Cartridge and method for quantifying same

Abstract

To provide a cartridge within which at least one quantification chamber is disposed, so as to achieve active quantification during transfer of liquid, thereby enhancing sensitivity and accuracy of screening by the cartridge.SOLUTION: An cartridge and a quantification method comprise a main cover, a container, a first pipette, and a rotary valve. The main cover has a first surface and a second surface opposite to each other. The first surface comprises a first quantification chamber, a first fluid tunnel, a first gas tunnel, and a storage chamber. A first end of the first quantification chamber is connected to the first fluid tunnel, a first end of the storage chamber is connected to the first gas tunnel, and a second end of the first quantification chamber is connected to a second end of the storage chamber. The first pipette is disposed on the main cover and protrudes from the second surface. The first pipette is connected to the second end of the first quantification chamber, and extends vertically in the container. A rotary valve is rotatably disposed on the second surface.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a cartridge and a method for quantifying the same, and more particularly to a cartridge for nucleic acid extraction and nucleic acid amplification and a method for quantifying the same. [Background technology]

[0002] Nucleic acid extraction and nucleic acid amplification are common techniques used in biomedical testing or diagnosis. Generally, nucleic acid extraction kits or nucleic acid extraction reagents are usually used in open routine laboratories for nucleic acid extraction, followed by nucleic acid amplification kits or nucleic acid amplification reagents to amplify or detect specific nucleic acid fragments. However, the above kits or reagents usually require manual operation, which is time-consuming and prone to sample or reagent contamination, thereby making them less efficient for use in mass testing or production line testing. Meanwhile, although some existing analytical cartridges can be operated mechanically, these cartridges usually have complex configurations and require expensive machines for operation, thereby increasing screening costs. Therefore, there is still a need to provide new and improved technologies for the relevant art to meet the actual needs of the relevant art. Summary of the Invention [Problem to be solved by the invention]

[0003] With this in mind, the present disclosure aims to provide a cartridge having at least one quantification chamber disposed therein, which achieves active quantification while transferring liquid, thereby increasing the sensitivity and accuracy of screening with the cartridge.

[0004] The present invention also aims to provide a method for quantification of cartridges that allows for synchronous quantification of liquid while the liquid is being transported by flowing through at least one quantification chamber in the cartridge, thereby simplifying operation and improving the sensitivity and accuracy of cartridge screening. [Means for solving the problem]

[0005] This is achieved by the cartridge and the quantification method thereof according to the independent claims. The dependent claims relate to corresponding further developments and improvements.

[0006] As will be more clearly understood from the detailed description below, the present disclosure provides a cartridge. The cartridge includes a main cover, a container, a first pipette, and a rotary valve. The main cover has a first surface and a second surface facing each other, the first surface including a first quantification chamber, a first fluid tunnel, a first gas tunnel, and a storage chamber extending in a plane, the first end of the first quantification chamber being connected to the first fluid tunnel, the first end of the storage chamber being connected to the first gas tunnel, and the second end of the first quantification chamber being connected to the second end of the storage chamber. The container is disposed on the second surface of the main cover and vertically overlaps the second end of the first quantification chamber. The first pipette is disposed on the main cover and partially protrudes from the second surface of the main cover. The first pipette is connected to the second end of the first quantification chamber and extends vertically into the at least one container. The rotary valve is rotatably disposed on the second surface of the main cover.

[0007] As will be more clearly understood from the detailed description below, the present disclosure provides a quantification method for a cartridge. The quantification method includes the following steps: First, a cartridge is provided, the cartridge including a main cover, a plurality of containers, a first pipette, and a rotary valve. The main cover includes a first quantification chamber, a first fluid tunnel, a first gas tunnel, and a storage chamber extending on a plane, the first end of the first quantification chamber being connected to the first fluid tunnel, the first end of the storage chamber being connected to the first gas tunnel, and the second end of the first quantification chamber being connected to the second end of the storage chamber. A first pipette is disposed on the main cover, and the first pipette is connected to the second end of the first quantification chamber. A plurality of containers are disposed on the main cover, one of the plurality of containers vertically overlapping the second end of the first quantification chamber. A rotary valve is rotatably disposed on the main cover. Next, a reagent is injected into the first quantification chamber from a first end of the first quantification chamber. After the reagent is injected into the first quantification chamber, a portion of the reagent is transferred from a second end of the first quantification chamber to a storage chamber. Then, by rotating the rotary valve, the remainder of the reagent in the first quantification chamber is transferred to one of the multiple containers. [Brief explanation of the drawings]

[0008] The present invention will now be further described by way of example with reference to the accompanying drawings, in which:

[0009] [Figure 1] 1 is a schematic view showing a cartridge according to a first embodiment of the present invention;FIG. 2 is an exploded view of the cartridge according to the first embodiment of the present invention;FIG. [Figure 2] 1 is a schematic view showing a cartridge according to a first embodiment of the present invention;

[0023] FIG. 2 is a side view of the cartridge according to the first embodiment of the present invention; [Figure 3] 1A and 1B are schematic diagrams illustrating a cartridge according to a first embodiment of the present invention;FIGS. 1A and 1B show cross-sectional views of a cartridge according to a first embodiment of the present disclosure;FIGS. [Figure 4]1A and 1B are schematic diagrams illustrating a cartridge according to a first embodiment of the present invention;FIGS. 1A and 1B show a cross-sectional view of a second quantification chamber according to a first embodiment of the present disclosure;FIGS. [Figure 5] 1A and 1B are schematic diagrams illustrating a cartridge according to a second embodiment of the present invention;FIGS. [Figure 6] 1 is a schematic diagram illustrating a cartridge according to a second embodiment of the present invention;FIG. 2 shows a side view of a cartridge according to a second embodiment of the present disclosure;FIG. [Figure 7] FIG. 1 is a schematic diagram showing a processing flow of a quantification method according to a preferred embodiment of the present disclosure. [Figure 8] FIG. 10 is a schematic view showing a cartridge according to a third embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0010] In order to provide a better understanding of the present disclosure, preferred embodiments will now be described in detail. Preferred embodiments of the present disclosure are illustrated in the accompanying drawings, which have numbered elements.

[0011] In this disclosure, forming a first feature over a second feature in the description may include embodiments in which the first and second features are formed in direct contact with each other, and may also include embodiments in which an additional feature may be formed between the first and second features such that the first and second features are not in direct contact with each other. Additionally, this disclosure may repeat reference numerals and / or letters in various examples. This repetition is for the purposes of brevity and clarity and does not, in itself, dictate a relationship between the various embodiments and / or configurations discussed. Furthermore, spatially relative terms such as "beneath," "below," "lower," "over," "above," and "upper" may be used herein for ease of description to describe the relationship of one element or feature to another element(s) or feature(s), as shown in the figures. Spatially relative terms are intended to encompass different orientations of the device during use or operation in addition to the orientation depicted in the figures. For example, if a device in the figures is inverted, elements described as "below" and / or "beneath" other elements or features would then be oriented "above" and / or "above" the other elements or features. The device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein may likewise be interpreted accordingly.

[0012] Terms such as "first," "second," and "third" may be used herein to describe various elements, components, regions, layers, and / or sections, but it should be understood that these elements, components, regions, layers, and / or sections should not be limited by these terms. These terms are used only to distinguish one element, component, region, layer, and / or section from another region, layer, and / or section. Terms such as "first," "second," and other numerical terms, when used herein, do not imply a sequence or order unless clearly indicated by context. Thus, a first element, component, region, layer, and / or section described below may be referred to as a second element, component, region, layer, and / or section without departing from the teachings of the embodiments.

[0013] As disclosed herein, the term "about" or "substantially" generally means within 20%, preferably within 10%, and more preferably within 5%, 3%, 2%, 1%, or 0.5% of a given value or range. Unless expressly stated otherwise, all numerical ranges, amounts, values, and percentages disclosed herein should be understood as being modified in all instances by the term "about" or "substantially." Accordingly, unless indicated to the contrary, the numerical parameters set forth in this disclosure and appended claims are approximations that may be varied as desired.

[0014] 1 to 4, a cartridge 100 according to a first embodiment of the present invention is shown. FIG. 1 is a schematic exploded view of the cartridge 100, FIG. 2 is a schematic side view of the cartridge 100, FIG. 3 is a schematic cross-sectional view of the cartridge 100, and FIG. 4 is a schematic cross-sectional view of a second quantification chamber 121. As shown in FIGS. 1 to 3, the cartridge 100 includes a main cover 102, a container 120a, a first pipette 112a, and a rotary valve. The main cover 102 includes a first surface 102a and a second surface 102b facing each other. The first surface 102a of the main cover 102 includes a first quantification chamber 108, a first fluid tunnel 104a, a first gas tunnel 106a, and a storage chamber 110. The container 120a is disposed on the second surface 102b of the main cover 102. The first pipette 112a has a hollow structure (as shown in FIG. 2 ), extends downward from the first surface 102a, partially protrudes from the second surface 102b, and extends into the container 120a. The rotary valve 116 is rotatably disposed on the second surface 102b. Note that the first end 108a of the first quantification chamber 108 is connected to the first fluid tunnel 104a, the first end 110a of the storage chamber 110 is connected to the first gas tunnel 106a, the second end 108b of the first quantification chamber 108 is connected to the second end 110b of the storage chamber 110, and the second ends 108b / 110b of the first quantification chamber 108 and the storage chamber 110 both connect to the first pipette 112a and overlap the lower container 120a in the vertical direction D3. Therefore, by locating the first quantification chamber 108, the cartridge 100 allows active quantification to be achieved while injecting reagent into the container 120a, thereby improving the sensitivity and accuracy of the cartridge 100 in screening.

[0015] To be precise, the main cover 102 extends along a plane defined by both directions D1 and D2 and may be formed by a plastic injection molding method using a suitable material selected from a group including, but not limited to, polypropylene (PP), polycarbonate (PC), polyimide (PI), polyethylene terephthalate (PET), and others having thermoplastic and biocompatible properties. The first fluid tunnel 104a and the first gas tunnel 106a are liquid or gas flow channels recessed downward from the first surface 102a. The first quantification chamber 108 and the storage chamber 110 are spaces recessed downward from the first surface 102a for liquid quantification or to accommodate necessary reagents. Those skilled in the art should fully understand that the spaces of the first quantification chamber 108 and / or the storage chamber 110 may include any possible shape or volume based on actual requirements and are not limited to those shown in FIG. 1 . 1, to prevent the reagent from remaining in the first quantification chamber 108 during reagent transfer and affecting the quantification results. In another embodiment, a hydrophobic film (not shown) may be further coated on the two side walls 108c of the first quantification chamber 108 to further prevent the reagent from remaining in the first quantification chamber 108, thereby facilitating the transfer of the reagent between each chamber and the container 120a.

[0016] 1 and 2, the rotary valve 116 is attached to the second surface 102b of the main cover 102 via a base 126 disposed below, and further includes a first portion 116a and a second portion 116b disposed from top to bottom. The first portion 116a partially protrudes to define a flow path 118a and an opening 118b, respectively. The opening 118b is used for gas circulation, and the flow path 118a is used for liquid circulation. The flow path 118a may include any suitable shape, such as, but not limited to, a linear shape as shown in FIG. 1. In one embodiment, the first portion 116a and the second portion 116b comprise different materials, for example, the first portion 116a comprises a material such as thermoplastic polyurethane (TPU), rubber, polyurethane material, polyethylene, polyethylene terephthalate (PET), thermoplastic polyester elastomer (TPEE), biocompatible resin, or a combination thereof for adhering to the second surface 102b, and the second portion 116b comprises a rigid material such as, but not limited to, polypropylene fiber, polycarbonate, etc. Through these arrangements, the second portion 116b of the rotary valve 116 may be externally connected to a motor (not shown), which drives and controls the rotary valve 116 within the cartridge 100 to rotate in a specific direction and angle, and then the opening 118b of the first portion 118 and the flow tunnel 116a are aligned with the first gas tunnel 106a and the first fluid tunnel 104a, respectively, thereby communicating the first fluid tunnel 104a, the first quantification chamber 108, the first pipette 112a, and the container 120a. In this way, while positive or negative pressure is provided by an external pump (not shown), the cartridge 100 is capable of delivering reagent from the first fluid tunnel 104a into the first quantification chamber 108 for liquid quantification.

[0017] 1, the main cover 102 further includes a second fluid tunnel 104b and a second gas tunnel 106b disposed on the first surface 102a, and a second pipette 112b. The second gas tunnel 106b is connected to the first fluid tunnel 104a, and the second fluid tunnel 104b is connected to the second pipette 112. The second pipette 112b has a hollow structure that extends downward from the first surface 102a, partially protrudes from the second surface 102b along the vertical direction D3, and extends into the container 120a, as shown in FIGS. Thus, while the motor drives and rotates the rotary valve 116 in the cartridge 100, the opening 116b on the rotary valve 118 is optionally aligned with the second gas tunnel 106b, so that the quantification reagent in the first quantification chamber can then be transferred to the container 120a via the first pipette 112a. Otherwise, the opening 116b of the rotary valve 118 is optionally aligned with the second fluid tunnel 104b, so that the second fluid tunnel 104b , the second pipette 112b, and the container 120a are in communication with each other. Then, while positive or negative pressure is provided to the cartridge 100 by an external pump, the quantification reagent in the container 120a can be further delivered to another container 120b also disposed on the second surface 102b through the second pipette 112b and the second fluid tunnel 104b. However, in another embodiment, the first pipette 112 may be omitted for the sake of simplicity of assembly, and only a through-hole (not shown) is connected to the second end 108b of the quantification chamber 108, and the through-hole penetrates the first surface 102a and the second surface 108b to simultaneously communicate the second end 108b / 110b of the first quantification chamber 102 with the storage chamber 110. Through this arrangement, the quantification reagent can be directly delivered into the container 120a through the through-hole.

[0018] Meanwhile, the containers 120a and 120b are each attached to the second surface 102b through a through-hole provided in the main cover 102. As shown in FIG. 3, the container 120a further includes a second quantification chamber 121 disposed between the top 122a and bottom 122b of the container 120a. The second quantification chamber 121 is a buffer region within the container 120a. The bottom surface V1 of the second quantification chamber 121 and the bottoms of the first pipette 112a and the second pipette 122b are preferably located on the same plane, and the top surface V2 of the second quantification chamber 121 is lower than the top surfaces of the first pipette 112a and the second pipette 112b. In one embodiment, the container 120a further includes two opposing sidewalls 124a inclined from the vertical direction D3 to prevent reagent from remaining in the second quantification chamber 121. Preferably, the inclined sidewall 124a extends from at least the top surface V2 to the bottom surface V1 of the second quantification chamber 121, as shown in FIG. 3, to effectively prevent liquid from remaining and affecting the quantification results. Also, the bottoms of the first pipette 112a and the second pipette 112b preferably have an inclined surface 113 so that they can easily extend into the container 122a without directly contacting the bottom surface 120b of the container 120a, thereby facilitating liquid absorption. While this embodiment is illustrated by disposing only two containers 120a and 120b in the cartridge 100, those skilled in the art should fully understand that the number of containers in the cartridge 100 may be adjusted based on actual requirements, for example, by further including other containers (not shown), each of which optionally includes the inclined sidewall 124 and / or the second quantification chamber 121 disposed therein, to facilitate the screening process within the cartridge 100.

[0019] 1 and 3 , the cartridge 100 further includes a sealing layer 128 and a package cover 130, which are sequentially attached to the first surface 102a of the main cover 102 by a suitable assembly process, such as, but not limited to, a thermal fusion process or an ultrasonic process. Specifically, the sealing layer 128 is disposed on the first surface 102a of the main cover 102 and seals the first fluid tunnel 104a, the second fluid tunnel 104b, the first gas tunnel 106a, the first gas tunnel 106b, the first quantification chamber 108, and the storage chamber 110, which are disposed on the main cover 102. The package cover 130 is disposed on the sealing layer 128 and also extends along a plane defined by directions D1 and D2. In one embodiment, the package cover 130 is also formed by a plastic injection molding process and includes, but is not limited to, a material selected from the group consisting of polypropylene, polycarbonate, polyimide, and polyethylene terephthalate. The package cover 130 and the main cover 102 preferably have corresponding contours, for example, but not limited to, a rectangular shape, as shown in Fig. 1. Those skilled in the art will readily understand that the contours of the package cover 130 and the main cover 102 shown in Fig. 1 are merely exemplary and may include other applicable shapes based on actual product requirements.

[0020] The package cover 130 includes a first surface 130a and a second surface 130b facing each other. The first surface 130a further includes a seal member 132, an airtight ring 133, and a container cover 134. The sealing film 132 includes, for example, aluminum foil and seals the opening of the container 120a. The airtight ring 133 is disposed on the sealing film 132, overlaps the underlying container 120a, and is covered and closed by the container cover 134 to strengthen the closure of the container 120a. However, in another embodiment, the sealing film 132 can be omitted to simplify assembly, especially when the container 120a does not contain a sample or reagent. The second surface 130b further includes at least one pin 136 and a plurality of assembly posts 138 disposed thereon, the pins 136 being disposed to protrude on the second surface 130b so as to pierce the sealing film 128 during attachment of the package cover 130 to the main cover 102 in order to introduce air into the cartridge 100. The assembly posts 138 are disposed around the periphery of the package cover 130, thus guiding the package cover 130 to seat and attach to the main cover 102 in a precise manner.

[0021] With the above configuration, the cartridge 100 of this embodiment can perform two-stage liquid quantification during reagent transfer, thereby improving the screening quality of the cartridge 100. To be precise, the required reagents may be first introduced into the cartridge 100 from an external area (not shown) or pre-loaded in a storage space (not shown) or other suitable container within the cartridge 100, and then the required reagents are transferred by rotating the rotary valve 116. During the transfer of the required reagents, the opening 116b of the rotary valve 118 is selectively aligned with the first gas tunnel 106a, and the flow path 118a is first selectively aligned with the first fluid tunnel 104a. Thus, the required reagents pass through the first fluid tunnel 104a and enter the first quantification chamber 108, allowing the volume of the required reagent to be accurately quantified, and the quantification reagents remain in the first quantification chamber 108. It should be noted that the opening 116b of the rotary valve 118, the first gas tunnel 106a, and the storage chamber 110 together form a gas circulation path, allowing the required reagent to pass quickly and smoothly through the first fluid tunnel 104a and the first quantification chamber 108. Then, the required reagent is first filled into the quantification chamber 108, and then an excess amount of the required reagent is transferred to the storage chamber 100 via the second end 108b of the first quantification chamber 108 (i.e., the second end 110b of the storage chamber 110). In this way, by arranging the first quantification chamber 108 and the storage chamber 110, a first-stage quantification of the required reagent is achieved, and a quantified reagent is obtained. Next, by controlling the external motor, the rotary valve 116 is rotated to selectively align the opening 116b of the rotary valve 118 with the second gas tunnel 106b, and the quantification reagent in the first quantification chamber 108 is pneumatically driven and injected into the container 120a via the first pipette 112a arranged in the vertical direction D3. Meanwhile, since the storage chamber 110 is not connected to any fluid tunnel, the excess amount of the required reagent still remains in the storage chamber 100 without flowing into the container 120a.These capabilities allow any reagent to be actively quantified before being injected into the container 120a by passing it through the first quantification chamber 108, rather than just being quantified by passively controlling an external motor.

[0022] Next, while the opening 116b of the rotary valve 118 is selectively aligned with the second fluid tunnel 104b, the quantification reagent in the container 120a can be further transferred through the second pipette 112b and the second fluid tunnel 104b to another space, such as another container 120b inside the cartridge 100. Referring to FIG. 4, during the transfer of the quantification reagent, only the quantification reagent in the second quantification chamber 121 can be delivered, and the quantification reagent positioned lower than the bottom surface V1 of the second quantification chamber 121 (i.e., lower than the bottom of the second pipette 112b) remains in the container 120a. That is, the transferred amount of the quantification reagent is precisely controlled to a predetermined value (Va1-Va2) obtained by subtracting the volume va2 from the volume va1. In one embodiment, a freeze-dried sample may be placed in container 120a in advance, and the amount of the mixed reagent transferred after mixing the freeze-dried sample with the quantification reagent may be controlled to a value (Va1-Va2). In this manner, in second quantification chamber 121, second-stage quantification can be performed synchronously and accurately while transferring the reagent in container 120a, allowing precise control of the amount of reagent transferred (Va1-Va2). Therefore, cartridge 100 of this embodiment can achieve active quantification while injecting any reagent into container 120a, and can also achieve further quantification while transferring the reagent from container 120a to container 120b. As a result, it can be used for screening with strict requirements for reaction volume, such as nucleic acid extraction reactions or nucleic acid amplification reactions, thereby improving reaction efficiency and screening sensitivity.

[0023] Those skilled in the art should fully understand that the cartridge of the present disclosure is not limited to the above-described types and may include other examples or modifications for effective use in any possible screening. Referring to FIGS. 5 and 6, a cartridge 200 according to a second embodiment of the present invention is shown. FIG. 5 is a schematic exploded view of the cartridge 200, and FIG. 6 is a schematic side view of the cartridge 200. The structure of the cartridge 200 of this embodiment is substantially the same as the structure of the cartridge 100 of the first embodiment described above, and the following description of all similarities will not be repeated. The difference between this embodiment and the first embodiment is that, as shown in FIG. 5, the cartridge 200 further includes a plurality of third fluid tunnels 104c and a plurality of third gas tunnels 106c arranged on the first surface 102a of the main cover 102, a plurality of containers 120a, 120b, and 220c, a plurality of third pipettes 112c, and a plurality of gas inlets 114 arranged on the second surface 102b of the main cover 102. 6, each of the third pipettes 112c and each of the gas inlets 114 includes a hollow structure extending along the vertical direction D3, extending downward from the first surface 102a and partially protruding from the second surface 102b. Note that each of the third pipettes 112c and each of the gas inlets 114 overlaps with one of the corresponding containers 120b, 220 in the vertical direction D3 and extends into one of the corresponding containers 120b, 220, and each of the third fluid tunnels 104c and each of the third gas tunnels 106c extends horizontally from the center to the periphery of the main cover 102 and connects to one of the corresponding third pipettes 112c and one of the corresponding gas inlets 114, respectively. In this way, the flow path 116a of the rotary valve 118 is selectively aligned with each of the third fluid tunnels 104c, communicating each of the third fluid tunnels 104c, a corresponding one of the third pipettes 112c, and a corresponding one of the containers 120b, 220, and the opening 116b of the rotary valve 118 is simultaneously selectively aligned with each of the third gas tunnels 106c for air circulation, so that the required reagent or sample can be randomly transferred between each of the containers 120a, 120b, 220.With this arrangement, the necessary reagents, samples, or freeze-dried biological materials can be first arbitrarily contained in each container 120a, 120b, and 220 according to the actual screening requirements, and then transferred to other containers via the third fluid tunnel 104c, the third pipette 112c, and the rotary valve 116. In other words, each of the containers 120a, 120b, and 220 can function as a sample container for containing any biological sample or biological material, an extraction container for containing a lysis solution or magnetic beads, a reaction container for containing a reaction reagent, and / or a washing container for containing a washing solution, buffer, etc., so that the aforementioned reagents, samples, or freeze-dried biological materials can be randomly mixed or transferred for subsequent screening. Those skilled in the art should fully understand that the specific number, size, and shape of the containers 120a, 120b, and 220 are not limited to those shown in FIG. 5 and can be further adjusted according to actual product requirements.

[0024] To facilitate understanding of the cartridge of the present disclosure for those skilled in the art, the quantification method of the cartridge of the present disclosure will be further described below. Referring to FIG. 7, FIG. 7 is a schematic diagram illustrating the process flow of a quantification method according to a preferred embodiment of the present disclosure. First, a cartridge 200 is provided (step S1). The cartridge 200 includes a main cover 102, a plurality of containers 120a, 120b, 220, a first pipette 112a, and a rotary valve 116. The main cover 102 further includes a first quantification chamber 108, a first fluid tunnel 104a, a first gas tunnel 106a, and a storage chamber 110 disposed thereon. The container 120a overlaps with the second end 108b of the first quantification chamber 108 in the vertical direction D3. The detailed structures and features of all of the above assemblies have been described in the preceding paragraphs and will not be described again below.

[0025] Next, a reagent is injected into the first quantification chamber 108 (step S2). To be precise, the reagent, which includes, for example, nucleic acid from a biological sample, is then transferred to the first quantification chamber 108 through the first fluid tunnel 104a for primary quantification by rotating the rotary valve 116 until the flow path 118a of the rotary valve 116 is aligned with the first fluid tunnel 104a and the opening 118b of the rotary valve 116 is aligned with the first gas tunnel 106a. In one embodiment, the containers 120a, 120b, 220 each contain a biological sample, a lysate, magnetic beads, an eluent, a washing solution, etc., and then the reagent (including nucleic acid) is delivered into the cartridge 200 through, for example, but not limited to, the following steps: First, the biological sample is transferred from one of the containers 220 containing the biological sample to another container 220 containing an extraction solution, causing cells in the biological sample to rupture or open, releasing substances such as nucleic acid. Next, the nucleic acid is transferred to one of the containers 220 containing magnetic beads, where the magnetic beads capture the nucleic acid. Subsequently, the magnetic beads and nucleic acid are transferred to another container 220 containing a wash solution, where the magnetic beads and nucleic acid are washed. The magnetic beads and nucleic acid are then transferred to one of the containers 220 containing an elution solution, where the nucleic acid is isolated from the magnetic beads. The isolated nucleic acid is then transferred to another container 220, thereby obtaining the necessary reagents. However, in another embodiment, the reagents (including the nucleic acid) can be placed directly in one of the containers 220, and then the reagents can be transferred from one of the containers 220 to the first quantification chamber 108 for quantification by rotating the rotary valve to a specific position, thereby simplifying the configuration and operational steps of the cartridge 200.

[0026] Then, a portion of the reagent (including nucleic acid) is transferred to the reservoir 110 (step S3). To be precise, while the reagent is being injected into the first quantification chamber 108, an excess portion of the reagent exceeding the capacity of the first quantification chamber 108 flows from the second end 108b of the first quantification chamber 108 into the storage chamber 110 adjacent to the first quantification chamber 108 due to the pressure difference between the first quantification chamber 108 and the storage chamber 110. The excess portion of the reagent is temporarily stored in the storage chamber 110, and the reagent remaining in the first quantification chamber 108 is synchronously quantified to obtain a quantified reagent having a volume equal to the capacity of the first quantification chamber 108. Therefore, through the arrangement of the first quantification chamber 108 and the storage chamber 110, the cartridge 200 is capable of accurately controlling the volume of the reagent during screening.

[0027] The reagent in the first quantification chamber 108 is then transferred to the container 120a (step S4). In other words, the rotary valve 116 is rotated again until the opening 118 of the rotary valve 116 is aligned with the second gas tunnel 106b to generate air pressure, so that the quantified reagent in the first quantification chamber 108 can be pushed into the container 120a by the air pressure, with the excess reagent still remaining in the storage chamber 110. In this way, the reagent (including nucleic acid) injected into the container 120a can be precisely controlled in volume. Note that in one embodiment, the container 120a can further include a freeze-dried sample disposed therein, so that the reagent injected into the container 120a can be mixed with the freeze-dried sample to form a mixture of nucleic acid and freeze-dried sample for subsequent screening. Therefore, by quantifying the reagent through the first quantification chamber 108, the mixing ratio between the reagent and the freeze-dried sample can also be precisely controlled, thereby improving the sensitivity and accuracy of subsequent screening.

[0028] A desired reaction, such as, but not limited to, nucleic acid amplification, can then be performed on the mixture of nucleic acids and lyophilized sample in container 120a under precisely controlled temperature conditions. However, in another embodiment, rotary valve 116 can be rotated again until channel 118 of rotary valve 116 is aligned with second fluid tunnel 104b, transferring the mixture of nucleic acids and lyophilized sample from container 120a to another container 120b for a desired reaction. While transferring the mixture of nucleic acids and lyophilized sample, the mixture of nucleic acids and lyophilized sample can be simultaneously quantified through second quantification chamber 121 and subsequently delivered to container 120b by passing through second pipette 112b and second fluid tunnel 104b to more precisely control the amount of reagent and increase reaction efficiency. In other words, according to the quantification method of the present disclosure, a two-stage active quantification process is performed through the first quantification chamber 108 and / or the second quantification chamber 121 disposed in the cartridge 200, respectively, so that the reagent is simultaneously quantified while being injected into or withdrawn from the container 120a. In this manner, the quantification method of this embodiment can achieve improved screening quantification of the cartridge 200 under simplified operation. While the quantification method of this embodiment has been described using the cartridge 200 of the above embodiment for screening, the present invention is not limited thereto. In another embodiment, the quantification method may be performed by using the cartridge 100 under actual requirements.

[0029] Those skilled in the art should also fully understand that the cartridge of the present disclosure may include other examples or modifications. The following description will detail different embodiments of the cartridge, and the following description will detail differences between the different embodiments, and identical features will not be described redundantly. Note that, in order to easily compare differences between the embodiments, identical components in the following embodiments are denoted by the same reference numerals.

[0030] 8, a cartridge 300 according to a third embodiment of the present invention is shown. The structure of the cartridge 300 of this embodiment is substantially the same as that of the cartridge 200 of the second embodiment described above, and all similarities will not be redundantly described below. The difference between this embodiment and the second embodiment is that the cartridge 300 further includes a bottom cover 240 for carrying the containers 120a, 120b, 220 and the rotary valve 116.

[0031] 8 , the bottom cover 240 may be disposed below the main cover 102 and attached to the second surface 102b of the main cover 102 by an appropriate assembly process, such as, but not limited to, a thermal fusion process or an ultrasonic process. The bottom cover 240 further includes a plurality of through-holes 242, 244 respectively penetrating two opposing surfaces of the bottom cover 240. Each of the through-holes 242, 244 may include any possible size for accommodating the containers 120a, 120b, 220 and the rotary valve 116, respectively. With this arrangement, the containers 120a, 120b, 220 and the rotary valve 116 are sandwiched between the main cover 102 and the bottom cover 240, thereby enhancing the structural reliability and sealing of the cartridge 300.

[0032] Overall, the cartridge of the present disclosure further includes at least one quantification chamber disposed therein to achieve active quantification of the reagent during reagent transfer, thereby simplifying cartridge operation and assembly and further improving cartridge screening sensitivity and accuracy.

Claims

1. A cartridge, a main cover having a first surface and a second surface facing each other, the first surface comprising a first quantification chamber, a first fluid tunnel, a first gas tunnel, a storage chamber, and a second gas tunnel extending on a plane, a first end of the first quantification chamber connected to the first fluid tunnel, a first end of the storage chamber connected to the first gas tunnel, a second end of the first quantification chamber connected to the second end of the storage chamber, and the second gas tunnel connected to the first fluid tunnel, the first quantification chamber and the storage chamber being spaces recessed downward from the first surface; a sealing layer covering the first surface of the main cover to seal the first fluid tunnel, the first gas tunnel, the second gas tunnel, the first quantification chamber, and the storage chamber, wherein each of the first quantification chamber and the storage chamber is sealed as a three-dimensional spindle shape; at least one container disposed on the second surface of the main cover, the at least one container vertically overlapping the second end of the first quantitation chamber; a first pipette disposed on the main cover and partially protruding from the second surface of the main cover, the first pipette connected to the second end of the first quantification chamber and extending vertically into the at least one container; a second fluid tunnel disposed on the first surface of the main cover and having one end vertically overlapping the at least one container; a second pipette disposed on the main cover and protruding from the second surface of the main cover, the second pipette connected to the second fluid tunnel and extending vertically into the at least one container; a rotary valve rotatably disposed on the second surface of the main cover, the rotary valve having a first portion and a second portion, the first portion including a flow passage and an opening disposed thereon; the rotary valve rotates, and when the flow channel and the opening are aligned with the first fluid tunnel and the first gas tunnel, respectively, pressure is provided to deliver reagent sequentially through the flow channel and the first fluid tunnel into the first quantification chamber for quantification; the reagent is then transferred to the at least one container via the first pipette by providing the pressure when the rotary valve rotates to align the opening with the second gas tunnel; The reagent is then transferred to another container via the second pipette and the second fluid tunnel by providing the pressure when the rotary valve rotates to align the opening with the second fluid tunnel.

2. 10. The cartridge of claim 1, further comprising a second quantification chamber disposed within the at least one container between the top and bottom of the at least one container, the second quantification chamber having a bottom surface flush with the bottom of the first pipette.

3. 3. A cartridge according to claim 1 or 2, characterized in that the first part and the second part are of different materials.

4. A package cover disposed on the sealing layer, the package cover comprising: an airtight ring disposed on a first surface of the package cover and vertically overlapping the at least one container; a container cover disposed on the sealing ring to seal the at least one container; and at least one pin protruding from the second surface of the package cover.

10. The cartridge of claim 1 further comprising:

5. a plurality of third fluid tunnels and a plurality of third gas tunnels arranged along different directions on the first surface of the main cover; a plurality of third pipettes and a plurality of gas inlets arranged on the main cover to protrude from the second surface of the main cover, each of the third pipettes being connected to a respective one of the third fluid tunnels, and each of the gas inlets being connected to a respective one of the third gas tunnels; and a plurality of containers respectively disposed on the second surface of the main cover, wherein each of the gas inlets and each of the third pipettes respectively extend into each of the containers; 10. The cartridge of claim 1 further comprising:

6. 1. A method of quantification, comprising: (a) providing a cartridge, said cartridge comprising: a main cover comprising a first quantification chamber, a first fluid tunnel, a first gas tunnel, a storage chamber, and a second gas tunnel extending on a plane, wherein a first end of the first quantification chamber is connected to the first fluid tunnel, a first end of the storage chamber is connected to the first gas tunnel, a second end of the first quantification chamber is connected to the second end of the storage chamber, and the second gas tunnel is connected to the first fluid tunnel, and the first quantification chamber and the storage chamber are spaces recessed downward from a first surface of the main cover; a sealing layer covering the first surface to seal the first fluid tunnel, the first gas tunnel, the second gas tunnel, the first quantification chamber, and the storage chamber, wherein each of the first quantification chamber and the storage chamber is sealed as a three-dimensional spindle shape; a first pipette disposed on the main cover, the first pipette connected to the second end of the first quantification chamber; a plurality of containers disposed on the main cover, one of the plurality of containers vertically overlapping the second end of the first quantitation chamber; a second fluid tunnel disposed on the first surface and having one end vertically overlapping the one of the plurality of containers; a second pipette disposed on the main cover and protruding from a second surface of the main cover, the second pipette connected to the second fluid tunnel and extending vertically into the one of the plurality of containers; providing a rotary valve rotatably disposed on the main cover, the flow passages and openings being disposed on a surface of the rotary valve; (b) rotating the rotary valve until the flow passage and the opening are aligned with the first fluid tunnel and the first gas tunnel, respectively; (c) applying pressure to inject reagent into the first quantification chamber through the flow path, the first fluid tunnel, and the first end of the first quantification chamber; (d) after the reagent is dispensed into the first quantification chamber, transferring a portion of the reagent through the second end of the first quantification chamber into the storage chamber; (e) rotating the rotary valve until the opening is aligned with the second gas tunnel to transfer another portion of the reagent quantified in the first quantification chamber from the first quantification chamber to the one of the plurality of containers; (f) rotating the rotary valve until the opening is aligned with the second fluid tunnel, applying the pressure, and transferring the other portion of the reagent from the one of the plurality of containers to another of the plurality of containers sequentially via the second pipette and the second fluid tunnel.

7. The cartridge comprises:

7. The method of claim 6, further comprising a second quantification chamber disposed in one of said plurality of containers between the top and bottom of said one of said plurality of containers.

8. Step (e) (e1) mixing the separate portion of the reagent with the biological sample disposed within the one of the plurality of containers to form a quantified mixture through the second quantification chamber and the second pipette; Step (f) (f1) rotating the rotary valve until the opening is aligned with the second fluid tunnel; (f2) applying the pressure to transfer the mixture from one of the plurality of containers to another of the plurality of containers sequentially via the second pipette and the second fluid tunnel.

9. 8. The quantification method of claim 7, wherein the bottom surface of the second quantification chamber and the bottom of the first pipette are flush with each other, and the top surface of the second quantification chamber is lower than the tops of the multiple containers.

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