized in that, The outer wall of the housing is provided with LED indicator lights for displaying test results, and the LED indicator lights are connected to the control module.

9. The portable isothermal amplification nucleic acid detection device according to claim 1, characterized in that, The optical path from the excitation source to the reaction tube is the excitation optical path, and the optical path from the reaction tube to the photoelectric sensor is the fluorescence detection optical path. The excitation optical path is perpendicular to the fluorescence detection optical path.

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