CN218620810U - Sample preprocessing device - Google Patents

Abstract

The utility model provides a sample preprocessing device, include the centrifuging tube and set up in the intraductal detachable filter column of centrifuging tube, from the top down is equipped with disperse phase adsorbent solid in the filter column in proper order, first sieve, solid phase adsorbent solid and second sieve, under the shock effect of filter column, disperse phase adsorbent solid forms the suspension with the liquid sample that adds the filter column, under the centrifugation of centrifuging tube or vacuum filtration effect, soluble thing passes through first sieve in the suspension, solid phase adsorbent solid and second sieve form the effluent that contains the nucleic acid, can get the whole removal of filter column behind the effluent liquid, consequently, inhibitor and adsorbent can be got rid of simultaneously, effectively reduced move the liquid number of times of operation, the operating efficiency is improved, avoid producing misoperation's possibility, and the setting up of solid phase adsorbent solid also can avoid the too early outflow of liquid sample before the centrifugation, when improving the operation convenience, the influence to the sample treatment effect has been reduced, the multiple advantage that efficiency promoted and effect has been fused.

Description

Sample preprocessing device

Technical Field

The utility model relates to a nucleic acid separation preliminary treatment technical field especially relates to a sample preprocessing device.

Background

Chinese patent CN 201280034773.0 discloses the isolation of nucleic acids, and the method provides a technology related to the isolation of nucleic acids. The technology relates to methods for extracting and purifying nucleic acids from exfoliated gut cells in fecal samples for quantitative and sensitive assays. In particular a novel method for the purification of specific DNA from stool using an inhibitor removal step and the direct capture of DNA from stool supernatant or a combination of these steps. The technology further provides a filtration device suitable for use with complex and viscous samples, such as fecal samples. Thus, provided herein are methods of isolating a target nucleic acid from a sample, the methods comprising removing a assay inhibitor, if present, from the sample to produce a clarified sample; capturing the target nucleic acid, if present, from the clarified sample with a capture reagent to form a capture complex; separating the capture complex from the clarified sample; and recovering the target nucleic acid, if present, from the capture complex in the nucleic acid solution. The method further comprises retaining the clarified sample after the capturing step; and repeating the separating and recovering steps using the retained clarified sample and the second capture reagent.

Chinese patent CN 202011493703.4 discloses a method for extracting DNA sample from feces and a method for detecting methylation of colorectal cancer related gene, comprising: a pretreatment step: centrifuging the fecal diluent sample to obtain a first supernatant; the stool in the stool dilution sample is taken from a subject in need of colorectal cancer detection; during centrifugation, the volume of the fecal diluent sample in a single centrifuge tube is no greater than 2mL, preferably 0.5-1.8mL, and more preferably 1.8mL. The method for extracting the DNA sample from the excrement is characterized in that the centrifugation parameters are set as follows: the rotating speed is 10000-13000rpm, and the time is 10s-5min. The method for extracting the DNA sample from the excrement further comprises the following steps: an adsorption step: uniformly mixing an adsorbent with the first supernatant to remove impurities in the supernatant to obtain a first mixed solution, and centrifuging the first mixed solution to obtain a second supernatant; preferably, the adsorbent is selected from cross-linked polyvinylpyrrolidone; preferably, the crospovidone is a crospovidone powder or a crospovidone tablet. The method for extracting the DNA sample from the excrement is characterized in that the mass-to-volume ratio of the adsorbent to the first supernatant is 1 (2-4), and preferably 1:4. The method for extracting the DNA sample from the excrement further comprises the following steps: a nucleic acid denaturation step: performing denaturation treatment on the second supernatant to enable the DNA double-stranded structure to form a single-stranded structure; preferably, the temperature of the denaturation treatment is 85-95 ℃ and the time is 5-15min, and more preferably 10min.

The above-mentioned inhibitor removing component added in the prior art, i.e., the adsorbent, is usually provided in a powder form or a tablet form, and the inhibitor removal is performed by utilizing the characteristic that the adsorbent is easily disintegrated and dispersed into a suspension. However, the above prior art has the following disadvantages:

first, the adsorbent in the suspension is removed by centrifugation after it has been activated, but this operation requires manual pipetting of the sample supernatant from which the inhibitor component has been removed. However, the content of human DNA in a stool sample is usually very low, so a large initial volume of a single sample, mostly a large volume of 2ml or more, is usually required for extracting human DNA in a stool sample, which causes requirements for multiple pipetting operations of a single sample, and causes poor throughput of sample treatment due to low efficiency.

Secondly, the sorbent needs to be added to the sample to be treated either by shaking, blending and pipetting, or by taking tablets or granules in solid form, depending on the formulation provided, so that this step of manual operation is required. If the centrifugal filtration mode is adopted to remove the adsorbent, the suspension sample containing the adsorbent needs to be transferred into the centrifugal filtration device by one or more times through a liquid transfer mode, and the inconvenience of operation is increased again for the initial single sample amount with large volume, so that the operation efficiency and the sample flux are influenced. And in order to prevent the small holes of the suction head from being blocked by particles when the suspension is sucked, a small section of the tip end of the suction head needs to be reduced, so that the manual operation is additionally added.

Thirdly, some strategies are removed by centrifugal filtration, but the treatment by centrifugal filtration usually causes the blockage of the sieve plate or the filter screen by fine particles, and once the blockage of the sieve plate or the filter screen occurs, human intervention operation is required, such as replacing a new centrifugal filtration device, or adding extra centrifugal time, thereby bringing inconvenience to the efficiency of sample treatment operation.

Therefore, it is necessary to design a better processing device to solve the above problems.

SUMMERY OF THE UTILITY MODEL

Problem to prior art existence, the utility model provides a reduce and move liquid operation number of times, improve operating efficiency, avoid producing misoperation's sample preprocessing device.

In order to achieve the above purpose, the utility model adopts the following technical scheme:

the utility model provides a sample preprocessing device, includes the centrifuging tube and sets up in the intraductal detachable filtration column of centrifuging tube, from the top down is equipped with disperse phase adsorbent solid, first sieve, fixed phase adsorbent solid and second sieve in proper order in the filtration column under the shock effect of filtration column, disperse phase adsorbent solid and addition the liquid sample of filtration column forms the suspension under the centrifugation of centrifuging tube or vacuum filtration effect, the thing process that can dissolve in the suspension first sieve the fixed phase adsorbent solid reaches the second sieve forms the effluent that contains nucleic acid, flows extremely the centrifuging tube bottom.

Further, the top of the centrifugal tube is provided with a tube cover, and when the filter column is arranged in the centrifugal tube, the top opening of the centrifugal tube is sealed through the tube cover.

Further, the tube cap inner wall with centrifuging tube top outer wall all is equipped with the screw thread, the tube cap pass through the screw thread spiro union in centrifuging tube top outer wall.

Further, the top of filtering the post is equipped with the flange, the flange overlap joint in the top terminal surface of centrifuging tube, so that filter the post hang in the centrifuging tube.

Further, stationary phase adsorbent solid prepackage in first sieve with between the second sieve, by first sieve with the second sieve centre gripping.

Further, the centrifuging tube bottom is equipped with the collection portion of back taper, and when effluent liquid got into collection portion, the filter column by the centrifuging tube takes out.

Further, the first sieve plate or/and the second sieve plate is/are a micropore sieve plate.

The utility model has the advantages that:

the method comprises the steps of adding a sample into a filter column pre-filled with dispersed phase adsorbent solids, a first sieve plate, fixed phase adsorbent solids and a second sieve plate, forming suspension by oscillating the dispersed phase adsorbent solids and the sample to realize removal of a first round of inhibitor, and allowing liquid containing required nucleic acid components to flow out through the first sieve plate, the fixed phase adsorbent solids and the second sieve plate through a centrifugal action to obtain effluent liquid containing nucleic acid for downstream nucleic acid extraction, so that removal of the second round of inhibitor is realized, and the filter column can be integrally removed after the effluent liquid is obtained, so that the inhibitor and the adsorbent can be simultaneously removed, the pipetting operation times are effectively reduced, the operation efficiency is improved, the possibility of operation errors is avoided, the fixed phase adsorbent solids are also arranged to avoid premature outflow of liquid samples before centrifugation, the operation convenience is improved, meanwhile, the influence on the sample treatment effect is reduced, and multiple advantages of efficiency improvement and effect improvement are fused.

Drawings

FIG. 1 is a schematic structural view of a filter column in the sample pretreatment device of the present invention;

FIG. 2 is a schematic view of the sample pretreatment apparatus before the filter column is loaded into the centrifuge tube;

FIG. 3 is a schematic view of the filter column of the sample pretreatment device according to the present invention being loaded into a centrifuge tube;

FIG. 4 is a schematic structural view of the installed sample pretreatment device of the present invention;

in the figure, 1-centrifuge tube, 11-tube cap, 12-screw thread, 13-collecting part, 2-filter column, 21-disperse phase adsorbent solid, 22-first sieve plate, 23-stationary phase adsorbent solid, 24-second sieve plate, 25-flange.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.

It should be noted that all the directional indicators (such as upper, lower, left, right, front, and rear … …) in the embodiments of the present invention are only used to explain the relative position relationship between the components, the motion situation, etc. in a specific posture (as shown in the drawings), and if the specific posture is changed, the directional indicator is changed accordingly.

As fig. 1 to fig. 4, the utility model provides a sample preprocessing device, include centrifuging tube 1 and set up detachable filter column 2 in centrifuging tube 1, from the top down is equipped with disperse phase adsorbent solid 21 in proper order in the filter column 2, first sieve 22, stationary phase adsorbent solid 23 and second sieve 24, under the shock effect of filter column 2, disperse phase adsorbent solid 21 forms the suspension with the liquid sample that adds filter column 2, under centrifuging tube 1's centrifugation or vacuum filtration, soluble thing is through first sieve 22 in the suspension, stationary phase adsorbent solid 23 and second sieve 24 form the effluent that contains nucleic acid, flow out and be used for low reaches nucleic acid to centrifuging tube 1 bottom and draw, disperse phase adsorbent solid 21 in the suspension will be held back by first sieve 22 and block.

The utility model discloses the sample is added and is equipped with disperse phase adsorbent solid 21 in advance, first sieve 22, stationary phase adsorbent solid 23, in the filtration post 2 of second sieve 24, form suspension with disperse phase adsorbent solid 21 and sample through the oscillation, realize that first round of inhibitor gets rid of, the liquid that contains required nucleic acid composition is via first sieve 22 to rethread centrifugal action, stationary phase adsorbent solid 23 and second sieve 24 flow, it is used for low reaches nucleic acid to extract to obtain the effluent that contains nucleic acid, realize that second round of inhibitor gets rid of, can get the whole removal of filtration post 2 behind the effluent, consequently, inhibitor and adsorbent can be got rid of simultaneously, effectively reduced and moved liquid operation number of times, the operating efficiency is improved, avoid producing misoperation's possibility, and the setting of stationary phase adsorbent solid 23 also can avoid the too early outflow of liquid sample before the centrifugation, in the time of improving the simple operation nature, the influence to the sample treatment effect has been reduced, efficiency promotion and the multiple advantage that the effect promoted have been fused.

Centrifuge tube 1 top is equipped with tube cap 11, when filter column 2 packs into centrifuge tube 1, through tube cap 11 sealed centrifuge tube 1 top opening, avoids intraductal liquid to leak when centrifugal action. In this embodiment, the inner wall of the tube cover 11 and the outer wall of the top of the centrifuge tube 1 are both provided with threads 12, and the tube cover 11 is screwed on the outer wall of the top of the centrifuge tube 1 through the threads 12.

Preferably, the top of filtering post 2 is equipped with flange 25 to the evagination, and flange 25 overlap joint in centrifuging tube 1's top end face to make filtering post 2 hang in centrifuging tube 1, through the setting of flange 25, can make the convenient centrifuging tube 1 of putting into of filtering post 2 or take out from centrifuging tube 1.

The utility model discloses in, stationary phase adsorbent solid 23 encapsulates between first sieve 22 and second sieve 24 in advance, by first sieve 22 and 24 centre grippings of second sieve, and first sieve 22 or second sieve 24 are the micropore sieve. The fixed phase adsorbent solid 23 is packaged in a sandwich mode, meanwhile, the dispersed phase adsorbent solid 21 is added above the first sieve plate 22 in advance, when a liquid sample is added into the filter column 2, the dispersed phase adsorbent solid 21 and the liquid sample are mixed to form turbid liquid in a vibration mode, and the first round of inhibitor removal is realized. Then the liquid containing the required nucleic acid components flows out through the first sieve plate 22, the stationary phase adsorbent solid 23 and the second sieve plate 24 by a centrifugal mode, and the effluent containing the nucleic acid is obtained for downstream nucleic acid extraction, so that the second round of inhibitor removal is realized. Meanwhile, the upper-layer dispersed phase adsorbent solid 21 can form a three-dimensional column state accumulation under the centrifugal action, so that the condition that the micropores of the sieve plate are blocked by large-particle impurities or adhesive impurities in a sample to cause unsmooth centrifugation is avoided. The lower stationary phase adsorbent solid 23 is added in the interlayer of the first sieve plate 22 and the second sieve plate 24, thereby avoiding the sample liquid from flowing out too early when the upper dispersed phase adsorbent solid 21 treatment is effective, and increasing the removal effect of an inhibitor. The utility model discloses the inhibitor is got rid of by simple disperse phase or stationary phase, becomes disperse phase and stationary phase and acts simultaneously, can block the adsorbent again when getting rid of the inhibitor, and further can prevent to block up first sieve 22 micropore at the condition that only uses stationary phase adsorbent solid 23, first sieve 22 and second sieve 24, adhesion large granule impurity in the sample liquid.

Centrifuge tube 1 bottom is equipped with the collection portion 13 of back taper, get into collection portion 13 when the effluent, it takes out by centrifuge tube 1 and abandons to filter post 2, it makes the consumptive material device to be about to filter post 2, get rid of the inhibitor and filter with the adsorbent and get rid of the integration and be one-step processing, can carry out the inhibitor simultaneously and get rid of and filter with the adsorbent and get rid of, effectively reduced and moved the liquid-transfering operation number of times, and the operation efficiency is improved, avoid producing misoperation's possibility, and the upper sample that needs to remain is brought into because of the too big sediment of imbibition dynamics when absorbing centrifugation back supernatant.

Encapsulate the adsorbent solid in advance in the used consumptive material device of centrifugation (filter column 2), avoided the manual operation inconvenience of taking and adding the adsorbent to bring, including the pretreatment operation that the tip was cut off to the suction head, suspension oscillation mixing operation, move the liquid operation to and the quantity error that may bring. Through the innovative design of consumables, the inhibitor removal treatment and the adsorbent removal treatment are combined into one operation. Meanwhile, premature outflow of sample liquid before centrifugation in the existing inhibitor removing operation is avoided, the influence on the sample treatment effect is reduced while the operation convenience is improved, and multiple advantages of efficiency improvement and effect improvement are combined.

In other embodiments, the removal of the inhibitor in the centrifuge tube 1 can be achieved by vacuum filtration, which can also reduce pipetting operations.

Although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (7)

1. A sample pretreatment device, comprising: centrifuging tube and set up detachable filter column in the centrifuging tube, from the top down is equipped with disperse phase adsorbent solid, first sieve, stationary phase adsorbent solid and second sieve in proper order in the filter column under the shock effect of filter column, disperse phase adsorbent solid and addition the liquid sample of filter column forms the suspension under the centrifugation of centrifuging tube or vacuum filtration effect, the thing process that can dissolve in the suspension first sieve the stationary phase adsorbent solid reaches the second sieve forms the effluent that contains nucleic acid, flows extremely the centrifuging tube bottom.

2. The sample pretreatment device according to claim 1, characterized in that: and a tube cover is arranged at the top of the centrifugal tube, and when the filter column is arranged in the centrifugal tube, the top opening of the centrifugal tube is sealed through the tube cover.

3. The sample pretreatment device according to claim 2, characterized in that: the tube cap inner wall with centrifuging tube top outer wall all is equipped with the screw thread, the tube cap pass through the screw thread spiro union in centrifuging tube top outer wall.

4. The sample pretreatment device according to claim 1, characterized in that: the top of filtering column is equipped with the flange, the flange overlap joint in the top terminal surface of centrifuging tube, so that filtering column hangs in the centrifuging tube.

5. The sample pretreatment device according to claim 1, characterized in that: the stationary phase adsorbent solid is pre-packaged between the first sieve plate and the second sieve plate and is clamped by the first sieve plate and the second sieve plate.

6. The sample pretreatment device according to claim 1, characterized in that: the centrifuging tube bottom is equipped with the collection portion of back taper, and when the effluent liquid got into collection portion, the filter column by the centrifuging tube takes out.

7. The sample pretreatment device according to claim 1, wherein: the first sieve plate or/and the second sieve plate is/are a micropore sieve plate.

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