Cartridge and quantification method therefor

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 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. In general, a nucleic acid extraction kit or nucleic acid extraction reagent is usually used in an open routine laboratory for nucleic acid extraction, and then a nucleic acid amplification kit or nucleic acid amplification reagent is used to amplify or detect a specific nucleic acid fragment. However, the above kit or reagent usually requires manual operation, which is time-consuming and prone to contamination of samples or reagents, thereby making it less efficient to use in mass testing or production line mode testing. Meanwhile, some existing analytical cartridges can be mechanically operated, but these cartridges usually have complex configurations and need to be matched with expensive machines for operation, thereby increasing screening costs. Therefore, it is still necessary to provide new and improved techniques for the related art to meet the actual demands of the related 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, achieving 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 a cartridge, which allows for synchronous quantification of liquid while it is transported by flowing through at least one quantification chamber in the cartridge, thereby simplifying operations 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, 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, and a second end of the first quantification chamber 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 cartridge quantification method. 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, 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 the second end of the storage chamber. The first pipette is disposed on the main cover, and the first pipette is connected to the second end of the first quantification chamber. The plurality of containers are disposed on the main cover, and one of the plurality of containers overlaps with the second end of the first quantification chamber in the vertical direction. The rotary valve is rotatably disposed on the main cover. A reagent is then 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. The remaining portion of the reagent in the first quantification chamber is then transferred to one of the containers by rotating the rotary valve. [Brief description of the drawings]

[0008] The 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 showing the cartridge according to the first embodiment of the present invention;FIG. [Diagram 2] BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a schematic view showing a cartridge according to a first embodiment of the present invention Fig. 2 is a side view of the cartridge according to the first embodiment of the present invention. [Diagram 3] 1 is a schematic diagram showing a cartridge according to a first embodiment of the present invention;FIG. 2 shows a cross-sectional view of a cartridge according to a first embodiment of the present disclosure;FIG. [Figure 4]1A-1C are schematic diagrams illustrating a cartridge according to a first embodiment of the present invention;FIGS. 1A-1C show cross-sectional views of a second quantification chamber according to a first embodiment of the present disclosure;FIGS. [Diagram 5] 1 shows a schematic diagram of a cartridge according to a second embodiment of the present invention, and FIGURE 2 shows an exploded view of a cartridge according to a second embodiment of the present disclosure. [Figure 6] 1 is a schematic diagram showing 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. 2 is a schematic diagram showing a process flow of a quantification method according to a preferred embodiment of the present disclosure. [Figure 8] FIG. 11 is a schematic view showing a cartridge according to a third embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] In order to provide a better understanding of the present disclosure, a preferred embodiment will now be described in detail. The preferred embodiment of the present disclosure is illustrated in the accompanying drawings having numbered elements.

[0011] In the present disclosure, forming a first feature on a second feature being described may include embodiments in which the first feature and the second feature are formed in direct contact, and may also include embodiments in which an additional feature may be formed between the first feature and the second feature such that the first feature and the second feature are not in direct contact. In addition, the present disclosure may repeat reference numbers and / or letters in various examples. This repetition is for brevity and clarity and does not, in itself, dictate a relationship between the various embodiments and / or configurations being discussed. Furthermore, spatially relative terms such as "beneath," "below," "lower," "over," "above," "upper," and the like 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. The spatially relative terms are intended to encompass different orientations of the device during use or operation in addition to the orientation shown 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, third, etc. 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 may only be used 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 the 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%, more preferably within 5%, 3%, 2%, 1%, or 0.5% of a given value or range. Unless expressly specified otherwise, all numerical ranges, amounts, values, and percentages disclosed herein should be understood in all instances as being modified by the term "about" or "substantially". Thus, unless indicated to the contrary, the numerical parameters set forth in this disclosure and the 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, in which FIG. 1 is a schematic diagram of an exploded view of the cartridge 100, FIG. 2 is a schematic diagram of a side view of the cartridge 100, FIG. 3 is a schematic diagram of a cross-sectional view of the cartridge 100, and FIG. 4 is a schematic diagram of a 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 quantification chamber 108, the first fluid tunnel 104a, the first gas tunnel 106a, and the storage chamber 110 are disposed on the first surface 102a of the main cover 102. 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 arranged on the second surface 102b. It should be noted 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. Thus, by positioning the first quantification chamber 108, the cartridge 100 allows active quantification to be achieved during injection of 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 the direction D1 and the direction 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 the actual requirements, and are not limited to those shown in FIG. 1. In one embodiment, the first quantification chamber 108 preferably includes two opposing side walls 108c inclined on a plane (the plane defined by directions D1 and D2) as shown in Fig. 1 to prevent the reagent from remaining in the first quantification chamber 108 during reagent transfer and affecting the quantification result. 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 part 116a and a second part 116b disposed from top to bottom. The first part 116a partially protrudes to define a flow passage 118a and an opening 118b, respectively, the opening 118b being used for gas circulation, and the flow passage 118a being used for liquid circulation. The flow passage 118a may include any suitable shape, such as, but not limited to, a straight shape as shown in FIG. 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, for example, thermoplastic polyurethane (TPU), rubber, polyurethane material, polyethylene, polyethylene terephthalate (PET), thermoplastic polyester elastomer (TPEE), biocompatible resin, or combinations thereof for adhering to the second surface 102b, and the second portion 116b comprises a rigid material such as, for example, 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 in the cartridge 100 to rotate in a particular 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, the cartridge 100 is capable of delivering reagents from the first fluid tunnel 104a into the first quantification chamber 108 for liquid quantification while a positive or negative pressure is provided by an external pump (not shown).

[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, thereby communicating the second fluid tunnel 104a, the second pipette 112b, and the container 120a. Then, while a positive or negative pressure is provided to the cartridge 100 by the external pump, the quantification reagent in the container 120a can be further delivered through the second pipette 112b and the second fluid tunnel 104b to another container 120b also disposed on the second surface 102b. However, in another embodiment, the first pipette 112 may be omitted under the requirement of simplifying the assembly, and only a through-hole (not shown) is connected to the second end 108b of the quantification chamber 108, which 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 these arrangements, the quantification reagent can be directly delivered into the container 120a through the through-hole.

[0018] Meanwhile, the containers 120a and 120b are respectively attached to the second surface 102b through through holes provided in the main cover 102. As shown in FIG. 3, the container 120a further includes a second quantification chamber 121 disposed inside between the top 122a and bottom 122b of the container 120a. The second quantification chamber 121 is a buffer area in the container 120a, and 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 in 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 side walls 124a inclined from the vertical direction D3 to prevent the 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, effectively preventing liquid from remaining and affecting the quantification result. Also, the bottoms of the first pipette 112a and the second pipette 112b preferably have an inclined surface 113, respectively, so as to easily extend into the container 122a without directly contacting the bottom 120b of the container 120a, thereby facilitating liquid absorption. Although this embodiment is illustrated by only disposing two containers 120a, 120b in the cartridge 100, the number of containers in the cartridge 100 may be adjusted based on actual requirements, for example, 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 process of screening in the cartridge 100, it should be fully understood by those skilled in the art.

[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 melting process or an ultrasonic process. To be precise, 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 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 the 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 include contours corresponding to each other, for example, but not limited to, both have a rectangular shape, as shown in Fig. 1. Those skilled in the art should easily 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 under the requirement of simplifying the assembly, especially when the container 120a does not contain a sample or a 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 protrudingly on the second surface 130b to pierce the sealing film 128 during attachment of the package cover 130 to the main cover 102 for introducing 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 the transfer of the reagent, and can improve the screening quality of the cartridge 100. To be precise, the required reagent may be first introduced into the cartridge 100 from an external area (not shown), or may be preloaded in a storage space (not shown) or other suitable container in the cartridge 100, and then the required reagent is transferred by rotating the rotary valve 116. During the transfer of the required reagent, the opening 116b of the rotary valve 118 is selectively aligned with the first gas tunnel 106a, and the flow path 118a is selectively aligned with the first fluid tunnel 104a first, so that the required reagent enters the first quantification chamber 108 through the first fluid tunnel 104a, and the volume of the required reagent can be accurately quantified, and the quantification reagent remains 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, so that the required reagent can pass through the first fluid tunnel 104a and the first quantification chamber 108 quickly and smoothly. 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 through 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, the first stage quantification of the required reagent is achieved, and the quantification reagent is obtained. Then, the rotary valve 116 is rotated by controlling the external motor 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] Then, 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 located lower than the bottom surface V1 of the second quantification chamber 121 (i.e., lower than the bottom of the second pipette 112b) is left in the container 120a. That is, the transfer amount of the quantification reagent is precisely controlled to a predetermined value (Va1-Va2) of the volume va1 minus the volume va2. In one embodiment, the freeze-dried sample may be placed in the container 120a in advance, and the amount of the mixed reagent transferred after the quantification reagent and the freeze-dried sample are mixed may be controlled to a value (Va1-Va2). In this way, in the second quantification chamber 121, the second stage quantification can be performed synchronously and accurately while transferring the reagent in the container 120a, and the amount of the reagent transferred (Va1-Va2) can be precisely controlled. Therefore, the cartridge 100 of this embodiment can achieve active quantification during the injection of any reagent into the container 120a, and can also achieve further quantification during the transfer of the reagent from the container 120a to the container 120b, so that it can be used for screening with strict requirements on the reaction volume, such as nucleic acid extraction reaction or nucleic acid amplification reaction, thereby improving the reaction efficiency and the sensitivity of the screening.

[0023] Those skilled in the art should also fully understand that the cartridge of the present disclosure is not limited to the above-mentioned type, and may include other examples or modifications to be effectively used in any possible screening. Referring to Fig. 5 and Fig. 6, a cartridge 200 according to a second embodiment of the present invention is shown, Fig. 5 is a schematic diagram of an exploded view of the cartridge 200, and Fig. 6 is a schematic diagram of a 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 all similarities will not be described repeatedly below. 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, 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. Each of the third pipettes 112c and each of the gas inlets 114 includes a hollow structure extending along a vertical direction D3, extending downward from the first surface 102a and partially protruding from the second surface 102b, as shown in Fig. 6. 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 of the main cover 102 to the periphery and connects to one of the corresponding third pipettes 112c and one of the corresponding gas inlets 114, respectively. In this manner, the flow passage 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 such an arrangement, the necessary reagents, samples or freeze-dried biological materials may be arbitrarily accommodated in each container 120a, 120b and 220 first 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, 220 can function as a sample container for accommodating any biological sample or biological material, an extraction container for accommodating a lysis solution or magnetic beads, a reaction container for accommodating a reaction reagent, and / or a washing container for accommodating a washing solution, a buffer solution, 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, 220 are not limited to those shown in FIG. 5, and can be further adjusted according to the actual product requirements.

[0024] In order to allow those skilled in the art to easily understand the cartridge of the present disclosure, the quantification method of the cartridge in the present disclosure will be further described below. Referring to FIG. 7, FIG. 7 is a schematic diagram showing the process flow of the 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, and 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. Note that the detailed structures and features of all the above assemblies are all described in the above 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 of a biological sample, is then transferred to the first quantification chamber 108 via 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 one of the containers 220 containing an extraction solution, and the cells in the biological sample are ruptured or opened to release substances such as their nucleic acid. The nucleic acid is then transferred to one of the containers 220 containing magnetic beads, which capture the nucleic acid, and then the magnetic beads and nucleic acid are transferred to another one of the containers 220 containing a washing solution, and the magnetic beads and nucleic acid are washed. Then, the magnetic beads and nucleic acid are further transferred to one of the containers 220 containing an elution solution, and the nucleic acid is isolated from the magnetic beads, and the isolated nucleic acid is then transferred to another one of the containers 220, thereby obtaining the required reagent. However, in another embodiment, the reagent (including the nucleic acid) can be placed directly in one of the containers 220, and then the reagent 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 as well as the operation steps of the cartridge 200.

[0026] Then, a part of the reagent (including nucleic acid) is transferred to the storage chamber 110 (step S3). To be precise, while the reagent is injected into the first quantification chamber 108, the excess part of the reagent exceeding the storage volume 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 part 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 storage volume 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 precisely controlling the volume of the reagent in screening.

[0027] Thereafter, the reagent in the first quantification chamber 108 is 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, and the excess part of the reagent still remains in the storage chamber 110. In this way, the reagent (including nucleic acid) injected into the container 120a can thus obtain a precisely controlled volume. It is noted 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 to perform subsequent screening. Thus, by quantifying the reagent through the first quantification chamber 108, the mixing ratio of the reagent and the freeze-dried sample can also be precisely controlled, thereby improving the sensitivity and accuracy of the subsequent screening.

[0028] Then, a desired reaction such as, but not limited to, nucleic acid amplification can be performed through the mixture of nucleic acid and freeze-dried sample in the container 120a under precisely controlled temperature conditions. However, in another embodiment, the rotary valve 116 can be rotated again until the flow path 118 of the rotary valve 116 is aligned with the second fluid tunnel 104b to transfer the mixture of nucleic acid and freeze-dried sample from the container 120a to another container 120b to perform the required reaction. Also, while transferring the mixture of nucleic acid and freeze-dried sample, the mixture of nucleic acid and freeze-dried sample can be further quantified simultaneously through the second quantification chamber 121 and subsequently delivered to the container 120b by passing through the second pipette 112b and the second fluid tunnel 104b to further precisely control the amount of reagent and increase the reaction efficiency. In other words, according to the quantification method of the present disclosure, a two-stage active quantification process is carried out through the first quantification chamber 108 and / or the second quantification chamber 121 disposed in the cartridge 200, respectively, so that the reagent is quantified simultaneously while the reagent is injected into the container 120a or sucked out from the container 120a. In this way, the quantification method of the present embodiment can obtain improved screening quantification of the cartridge 200 under simplified operation. It should be noted that the quantification method of the present embodiment has been described by taking the cartridge 200 of the above embodiment as an example for performing screening, but the present invention is not limited thereto. In another embodiment, the quantification method may be carried out by using the cartridge 100 under practical 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 details different embodiments of the cartridge, and the following description details the differences between the different embodiments, and the same features are not described redundantly. In addition, in order to easily compare the differences between the embodiments, the same components are given the same reference numerals in the following embodiments.

[0030] Referring to Fig. 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] To be precise, as shown in FIG. 8, the bottom cover 240 may be disposed under the main cover 102 and attached to the second surface 102b of the main cover 102 by a suitable assembly process, such as, but not limited to, a thermal melting process or an ultrasonic process. The bottom cover 240 further includes a plurality of through holes 242, 244 respectively penetrating the two opposing surfaces of the bottom cover 240. Each of the through holes 242, 244 may include any possible size for accommodating each of the containers 120a, 120b, 220 and the rotary valve 116. With these arrangements, 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 the operation and assembly of the cartridge and further improving the sensitivity and accuracy of cartridge screening.

Claims

1. A cartridge comprising: 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, and a storage chamber extending in a plane, a first end of the first quantification chamber being connected to the first fluid tunnel, a first end of the storage chamber being connected to the first gas tunnel, and a second end of the first quantification chamber being connected to the second end of the storage chamber; 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 quantification chamber; a first pipette disposed on the main cover and partially protruding from the second surface of the main cover, the first pipette being connected to the second end of the first quantification chamber and extending vertically into the at least one container; a rotary valve rotatably disposed on the second surface of the main cover.

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

3. 3. The cartridge of claim 1 or 2, wherein the rotary valve further comprises a first portion and a second portion having different materials, the flow passage and the opening being located on the first portion.

4. 4. The cartridge of claim 3, wherein the flow path and the opening are aligned with the first fluid tunnel and the first gas tunnel, respectively.

5. a second gas tunnel disposed on the first surface of the main cover and connected to the first fluid tunnel, the opening of the rotary valve being optionally aligned with the second gas tunnel; 4. The cartridge of claim 3, further comprising: a second fluid tunnel disposed on the first surface of the main cover, one end of the second fluid tunnel vertically overlapping the at least one container, and the flow path of the rotary valve being aligned with the second fluid tunnel.

6. 6. The cartridge of claim 5, further comprising a second pipette disposed on the main cover and protruding from the second surface of the main cover, the second pipette connected to the fluid tunnel and extending vertically into the at least one container.

7. a sealing layer covering the first surface of the main cover; a package cover disposed on the sealing layer, the package cover comprising: a sealing 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 for sealing the at least one container; a package cover, further comprising at least one pin protruding from a second surface of the package cover; 10. The cartridge of claim 1 further comprising:

8. 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 third 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 each of the fluid tunnels and each of the gas inlets being connected to each of the third gas tunnels; a plurality of receptacles respectively disposed on the second surface of the main cover, each of the gas inlets and each of the pipettes respectively extending into each of the receptacles; The cartridge of claim 1 further comprising:

9. A method of quantification, the method comprising: A cartridge is provided, the cartridge comprising: a main cover including a first quantification chamber, a first fluid tunnel, a first gas tunnel, and a storage chamber extending on a plane, a first end of the first quantification chamber being connected to the first fluid tunnel, a first end of the storage chamber being connected to the first gas tunnel, and a second end of the first quantification chamber being connected to the second end of the storage chamber; 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 quantification chamber; a rotary valve rotatably disposed on the main cover, the flow passages and the openings being disposed on a surface of the rotary valve; injecting a reagent into the first quantification chamber from the first end of the first quantification chamber; after the reagent is dispensed into the first quantification chamber, transferring a portion of the reagent from the second end of the first quantification chamber to the storage chamber; and transferring, by a rotary valve, another portion of said reagent from said first quantification chamber to one of said plurality of containers.

10. 10. The method of claim 9, further comprising rotating the rotary valve until the flow passage of the rotary valve is aligned with the first fluid tunnel and the opening of the rotary valve is aligned with the first gas tunnel.

11. a second gas tunnel disposed on the main cover and connected to the first fluid tunnel; a second fluid tunnel disposed on the main cover; a second pipette disposed on the main cover and connected to the second fluid tunnel, the second pipette extending vertically into one of the plurality of vessels; 11. The quantification method according to claim 9 or 10, further comprising: a second quantification chamber disposed in one of the plurality of containers between the top and bottom of one of the plurality of containers.

12. 12. The quantification method of claim 11, further comprising rotating the rotary valve until the opening of the rotary valve is aligned with the second gas tunnel while transferring the other portion of the reagent from the first quantification chamber to one of the plurality of containers.

13. mixing the reagent and the biological sample within one of a plurality of containers to form a quantified mixture through the second quantification chamber and the second pipette; 12. The method of claim 11, further comprising: transferring the mixture from the second fluid tunnel to another one of the plurality of vessels.

14. 14. The method of claim 13, further comprising rotating the rotary valve until the flow passage of the rotary valve is aligned with the second fluid tunnel.

15. 12. The quantification method of claim 11, 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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