CN214422610U

CN214422610U - Detection cassette and nucleic acid detection device

Info

Publication number: CN214422610U
Application number: CN202120103094.0U
Authority: CN
Inventors: 盛广济; 孙筱
Original assignee: Suzhou Sinafu Medical Technology Co ltd
Current assignee: Suzhou Sinafu Medical Technology Co ltd
Priority date: 2021-01-14
Filing date: 2021-01-14
Publication date: 2021-10-19
Anticipated expiration: 2031-01-14

Abstract

The utility model relates to a detect box and nucleic acid detection device. The detection kit comprises: a runner assembly including a transit runner; a plurality of reagent reservoirs; at least one sample cell; a fixing member for fixing an effective substance in a sample; and the reagent pool, the sample pool and the transfer area are all connected with the transfer flow channel, the reagent pool or the sample pool can be communicated with the transfer area through the transfer flow channel, and when pressure difference exists at two ends of the transfer flow channel, material transfer can be realized. The nucleic acid detecting apparatus includes the above-described cartridge. The detection box can be used for carrying out multi-step processing, the number of parts is small, the structure is relatively simple, and the manufacturing difficulty and the manufacturing cost can be reduced to a certain extent.

Description

Detection cassette and nucleic acid detection device

Technical Field

The utility model relates to the technical field of medical equipment, especially, relate to a detect box and nucleic acid detection device.

Background

In many chemical or biological assays and tests, it is often necessary to subject the sample to multiple processing steps. For example, in the case of nucleic acid detection, it is necessary to perform a multi-step process such as lysis, binding, washing, and elution on an obtained sample. For these tests, special test cartridges are usually designed to perform the tests. However, some of the cartridges in the related art can implement various processes, but have complicated structures, high difficulty in manufacturing processes, and high manufacturing costs.

SUMMERY OF THE UTILITY MODEL

Based on this, the utility model provides a detect box, this detect box can carry out multistep processing, and its part quantity is less, and the structure is simple relatively, can reduce to a certain extent and make the degree of difficulty and manufacturing cost.

A cartridge, comprising:

a runner assembly including a transit runner;

a plurality of reagent reservoirs;

at least one sample cell;

a fixing member for fixing an effective substance in a sample;

and the reagent pool, the sample pool and the transfer area are all connected with the transfer flow channel, the reagent pool or the sample pool can be communicated with the transfer area through the transfer flow channel, and when pressure difference exists at two ends of the transfer flow channel, material transfer can be realized.

In one embodiment, the transfer area includes a first transfer portion and a plurality of second transfer portions, the transfer channel includes a main channel, a plurality of first branch channels and a plurality of second branch channels, a first end of each of the first branch channels is connected to the main channel, the first transfer portion and each of the second transfer portions are connected to a second end of each of the first branch channels in a one-to-one correspondence manner, a first end of each of the second branch channels is connected to the main channel, and the sample wells and each of the reagent wells are connected to a second end of each of the second branch channels in a one-to-one correspondence manner.

In one embodiment, the cartridge further comprises a magnetic member capable of fixing the position of the fixing member by a magnetic attraction force.

In one embodiment, the cartridge includes a main body member including a first plate and a second plate, the first plate is connected with the top of the second plate, the reagent pool and the sample pool are both connected with the first plate, the transfer area is connected with the second plate, the first branch flow channel comprises a first section of the first branch flow channel and a second section of the first branch flow channel, the first section of the first branch flow channel is connected with the second section of the first branch flow channel, the second branch flow channel comprises a first section of a second branch flow channel and a second section of the second branch flow channel, the first section of the second branch flow channel is connected with the second section of the second branch flow channel, the bottom surface of the first plate is provided with the main flow channel, the first section of the first branch flow channel and the first section of the second branch flow channel, and the second plate is provided with a first branch flow channel second section and a second branch flow channel second section which penetrate through the second plate.

In one embodiment, the detection box further comprises a plurality of reagent temporary storage areas corresponding to the reagent pools in a one-to-one manner, the flow channel assembly further comprises a plurality of reagent temporary storage flow channels, each reagent pool and one reagent temporary storage area can be communicated through the corresponding reagent temporary storage flow channel, when a pressure difference exists between the reagent pool and the reagent temporary storage area, material transfer can be realized, and the reagent temporary storage areas are connected with the transfer flow channels;

the detection box further comprises a sample temporary storage area, the flow channel assembly further comprises a sample temporary storage flow channel, the sample pool and the sample temporary storage area can be communicated with each other through the sample temporary storage flow channel, when the sample pool and the sample temporary storage area have pressure difference, material transfer can be achieved, and the sample temporary storage area is connected with the transfer flow channel.

In one embodiment, the testing cassette comprises a main body member, the reagent reservoir and the sample reservoir are both connected with the top of the main body member, the reagent buffer and the sample buffer are both connected with the bottom of the main body member, and the reagent buffer flow passage and the sample buffer flow passage are both communicated with each other and penetrate through the main body member.

In one embodiment, the detection box comprises a main body part, wherein a plurality of elastic membranes are connected to the main body part to form a reagent temporary storage area, a sample temporary storage area and the transfer area at corresponding positions respectively, the elastic membranes are connected with the transfer flow channel, the elastic membranes at corresponding positions can expand under the action of external force to form a reagent temporary storage cavity, a sample temporary storage cavity and a transfer cavity respectively, and when a pressure difference exists between one side of the elastic membranes, which is far away from the transfer flow channel, and one side of the elastic membranes, which is close to the transfer flow channel, material transfer can be realized.

In one embodiment, the transfer channel is provided with a protrusion at a region contacting with the reagent buffer, the sample buffer and the transfer region on the elastic membrane, and the protrusion abuts against the elastic membrane.

In one embodiment, the bottom of the elastic membrane is provided with a propping piece, and the propping piece can prop the elastic membrane tightly towards the main body piece.

According to the detection box, the reagent pool, the sample pool and the transfer area are connected with the transfer flow channel, and the reagent pool or the sample pool can be communicated with the transfer area through the transfer flow channel, so that pressure difference exists at two ends of the transfer flow channel, and the transfer of materials in the area of the two ends of the transfer flow channel can be realized. When the reaction is carried out, the sample in the sample cell firstly reaches the transfer area through the transfer flow channel, in the process, the fixing piece fixes the effective substances in the sample, and the rest waste liquid can return to the sample cell in the original way. The reagent in the reagent pool can contact with the effective substances in the transfer flow channel in the process of reaching the transfer area through the transfer flow channel, and the contacted waste liquid can also return to the reagent pool in the same way. The reagents in each reagent pool are sequentially contacted with the effective substances in the transfer flow channel according to the mode, so that multi-step processing can be completed in one detection box. Moreover, the detection box has fewer parts and a relatively simple structure, so that the manufacturing difficulty and the manufacturing cost can be reduced to a certain extent.

The utility model discloses still provide a nucleic acid testing arrangement, wherein supporting detection box can carry out multistep processing, and its part quantity is less, and the structure is simple relatively, can reduce to a certain extent and make the degree of difficulty and manufacturing cost.

A nucleic acid detecting apparatus includes the above-described cartridge.

According to the nucleic acid detection device, in the matched detection box, the reagent pool, the sample pool and the transfer area are connected with the transfer flow channel, and the reagent pool or the sample pool can be communicated with the transfer area through the transfer flow channel, so that pressure difference exists at two ends of the transfer flow channel, and the transfer of materials in the areas at the two ends of the transfer flow channel can be realized. When the reaction is carried out, the sample in the sample cell firstly reaches the transfer area through the transfer flow channel, in the process, the fixing piece fixes the effective substances in the sample, and the rest waste liquid can return to the sample cell in the original way. The reagent in the reagent pool can contact with the effective substances in the transfer flow channel in the process of reaching the transfer area through the transfer flow channel, and the contacted waste liquid can also return to the reagent pool in the same way. The reagents in each reagent pool are sequentially contacted with the effective substances fixed in the transfer flow channel according to the mode, so that multi-step processing can be completed in one detection box. Moreover, the detection box has fewer parts and a relatively simple structure, so that the manufacturing difficulty and the manufacturing cost can be reduced to a certain extent.

Drawings

Fig. 1 is a schematic view of an overall structure of a detection box according to an embodiment of the present invention;

FIG. 2 is a perspective view of the cartridge of FIG. 1;

FIG. 3 is a perspective view showing an exploded structure of the cartridge of FIG. 1;

FIG. 4 is a sectional view of the cartridge of FIG. 1;

FIG. 5 is a top view of a first plate of the cartridge of FIG. 1;

FIG. 6 is a bottom view of the first plate of the cartridge of FIG. 1;

FIG. 7 is a plan view of a second plate of the cartridge of FIG. 1.

Reference numerals:

a sample cell 100;

a first reagent cell 210, a second reagent cell 220, a third reagent cell 230, and a fourth reagent cell 240;

a first cover 310, a second cover 320, a third cover 330, and a fourth cover 340;

a first plate 400, a main channel 410, a first branch channel first section 420, a first branch channel first section first region 421, a first branch channel first section second region 422, a first branch channel first section third region 423, a first branch channel first section fourth region 424, a first branch channel first section fifth region 425, a second branch channel first section 430, a second branch channel first section first region 431, a second branch channel first section second region 432, a second branch channel first section third region 433, a second branch channel first section fourth region 434, a second branch channel first section fifth region 435, a sample temporary storage channel first section 440, a reagent temporary storage channel first section first region 451, a reagent temporary storage channel first section second region 452, a reagent temporary storage channel third section 453, a reagent temporary storage channel first section fourth region 454, a first amplification channel second section 460, a second amplification channel second section 470, and an amplification pool 480;

a second plate 500, a first branched flow channel second section 510, a first branched flow channel second section first region 511, a first branched flow channel second section second region 512, a first branched flow channel second section third region 513, a first branched flow channel second section fourth region 514, a first branched flow channel second section fifth region 515, a second branched flow channel second section 520, a second branched flow channel second section first region 521, a second branched flow channel second section second region 522, a second branched flow channel second section third region 523, a second branched flow channel second section fourth region 524, a second branched flow channel second section fifth region 525, a sample temporary storage flow channel second section 530, a reagent temporary storage flow channel second section first region, a reagent temporary storage flow channel second section second region 542, a reagent temporary storage flow channel second section third region 543, a reagent temporary storage flow channel second section fourth region 544, a first amplification flow channel first section 550, a first amplification flow channel third section 560, and a second amplification flow channel first section 570;

the third plate 600, the transition zone 610, the first transition part 611, the second transition part first zone 6121, the second transition part second zone 6122, the second transition part third zone 6123, the second transition part fourth zone 6124, the sample buffer 620, the reagent buffer first zone 631, the reagent buffer second zone 632, the reagent buffer third zone 633, the reagent buffer fourth zone 634, and the amplification reaction zone 640.

Detailed Description

In order to make the above objects, features and advantages of the present invention more comprehensible, embodiments of the present invention are described in detail below with reference to the accompanying drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, as those skilled in the art will be able to make similar modifications without departing from the spirit and scope of the present invention.

In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", and the like, indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for convenience of description and simplicity of description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore, should not be construed as limiting the present invention.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.

In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," and "fixed" are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meaning of the above terms in the present invention can be understood according to specific situations by those skilled in the art.

In the present application, unless expressly stated or limited otherwise, the first feature may be directly on or directly under the second feature or indirectly via intermediate members. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.

It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "upper," "lower," "left," "right," and the like as used herein are for illustrative purposes only and do not denote a unique embodiment.

Referring to fig. 1 to 3, a schematic diagram of an overall structure of the detecting box, a perspective view of the detecting box in fig. 1, and an exploded perspective view of the detecting box in fig. 1 according to an embodiment of the present invention are respectively shown. An embodiment of the utility model provides a detect box includes parts such as at least one sample cell 100, a plurality of reagent pond, runner assembly, mounting and transfer area 610. The sample cell 100 is used for storing samples, and each reagent cell can store different reagents. The flow channel assembly comprises a transfer flow channel, the sample cell 100 and the transfer area 610 can be communicated through the transfer flow channel, and when a pressure difference exists between the sample cell 100 and the transfer area 610 at two ends of the transfer flow channel, substances can be transferred through the transfer flow channel. Each reagent pool can be communicated with the transfer area 610 through a transfer flow channel, and when pressure difference exists between the reagent pools at the two ends of the transfer flow channel and the transfer area 610, substances can be transferred through the transfer flow channel.

The sample in the sample cell 100 reaches the transfer region 610 through the transfer flow channel, in the process, the fixing member fixes the effective substance in the sample, and the rest waste liquid can return to the sample cell 100. In the process that the reagent in the reagent pool reaches the transfer area 610 through the transfer flow channel, the reagent can contact with the effective substances in the transfer flow channel, and the contacted waste liquid can also return to the reagent pool in the same way. The reagents in each reagent pool are sequentially contacted with the effective substances fixed in the transfer flow channel according to the mode, so that multi-step processing can be completed in one detection box.

The cartridge may be used to perform chemical or biological assay, for example, it may be used to perform nucleic acid detection. In this example, the cartridge will be described by taking nucleic acid detection as an example.

The sample cell 100 contains a cell lysis solution for lysing a cell virus or other sample to release a nucleic acid substance. The top of the sample cell 100 has a first cover 310, and the first cover 310 is clamped to the sample cell 100. The first cover 310 is opened to allow the extracted specimen to be placed therein. Four reagent wells are provided, namely a first reagent well 210, a second reagent well 220, a third reagent well 230 and a fourth reagent well 240. The second cover 320 is arranged on the top of the first reagent pool 210 and the second reagent pool 220, the third cover 330 is arranged on the top of the third reagent pool 230 and the fourth reagent pool 240, and both the second cover 320 and the third cover 330 are not detachable. The reagent in the reagent tank is sealed by the cover bodies so as to prevent the reagent from leaking.

Cleaning solutions are placed in the first reagent reservoir 210 and the second reagent reservoir 220, and are used for cleaning the cleaved nucleic acids and removing impurities in the nucleic acids. The fixing member in the transfer flow channel is a magnetic particle, and can adsorb the cleaved nucleic acid, so that the nucleic acid is fixed on the surface of the magnetic particle. An elution solution is placed in the third reagent reservoir 230 for eluting and separating the nucleic acid adsorbed on the magnetic particles. The fourth reagent pool 240 contains a first PCR reaction solution containing various enzymes and primers required by the nested PCR. It should be noted that the positional relationship between the sample cell 100 and each reagent cell is not limited, and the drawing only provides one of the positional arrangements, and other manners may also be adopted, for example, the positions of the second reagent cell 220 for holding the cleaning solution and the third reagent cell 230 for holding the eluent are interchanged. The number of the reagent pools and the types of the reagents in the reagent pools are not limited, and can be selected according to the experiment requirements, for example, reagent pools such as a diluent pool are added.

When the detection box is used for detecting nucleic acid, the multi-step treatment can be completed in one detection box only by enabling a sample to enter the transfer flow channel, fixing the cracked nucleic acid on the surface of the magnetic particles and enabling reagents in each reagent pool to sequentially pass through the transfer flow channel. Moreover, the number of parts of the whole detection box is small, the structure is simple, the manufacturing cost can be reduced to a certain extent, and the market competitiveness is improved.

Preferably, each part is made of plastic, so that the whole detection box is light in weight and low in cost. The parts may be integrally injection molded or may be separately molded and then fixed by bonding or the like.

Preferably, in some embodiments, the transition zone 610 includes a first transition portion 611 and a plurality of second transition portions. The transfer flow channel includes a main flow channel 410, a plurality of first branch flow channels and a plurality of second branch flow channels. The first end of each first branch flow channel is connected with the main flow channel 410, and the first transit parts 611 and the second transit parts are connected to the second ends of the first branch flow channels in a one-to-one correspondence manner. The first end of each second branch flow channel is connected to the main flow channel 410, and the sample wells 100 and each reagent well are connected to the second ends of the second branch flow channels in a one-to-one correspondence.

Specifically, the plurality of second transition portions are a first transition portion zone 6121, a second transition portion zone 6122, a second transition portion zone 6123 and a second transition portion zone 6124, respectively. There are five first branch flow passages, and each of the five first branch flow passages is connected to one of the first transit section 611, the second transit section first zone 6121, the second transit section second zone 6122, the second transit section third zone 6123, and the second transit section fourth zone 6124. There are five second branched flow channels, and each of the five second branched flow channels is connected to one of the cuvette 100, the first reagent cuvette 210, the second reagent cuvette 220, the third reagent cuvette 230, and the fourth reagent cuvette 240.

The transition area 610 is divided into a plurality of branch runners, and a plurality of first branch runners and a plurality of second branch runners are correspondingly arranged, and the plurality of first branch runners and the plurality of second branch runners only intersect at the main runner 410. Thus, the sample cell 100, a plurality of reagent cells, each of which corresponds to one flow path, can be made. For example, the sample in the sample cell 100 flows to the main channel 410 through one of the second branch channels, and flows to one of the second transitions through one of the first branch channels. The reagent in the first reagent reservoir 210 flows into the main channel 410 through another second branch channel, and flows into another second transition portion through another first branch channel. Thus, when the sample in the sample cell 100 and the reagents in the respective reagent cells flow in the transfer flow channel, only the main flow channel 410 is overlapped, so that the mutual influence between the reagents remaining in the flow channel can be reduced, and the reaction result can be more accurate.

It should be noted that the shapes of the first branch flow channel and the second branch flow channel are not limited, and can be adjusted according to the specific shape and size of the detection box. The number of the reagent wells can be adjusted according to the number of the sample wells 100.

In some embodiments, a magnetic member is also provided. The electromagnet is selected as the magnetic part, the electromagnet has magnetism when being electrified, the magnetic particles are fixed, the electromagnet has no magnetism when being powered off, and the magnetic particles are dissociated in the transfer flow channel. When the waste liquid is discharged through the transfer channel, the magnetic particles are fixed by the magnetic member, so that the cleaved nucleic acid is fixed in the transfer channel and is not discharged together with the waste liquid.

In some embodiments, the cartridge includes a first plate 400 and a second plate 500. The first plate 400 is fixed to the top of the second plate 500. The first plate 400 and the second plate 500 may be manufactured separately and then fixedly connected, or may be integrally formed. If the two parts are connected after being manufactured separately, a sealing element can be arranged between the two parts to enhance the sealing property of the two parts. The sample wells 100 and the respective reagent wells are connected to the first plate 400.

The staging area 610 is coupled to the second plate 500. The first branched flow path includes a first branched flow path first section 420 and a first branched flow path second section 510. The second branched flow channel includes a second branched flow channel first section 430 and a second branched flow channel second section 520. The main channel 410, the first branch channel first section 420 and the second branch channel first section 430 are disposed on the bottom surface of the first plate 400. The first branched flow channel second section 510 and the second branched flow channel second section 520 are disposed on the second plate 500 and penetrate the second plate 500. The first branched flow path first section 420 corresponds to the first branched flow path second section 510, and is communicated with the first branched flow path first section. The first section 430 of the second branch flow channel corresponds to the second section 520 of the second branch flow channel, and the two sections are communicated with each other.

The substances in the sample cell 100 or each reagent cell can sequentially flow through the second branch flow channel second section 520, the second branch flow channel first section 430, the main flow channel 410, the first branch flow channel first section 420, and the first branch flow channel second section 510, and finally enter the corresponding first transition part 611 or second transition part.

In this embodiment, because each runner all sets up in the inside of part, has better leakproofness, can isolate sample or reagent with the external world to avoid the foreign matter to get into, thereby can improve the accuracy of reaction. The structure is simple, the manufacture is easy, only two plates are needed to be arranged, and each flow channel is hollowed out on each plate. If the split-type structure is manufactured, only the bottom surface of the first plate 400 needs to be dug with grooves to form the

first sections

420 and 430 of the first branched runners, and the second plate 500 needs to be dug with a plurality of through holes to form the

second sections

510 and 520 of the first branched runners. The shapes of the first section 420 of the first branch flow passage and the first section 430 of the second branch flow passage are not limited, and can be adjusted according to the specific shape and size of the detection box.

In some embodiments, the cartridge further comprises a sample buffer 620 and a plurality of reagent buffers. The transfer flow channel also comprises a sample temporary storage flow channel and a plurality of reagent temporary storage flow channels. The sample buffer 620 corresponds to the position of the sample cell 100, and each reagent cell corresponds to a reagent buffer position.

The sample cell 100 and the sample buffer 620 can be communicated through the sample buffer flow channel, and if the pressure of the sample buffer 620 on the side far away from the sample cell 100 is lower than the pressure on the side close to the sample cell 100, the sample can be transferred from the sample cell 100 to the sample buffer 620. The sample buffer 620 is connected to the transfer channel, and specifically, the sample buffer 620 is connected to the second section 520 of the second branch channel at the corresponding position. That is, the sample in the sample cell 100 first passes through the sample temporary storage channel from the interior of the sample cell 100 to the sample temporary storage area 620, and then sequentially passes through the second branch channel second section 520, the second branch channel first section 430, the main channel 410, the first branch channel first section 420, and the first branch channel second section 510. If the pressure in the sample buffer 620 is greater on the side away from the sample cell 100 than on the side closer to the sample cell 100, the sample is transferred to the sample cell 100 in the opposite direction of the path described above.

Each reagent pond can communicate through the runner of keeping in of corresponding reagent with the reagent buffer of corresponding position between, if the pressure that reagent buffer kept away from reagent pond one side is less than the pressure that is close to reagent pond one side, reagent can transfer to in the reagent buffer from the reagent pond. Each reagent buffer is connected to the transfer channel at the corresponding position, and specifically, each reagent buffer is connected to the second branch channel second section 520 at the corresponding position. That is, the reagent in the reagent pool first passes through the temporary reagent storage channel from the reagent pool to the temporary reagent storage area, and then sequentially passes through the second branch channel second section 520, the second branch channel first section 430, the main channel 410, the first branch channel first section 420, and the first branch channel second section 510. The waste liquid is recovered by the reverse path. If the pressure of the reagent temporary storage area at the side far away from the reagent pool is higher than the pressure of the reagent temporary storage area at the side near to the reagent pool, the reagent is transferred to the reagent pool in the opposite direction of the path.

When a pressure difference exists between the sample buffer 620 or the reagent buffer and the transfer area 610, the transfer of the sample or the reagent between the sample buffer 620 and the transfer area 610 or between the reagent buffer and the transfer area 610 can be realized.

The detection box is independent when carrying out each reaction. For example, the sample is transferred from the sample cell 100 to the buffer zone 620 and then transferred to one of the transfer zones 610, so that the nucleic acid is immobilized in the transfer channel and then returned to the sample cell 100. Thereafter, the reagents in any one of the reagent wells may reach the corresponding reagent buffer and be transferred to another reagent buffer in the transfer zone 610 through the transfer flow channel. In this process, the reagent reacts with the nucleic acid in the transfer flow channel. Thereafter, the waste liquid is returned to the original reagent tank. The reagents in the other reagent reservoirs are then also reacted with the nucleic acids in the manner described above. In the process, when the reagent in one of the reagent pools participates in the reaction, the reagents in the other reagent pools and the corresponding reagent temporary storage area can not flow into the transfer flow channel again as long as no pressure difference exists, all reactions can be relatively independent, mutual influence is not easy to happen, and the reaction result can be more accurate.

Further, a third plate 600 is disposed at the bottom of the second plate 500, and a sample buffer 620, a reagent buffer and a transfer area 610 are disposed at corresponding positions on the third plate 600. Specifically, the third plate 600 may be hollowed at a corresponding position, and an elastic membrane may be fixed at the hollowed hole to form a sample buffer 620, a reagent buffer and a transfer region 610.

The sample cell 100 and the reagent cell are disposed on the top of the first plate 400, the temporary sample storage area 620, the temporary reagent storage area and the transfer area 610 are disposed on the bottom of the second plate 500, and the temporary reagent storage flow channel and the temporary sample storage flow channel penetrate through the first plate 400 and the second plate 500, so that the sample can be transferred between the sample cell 100 and the temporary sample storage area 620, and the reagent can be transferred between the reagent cell and the corresponding temporary reagent storage area.

In this embodiment, the reagent temporary storage channel and the sample temporary storage channel may be through holes extending in the vertical direction, and certainly, other inclined through holes may be provided.

The elastic membrane can deform when being adsorbed downwards, deforms and expands downwards to form a cavity, and sucks the sample in the sample cell 100. When the elastic membrane at the temporary reagent storage area deforms and expands downwards to form a cavity, the reagent in the reagent pool can be sucked. When the adsorption to the elastic membrane is removed, the elastic membrane is reset and tightly attached to the bottom of the second plate 500, and the waste liquid in the cavity can be pressed upwards into the sample cell 100 or the reagent cell. Or, the elastic membrane at the transit area 610 is adsorbed and expanded downward to form a cavity, and at the same time, the elastic membrane at the sample buffer 620 or a certain reagent buffer rebounds upward to reset, so that the substance can be transferred between the transit area 610 and the corresponding sample buffer or reagent buffer.

Specifically, the method comprises the following steps. The bottom of the elastic membrane can be connected with a drawer, an air pump and other similar pneumatic elements to absorb the deformation and expansion of the elastic membrane or make the elastic membrane rebound and reset.

In the above-mentioned structure of inflation, when the waste liquid returns the back, as long as no longer adsorb elastic membrane downward deformation, the waste liquid just can be stable be stored in sample cell 100 or reagent pond, can not influence subsequent other reactions. Moreover, the structure is simple, and the elastic film is fixed at the corresponding position of the bottom of the second plate 500. The elastic film is also easy to obtain and has low cost.

Preferably, the position corresponding to each elastic membrane on the transfer flow channel of the second plate 500 is provided with a downward convex protruding part, so that the elastic membrane is elastically deformed, and under the action of the resilience force, the bottom of the second plate 500 can be stably attached to the transfer flow channel, subsequent other processing steps are not affected, and the stability of the detection box during use is further improved. And the reagent can not be shifted in position during transportation.

Preferably, the bottom of the elastic membrane can be provided with a supporting member to tightly support the elastic membrane against the second plate 500, so that the elastic membrane can stably adhere to the bottom of the second plate 500 without deformation, subsequent other reactions cannot be influenced, and the stability of the detection box in use is further improved.

Preferably, the bottom of the elastic membrane is provided with a holding piece, and the holding piece is provided with an accommodating groove for accommodating a cavity formed after the elastic membrane deforms so as to support the cavity. Preferably, a sealing ring can be arranged between the elastic membrane and the abutting piece to enhance the sealing performance.

Referring to FIGS. 3 to 7, FIGS. 4 to 7 respectively show a sectional view of the cartridge of FIG. 1, a plan view of a first plate of the cartridge of FIG. 1, a bottom view of the first plate of the cartridge of FIG. 1, and a plan view of a second plate of the cartridge of FIG. 1. The following detailed description is made with reference to the accompanying drawings.

Specifically, the sample buffer 620 is located right below the sample cell 100, and the plurality of reagent buffers are a first reagent buffer 631, a second reagent buffer 632, a third reagent buffer 633 and a fourth reagent buffer 634. The first reagent buffer zone 631 is located below the first reagent bath 210, the second reagent buffer zone 632 is located below the second reagent bath 220, the third reagent buffer zone 633 is located below the third reagent bath 230, and the fourth reagent buffer zone 634 is located below the fourth reagent bath 240.

Referring to fig. 3, 4 and 6, the sample buffer flow path includes a first section 440 of the sample buffer flow path extending through the first plate 400 and a second section 530 of the sample buffer flow path extending through the second plate 500. The bottom end of the sample buffer channel first section 440 is aligned with and communicates with the top end of the sample buffer channel second section 530, both of which are located within the sample buffer 620. When there is a pressure difference between the sample cell 100 and the sample buffer 620, the sample can flow between the sample cell 100 and the sample buffer 620 via the first section 440 of the sample buffer flow channel and the second section 530 of the sample buffer flow channel. The second branch flow channel second section three region 523 is located at one side of the sample temporary storage flow channel second section 530, and penetrates through the second plate 500, and the second branch flow channel second section three region 523 is located within the range of the sample temporary storage region 620.

The first plate 400 is further provided with a first section 451 of a reagent temporary storage flow channel, the second plate 500 is provided with a second section 541 of the reagent temporary storage flow channel, the bottom end of the first section 451 of the reagent temporary storage flow channel is aligned with the top end of the second section 541 of the reagent temporary storage flow channel, and the two are communicated and both are positioned within the range of the first section 631 of the reagent temporary storage flow channel. Reagent is able to flow between the first reagent reservoir 210 and the reagent buffer zone one zone 631 via the reagent buffer flow path first section first zone 451 and the reagent buffer flow path second section first zone 541 when a pressure differential exists between the first reagent reservoir 210 and the reagent buffer zone one zone 631. The second branch flow channel first section 521 is located at one side of the reagent temporary storage flow channel second section first section 541, and penetrates through the second plate 500, and the second branch flow channel second section first section 521 is located within the range of the reagent temporary storage area first section 631.

Similarly, the second reagent reservoir 220 is connected to the reagent temporary storage flow channel second section second region 632 through the reagent temporary storage flow channel first section second region 452, and a second branch flow channel second section second region 522 is disposed at one side of the reagent temporary storage flow channel second section second region 542.

The third reagent pool 230 is communicated with the reagent temporary storage area three zone 633 through the reagent temporary storage flow channel first section three zone 453 and the reagent temporary storage flow channel second section three zone 543, and a second branch flow channel second section four zone 524 is arranged on one side of the reagent temporary storage flow channel second section three zone 543.

The fourth reagent pool 240 is communicated with the reagent temporary storage region fourth region 634 through the reagent temporary storage flow channel first section fourth region 454 and the reagent temporary storage flow channel second section fourth region 544, and a second branch flow channel second section fifth region 525 is arranged on one side of the reagent temporary storage flow channel second section fourth region 544.

Referring to fig. 3 and 6, five second branched flow paths arranged on the bottom surface of the first plate 400 are a second branched flow path first section first area 431, a second branched flow path first section second area 432, a second branched flow path first section third area 433, a second branched flow path first section fourth area 434, and a second branched flow path first section fifth area 435, respectively. One end of the first section 431 of the second branch flow channel is connected to the main flow channel 410, and the other end is located on the top of the first section 521 of the second branch flow channel on the second plate 500, and the two are connected. One end of the second branch flow channel first section second area 432 is connected to the main flow channel 410, and the other end is located on the top of the second branch flow channel second section second area 522 on the second plate 500, and the two are connected. One end of the second branch flow channel first section three region 433 is communicated with the main flow channel 410, and the other end is located on the top of the second branch flow channel second section three region 523 on the second plate 500, and the two are communicated with each other. One end of the second branch flow channel first section fourth region 434 is connected to the main flow channel 410, and the other end is located on the top of the second branch flow channel second section fourth region 524 on the second plate 500, and the two are connected. One end of the second branch flow channel first section fifth area 435 is communicated with the main flow channel 410, and the other end is located on the top of the second branch flow channel second section fifth area 525 on the second plate 500, and the two are communicated.

The five first branch flow channel first sections 420 disposed on the bottom surface of the first plate 400 are a first branch flow channel first section first region 421, a first branch flow channel first section second region 422, a first branch flow channel first section three region 423, a first branch flow channel first section fourth region 424, and a first branch flow channel first section fifth region 425, respectively. One end of the first branch flow channel first section area 421 is communicated with the main flow channel 410, and the other end is located on the top of the first branch flow channel second section area 511 on the second plate 500, and the two are communicated with each other. One end of the first branched flow passage first section second region 422 is communicated with the main flow passage 410, and the other end is positioned on the top of the first branched flow passage second section second region 512 on the second plate 500, and the two are communicated. One end of the first branch flow channel third section 423 is communicated with the main flow channel 410, and the other end is located at the top of the first branch flow channel second section 513 on the second plate 500, and the two are communicated with each other. One end of the first branched flow passage first section fourth area 424 is communicated with the main flow passage 410, and the other end is positioned on the top of the first branched flow passage second section fourth area 514 on the second plate 500, and the two are communicated with each other. One end of the first branched flow passage first section fifth area 425 is communicated with the main flow passage 410, and the other end is positioned on the top of the first branched flow passage second section fifth area 515 on the second plate 500, and the two are communicated.

The respective reaction processes are explained with reference to the entire drawings. The first cover 310 is opened, the extracted sample is placed in the cuvette 100, and then the first cover 310 is closed. The sample is lysed within the sample cell 100, releasing the nucleic acids. In the process, preferably, a stirrer may be provided to stir it to accelerate the cracking. Alternatively, an ultrasonic device may be provided to accelerate the lysis by ultrasonic waves. Alternatively, heating means may be provided to accelerate the cracking by increasing the temperature. Alternatively, the above-described modes may be combined.

After the cell lysis is completed, the elastic membrane at the sample buffer 620 is deformed and expanded downward, and the sample lysate in the sample cell 100 sequentially passes through the first section 440 and the second section 530 of the sample buffer channel and enters the sample buffer 620.

The elastic membrane at the second zone 6122 of the second transfer section is then deformed and expanded downward while the elastic membrane at the sample buffer 620 is slowly repositioned upward. In the process, the sample and the lysis solution sequentially flow through the second branch flow channel second section three region 523, the second branch flow channel first section three region 433, the main flow channel 410, the first branch flow channel first section two region 422, and the first branch flow channel second section two region 512 to reach the second transition section second region 6122. When the sample and the lysis solution flow through the main channel 410, the nucleic acid in the sample is adsorbed by the magnetic particles. The elastic membrane at the sample buffer 620 is then deformed and expanded downward while the elastic membrane at the second zone 6122 of the second transfer section is slowly repositioned upward. The material in the second zone 6122 of the second transfer section will be transferred in the opposite direction of the path described above, back again into the sample buffer 620. The above steps are repeated to make the sample lysate oscillate between the second zone 6122 of the second transit section and the temporary sample storage zone 620 for many times, so that the nucleic acid is adsorbed on the magnetic particles as completely as possible. Finally, the magnetic part circular telegram, through the fixed magnetic particle of magnetic attraction, and then fixed nucleic acid position, sample schizolysis waste liquid is stayed in sample buffer 620, and the elastic membrane of sample buffer 620 department is in the expanded state that warp downwards. Then, the elastic membrane at the temporary sample storage area 620 is slowly restored upwards, and the waste liquid is pressed into the sample cell 100.

The elastic membrane at the reagent buffer zone 631 is deformed and expanded downwards, and the cleaning solution in the first reagent pool 210 flows through the reagent buffer flow path first section 451 and the reagent buffer flow path second section 541 sequentially into the reagent buffer zone 631.

The elastic membrane at the third zone 6123 of the second transfer section is deformed and expanded downwards, and at the same time, the elastic membrane at the first reagent buffer zone 631 is slowly restored upwards. In this process, the cleaning solution flows through the second branch flow channel second section first area 521, the second branch flow channel first section first area 431, the main flow channel 410, the first branch flow channel first section fourth area 424, and the first branch flow channel second section fourth area 514 in sequence to reach the second transition section third area 6123. When the washing solution flows through the main channel 410, the nucleic acid adsorbed on the magnetic particles can be washed to remove other impurities. Then, the elastic membrane at the first reagent buffer zone 631 is deformed and expanded downward, and at the same time, the elastic membrane at the third zone 6123 of the second transfer portion is slowly restored upward. The substance in the third zone 6123 of the second transfer section will be transferred in the opposite direction of the path described above and again back into the reagent buffer zone one zone 631. The above steps are repeated, so that the cleaning liquid vibrates for many times between the third zone 6123 of the second transfer part and the first zone 631 of the reagent temporary storage region, impurities can be thoroughly removed, and the cleaning effect is optimized. Finally, the magnetic member is energized to fix the magnetic particles, the waste cleaning solution is retained in the first reagent buffer area 631, and the elastic membrane at the first reagent buffer area 631 is in a downward deformed and expanded state. Then, the elastic membrane at the first reagent buffer zone 631 is returned to the upward position, and the waste liquid is pressed into the first reagent tank 210.

The elastic membrane at the second reagent temporary storage area 632 deforms and expands downward, and the cleaning solution in the second reagent pool 220 flows through the first reagent temporary storage flow channel section 452 and the second reagent temporary storage flow channel section 542 in sequence and enters the second reagent temporary storage area 632.

The elastic membrane in the fourth zone 6124 of the second transit section deforms and expands downward, and the elastic membrane in the second reagent buffer zone 632 is gradually restored upward. In this process, the cleaning solution flows through the second branch flow channel second section second area 522, the second branch flow channel first section second area 432, the main flow channel 410, the first branch flow channel first section fifth area 425, and the first branch flow channel second section fifth area 515 in sequence to reach the fourth area 6124 of the second transition part. When the washing solution flows through the main channel 410, the nucleic acid adsorbed on the magnetic particles can be washed to remove other impurities. Then, the elastic film at the second zone 632 of the reagent temporary storage zone is deformed and expanded downward, and at the same time, the elastic film at the fourth zone 6124 of the second transfer part is slowly restored upward. The material in the fourth zone 6124 of the second turnaround section will be transferred in the opposite direction of the path described above back into the second reagent buffer zone 632 again. The above steps are repeated to make the cleaning solution vibrate between the fourth zone 6124 of the second transit part and the second zone 632 of the reagent temporary storage zone for many times, so that the impurities can be removed more thoroughly, and the cleaning effect is optimized. Finally, the magnetic member is energized to fix the magnetic particles, the waste cleaning solution is retained in the second reagent buffer area 632, and the elastic membrane at the second reagent buffer area 632 is in a downward deformed and expanded state. Then, the elastic membrane at the second reagent buffer area 632 is gradually restored upward, and the waste liquid is pressed into the second reagent pool 220.

So that the elastic membrane at the third zone 633 of the temporary reagent storage area deforms and expands downward, and the eluent in the third reagent pool 230 sequentially flows through the first section 453 and the second section 543 of the temporary reagent storage flow channel and enters the third zone 633 of the temporary reagent storage area.

The elastic film at the first zone 6121 of the second transfer part is deformed and expanded downwards, and meanwhile, the elastic film at the third zone 633 of the reagent temporary storage zone is slowly reset upwards. In this process, the cleaning solution flows through the second branch flow channel second section fourth area 524, the second branch flow channel first section fourth area 434, the main flow channel 410, the first branch flow channel first section first area 421, and the first branch flow channel second section first area 511 in sequence to reach the second transition portion first area 6121. When the eluent flows through the main channel 410, the nucleic acids can be separated from the magnetic particles, so that the nucleic acids are dissociated in the eluent and enter the first zone 6121 of the second intermediate transfer portion together. Then, the elastic film at the third zone 633 of the reagent temporary storage zone is deformed and expanded downwards, and at the same time, the elastic film at the first zone 6121 of the second transfer part is slowly restored upwards. The material in the first zone 6121 of the second transfer section will be transferred in the opposite direction of the path described above, back again into the reagent staging zone three zone 633. The above steps are repeated to make the eluent oscillate between the first zone 6121 of the second transit part and the three zone 633 of the reagent temporary storage area for many times, so as to fully elute the nucleic acid. Finally, the magnetic part is electrified to fix the magnetic particles, and the elastic membrane at the three zones 633 of the temporary reagent storage zone is in a downward deformation and expansion state.

Then, the magnetic member is powered off, and the magnetic particles can move freely, so that the elastic film at the first transit part 611 deforms and expands downwards, and meanwhile, the elastic film at the reagent temporary storage area 633 is reset slowly upwards. In this process, the eluent flows through the second branch flow channel second section fourth region 524, the second branch flow channel first section fourth region 434, the main flow channel 410, the first branch flow channel first section third region 423, and the first branch flow channel second section third region 513 in this order into the first transit section 611.

The elastic membrane at the reagent temporary storage region fourth region 634 deforms and expands downward, and the PCR reaction solution in the fourth reagent pool 240 flows through the reagent temporary storage flow channel first section fourth region 454 and the reagent temporary storage flow channel second section fourth region 544 in sequence and enters the reagent temporary storage region fourth region 634.

The elastic membrane at the first transit part 611 is deformed and expanded downwards continuously, and at the same time, the elastic membrane at the reagent buffer zone four zone 634 is reset upwards slowly. In this process, the first PCR reaction solution in the reagent temporary storage region fourth region 634 flows through the second branch flow channel second section fifth region 525, the second branch flow channel first section fifth region 435, the main flow channel 410, the first branch flow channel first section third region 423, and the first branch flow channel second section third region 513 in sequence to reach the first transit part 611. The first PCR reaction solution is mixed with the nucleic acid eluting solution in the first transit part 611 to perform the nested first PCR reaction.

Preferably, a plurality of amplification cells 480 are further disposed in the first plate 400, and a second PCR reaction solution is stored in the amplification cells 480, and the second PCR reaction solution can provide primers, enzymes, magnesium ions, DNTP, and the like required for the second PCR reaction. The primers can specifically amplify nucleic acid, the same primers or different primers can be placed in each amplification pool 480, multiple primers can be placed in a single amplification pool 480, and the primers can be in a freeze-dried or liquid form. Because the plurality of amplification pools 480 are arranged, a plurality of PCR reactions can be completed on the same detection box only by extracting a sample once, and the use is convenient.

The top of each amplification pool 480 is provided with a fourth cover 340, and the substances in each amplification pool 480 are sealed by the fourth cover 340 so as to prevent the foreign substances from entering and affecting the performance of the amplification pools.

A plurality of amplification reaction regions 640 are provided on the third plate 600, and the amplification reaction regions 640 are also formed by an elastic membrane. The amplification reaction region 640 and the first transition part 611 can communicate with each other via a first amplification flow channel, and the amplification cell 480 and the amplification reaction region 640 can communicate with each other via a second amplification flow channel. The elastic membrane at the amplification reaction region 640 is deformed and expanded downward, and at the same time, the elastic membrane at the first transit portion 611 is slowly restored upward. In this process, the reaction solution of the first PCR can enter the amplification reaction region 640. The second PCR reaction solution in the amplification chamber 480 also enters the amplification reaction region 640. After the second PCR reaction solution is mixed with the first PCR reaction solution, the PCR reaction is completed in the amplification reaction region 640.

In the embodiment, the steps of cracking, cleaning and the like before PCR reaction and the PCR reaction can be integrated on one detection box for completion, so that the method is very quick and convenient. Before the amplification reaction region 640 is deformed and expanded downward, the PCR reaction solution II is separated from the other various reagents, and the previous steps can be performed more stably without being affected by each other. And a nested PCR step is arranged, so that the specificity and the sensitivity of detection are enhanced.

Specifically, the first amplification flow channel includes a first amplification flow channel first section 550, a first amplification flow channel second section 460, and a first amplification flow channel third section 560, which are connected in sequence. The second amplification flow channel includes a second amplification flow channel first section 570 and a second amplification flow channel second section 470. The first amplification flow channel first section 550, the first amplification flow channel third section 560, and the second amplification flow channel first section 570 are all disposed on the second plate 500, and penetrate the second plate 500. The first amplification flow channel second section 460 and the second amplification flow channel second section 470 are both disposed at the bottom surface of the first plate 400.

The elastic membrane at the amplification reaction region 640 is deformed and expanded downward, and at the same time, the elastic membrane at the first transit portion 611 is slowly restored upward. In this process, the reaction solution of the first PCR sequentially flows through the first section 550 of the first amplification flow channel, the second section 460 of the first amplification flow channel, and the third section 560 of the first amplification flow channel, and enters the amplification reaction region 640. And, the second PCR reaction solution in the amplification pool 480 flows through the second amplification flow channel section 470 and the first amplification flow channel section 570 in sequence and enters the amplification reaction region 640.

In the above steps, some of the sample and the reagent may remain in the flow channel, and cannot reach the predetermined area, so that a certain loss exists. Therefore, in the initial calculation of the amount, the various substances are kept in balance to ensure that a sufficient amount of the reaction is carried out.

In some embodiments, there is also provided a nucleic acid detecting apparatus including the above-described cartridge. The detection cassette is mounted on the main body of a nucleic acid detection device, and the nucleic acid detection device is connected with a pneumatic element, a temperature control module and an optical module. The elastic membrane is sucked by the pneumatic piece to be deformed, expanded or reset, the reaction temperature is adjusted by the temperature control module, and fluorescence detection is performed by the optical module. The pneumatic element, the temperature control module and the optical module can be realized by adopting the prior art, and the details are not repeated herein.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only represent some embodiments of the present invention, and the description thereof is specific and detailed, but not to be construed as limiting the scope of the present invention. It should be noted that, for those skilled in the art, without departing from the spirit of the present invention, several variations and modifications can be made, which are within the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the appended claims.

Claims

1. A cartridge, comprising:

a runner assembly including a transit runner;

a plurality of reagent reservoirs;

at least one sample cell;

a fixing member for fixing an effective substance in a sample;

2. The testing cassette of claim 1, wherein the transition area comprises a first transition portion and a plurality of second transition portions, the transition flow channel comprises a main flow channel, a plurality of first branch flow channels and a plurality of second branch flow channels, a first end of each of the first branch flow channels is connected to the main flow channel, the first transition portions and the second transition portions are connected to a second end of each of the first branch flow channels in a one-to-one correspondence manner, a first end of each of the second branch flow channels is connected to the main flow channel, and the sample wells and the reagent wells are connected to a second end of each of the second branch flow channels in a one-to-one correspondence manner.

3. The cartridge according to claim 2, further comprising a magnetic member capable of fixing a position of the fixing member by a magnetic attraction force.

4. The cartridge according to claim 2, wherein the cartridge comprises a main body member, the main body piece comprises a first plate and a second plate, the first plate is connected with the top of the second plate, the reagent pool and the sample pool are both connected with the first plate, the transfer area is connected with the second plate, the first branch flow channel comprises a first branch flow channel first section and a first branch flow channel second section, the first branch flow channel first section is connected with the first branch flow channel second section, the second branch flow channel comprises a first section of the second branch flow channel and a second section of the second branch flow channel, the first section of the second branch flow channel is connected with the second section of the second branch flow channel, the bottom surface of the first plate is provided with the main runner, the first section of the first branch runner and the first section of the second branch runner, and the second plate is provided with a first branch flow channel second section and a second branch flow channel second section which penetrate through the second plate.

5. The detecting box of claim 1, wherein the detecting box further comprises a plurality of reagent temporary storage areas corresponding to the reagent pools in a one-to-one manner, the flow channel assembly further comprises a plurality of reagent temporary storage flow channels, each reagent pool and one reagent temporary storage area can be communicated through the corresponding reagent temporary storage flow channel, when a pressure difference exists between the reagent pool and the reagent temporary storage area, material transfer can be realized, and the reagent temporary storage area is connected with the transfer flow channel;

6. The testing cassette of claim 5, wherein the testing cassette comprises a main body member, the reagent reservoir and the sample reservoir are connected to a top portion of the main body member, the reagent buffer and the sample buffer are connected to a bottom portion of the main body member, and the reagent buffer flow passage and the sample buffer flow passage are communicated to each other through the main body member.

7. The detecting box according to claim 1, wherein the detecting box comprises a main body member, a plurality of elastic membranes are connected to the main body member to form a reagent temporary storage area, a sample temporary storage area and the transfer area at corresponding positions, respectively, the elastic membranes are connected to the transfer flow channel, the elastic membranes at corresponding positions can expand under the action of external force to form a reagent temporary storage cavity, a sample temporary storage cavity and a transfer cavity, respectively, and when a pressure difference exists between one side of the elastic membranes away from the transfer flow channel and one side of the elastic membranes close to the transfer flow channel, material transfer can be achieved.

8. The detecting box of claim 7, wherein the transferring channel has a protrusion portion at a contact area with the reagent buffer, the sample buffer and the transferring region of the elastic membrane, and the protrusion portion abuts against the elastic membrane.

9. The detecting box according to claim 7, wherein a holding member is provided at a bottom of the elastic membrane, and the holding member can hold the elastic membrane against the main body member.

10. A nucleic acid detecting apparatus comprising the cartridge according to any one of claims 1 to 9.