Nucleic acid analysis apparatus
11426735 ยท 2022-08-30
Assignee
Inventors
Cpc classification
B01L2300/1805
PERFORMING OPERATIONS; TRANSPORTING
B01L2200/14
PERFORMING OPERATIONS; TRANSPORTING
B01L2300/08
PERFORMING OPERATIONS; TRANSPORTING
B01L7/04
PERFORMING OPERATIONS; TRANSPORTING
B01L2200/16
PERFORMING OPERATIONS; TRANSPORTING
B01L2200/025
PERFORMING OPERATIONS; TRANSPORTING
B01L3/502715
PERFORMING OPERATIONS; TRANSPORTING
B01L7/525
PERFORMING OPERATIONS; TRANSPORTING
International classification
B01L3/00
PERFORMING OPERATIONS; TRANSPORTING
B01L7/04
PERFORMING OPERATIONS; TRANSPORTING
Abstract
A nucleic acid analysis apparatus includes a casing, a main frame, a fluid delivery unit, a thermal unit, a driving unit, and at least one optical unit. The casing has an upper casing and a lower casing. The main frame is disposed in the lower casing and has a chamber for mounting a cartridge therein. The fluid delivery unit is adapted to transport reagents within the cartridge for sample purification and/or nucleic acid extraction. The thermal unit is adapted to provide a predefined temperature for nucleic acid amplification. The driving unit is disposed in the lower casing and connected with the main frame, and includes a motion control unit capable of pressing the cartridge during sample purification and/or nucleic acid extraction and rotating the cartridge with a predefined program during nucleic acid amplification and/or detection. The optical unit includes plural optical components for detection.
Claims
1. A nucleic acid analysis apparatus, comprising: a casing having an upper casing and a lower casing; a main frame disposed in the lower casing and having a chamber for mounting a cartridge therein; a fluid delivery unit disposed in the lower casing and connected with the main frame, being adapted to transport reagents within the cartridge for sample purification and/or nucleic acid extraction, wherein the fluid delivery unit comprises a plurality of microchannels for connecting with the cartridge and a pump adapted to provide pneumatic force to transport the reagents stored within the cartridge; a thermal unit disposed on the main frame of the lower casing and comprising a heater to provide a predefined temperature for nucleic acid amplification; a driving unit disposed in the lower casing and connected with the main frame, the driving unit comprising a motor, a shaft, a driven gear, and a motion control gear, wherein the motion control gear is engaged with the driven gear mounted on the shaft, and the motor drives rotations of the driven gear and the motion control gear, and thus drives movement and rotation of the cartridge clamped by the motion control gear, wherein the motion control gear comprises at least one protrusion, the cartridge comprises at least one guiding groove having a vertical groove and an inclined groove, and the protrusion is able to slide in the guiding groove, so that the motion control gear is capable of pressing the cartridge toward the fluid delivery unit during sample purification and/or nucleic acid extraction and rotating the cartridge during nucleic acid amplification and/or detection; and at least one optical unit disposed on the main frame of the lower casing and comprising at least one light source and at least one optical detector for detection.
2. The nucleic acid analysis apparatus according to claim 1, wherein the motor comprises a stepper motor, which actuates a rotation of the motion control gear through gear transmission.
3. The nucleic acid analysis apparatus according to claim 2, wherein the motion control gear comprises a plurality of rollers, which are accommodated in an annular groove of the main frame.
4. The nucleic acid analysis apparatus according to claim 1, wherein the nucleic acid analysis apparatus further comprises a fluid connector located between the main frame and the fluid delivery unit, and a flat spring equipped on the fluid connector.
5. The nucleic acid analysis apparatus according to claim 4, wherein a forward rotation of the motion control gear allows the cartridge to be locked and in contact with the fluid connector.
6. The nucleic acid analysis apparatus according to claim 5, wherein a reverse rotation of the motion control gear allows the cartridge to be bounced up by the flat spring, detach the fluid connector, and rotate according to the predefined program.
7. The nucleic acid analysis apparatus according to claim 1, wherein the nucleic acid analysis apparatus further comprises a sensor to recognize a position of the cartridge.
8. The nucleic acid analysis apparatus according to claim 1, wherein the nucleic acid analysis apparatus further comprises an indicator, which is shining when the cartridge is pressed to the end to inform a user to release his hand.
9. The nucleic acid analysis apparatus according to claim 1, wherein the optical unit comprises a light source and an optical detector.
10. The nucleic acid analysis apparatus according to claim 1, wherein the nucleic acid analysis apparatus comprises multiple optical units, and each optical unit offers a unique wavelength of illumination to detect multiple targets.
11. The nucleic acid analysis apparatus according to claim 1, wherein the nucleic acid analysis apparatus further comprises a touch screen disposed on the lower casing with adjustable operation angles.
12. The nucleic acid analysis apparatus according to claim 1, wherein the cartridge comprises a reaction chip and a cartridge body, and the reaction chip is disposed on one side of the cartridge body.
13. The nucleic acid analysis apparatus according to claim 12, wherein the main frame further comprises at least one positioning pin, and the reaction chip comprises at least one alignment slot capable of being aligned with the at least one positioning pin on the main frame.
14. The nucleic acid analysis apparatus according to claim 12, wherein the cartridge body comprises plural chambers used to store reagents for sample purification and nucleic acid extraction.
15. The nucleic acid analysis apparatus according to claim 12, wherein the reaction chip further comprises at least one sample loading hole at a top surface of the reaction chip for adding sample into the cartridge.
16. The nucleic acid analysis apparatus according to claim 12, wherein the thermal unit carries the reaction chip thereon to provide heating.
17. The nucleic acid analysis apparatus according to claim 12, wherein the reaction chip is a planar fluidic chip and comprises a plurality of detection wells and at least one microchannel connected with the detection wells.
18. The nucleic acid analysis apparatus according to claim 17, wherein the shape of the reaction chip is substantially a regular polygon, and each of the detection wells has at least one planar surface.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1)
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DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
(9) The present invention will now be described more specifically with reference to the following embodiments. It is to be noted that the following descriptions of the embodiments of this invention are presented herein for purpose of illustration and description only; it is not intended to be exhaustive or to be limited to the precise form disclosed.
(10) The present invention provides an all-in-one nucleic acid analysis apparatus, which integrates a fluid delivery unit, a thermal unit, a driving unit, and at least one optical unit on one single device, so that the processes of sample purification, nucleic acid extraction, nucleic acid amplification and nucleic acid detection can be performed on the all-in-one apparatus to realize nucleic acid analysis in real time.
(11)
(12) In an embodiment, the nucleic acid analysis apparatus 10 further includes a controller, such as micro control unit (MCU), which controls the operations of the fluid delivery unit 3, the thermal unit 4, the driving unit 5 and the optical unit 6.
(13) In an embodiment, the upper casing 11 and the lower casing 12 are connected through a hinge, but not limited thereto. The upper casing 11 can be opened, so that the cartridge 7 is able to be placed into the chamber 21 of the main frame 2. When the upper casing 11 is closed, a confined space is formed in the casing 1.
(14)
(15) The number of the detection wells 721 is not limited, and may be up to 40 or even more, and the apparatus could perform multiplexing nucleic acid analysis. In an embodiment, the shape of the reaction chip 72 is substantially a regular polygon, so that the reaction chip 72 has plural planar side surfaces to be in line with the optical unit 6 to facilitate light focusing. Certainly, the shape of the reaction chip 72 is not limited to the regular polygon and it may also be circular or other shape, since the light could be focused on the sample in the detection well 721 by the arrangement of optical components of the optical unit 6.
(16) In an embodiment, the reaction chip 72 further includes at least one alignment slot 723, and the main frame 2 further includes at least one positioning component 22 (as shown in
(17) The cartridge body 71 includes plural chambers 711 used to store reagents for sample purification and/or nucleic acid extraction. The cartridge body 71 also includes plural channels connected with the chambers 711 for fluid delivery. In an embodiment, the cartridge body 71 is but not limited to a cylindrical body. The cartridge body 71 further includes plural openings 712 at the bottom surface of the cartridge body 71, and the openings 712 are communicated with the chambers 711 through the channels. The shape of the openings 712 may be but not limited to circular, linear or other regular or irregular shape.
(18) The reaction chip 72 further includes at least one sample loading hole 724 at the top surface of the reaction chip 72, and the sample loading hole 724 aligns and communicates with at least one chamber 711 of the cartridge body 71 for adding sample into the cartridge 7.
(19) During the operation, once the sample is loaded, the sample loading hole 724 is sealed and the cartridge 7 is placed into the chamber 21 of the nucleic acid analysis apparatus 10 and is forced to tightly contact the fluid delivery unit 3 without leakage, and then the flow processing is carried out by the fluid delivery unit 3. The fluid delivery unit 3 works concurrently with the cartridge 7 to carry out sample purification, nucleic acid extraction and fluid delivery so as to have a fully automatic device. The fluid delivery could be realized by pneumatic, vacuum, plunger, chamber deformation, thermal-induced expansion, acoustics, centrifugal force or other methods as long as the sample processing is completed within the cartridge body 71.
(20) In an embodiment, the flow is driven pneumatically through microchannels and holes. For example, the fluid delivery unit 3 is similar to the integrated fluidic module of U.S. Pat. No. 10,124,335 B2 filed by the applicant of the present invention, the entire contents of which are incorporated herein by reference and are not redundantly described here. In brief, the fluid delivery unit 3 of the present invention includes the fluid manifold, the valve stator, the valve rotor, the valve housing and the fluid sources as disclosed in U.S. Pat. No. 10,124,335 B2. The fluid manifold includes plural microchannels for connecting with the chambers 711 of the cartridge 7 through the bottom openings 712. By the alignments of the through holes and/or grooves of the valve stator and the valve rotor, multi-way fluid path switching is realized when the valve rotor is rotated to different positions, so as to regulate the fluid operations in the cartridge 7. Thereby, the reagents stored within the cartridge 7 are able to be transported to desired locations through pneumatic force from pumps of the fluid delivery unit 3, so as to automatically perform the sample purification and the nucleic acid extraction. Certainly, the fluid delivery unit 3 is not limited to the above-mentioned design, and can be any other type as long as it is able to realize multiple fluid delivery and multi-way fluid path switching in the cartridge 7.
(21)
(22) In an embodiment, the nucleic acid analysis apparatus 10 is designed to amplify nucleic acid based on isothermal method and therefore only a constant temperature instead of thermal cycling among three different temperature zones is needed. As a result, the thermal unit 4 is significantly simplified. In addition, the chamber 21 of the nucleic acid analysis apparatus 10 is designed with superior thermal insulation and therefore the inner temperature is easily maintained. Once the chamber 21 is in a uniform temperature environment, heat loss from the detection wells 721 and sample towards the environment could be minimized. At the amplification and/or detection processes, the whole closed chamber 21 and the sample at each detection well 721 are substantially in the same temperature, regardless the cartridge 7 is in motion or in stationary.
(23) The thermal unit 4 provides the required temperature within the chamber 21 during the operation, wherein the temperature control is independent of the number and shape of detection wells 721. In an embodiment, the thermal unit 4 further includes a temperature sensor to control the accuracy of the temperature.
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(25) The motion control unit 52 may be actuated by the stepper motor 51 through gear, belt, chain, rack, worm or other mechanical transmission mechanism. In an embodiment, the motion control unit 52 is a motion control gear, and the driving unit 5 further includes a driven gear 53 connected with the stepper motor 51 through a shaft 54. The motion control unit 52 is engaged with the driven gear 53, and the stepper motor 51 drives the rotations of the driven gear 53 and the motion control unit 52, and thus drives the movement and rotation of the cartridge 7 clamped by the motion control unit 52.
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(27) As shown in
(28) In an embodiment, the motion control unit 52 includes two symmetric protrusions 522, and correspondingly, the cartridge 7 also includes two symmetric guiding grooves 73. Certainly, the numbers of the protrusions 522 and the guiding grooves 73 are not limited to two.
(29)
(30) In an embodiment, a sensor is embedded at the bottom of the chamber 21 to recognize the position of the cartridge 7 and is able to feedback as long as the cartridge 7 is pressed to contact the fluid connector 82. At the moment, the protrusion 522 of the motion control unit 52 reaches the highest point of the vertical groove 731, i.e. the cartridge 7 is presses to the end and after which the motion control unit 52 is actuated to lock the cartridge 7, and an indicator, such as LED, is shining to inform the user to release his hand. By this design, the cartridge 7 is installed by user's hand, and thus the driven motor torque is minimized.
(31) When the cartridge 7 is pressed down to the end, a signal is sent to the MCU, and then the stepper motor 51 is triggered so that the motion control unit 52 can rotate forwardly and the protrusion 522 can slide along the inclined groove 732 on the cartridge 7 (as shown in
(32) After the sample with extracted nucleic acid is dispensed to the detection wells 721 of the cartridge 7 and heated to a pre-defined temperature, the motion control unit 52 rotates reversely and the protrusion 522 moves backwardly along the inclined groove 732 (as shown in
(33) At the nucleic acid amplification and detection stage, since the cartridge 7 is constrained by the protrusion 522 at this moment, the further reverse rotation of the motion control unit 52 will drive the rotation of the cartridge 7. Therefore, the cartridge 7 can be rotated with the predefined programs by the motion control unit 52 to align the detection wells 721 with each optical unit 6 sequentially for nucleic acid detection until the completion of the detection.
(34) In an embodiment, the nucleic acid analysis apparatus 10 includes multiple optical units 6. The optical unit 6 has optical components such as light source, lens, filter and optical detector to realize the optical detection so that the sample could be detected in real time during the nucleic acid amplification. The optical unit 6 may include at least one light source 61 and at least one optical detector 62 (as shown in
(35) By utilizing the isothermal based amplification, the thermal unit 4 is significantly simplified, and thus, the nucleic acid analysis apparatus 10 can be compact designed and is even smaller than a common teacup. In an embodiment, the nucleic acid analysis apparatus 10 has a height ranged between 100 mm and 200 mm and a width ranged between 80 mm and 120 mm. Since the nucleic acid analysis apparatus 10 is cup sized, it is portable and suitable for POC diagnostics.
(36) Further, by rearranging the driving unit 5 and the motion control unit 52 to the lower casing 12, both size and weight of the top part of the nucleic acid analysis apparatus 10 are reduced.
(37) In an embodiment, the nucleic acid analysis apparatus 10 is designed for isothermal based amplification, and thus can be used to perform all isothermal amplification methods, such as nucleic acid sequence-based amplification (NASBA), strand displacement amplification (SDA), helicase-dependent amplification (HDA), loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA) and nicking enzyme amplification reaction (NEAR).
(38) In another embodiment, the nucleic acid analysis apparatus 10 may also be designed for PCR-based amplification. For example, the spring-supported component 81 may be equipped with a heater therein to provide another temperature zone.
(39) In conclusion, the present invention provides an all-in-one nucleic acid analysis apparatus, which integrates the fluid delivery unit, the thermal unit, the driving unit and the optical unit on one single device, so that the processes of sample purification, nucleic acid extraction, nucleic acid amplification and nucleic acid detection can be performed on the all-in-one apparatus to realize nucleic acid analysis in real time. Therefore, the nucleic acid analysis apparatus provides an easy and fast nucleic acid analysis. In addition, since the thermal unit is significantly simplified, the nucleic acid analysis apparatus can be compact designed, so it is portable and suitable for POC diagnostics. Moreover, since the cartridge is pressed down by user's hand, the nucleic acid analysis apparatus has reduced motor size for motion control unit. Besides, by rearranging the driving unit and the motion control unit to the lower casing, both size and weight of the top part of the nucleic acid analysis apparatus are reduced. Further, the touch screen disposed on the lower casing could be enlarged and could have adjustable operation angles to facilitate user's viewing and operation.
(40) While the invention has been described in terms of what is presently considered to be the most practical and preferred embodiments, it is to be understood that the invention needs not be limited to the disclosed embodiment. On the contrary, it is intended to cover various modifications and similar arrangements included within the spirit and scope of the appended claims which are to be accorded with the broadest interpretation so as to encompass all such modifications and similar structures.