Device for testing genetic samples
The device addresses incomplete sample preparation and temperature control issues in LAMP testing by integrating a vortex-centrifugation unit and temperature control, facilitating accurate and cost-effective field testing with visual and photometric/fluorescent detection.
Patent Information
- Authority / Receiving Office
- AU · AU
- Patent Type
- Applications
- Current Assignee / Owner
- ÁLLATORVOSTUDOMÁNYI EGYETEM
- Filing Date
- 2024-12-02
- Publication Date
- 2026-07-16
Smart Images
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Abstract
Description
The present invention relates to a device for testing genetic samples, which is suitable for performing LAMP (Loop mediated isothermal AMPlification) based tests. The LAMP technique, i.e. loop mediated isothermal amplification technique, has various application possibilities in veterinary sciences, especially in diagnosing of infectious diseases. The test relies on the fact that an easily assessable DNA product is formed during a reaction occurring in a short time in a sample to be tested placed in an isothermal, i.e. constant temperature environment. This type of test can be performed in a thermostat. The document CN203720089U discloses a fluorescence intensity analyzer for LAMP testing. The analyzer comprises transparent, cylindrical sample holding elements for receiving reaction tubes, which are arranged on an insulated sample holding plate provided with a heating plate. A UV light source is arranged in front of each sample holding seat to illuminate the reaction product formed in the samples. The intensity of the light emitted by the illuminated samples is detected by a sensor connected to a data processing unit. This analyzer has the drawback that it does not allow for complete sample preparation. In addition, the samples are not tempered evenly. It is an object of the invention to provide a device for testing genetic samples that allows for simple and accurate preparation of samples tested in the framework of LAMP-based tests, as well as for accurate and cost-effective implementation and evaluation of the results of such tests. The above-mentioned objects are achieved by providing a device for testing genetic samples comprising a housing, and a control unit, a temperature control unit with a heating element, an illumination unit and a detection means, which are arranged inside the housing, and a sample holding elements arranged in the housing. The device further comprises a vortex-centrifugation unit having a drive unit and a rotatable sample holding tray connected to the drive unit and provided with the sample holding elements. A shaking element is provided on the centre of the sample holding tray. The housing consists of a first housing part and a second housing part separated by a partition wall. Various preferred embodiments of the invention are specified by the dependent claims. The present invention will now be described, by way of examples, with reference to the accompanying drawings, in which: Fig.l is an exploded perspective view of the device according to a first embodiment of the invention. Fig. 2 is a perspective front view of the device according to the first embodiment of the invention in a closed state. Fig. 3 is a perspective front view of the device according to the first embodiment of the invention in a closed state without the second housing part. Fig. 4 is a perspective front view of the vortex-centrifugation unit, the temperature control unit and the control unit of the device according to the first embodiment of the invention. Fig. 5A is a perspective top view of the partition wall and the temperature control unit of the device according to the first embodiment of the invention. Fig. 5B is a perspective bottom view of the partition wall and the temperature control unit of the device according to the first embodiment of the invention. Fig. 6 is an exploded perspective view of the device according to a second embodiment of the invention. Fig. 7 is an exploded perspective view of the vortex-centrifugation unit of the device according to the second embodiment of the invention. Fig. 8 is a perspective view of the device according to the second embodiment of the invention, in an open state, during use. Fig. 1 and Fig. 2 show the arrangement of the main units of a device 1 according to a first embodiment of the invention. The device 1 comprises a housing 10, which consists of a first housing part 11 and a second housing part 12 separated by a partition wall 15. The housing 10 may have a wall structure with a thickness of 10 mm and being provided with a gyroid type filling preferably formed by 3D printing from an opaque and thermally insulating polymer material. The edge 16 of the partition wall 15 may be provided with a seal 17 made of an elastic polymer, such as TPU material, which abuts against the edge 5 of the second housing part 12 and contacts the edge 6 of the first housing part 11 in the closed state of the housing 10. The closing of the housing 10 may be ensured, for example, by magnetic elements placed along the edges 5, 6 and 16. The second housing part 12 and the first housing part 11 may be connected to each other via hinges 3 arranged at the rear parts of their edges 5, 6. The housing 10 is preferably provided as a cube with an edge length of 20 cm. The power supply of the device 1 may be an uninterruptible power supply. The power requirement of the device 1 is preferably at most 65 W. The device 1 comprises a control unit 20, a temperature control unit 30 with a heating element, an illumination unit 40 for illuminating the genetic samples, a detection means, sample holding elements and a vortex-centrifugation unit 50 for centrifuging and shaking a sample. The illumination unit 40 and the vortex-centrifugation unit 50 are preferably located in the first housing part 11 and the control unit 20 and the temperature control unit 30 are preferably located in the second housing part 12. The illumination unit 40 is preferably arranged in a groove 18 defined by an opening formed on the partition wall 15 along the edge on the side of the front surface 2 of the housing 10. In this embodiment, the illumination unit 40 consists of a UV light source 41 and a blue light source 42 producing blue light in the visible spectrum, which are for example formed as a LED array. The control unit 20 is preferably a microprocessor for controlling the temperature control unit 30, the illumination unit 40 and the vortex-centrifugation unit 50. The control unit 20 is preferably connected to a control panel 22, which is placed, for example on the second housing part 12 on the front surface 2 of the housing 10. As shown in Fig. 2, the control panel 22 may be provided with an LCD display 23 and control buttons 71, 72, 73, 74, 75 and 76 for switching on and off the UV light source 41, the blue light source 42 and the temperature control unit 30, as well as for setting the tempering temperature, the rotation speed and time range of the vortexcentrifugation unit 50. The detection means may be a window 60 for observing genetic samples that can be placed in the device 1, the window 60 may be formed in this case in the first housing part 11 and may be provided with a light filter for filtering the light emitted by the light sources of the illumination unit 40, i.e. the UV light source 41 and the blue light source 42. The window 60 may be formed for example from an orange-coloured transparent polycarbonate sheet. The window 60 is preferably provided on the region of the front surface 2 of the first housing part 11. Moreover, a detector (not shown in Fig. 1) may be arranged in the first housing part 11 on the dividing wall 15, which is suitable for photometric and / or fluorescent detection. The detector can thus be suitable for detecting the light emitted by a sample placed in the device 1 and illuminated by the illumination unit 40. Fig. 1 and Fig. 3 show the configuration of the vortex-centrifugation unit 50 for preparing the genetic sample to be examined. The vortex-centrifugation unit 50 comprises a drive unit and a rotatable sample holding tray 55 connected to the drive unit. The drive unit may be formed as a motor 53 placed in a recess 52 provided on the partition wall 15, which enables the rotation of the sample holding tray 55 with a rotation speed of 600-2200 rpm preferably. The sample holding tray 55 comprises sample holding elements and a shaking element 57 provided on the centre of the sample holding tray 55. In the embodiment shown in Fig. 3, the sample holding tray 55 formed as a folded metal plate may comprise a square base plate 81 and four side walls 82 protruding from the edges of the base plate 81 and inclined outwardly with respect to the surface of the base plate 81. The sample holding elements may be cylindrical sample holding seats 54 having a central axis inclined with respect to the surface of the base plate 81, which are typically suitable for receiving Eppendorf tubes of 1,5 ml or 2 ml. The sample holding elements may also be formed with circular cutouts 58 arranged in a row on each side wall 82, preferably at the same distance from each other for inserting a string of PCR tubes of 0,2 ml. The shaking element 57 is preferably a cylindrical block having one or two conical eccentric recesses 83, which are suitable for receiving Eppendorf tubes of 1,5 ml and 2 ml respectively. The sample holding seats 53 and the shaking element 57 may be secured to the sample holding tray 55 by welding. Fig. 4, Fig. 5A and Fig. 5B show the configuration of the temperature control unit 30 according to the first embodiment of the invention. In the present embodiment the temperature control unit 30 comprises a cold air channel 31, a hot air channel 32 having a heating element and a temperature sensor and a fan 34 for homogeneous tempering. The tempering temperature is preferably set in the range of 30-85°C + / -0.5°C. In this case the outlet opening 35 of the cold air channel 31 and the outlet opening 36 of the hot air channel 32 are formed on the partition wall 15, preferably along two adjacent edges of the partition wall 15. As can be seen in the embodiment shown in Fig. 5B, the cold air channel 31 and the hot air channel 32 formed by a folded sheet may be surrounded at least partially by a heat shielding element 37 formed for example by a shield-like sheet ensuring thermal insulation between the temperature control unit 30 and the drive unit 51. Openings 9 may be formed preferably on the side walls of the second housing part 12 for the fan 34 attached to the second housing part 12. The control unit 20 may control the heating element by a PWM (Pulse-Width Modulation) control based on the command given by the user and the temperature values measured by the temperature sensor. The temperature control is preferably based on the PTC (Positive Temperature Coefficient) principle. Fig. 6 shows a device 101 according to a second embodiment of the invention. The device 101 comprises a housing 110, which consists of a first housing part 111 and a second housing part 112 separated by a partition wall 115. The housing 110 may have a heat-insulating opaque wall structure. In the present embodiment, the first housing part 111 is provided with a recess 113 and the second housing part 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 the figure), in which the recess 113 and the recess 114 are placed separated by a partition wall 115. The overall dimensions of the housing 110 is for example 20x20x20 cm. The power supply of the device 101 shown in Fig. 6 may be ensured by connection to the grid at a voltage ranging from 100 V to 240 V. The device 101 preferably includes a power supply consisting of a battery providing a DC output voltage of 12 V ensuring the power supply in the case of field use. Similarly to the device 1 according to the first embodiment of the invention described above, the device 101 comprises a control unit 120, a temperature control unit 130 with a heating element, an illumination unit 140 for illuminating genetic samples, a detection means, as well as sample holding elements and a vortex-centrifugation unit 150 for centrifuging and shaking the sample. In the second embodiment, the vortex-centrifugation unit 150 is preferably arranged in the first housing part 111 and the control unit 120, the temperature control unit 130, the illumination unit 140 are preferably arranged in the second housing part 112. As shown in Fig. 6, the temperature control unit 130 is advantageously configured as a block having sample holding elements arranged in the recess 114. Sample holding elements suitable for receiving the samples to be tested may be formed for example as through holes 135 and 138 arranged in the block, which are typically suitable for receiving respectively Eppendorf tubes of 1,5 ml and 2 ml and PCR tubes of 0,2 ml. The tempering temperature of the block part provided with holes 135 suitable for receiving Eppendorf tubes may be set set in the range of 30-100°C +1- 1°C and the tempering temperature of the block part provided with holes 138 suitable for receiving PCR tubes may be set in the range of 30-70°C +1- 1°C. The illumination unit 140 may be arranged in the second housing part 112 below the sample holding elements of the temperature control unit 130. In the present embodiment, the illumination unit 140 preferably consists of a UV light source, a blue light source producing blue light within the visible spectrum, and a white light source producing white light, which are typically designed as a LED array. In this embodiment, the detection means is a detector 165, which is arranged in the second housing part 112 below the sample holding elements of the tempering unit 130. The detector 165 is suitable for photometric and fluorescent detection. The detector 165 may be a camera or a sensor capable of photometrically detecting light in the visible wavelength range of 350-650 nm emitted by the sample placed in the tempering unit 130 and illuminated by the illumination unit 140. The detector 165 may also be suitable for detecting fluorescent light, such as FAM™ and HEX™ or other fluorophores within a similar spectrum, or even simultaneously detecting two fluorophores, emitted by the sample placed in the tempering unit 130 and illuminated by the illumination unit 140. The detector 165 is preferably not sensitive to measurement errors resulting from changes in the intensity of the illumination unit 140. Similar to the device 1 according to the first embodiment of the invention, the second housing part 112 of the device 101 may include a window for observing genetic samples that can be placed in the device 101, which may be provided with a light filter for filtering the light emitted by the light sources of the illumination unit 140, i.e. the UV light source, the blue light source and the white light source. The window (not shown in Fig. 6) may be formed for example from an orange-coloured transparent polycarbonate sheet. The control unit 120 may be a microprocessor for controlling the temperature control unit 130, the illumination unit 140, the vortex-centrifugation unit 150 and optionally the detector 165. In this embodiment, the control unit 120 is preferably placed in the second housing part 112, above the temperature control unit 130. As shown in Fig. 6, the control unit 120 is preferably connected to a control panel 122, which is arranged, for example, 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 a control button for turning on and off the light sources, i.e. in this case the UV light source, the blue light source and the white light source, as well as the temperature control unit 130 and for setting the tempering temperature. In addition, the control panel 122 may also be suitable for selecting the operating mode of the vortex-centrifugation unit 150 and for setting the rotation speed and rotation time range. The vortex-centrifugation unit 150 may be operated in two types of operating mode: a vortexing mode including a fast rotation operation and a centrifugation mode. The first mode may be switched on and off using the control button and the parameters of the centrifugation mode can be set using the LCD touch screen control and the display interface 123. The control unit 120 may further include a data storage unit for storing, for example, the settings of the individual modes of the vortex-centrifugation unit 150 and the illumination unit 140, as well as for analysing the measurements performed by the detector 165 and storing the measurement results. The control unit 120 may be connected via the internet to a central server, which stores, for example, the above-mentioned data, and which may be connected to any selected application. The control unit 120 may also have a USB type connection. As shown in Fig. 6 and Fig. 7, the configuration of the vortex-centrifugation unit 150 of the device 101 according to the second embodiment of the invention may be similar to the configuration of the vortex-centrifugation unit 50 of the device 1 according to the first embodiment of the invention shown in Fig. 3. The vortex-centrifugation unit 150 includes a drive unit and a rotatable sample holding tray 155 connected to the drive unit. The drive unit (not shown in the figure) may consist of a motor arranged in the first housing part 111, which enables the rotation of the sample holding tray 155 with a centrifugal force of preferably 0-20000 x g. The sample holding tray 155 includes sample holding elements and a shaking element 157 placed on the centre of the sample holding tray 155. In the embodiment shown in Fig. 7, the sample holding tray 155 formed as a folded metal plate may comprise a square base plate 181 and preferably four side walls 182 protruding from the edges of the base plate 181 and inclined outwardly with respect to the surface of the base plate 181. The sample holding elements may be, for example, sample holding tabs 154 cut out of the base plate 181, inclined with respect to its surface and provided with a corresponding circular opening, which may be suitable for receiving Eppendorf tubes of 1,5 ml or 2 ml. The sample holding elements may be seats fitted to the side of the tubes, which are suitable for supporting the tubes when rotating at high speeds. The sample holding elements may further be formed on each side wall 182, preferably with circular cutouts 158 arranged in a row at equal distances from each other for inserting a string of 0,2 ml PCR tubes. The shaking element 157 may be a cylindrical block having a conical eccentric recess 183, which is suitable for receiving 1,5 ml and 2 ml Eppendorf tubes respectively. The shaking element 157 may be secured to the sample holding tray 155, for example, by force-fitted or form-fitting connection or welding. The operation of the device 1 and the device 101 according to the first and second embodiment of the invention respectively will be described below with reference to Figs. 2, 3 and 8. The LAMP-based tests that can be performed with the device 1, 101 basically consist of vortexing and centrifuging genetic samples, tempering the samples, illuminating the samples, detecting the light emitted by the illuminated samples and evaluating the results of the measurement. For performing the vortexing operation, the Eppendorf tube having typically a volume of 1,5 ml or 2 ml containing the sample with the desired reaction solution is placed in the eccentric recess 83, 183 of the shaking element 57, 157 in the open state of the housing 10, 110 and is held therein by hand. The rotation of the sample holding tray 55, 155 may be started at a slow speed using the control button 75 of the control panel 22 or the control button of the control panel 122 (not shown in the figures). The speed of the vortexing mode may be set to the desired value. Subsequently, the 1,5 ml and 2 ml Eppendorf tubes and the string of 0,2 ml PCR tubes containing the samples may be placed respectively in the sample holding seats 54 and cutouts 58 of the sample holding tray 55, or in the sample holding tabs 154 and cutouts 158 of the sample holding tray 155 in the resting position. The housing 10, 110 is then closed. In the case of the device 1 according to the first embodiment of the invention, centrifugation may be performed at a fast rotation speed. The device 101 according to the second embodiment of the invention may be operated in two types of operating mode: a vortexing mode including a fast rotation operation and a centrifugation mode. The vortexing, i.e. mixing mode may be controlled by the control button of the control unit 122. The rotation speed of the vortexing operation may also be adjusted. Similarly, the fast rotation mode may be controlled by the control button of the control unit 122. In the fast rotation mode, the tubes are placed respectively in the sample holding tabs 154 and the cutouts 158 of the sample holding tray 155. The time duration of the fast rotation operation corresponds to the time duration of holding the control button in the pressed position. During this operation, the sample holding tray 155 may be rotated with a centrifugal force ranging from 0 to 6000 x g. The centrifugation mode may be controlled by the LCD touch screen control and display interface 123 of the control unit 122. In this mode, the sample holding tray 155 may be continuously rotated or rotated for a set period of time ranging from 0 to 30 minutes for example with a centrifugal force of 14000-20000 x g. The tempering operation is used for isothermal amplification of any genetic marker in the genetic samples. In the case of the device 1 according to the first embodiment of the invention as shown in Figs. 1-5B, the string of 8 PCR tubes containing previously shaken and centrifuged genetic samples is placed in the cutouts 58, the housing 10 is closed and the temperature control unit 30 is switched on with the control button 73. The desired tempering temperature and the slow rotation speed required for this operation may be set with the control buttons 75 and 74, respectively. In the case of the device 101 according to the second embodiment of the invention as shown in Figs. 6-8, the genetic samples previously shaken and centrifuged in the vortex-centrifugation unit 150 in the first housing part 111 are transferred to the holes 135 for accommodating Eppendorf tubes and to the holes 138 for accommodating PCR tubes of the temperature control unit 130 located in the second housing part 112. The block part provided with the holes 135 and the block part provided with the holes 138 are heated to the adequate temperature. In both cases, the time duration for performing the above-mentioned operations may be selected by the control panel 22, 122 of the control unit 20, 120, for example within a time interval ranging from 1 to 120 minutes. In the case of the device 1 according to the first embodiment of the invention shown in Figs. 15B, the analysis of the product formed during the LAMP reaction may be carried out by visual assessment of the adequately illuminated samples. The PCR string placed on the sample holding tray 55 is pivoted into a position parallel to the illumination unit 40. In the closed state of the housing 10 depending on the detection marker such as the fluorescent dye used in the samples, the UV light source 41 or the blue light source 42 illuminating the sampled in the tubes may be selected using the buttons 71 and 72, respectively. The colour change of the illuminated samples may be observed through the window 60. The amount of DNA formed during the reaction in the PCR tubes may be detected by visual observation. The illumination and analysis of the tested samples may be carried out after the tempering step or, if desired, simultaneously. In the case of the device 1 according to the first embodiment of the invention, the analysis of the product formed during the LAMP reaction may be further carried out using a detector, which will be described in more detail below referring to the second embodiment of the invention. In the case of the device 101 according to the second embodiment of the invention shown in Figs. 6-8, the analysis of the product formed during the LAMP reaction may be carried out by analysing the measurements performed by the detector 165. However, the analysis of the product formed during the LAMP reaction may be carried out in a similar manner to the device 1 according to the first embodiment of the invention, by visually assessing the adequately illuminated samples through a window formed on the housing 101. In the closed state of the housing 110, depending on the detection marker such as the fluorescent dye potentially applied in the samples, the adequate light source of the illumination unit 140 illuminating the samples in the tubes, i.e. the UV light source, the blue light source or the white light source, may be selected using the LCD touch screen control and display interface 123. Photometric and / or fluorescent detection may be performed with the detector 165. In the case of photometric detection, the detected light emitted by the illuminated sample falls within the visible wavelength range ranging from 350 nm to 650 nm. In the case of fluorescent detection, the fluorescent light such as FAM™ and HEX™ or other fluorophores or even a combination of two fluorophores falling within a similar spectrum emitted by the illuminated sample may be detected. In the present case, the analysis of the obtained measurement data is carried out by the control unit 120. The illumination of the sample with the illumination unit 140 and the detection of the light emitted by the sample with the detector 165 and the evaluation of the results of the measurement may be performed after the tempering step or simultaneously. It should be noted that the above-mentioned embodiments may be combined by the skilled person. The advantage of the device according to the invention is that it allows for extensive field testing of genetic samples without requiring any modem laboratory equipment. Thanks to the compact configuration of the device, it is not necessary to transport the samples to a central laboratory, thereby minimizing the risk of sample degradation. Another advantage of the device according to the invention is that it allows for visual assessment of the tests.
Claims
1. A device (1, 101) for testing genetic samples, said device 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, which are arranged inside the housing (10, HO),- sample holding elements arranged in the housing (10, 110), characterized in that- 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 elements, wherein a shaking element (57, 157) is provided on the centre of the sample holding tray (55, 155), and- wherein the housing (10, 110) consists of a first housing part (11, 111) and a second housing part (12, 112) separated by a partition wall (15, 115).
2. The device (1, 101) according to claim 1, characterized in that the illumination unit (40, 140) comprises at least one of the following light sources: a UV light source (41), a blue light source (42), a white light source.
3. The device (1, 101) according to claim 1 or 2, characterized in that the illumination unit (40) and the vortex-centrifugation 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 vortexcentrifugation unit (150) is arranged in the first housing part (111), and the illumination unit (140), the control unit (120) and the temperature control unit (130) are arranged in the second housing part (112).
5. The device (1) according to any of claims 1 to 3, characterized in that the temperature control unit (30) comprises 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 thehot air channel (32), wherein the outlet opening (35) of the cold air channel (31) and the outlet opening (36) of the hot air channel (32) are provided on the partition wall (15).
6. The device according to claim 5, characterized in that the cold air channel (31) and the hot air channel (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 provided as a block with sample holding elements.
8. The device (1, 101) according to any one of the preceding claims, characterized in that the detection means is a window (60) having a light filter provided in the first housing part (11, 111) or in the second housing part (12, 112) and / or a detector (165) located in the first housing part (11) or in the second housing part (112) suitable for photometric and / or fluorescent detection.
9. The device (1, 101) according to claim 8, characterized in that the detector (165) is suitable for detecting light emitted by a sample inserted in 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) comprises 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, thermally insulating wall structure.