Apparatus for detecting a genetic sample

Abstract

The invention relates to a device (1, 101) for detecting a genetic sample, comprising a housing (10, 110), a control unit (20, 120), a temperature control unit (30, 130) with a heating element, an illumination unit (40, 140) and a detection means, the control unit, the temperature control unit, the illumination unit and the detection means being arranged within the housing (10, 110), and a sample holding element being arranged within the housing (10, 110). The device (1, 101) further comprises a vortex centrifugation unit (50, 150) having a drive unit and a rotatable sample holding tray (55, 155) connected to the drive unit and provided with the sample holding element, wherein an oscillation element (57, 157) is provided in the center of the sample holding tray (55, 155). The housing (10, 110) consists of a first housing part (11, 111) and a second housing part (12, 112), which are separated by a partition wall (15, 115).

Description

[0001] This invention relates to a device for detecting gene samples, which is suitable for performing detection based on loop-mediated isothermal amplification (LAMP).

[0002] LAMP technology (i.e., loop-mediated isothermal amplification) has a variety of applications in veterinary medicine, especially in the diagnosis of infectious diseases. The detection relies on the fact that an easily assessable DNA product is formed during the short reaction period of a sample placed in an isothermal (i.e., constant temperature) environment. This type of detection can be performed in a thermostat.

[0003] Document CN203720089U discloses a fluorescence intensity analyzer for LAMP detection. This analyzer includes transparent cylindrical sample holding elements for receiving reaction tubes, arranged on an insulating sample holding plate equipped with a heating plate. A UV light source is positioned in front of each sample holder to irradiate the reaction products formed in the sample. The intensity of the light emitted by the irradiated sample is detected by a sensor connected to a data processing unit. A drawback of this analyzer is that it does not allow for complete sample preparation. Furthermore, the samples are not uniformly heated.

[0004] The purpose of this invention is to provide a device for detecting gene samples that allows for the simple and accurate preparation of the sample to be detected within the framework of LAMP-based detection, and to enable the accurate and cost-effective implementation and evaluation of the results of such detection.

[0005] The above objective is achieved by providing a device for detecting gene samples, the device comprising: a housing; a control unit, a temperature control unit having a heating element, an illumination unit and a detection device, the components being arranged within the housing; and a sample holding element arranged within the housing.

[0006] The device also includes a vortex centrifuge unit with a drive unit and a rotatable sample holding tray connected to the drive unit and equipped with a sample holding element. The oscillation element is located at the center of the sample holding tray. The housing consists of a first housing section and a second housing section, which are separated by a partition wall.

[0007] Various preferred embodiments of the present invention are specifically described in the dependent claims.

[0008] The invention will now be described by way of example with reference to the accompanying drawings, in which: Figure 1 This is an exploded perspective view of the device according to the first embodiment of the present invention.

[0009] Figure 2 This is a three-dimensional front view of the device in a closed state according to the first embodiment of the present invention.

[0010] Figure 3 This is a perspective front view of the device according to the first embodiment of the present invention in the closed state without the second housing portion.

[0011] Figure 4 This is a perspective front view of the vortex centrifugal unit, temperature control unit, and control unit of the device according to the first embodiment of the present invention.

[0012] Figure 5A This is a perspective top view of the partition wall and temperature control unit of the device according to the first embodiment of the present invention.

[0013] Figure 5B This is a perspective bottom view of the partition wall and temperature control unit of the device according to the first embodiment of the present invention.

[0014] Figure 6 This is an exploded perspective view of the device according to a second embodiment of the present invention.

[0015] Figure 7 This is an exploded perspective view of the vortex centrifugal unit of the device according to the second embodiment of the present invention.

[0016] Figure 8 This is a perspective view of the device in the open state during use according to the second embodiment of the present invention.

[0017] Figure 1 and Figure 2 The arrangement of the main units of device 1 according to a first embodiment of the present invention is shown. Device 1 includes a housing 10, which is composed of a first housing portion 11 and a second housing portion 12, separated by a partition wall 15. The housing 10 may have a wall structure with a thickness of 10 mm and is provided with a helical gyroid-type filler, which is preferably formed by 3D printing from an opaque and heat-insulating polymer material. The edge 16 of the partition wall 15 may be provided with a seal 17 made of an elastic polymer (e.g., TPU material), which abuts against the edge 5 of the second housing portion 12 and contacts the edge 6 of the first housing portion 11 in the closed state of the housing 10. For example, the closure of the housing 10 may be ensured by magnetic elements placed along edges 5, 6, and 16. The second housing portion 12 and the first housing portion 11 may be connected to each other via hinges 3 arranged at the rear portions of their edges 5 and 6. The housing 10 is preferably configured as a cube with a side length of 20 cm.

[0018] The power supply for device 1 can be an uninterruptible power supply. The preferred power requirement for device 1 is a maximum of 65 W.

[0019] The device 1 includes a control unit 20, a temperature control unit 30 with a heating element, an illumination unit 40 for irradiating gene samples, a detection device, a sample holding element, and a vortex centrifuge unit 50 for centrifuging and shaking the samples. The illumination unit 40 and the vortex centrifuge unit 50 are preferably located in the first housing portion 11, and the control unit 20 and the temperature control unit 30 are preferably located in the second housing portion 12.

[0020] The lighting unit 40 is preferably arranged in a recess 18, which is defined by an opening formed on the edge of the partition wall 15 along one side of the front surface 2 of the housing 10. In this embodiment, the lighting unit 40 consists of a UV light source 41 and a blue light source 42 that generates blue light in the visible spectrum; for example, the UV light source and the blue light source form an LED array.

[0021] The control unit 20 is preferably a microprocessor used to control the temperature control unit 30, the lighting unit 40, and the vortex centrifugal unit 50. The control unit 20 is preferably connected to a control panel 22, which is, for example, positioned on the front surface 2 of the housing 10 on a second housing portion 12. Figure 2 As shown, the control panel 22 may be equipped with an LCD display 23 and control buttons 71, 72, 73, 74, 75 and 76. These control buttons are used to turn the UV light source 41, the blue light source 42 and the temperature control unit 30 on and off, and to set the temperature, rotation speed and time range of the vortex centrifugal unit 50.

[0022] The detection device can be a window 60 for observing a gene sample that can be placed in the device 1. In this case, the window 60 can be formed in the first housing portion 11 and can be provided with a filter for filtering light emitted by the light source of the illumination unit 40 (i.e., UV light source 41 and blue light source 42). The window 60 can be made of, for example, a transparent orange polycarbonate sheet. The window 60 is preferably disposed on the front surface 2 of the first housing portion 11. In addition, the detector ( Figure 1 (Not shown) can be arranged on the partition wall 15 in the first housing portion 11. This detector is suitable for photometric detection and / or fluorescence detection. Thus, the detector can be adapted to detect the light emitted by a sample placed in the device 1 and illuminated by the illumination unit 40.

[0023] Figure 1 and Figure 3The configuration of a vortex centrifuge unit 50 for preparing gene samples to be tested is shown. The vortex centrifuge unit 50 includes a drive unit and a rotatable sample holding tray 55 connected to the drive unit. The drive unit may be configured as a motor 53, which is positioned in a recess 52 provided on a partition wall 15, enabling the sample holding tray 55 to rotate at a speed preferably between 600 rpm and 2200 rpm. The sample holding tray 55 includes a sample holding element and an oscillating element 57 disposed at the center of the sample holding tray 55.

[0024] exist Figure 3 In the illustrated embodiment, the sample holding tray 55, formed as a folded metal plate, may include a square base plate 81 and four sidewalls 82 that project from the edges of the base plate 81 and are inclined outwards relative to the surface of the base plate 81. These sample holding elements may be cylindrical sample holders 54 having a central axis inclined relative to the surface of the base plate 81, typically suitable for receiving 1.5 ml or 2 ml Eppendorf tubes. These sample holding elements may also be formed with circular cutouts 58 arranged in rows on each sidewall 82, preferably spaced equidistant from each other, to insert a series of 0.2 ml PCR tubes. The oscillation element 57 is preferably a cylindrical block having one or two conical eccentric recesses 83 suitable for receiving 1.5 ml and 2 ml Eppendorf tubes, respectively. The sample holders 53 and the oscillation element 57 may be welded to the sample holding tray 55.

[0025] Figure 4 , Figure 5A and Figure 5B The configuration of a temperature control unit 30 according to a first embodiment of the present invention is shown. In this embodiment, the temperature control unit 30 includes a cold air passage 31, a hot air passage 32 having a heating element and a temperature sensor, and a fan 34 for uniform temperature regulation. The temperature regulation temperature is preferably set in the range of 30-85 °C + / - 0.5 °C.

[0026] In this configuration, the outlet opening 35 of the cold air passage 31 and the outlet opening 36 of the hot air passage 32 are formed on the partition wall 15, preferably along two adjacent edges of the partition wall 15. Figure 5B As can be seen in the illustrated embodiment, the cold air passage 31 and hot air passage 32 formed by the folded sheet can be at least partially surrounded by a heat shielding element 37, which is formed, for example, by a shield-shaped sheet, to ensure thermal insulation between the temperature control unit 30 and the drive unit 51. Openings 9 can preferably be formed on the sidewall of the second housing portion 12; these openings are for attaching to the fan 34 of the second housing portion 12.

[0027] The control unit 20 can control the heating element through pulse-width modulation (PWM) control based on user-provided commands and temperature values ​​measured by a temperature sensor. Temperature control is preferably based on the positive temperature coefficient (PTC) principle.

[0028] Figure 6 A device 101 according to a second embodiment of the present invention is shown. Device 101 includes a housing 110, which is composed of a first housing portion 111 and a second housing portion 112, separated by a partition wall 115. The housing 110 may have a heat-insulating, opaque wall structure. In this embodiment, the first housing portion 111 is provided with a recess 113, and the second housing portion 112 is provided with a recess 114. The housing 110 may be configured as a block made of plastic with a protective cover (not shown), in which the recesses 113 and 114 are separated by the partition wall 115. The overall dimensions of the housing 110 are, for example, 20 cm × 20 cm × 20 cm.

[0029] Figure 6 The power supply for the illustrated device 101 can be ensured by connecting it to a power grid with a voltage range of 100 V to 240 V. Device 101 preferably includes a power source consisting of a battery that provides a 12 V DC output voltage to ensure power supply during field use.

[0030] Similar to the device 1 described in the first embodiment of the present invention, device 101 includes a control unit 120, a temperature control unit 130 with a heating element, an illumination unit 140 for irradiating gene samples, a detection device, a sample holding element, and a vortex centrifuge unit 150 for centrifuging and shaking samples. In the second embodiment, the vortex centrifuge unit 150 is preferably arranged in the first housing portion 111, and the control unit 120, the temperature control unit 130, and the illumination unit 140 are preferably arranged in the second housing portion 112.

[0031] like Figure 6As shown, the temperature control unit 130 is advantageously configured as a block with sample holding elements disposed in the recess 114. The sample holding elements suitable for receiving samples to be tested can be formed, for example, as well as wells 135 and 138 disposed in the block, typically suitable for receiving 1.5 ml and 2 ml Eppendorf tubes and 0.2 ml PCR tubes, respectively. The temperature control portion of the block with wells 135 for receiving Eppendorf tubes can be set in the range of 30-100°C + / - 1°C, and the temperature control portion of the block with wells 138 for receiving PCR tubes can be set in the range of 30-70°C + / - 1°C.

[0032] The illumination unit 140 may be arranged in the second housing portion 112, below the sample holding element of the temperature control unit 130. In this embodiment, the illumination unit 140 preferably consists of a UV light source, a blue light source that generates blue light in the visible spectrum, and a white light source that generates white light, which are typically designed as an LED array.

[0033] In this embodiment, the detection device is detector 165, which is arranged in the second housing portion 112, below the sample holding element of the temperature-regulating unit 130. Detector 165 is suitable for photometric and fluorescence detection. Detector 165 can be a camera or sensor capable of photometrically detecting light emitted in the visible wavelength range of 350 nm to 650 nm by a sample placed in the temperature-regulating unit 130 and illuminated by the illumination unit 140. Detector 165 can also be used to detect fluorescence, such as FAM™ and HEX™ or other fluorophores in similar spectra, or even simultaneously detect two fluorophores emitted by a sample placed in the temperature-regulating unit 130 and illuminated by the illumination unit 140. Detector 165 is preferably insensitive to measurement errors caused by intensity variations in the illumination unit 140.

[0034] Similar to device 1 according to the first embodiment of the present invention, the second housing portion 112 of device 101 may include a window for observing gene samples that can be placed in device 101. This window may be equipped with a filter for filtering light emitted by the light source (i.e., UV light source, blue light source, and white light source) of illumination unit 140. (Window) Figure 6 (Not shown) can be made of, for example, an orange transparent polycarbonate sheet.

[0035] The control unit 120 may be a microprocessor for controlling the temperature control unit 130, the illumination unit 140, the vortex centrifuge unit 150, and the optional detector 165. In this embodiment, the control unit 120 is preferably placed in the second housing portion 112, above the temperature control unit 130.

[0036] like Figure 6 As shown, the control unit 120 is preferably connected to the control panel 122, which is, for example, arranged on the front surface of the housing 110. In this embodiment, the control panel 122 is provided with an LCD touch screen control and display interface 123 and control buttons. The LCD touch screen control and display interface and the control buttons are used to turn the light source (i.e., in this case, the UV light source, blue light source, and white light source) and the temperature control unit 130 on and off, and to set the temperature. In addition, the control panel 122 can also be used to select the operating mode of the vortex centrifuge unit 150 and to set the rotation speed and rotation time range.

[0037] The vortex centrifuge unit 150 can operate in two modes: a vortex mode with rapid rotation, and a centrifuge mode. The first mode can be turned on and off using control buttons, and the parameters for the centrifuge mode can be set using the LCD touchscreen control and display interface 123.

[0038] The control unit 120 may also include a data storage unit, for example, for storing settings of various modes of the vortex centrifugal unit 150 and settings of the illumination unit 140, as well as for analyzing and storing measurement results performed by the detector 165. The control unit 120 may be connected via the Internet to a central server that stores, for example, the data described above, and this central server may be connected to any selected application. The control unit 120 may also have a USB-type connection.

[0039] like Figure 6 and Figure 7 As shown, the configuration of the vortex centrifuge unit 150 of the device 101 according to the second embodiment of the present invention can be consistent with... Figure 3 The configuration of the vortex centrifuge unit 50 of the device 1 shown according to the first embodiment of the present invention is similar.

[0040] The vortex centrifuge unit 150 includes a drive unit and a rotatable sample holding tray 155 connected to the drive unit. The drive unit (not shown) may consist of a motor arranged in the first housing portion 111, which enables the sample holding tray 155 to rotate with a centrifugal force preferably 0-20000 × g. The sample holding tray 155 includes a sample holding element and an oscillating element 157 located at the center of the sample holding tray 155.

[0041] exist Figure 7In the illustrated embodiment, the sample holding tray 155, formed as a folded metal plate, may include a square base plate 181 and preferably four sidewalls 182 that project from the edge of the base plate 181 and slope outwards relative to the surface of the base plate 181. These sample holding elements may be, for example, sample holding lugs 154 cut from the base plate 181, sloped relative to the surface of the base plate, and provided with corresponding circular openings suitable for receiving 1.5 ml or 2 ml Eppendorf tubes. The sample holding elements may be seats mounted to one side of the tubes, suitable for supporting the tubes when they are rotated at high speeds. These sample holding elements may also be formed on each sidewall 182, preferably with circular cutouts 158 arranged in rows at equal intervals to insert a series of 0.2 ml PCR tubes. The oscillation element 157 may be a cylindrical block with a tapered eccentric recess 183 suitable for receiving 1.5 ml and 2 ml Eppendorf tubes, respectively. For example, the oscillating element 157 can be fixed to the sample holding tray 155 by interference fit or form fit connection or welding.

[0042] The following will refer to Figure 2 , Figure 3 and Figure 8 The operation of device 1 and device 101 according to the first and second embodiments of the present invention will be described respectively.

[0043] The LAMP-based assays that can be performed using devices 1 and 101 consist essentially of vortexing and centrifuging the gene sample, temperature conditioning the sample, irradiating the sample, detecting the light emitted by the irradiated sample, and evaluating the measurement results.

[0044] To perform the vortex operation, with housings 10 and 110 in the open position, Eppendorf tubes containing samples and desired reaction solutions, typically 1.5 ml or 2 ml in volume, are placed in the eccentric recesses 83 and 183 of the oscillating elements 57 and 157, and the Eppendorf tubes are held in the eccentric recesses by hand. The rotation of the sample holding trays 55 and 155 can be started slowly using control buttons 75 of control panel 22 or control buttons of control panel 122 (not shown). The speed of the vortex mode can be set to a desired value. Subsequently, 1.5 ml and 2 ml Eppendorf tubes containing samples, along with a series of 0.2 ml PCR tubes, can be placed in the sample holding seats 54 and cutouts 58 of sample holding tray 55 in their resting positions, or in the sample holding lugs 154 and cutouts 158 of sample holding tray 155 in their resting positions. Housings 10 and 110 are then closed. In this case, the device 1 according to the first embodiment of the invention can perform centrifugation at a high rotational speed.

[0045] The device 101 according to the second embodiment of the present invention can operate in two operating modes: a vortex mode including rapid rotation; and a centrifugation mode. The vortex (i.e., mixing) mode can be controlled via a control button on the control unit 122. The rotation speed of the vortex operation can also be adjusted. Similarly, the rapid rotation mode can be controlled via a control button on the control unit 122. In the rapid rotation mode, the tubes are placed in the sample holding lugs 154 and the slits 158 of the sample holding tray 155, respectively. The duration of the rapid rotation operation corresponds to the duration for which the control button is held in the pressed position. During this operation, the sample holding tray 155 can rotate with a centrifugal force ranging from 0 to 6000 × g. The centrifugation mode can be controlled via the LCD touchscreen and display interface 123 of the control unit 122. In this mode, the sample holding tray 155 can rotate continuously with a centrifugal force ranging from 14000 × g to 20000 × g, or rotate for a set time period (e.g., within the range of 0 to 30 minutes).

[0046] Temperature control is used for isothermal amplification of any gene marker in a gene sample.

[0047] In such Figures 1 to 5B In the case of the device 1 according to the first embodiment of the present invention, a series of eight PCR tubes containing previously shaken and centrifuged gene samples are placed in the cut 58, the housing 10 is closed, and the temperature control unit 30 is turned on using the control button 73. The desired temperature and the required slow rotation speed for this operation can be set using the control buttons 75 and 74, respectively.

[0048] In such Figures 6 to 8 In the case of the device 101 according to the second embodiment of the present invention, the gene sample previously vortexed and centrifuged in the vortex centrifuge unit 150 in the first housing portion 111 is transferred to the orifice 135 for receiving Eppendorf tubes and the orifice 138 for receiving PCR tubes in the temperature control unit 130 located in the second housing portion 112. The block portion provided with orifice 135 and the block portion provided with orifice 138 are heated to an appropriate temperature.

[0049] In both cases, the duration for performing the above operations can be selected via the control panels 22 and 122 of the control units 20 and 120, for example, within a time interval range of 1 minute to 120 minutes.

[0050] In such Figures 1 to 5B In the case of the apparatus 1 according to the first embodiment of the present invention, analysis of the products formed during the LAMP reaction can be performed by visual evaluation of the fully irradiated sample. The PCR series placed on the sample holding tray 55 is pivoted to a position parallel to the illumination unit 40.

[0051] With the housing 10 closed, depending on the detection marker used in the sample (e.g., a fluorescent dye), buttons 71 and 72 can be used to select either the UV light source 41 or the blue light source 42 to irradiate the sample in the tube. The color change of the irradiated sample can be observed through window 60. The amount of DNA formed during the reaction in the PCR tube can be detected by visual observation. Irradiation and analysis of the sample can be performed after the temperature setting step, or, if desired, simultaneously with the temperature setting step. In the case of the device 1 according to the first embodiment of the invention, a detector can be used to further perform analysis of the products formed during the LAMP reaction, which will be described in more detail below with reference to the second embodiment of the invention.

[0052] In such Figures 6 to 8 In the case of the device 101 according to the second embodiment of the present invention, the analysis of the products formed during the LAMP reaction can be performed by the measurement performed by the analytical detector 165. However, the analysis of the products formed during the LAMP reaction can be performed in a manner similar to that of the device 1 according to the first embodiment of the present invention, by visually evaluating the fully illuminated sample through a window formed on the housing 101.

[0053] With the housing 110 closed, depending on the potential detection marker (e.g., fluorescent dye) in the sample, the appropriate light source (i.e., UV, blue, or white light) of the illumination unit 140 can be selected using the LCD touchscreen control and display interface 123 to illuminate the sample in the tube. Photometric and / or fluorescence detection can be performed using the detector 165.

[0054] In photometric detection, the light emitted by the irradiated sample is detected within the visible wavelength range of 350 nm to 650 nm. In fluorescence detection, the fluorescence emitted by the irradiated sample can be detected, such as FAM™ and HEX™ or other fluorophores, or even a combination of two fluorophores falling within a similar spectrum.

[0055] In the current situation, the control unit 120 performs the analysis of the acquired measurement data. Illumination of the sample by the illumination unit 140, detection of the light emitted by the sample by the detector 165, and evaluation of the measurement results can be performed after or simultaneously with the temperature adjustment step.

[0056] It should be noted that the above embodiments can be combined by a person skilled in the art.

[0057] The advantage of the device according to the invention is that it allows for extensive on-site testing of genetic samples without the need for any modern laboratory equipment. Due to the compact configuration of the device, there is no need to transport samples to a central laboratory, thus minimizing the risk of sample degradation. Another advantage of the device according to the invention is that it allows for visual evaluation of the tests.

Claims

1. A device (1, 101) for detecting gene samples, said device comprising: - Casing (10, 110) - A control unit (20, 120), a temperature control unit (30, 130) with a heating element, an illumination unit (40, 140), and a detection device, wherein the control unit, the temperature control unit, the illumination unit, and the detection device are arranged within the housing (10, 110); - Sample holding element, the sample holding element being arranged within the housing (10, 110), Its features The device (1, 101) further includes a vortex centrifuge unit (50, 150), which has a drive unit and a rotatable sample holding tray (55, 155). The sample holding tray is connected to the drive unit and is provided with the sample holding element, wherein the oscillation element (57, 157) is located at the center of the sample holding tray (55, 155), and -The housing (10, 110) is composed of a first housing part (11, 111) and a second housing part (12, 112), and the first housing part and the second housing part are separated by a partition wall (15, 115).

2. The device (1, 101) according to claim 1, characterized in that, The lighting unit (40, 140) includes at least one of the following light sources: UV light source (41), blue light source (42), and white light source.

3. The device (1, 101) according to claim 1 or 2, characterized in that, The lighting unit (40) and the vortex centrifugal unit (50) are arranged in the first housing part (11), and the control unit (20) and the temperature control unit (30) are arranged in the second housing part (12).

4. The device (1, 101) according to claim 1 or 2, characterized in that, The vortex centrifugal unit (150) is arranged in the first housing portion (111), and the lighting unit (140), the control unit (120) and the temperature control unit (130) are arranged in the second housing portion (112).

5. The device (1) according to any one of claims 1 to 3, characterized in that, The temperature control unit (30) includes a cold air channel (31), a hot air channel (32), a fan (34), and a temperature sensor, wherein the heating element and the temperature sensor are attached to the hot air channel (32), and the outlet opening (35) of the cold air channel (31) and the outlet opening (36) of the hot air channel (32) are disposed on the partition wall (15).

6. The device according to claim 5, characterized in that, The cold air passage (31) and the hot air passage (32) are surrounded by a heat shielding element (37).

7. The device (101) according to claim 1, 2 or 4, characterized in that, The temperature control unit (130) is configured as a block having the sample holding element.

8. The device (1, 101) according to any one of the preceding claims, characterized in that, The detection device is a window (60) and / or a detector (165), the window having a filter disposed in the first housing portion (11, 111) or the second housing portion (12, 112), the detector being located in the first housing portion (11) or the second housing portion (112) and being suitable for photometric detection and / or fluorescence detection.

9. The device (1, 101) according to claim 8, characterized in that, The detector (165) is adapted to detect light emitted from a sample inserted into the device (1, 101) and illuminated by the illumination unit (40, 140).

10. The device (1, 101) according to any one of the preceding claims, characterized in that, The housing (10, 110) includes a control panel (22, 122) connected to the control unit (20, 120).

11. The device (1, 101) according to any one of the preceding claims, characterized in that, The housing (10, 110) has an opaque heat-insulating wall structure.

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