CN216908001U - Simple device for extracting nucleic acid from pharyngeal swab or nasopharyngeal swab sample - Google Patents
Simple device for extracting nucleic acid from pharyngeal swab or nasopharyngeal swab sample Download PDFInfo
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- CN216908001U CN216908001U CN202122341236.XU CN202122341236U CN216908001U CN 216908001 U CN216908001 U CN 216908001U CN 202122341236 U CN202122341236 U CN 202122341236U CN 216908001 U CN216908001 U CN 216908001U
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Abstract
The utility model discloses a simple device for extracting nucleic acid from a pharyngeal swab or a nasopharyngeal swab sample, which integrates functions of lysate storage, sample cracking, fixed liquid drainage, impurity removal and the like by expanding one end of an accommodating cavity in a main body to form a cracking chamber and arranging a filter element in a liquid outlet nozzle detachably connected with the main body, and is simple in structure, convenient to operate and quite suitable for on-site nucleic acid extraction; this filter piece is including the activated carbon layer, magnetic bead layer and the filtration rete that stack gradually the connection, the magnetic bead on magnetic bead layer does not adsorb nucleic acid, and various inorganic matters such as macromolecular impurity, colloid, heavy metal in the sample can be got rid of on the activated carbon layer, and the magnetic bead layer can adsorb protein and lipid, and the filtration rete further adsorbs protein and gets rid of, and the three uses in coordination obtains the nucleic acid product that is fit for the low reaches and detects.
Description
Technical Field
The utility model relates to the technical field of biological sample processing, in particular to a simple device for extracting nucleic acid from a pharyngeal swab or a nasopharyngeal swab sample.
Background
The separation and extraction of nucleic acids for effective detection from various specimens/samples is the basis for molecular biological detection, and the method for separating and purifying nucleic acids generally comprises the main steps of cell lysis, separation of nucleic acids from other biomacromolecules, nucleic acid purification and the like, and the cell lysis can be realized by methods of mechanical action, chemical action, enzyme action and the like.
The separation and purification of nucleic acid commonly used in the market at present mainly comprises a centrifugal column method and a magnetic bead method, wherein the centrifugal column method has high extraction efficiency and good effect, but needs repeated centrifugation, and is not beneficial to high-flux and automatic operation; the magnetic bead method uses superparamagnetic silicon oxide nano-microspheres (magnetic beads for short) with specific adsorption on nucleic acid and adsorbs nucleic acid calculation, and then nucleic acid is eluted from the magnetic beads. The two methods are complicated to operate, and need to be matched with more instruments and devices (particularly centrifugation or elution devices, the devices cannot leave a laboratory environment, are not suitable for an outdoor environment, are difficult to deal with emergencies, and are not beneficial to the realization of the instant nucleic acid detection (POCT).
SUMMERY OF THE UTILITY MODEL
The utility model aims to provide a simple device for extracting nucleic acid from a pharyngeal swab or a nasopharyngeal swab sample, which has the advantages of small volume, convenient carrying, simple and convenient operation, no need of a centrifugal machine, elution equipment and the like, and is suitable for outdoor or indoor field nucleic acid instant detection.
In order to achieve the purpose, the utility model adopts the technical scheme that:
a simple device for extracting nucleic acid from a pharyngeal swab or a nasopharyngeal swab sample comprises a main body and a liquid outlet nozzle detachably connected with the main body, wherein one end of the main body is closed, the other end of the main body is open, the open end of the main body is communicated with the inside of the closed end to form an accommodating cavity, and one end, close to the closed end, of the accommodating cavity expands to form a cracking chamber; the liquid outlet nozzle with the connection can be dismantled to the open end, liquid outlet channel has in the liquid outlet nozzle, liquid outlet channel holds and has filtered the piece, filter the piece including the activated carbon layer, magnetic bead layer and the filter membrane layer of range upon range of connection in proper order, the magnetic bead on magnetic bead layer does not adsorb nucleic acid.
Furthermore, the holding cavity can hold a cotton swab, the absorbent cotton end of the cotton swab extends into the cracking chamber, the absorbent cotton end collects a sample to be tested, and cell lysis solution is stored in the cracking chamber.
Furthermore, the cotton swab is the double-end cotton swab, the inner wall of open-ended end has the first fender edge of inside extension, another cotton wool end overlap joint of double-end cotton swab first fender edge, another cotton wool end has not gathered the sample that awaits measuring.
Furthermore, the liquid outlet device also comprises a connecting piece, wherein one end of the connecting piece is detachably connected with the open end, and the other end of the connecting piece is detachably connected with the liquid outlet nozzle.
Further, connecting piece one end is equipped with the external screw thread, and the other end is equipped with the internal thread, the external screw thread with the inner wall threaded connection of liquid outlet nozzle, the internal thread with the outer wall threaded connection of open end.
Further, the connecting piece is including opening end connecting portion and the drain nozzle connecting portion that fuse, opening end connecting portion with the drain nozzle connecting portion are cylindricly, the diameter of opening end connecting portion is greater than the diameter of drain nozzle connecting portion.
Furthermore, the end of the double-end cotton swab, which is not collected with the sample to be detected, is lapped on the inner wall of the liquid outlet nozzle connecting part.
Further, the filter member is tightly connected with the inner wall of the liquid outlet nozzle.
Furthermore, the liquid outlet nozzle comprises a connecting section, a filtering section and a liquid outlet section which are connected into a whole, the liquid outlet section is in a conical shape, the filtering section is in a cylindrical shape, the inner diameter of the filtering section is larger than the maximum inner diameter of the liquid outlet section, a second blocking edge is formed at the joint of the filtering section and the liquid outlet section, one end of the filtering piece is abutted to the second blocking edge, and the filtering piece is tightly connected with the inner wall of the filtering section.
Furthermore, the filter membrane layer is a nitrocellulose membrane layer, and the magnetic beads of the magnetic bead layer are amino magnetic beads.
Compared with the prior art, the utility model has the following beneficial effects:
1. the simple device for extracting nucleic acid from the pharyngeal swab or the nasopharyngeal swab sample provided by the utility model has the advantages that one end of the accommodating cavity in the main body is expanded to form the cracking chamber, and the liquid outlet detachably connected with the main body is internally provided with the filtering piece; this filter piece is including the activated carbon layer, magnetic bead layer and the filtration rete that stack gradually the connection, the magnetic bead on magnetic bead layer does not adsorb nucleic acid, and various inorganic matters such as macromolecular impurity, colloid, heavy metal in the sample can be got rid of on the activated carbon layer, and the magnetic bead layer can adsorb protein and lipid, and the filtration rete further adsorbs protein and gets rid of, and the three uses in coordination obtains the nucleic acid product that is fit for the low reaches and detects.
2. The accommodating cavity of the simple device main body can accommodate a cotton swab, the absorbent cotton end of the cotton swab extends into the cracking chamber, a sample to be detected is collected by the absorbent cotton end, and cell lysis solution is stored in the cracking chamber; therefore, after the sample is collected, the cotton swab can be directly placed in the containing cavity for a cracking reaction, dissolution and transfer are not needed, the speed and the efficiency are high, and sample pollution caused by multiple contact is reduced;
3. the cotton swab capable of being accommodated by the simple device provided by the utility model is a double-head cotton swab, and one absorbent cotton end of the cotton swab is used for collecting a sample to be detected; another absorbent cotton end is arranged in adsorbing some great impurities in the sample solution after cell lysis solution schizolysis, avoids in it gets into the liquid outlet mouth, improves the adsorption efficiency and the adsorption effect of follow-up filtration piece.
Drawings
FIG. 1: the simple device provided by the utility model has a cross section in a use state of sample cell lysis;
FIG. 2: the simple device provided by the utility model is in a cross section under a filtering state;
FIG. 3: fig. 2 is an enlarged schematic view of region a.
In the figure:
1. the device comprises a main body, 11, an accommodating cavity, 111, a cracking chamber, 12, a cotton swab, 121, a first absorbent cotton end, 122 and a second absorbent cotton end;
2. the liquid outlet nozzle 21, the filter piece 211, the activated carbon layer 212, the magnetic bead layer 213, the filter membrane layer 22, the connecting section 23, the filter section 24, the liquid outlet section 25 and the second blocking edge;
3. the connecting piece 31, the open end connecting part 32, the liquid outlet nozzle connecting part 33 and the first blocking edge.
Detailed Description
The utility model provides a new scheme, which is shown more clearly and is described in detail below by combining the accompanying drawings and specific embodiments.
Referring to fig. 1-3, the present invention discloses a simple device for extracting nucleic acid from a pharyngeal swab or a nasopharyngeal swab sample, the device includes a main body 1 and a liquid outlet 2, the main body 1 and the liquid outlet 2 can be connected by a common detachable connection manner such as screwing, clipping, etc. In a particular embodiment the body 1 and the spout 2 are connected by a separate connector 3, in other embodiments the connector 3 may be integral with the body 1 or with the spout 2.
Referring to fig. 1 and 2, one end of the main body 1 is closed, the other end is open, the open end of the main body 1 is communicated with the inside of the closed end to form an accommodating cavity 11, and one end of the accommodating cavity 11 close to the closed end is expanded to form a cracking chamber 111; holding chamber 11 can hold cotton swab 12, and the cotton swab end of cotton swab 12 stretches into lysis chamber 111, the cotton swab end is gathered there is the sample that awaits measuring, the sample that awaits measuring can be for nose swab sample, pharynx swab sample or nasopharynx swab sample, and the cell lysis liquid has been stored in lysis chamber 111. The cotton swab 12 may be a single-head cotton swab or a double-head cotton swab, in a specific embodiment, the cotton swab 12 is a double-head cotton swab, the first cotton swab end 121 collects a sample to be tested, the first cotton swab end 121 extends into or is accommodated in the lysis chamber 111, and the second cotton swab end 122 is configured to adsorb some large impurities (e.g., nasal swabs, pharyngeal swabs, or other nasal cavities or other secretions or other insoluble particle foreign matters adhered to the sample solution after cell lysate lysis, so as to prevent the large impurities from entering the liquid outlet 2, and improve the adsorption efficiency and the adsorption effect of the subsequent filter 21.
In the above-described embodiment, the main body 1 is connected to the spout 2 by a separate connecting member 3, one end of the connecting member 3 is detachably connected to the open end, and the other end is detachably connected to the spout 2. Specifically, referring to fig. 2 and 3, one end of the connecting member 3 is provided with an external thread (not labeled in the figures), and the other end is provided with an internal thread (not shown in the figures), the external thread is in threaded connection with the inner wall of the liquid outlet nozzle 2, and the internal thread is in threaded connection with the outer wall of the open end. The opening end of the main body 1 is connected with the liquid outlet 2 through the two threads of the connecting piece 3, so that the disassembly is convenient, the sealing performance is good, and liquid leakage is not easy to occur. In other embodiments, the connecting member 3 may also be connected to the main body 1 and the liquid outlet 2 by other common methods such as clamping, as long as the effects of convenient detachment and liquid leakage prevention can be achieved.
Specifically, referring to fig. 2, the connecting member 3 includes an open-end connecting portion 31 and a liquid outlet nozzle connecting portion 32 connected integrally, the open-end connecting portion 31 and the liquid outlet nozzle connecting portion 32 are both cylindrical, and the diameter of the open-end connecting portion 31 is larger than that of the liquid outlet nozzle connecting portion 32; the end (i.e. the second absorbent cotton end 122) of the double-ended cotton swab, which is not collected with or is not collected with a nasopharyngeal swab sample, is lapped on the inner wall of the liquid outlet nozzle connecting part 32. Since the diameter of the opening-end connecting part 31 is larger than that of the liquid outlet nozzle connecting part 32, a blocking edge is formed at the joint of the opening-end connecting part and the liquid outlet nozzle connecting part, the blocking edge is a first blocking edge 33, and the first blocking edge 33 extends towards the liquid outlet nozzle connecting part 32 to form the inner wall of the liquid outlet nozzle connecting part 32. The second absorbent cotton end 122 is overlapped on the first blocking edge 33, so that the sample solution after the cell lysate is cracked can be prevented from directly flowing into the liquid outlet nozzle 2, and an initial filtering effect is provided. In other embodiments, the first blocking edge 33 may be formed by extending the open end inward.
The liquid outlet nozzle 2 is used for discharging the extracted nucleic acid, a liquid outlet channel is arranged in the liquid outlet nozzle 2, and a filter piece 21 is arranged in the liquid outlet channel. Referring to fig. 2 and 3, the filter member 21 includes an activated carbon layer 211, a magnetic bead layer 212, and a filter membrane layer 213, which are sequentially stacked and connected, the magnetic bead of the magnetic bead layer 212 does not adsorb nucleic acid, the activated carbon layer 211 can remove macromolecular impurities, colloids, heavy metals and other inorganic substances in the sample, the magnetic bead layer 212 can adsorb protein and lipids without adsorbing nucleic acid, the filter membrane layer 213 further adsorbs and removes protein, the three are used in cooperation to obtain a nucleic acid product suitable for downstream detection, and the nucleic acid product is matched with the second absorbent cotton end 122, so that the efficiency is higher, and the effect is better. Specifically, the filtration membrane layer 213 may be one or more layers of nitrocellulose membrane, and the magnetic bead layer 212 may be one or more layers of amino magnetic beads (PuriMag G-NH)2) Is of the formula-NH2Superparamagnetic microspheres of surface functional groups, from Fe3O4Core and GMA coating for adsorbing protein and lipidThe surface thereof. The activated carbon layer 211 may be formed by laying activated carbon on the amino magnetic bead layer, and the filter element 21 with a three-layer structure may be manufactured by a common pressing method, and may be formed by pressing two layers first, and then laying activated carbon to press the last layer.
The liquid outlet nozzle 2 can comprise a connecting section 22, a filtering section 23 and a liquid outlet section 24 which are connected into a whole, the liquid outlet section 24 is conical, the filtering section 23 is cylindrical, the inner diameter of the filtering section 23 is larger than the maximum inner diameter of the liquid outlet section 24, a second blocking edge 25 is formed at the joint of the filtering section 23 and the liquid outlet section 24, one end (one end of a filtering film layer) of the filtering piece 21 is abutted against the second blocking edge 25, and the filtering piece 21 and the inner wall of the filtering section 23 are preferably tightly connected so as to prevent unfiltered solution from flowing out of gaps. The outer part of the liquid outlet nozzle 2 can be detachably connected with a protective cap.
In the embodiment of the present invention, the first blocking edge 33 and the second blocking edge 25 are disposed to respectively fix the second absorbent cotton end 122 and the filtering member 21 at corresponding positions, so as to prevent the second absorbent cotton end and the filtering member from moving easily, thereby ensuring the absorption effect. The cylindrical design of the connecting piece 3 is matched with the short open cylinder and the conical shape of the liquid outlet nozzle, so that the device has a more regular, compact and beautiful appearance. The length of the main body of the utility model is adapted to the length of the cotton swab, the closed end is expanded to form a cracking chamber, not only can lysate be stored to provide a reaction site for sample cell cracking, but also the first absorbent cotton end of the cotton swab can be accommodated, and the air in the cracking chamber can be squeezed, so that convenient and accurate liquid drainage can be realized.
In the utility model, the main body 1, the liquid outlet 2 and the connecting piece 3 can be transparent or semitransparent, so that the observation is convenient. The cracking chamber 111 of the main body 1 may be made of plastic or rubber, etc. having a property of being deformable by extrusion, so as to facilitate the extrusion of air.
In the original sample cell cracking process of the nucleic acid to be detected, inhibitors such as package impurities and adsorption enzymes can be released, downstream molecular detection is influenced, amplification reaction is inhibited, Bst DNA polymerase is used in LAMP reaction, and the tolerance of the enzyme is good, so that the requirement on a template during LAMP amplification is low, the requirement on the purification degree of the nucleic acid of the sample to be detected is lower than that of other detection methods, and the nucleic acid sample which is roughly extracted can participate in the LAMP reaction. The simple device provided by the utility model is very suitable for being used together with LAMP. The simple device can be used for field detection and field sampling field detection (can be matched with the existing small portable LAMP instruments in the market), and the problems that in methods such as PCR (polymerase chain reaction) and the like, the detection result caused by degradation of samples in transportation (RNA viruses are easily degraded in the transportation process and the extraction process) is not accurate enough, time and labor are consumed and the like are solved.
The use method of the utility model can be as follows:
(1) obtaining a double-head cotton swab with a sample to be detected (such as a nasal swab or a pharyngeal swab or a nasopharyngeal swab sample) at one end for collection (field immediate collection can be realized, and a sample stored after centralized collection can also be sucked or stained);
(2) referring to fig. 1, initially, the closed end of the main body 1 of the simple device is located below, and the liquid outlet 2 is located above; a user opens the simple device through the connecting piece 3, stretches the first absorbent cotton end 121 of the double-end cotton swab into the cracking chamber 111 storing cell lysis solution to react (the cell lysis solution and a sample can be fully mixed by shaking or shaking), and then connects (locks) the main body 1 and the liquid outlet nozzle 2 through the connecting piece 3 to prevent subsequent leakage;
(3) referring to fig. 2, the device after the cleavage reaction in step (2) is turned upside down, the liquid outlet 2 is located below the liquid outlet 1, the main body 1 is located above the liquid outlet, the reacted sample solution passes through the second absorbent cotton end 122 of the double-ended cotton swab, the activated carbon layer 211 of the filter element 21, the magnetic bead layer 212 and the filter membrane layer 213 in sequence, impurities, insoluble particles and the like in the sample and the lysate are filtered to reach the liquid outlet 2, the lysis chamber 111 is slightly squeezed from the outside, and the filtered nucleic acid extract is conveniently and accurately discharged from the liquid outlet 2 for subsequent detection.
Although the present invention has been described in detail in the foregoing embodiments, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the spirit and scope of the utility model.
Claims (10)
1. The utility model provides a simple and easy device that is used for nasopharyngeal swab or nasopharyngeal swab sample nucleic acid to draw, includes the main part and with the liquid outlet mouth of main part detachable connection, its characterized in that: one end of the main body is closed, the other end of the main body is open, the open end of the main body is communicated with the inside of the closed end to form an accommodating cavity, and one end of the accommodating cavity, which is close to the closed end, is expanded to form a cracking chamber; the liquid outlet nozzle with the connection can be dismantled to the open end, liquid outlet channel has in the liquid outlet nozzle, liquid outlet channel holds and has filtered the piece, filter the piece including the activated carbon layer, magnetic bead layer and the filter membrane layer of range upon range of connection in proper order, the magnetic bead on magnetic bead layer does not adsorb nucleic acid.
2. The simplified device for extraction of nucleic acid from pharyngeal swab or nasopharyngeal swab samples as recited in claim 1, wherein: the holding chamber can hold the cotton swab, the cotton wool end of cotton swab stretches into the lysis chamber, the cotton wool end is gathered there is the sample that awaits measuring, the indoor storage of lysis has the cell lysis liquid.
3. The simplified device for extraction of nucleic acid from pharyngeal swab or nasopharyngeal swab samples as recited in claim 2, wherein: the cotton swab is the double-end cotton swab, the inner wall of open-ended end has the first fender edge of inside extension, another cotton wool end overlap joint of double-end cotton swab first fender edge, another cotton wool end has not gathered the sample that awaits measuring.
4. The simplified device for extraction of nucleic acid from pharyngeal swab or nasopharyngeal swab samples as recited in claim 3, wherein: still include the connecting piece, the one end detachable of connecting piece is connected open end, other end detachable connects the drain nozzle.
5. The simplified device for pharyngeal swab or nasopharyngeal swab sample nucleic acid extraction as recited in claim 4, wherein: connecting piece one end is equipped with the external screw thread, and the other end is equipped with the internal thread, the external screw thread with the inner wall threaded connection of liquid outlet nozzle, the internal thread with the outer wall threaded connection of open end.
6. The simplified device for nasopharyngeal swab or nasopharyngeal swab sample nucleic acid extraction as recited in claim 4 or 5, wherein: the connecting piece includes integrative opening end connecting portion and liquid outlet nozzle connecting portion even, opening end connecting portion with liquid outlet nozzle connecting portion are cylindricly, the diameter of opening end connecting portion is greater than the diameter of liquid outlet nozzle connecting portion.
7. The simplified device for extraction of nucleic acid from pharyngeal swab or nasopharyngeal swab samples as recited in claim 6, wherein: the double-end cotton swab is not connected with the inner wall of the liquid outlet nozzle connecting part in a lap joint mode, wherein the end, not collected with a sample to be detected, of the double-end cotton swab is connected with the inner wall of the liquid outlet nozzle connecting part in a lap joint mode.
8. The simplified device for extraction of nucleic acid from pharyngeal swab or nasopharyngeal swab samples as recited in claim 1, wherein: the filtering piece is tightly connected with the inner wall of the liquid outlet nozzle.
9. The simplified apparatus for pharyngeal swab or nasopharyngeal swab sample nucleic acid extraction as recited in claim 8, wherein: the liquid outlet nozzle comprises a connecting section, a filtering section and a liquid outlet section which are connected into a whole, the liquid outlet section is conical, the filtering section is cylindrical, the inner diameter of the filtering section is larger than the maximum inner diameter of the liquid outlet section, the filtering section and the joint of the liquid outlet section form a second blocking edge, one end of the filtering piece is abutted against the second blocking edge, and the filtering piece is tightly connected with the inner wall of the filtering section.
10. The simplified device for nucleic acid extraction from pharyngeal swab or nasopharyngeal swab samples according to claim 1, wherein: the filter membrane layer is a nitrocellulose membrane layer, and the magnetic beads of the magnetic bead layer are amino magnetic beads.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202122341236.XU CN216908001U (en) | 2021-09-26 | 2021-09-26 | Simple device for extracting nucleic acid from pharyngeal swab or nasopharyngeal swab sample |
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| CN202122341236.XU CN216908001U (en) | 2021-09-26 | 2021-09-26 | Simple device for extracting nucleic acid from pharyngeal swab or nasopharyngeal swab sample |
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| CN216908001U true CN216908001U (en) | 2022-07-08 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114045207A (en) * | 2021-09-26 | 2022-02-15 | 厦门健康工程与创新研究院 | Nucleic acid extraction and detection system and method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114045207A (en) * | 2021-09-26 | 2022-02-15 | 厦门健康工程与创新研究院 | Nucleic acid extraction and detection system and method |
| CN114045207B (en) * | 2021-09-26 | 2024-02-06 | 厦门健康工程与创新研究院 | Nucleic acid extraction detection system and method |
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