CN217103880U - Reaction device for rapid nucleic acid detection - Google Patents

Abstract

The utility model discloses a reaction unit for quick nucleic acid detects aims at overcoming and lacks an instrument that can satisfy sampling, nucleic acid extraction and nucleic acid amplification detection function among the prior art. The device comprises a reaction container and a sample collector, wherein the reaction container is provided with a nucleic acid extraction area and a nucleic acid amplification detection area, the sample collector comprises a sampling head and a piston, and the piston is slidably arranged in the nucleic acid extraction area so as to inject substances in the nucleic acid extraction area into the nucleic acid amplification detection area.

Description

Reaction device for rapid nucleic acid detection

Technical Field

The utility model belongs to a biochemical or molecular biology device, in particular to a reaction device for rapid nucleic acid detection.

Background

Currently, methods for diagnosing microbial infections include the steps of: (1) collecting a sample from a patient; (2) extracting nucleic acids from a patient sample; (3) taking the extracted nucleic acid as a template to carry out amplification reaction; and (4) carrying out sequence analysis or gene structure function analysis on the nucleic acid generated by the amplification reaction.

For example, a method for detecting and diagnosing whether a patient is infected with a novel coronavirus includes the steps of: (1) collecting a saliva sample of a patient by using a cotton swab; (2) inserting the cotton swab into a test tube with a virus preservation reagent, shaking up and down for several times to ensure that the preservation reagent and the saliva sample are fully mixed, and then packaging, disinfecting, inactivating and preserving the test tube; (3) in a clean laboratory, a saliva sample in a test tube is dripped into the test tube with the new coronavirus lysate to carry out a lysis reaction, so that a series of RNAs are obtained; (4) adding the RNA into a test tube containing a new coronavirus amplification solution, carrying out amplification reaction, and carrying out specificity detection on the RNA in the test tube by using a fluorescence detection device so as to determine whether the patient is infected with the new coronavirus.

However, the prior art is complicated in process operation, the steps of the method for diagnosing microbial infection need to be completed step by step on a plurality of devices, if a plurality of samples need to be detected, even tens of thousands of samples need to be detected, a large amount of manpower and material resources are needed to share the work of each step, the cost and the detection period are undoubtedly increased, and complicated, single and tedious steps easily cause operation errors. Although the above steps can be completed completely by fully automatic equipment, the high price makes it impossible to popularize them all-round.

For colloidal gold test strip products, early pregnancy detection can be said to be an outstanding representative of colloidal gold chromatography, and various advantages of chromatography detection are reflected to the full extent. And for infectious disease detection, drug screening and myocardial infarction detection, including recently attracting new coronavirus detection, the method shows the distinctive characteristics of the detection without exception, is simple and convenient to operate, has a quick result, and is suitable for field detection operation. Particularly, in the global range, as Q-PCR devices and operators cannot be popularized, colloidal gold or molecular diagnosis POCT products become mainstream test products, and even a plurality of rapid domestic detection products of new coronavirus are realized.

Therefore, the method has important practical significance for developing a more convenient rapid nucleic acid detection mode.

SUMMERY OF THE UTILITY MODEL

In order to overcome the problems of the prior art, the utility model provides a reaction device for rapid nucleic acid detection.

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

a reaction device for rapid nucleic acid detection comprises a reaction container and a sample collector, wherein the reaction container is provided with a nucleic acid extraction area and a nucleic acid amplification area, the sample collector comprises a sampling head and a piston, and the piston is slidably arranged in the nucleic acid extraction area so as to inject substances in the nucleic acid extraction area into the nucleic acid amplification area.

In some embodiments, the sample collector has a first position on the reaction vessel, and if the sample collector is disposed in the first position, the sampling head is located in the nucleic acid extraction region, and the nucleic acid extraction region and the nucleic acid amplification region are not in communication.

In some embodiments, the plunger seals the nucleic acid extraction region when the sample collector is disposed in the first position.

In some embodiments, the sample collector has a second position on the reaction vessel, and the piston is slidably disposed within the nucleic acid extraction region if the sample collector is disposed in the second position, the nucleic acid extraction region being in communication with the nucleic acid amplification region.

In some embodiments, the plunger seals the nucleic acid extraction region when the sample collector is disposed in the second position.

In some embodiments, the reaction vessel has a pierceable separator for separating the nucleic acid extraction zone and the nucleic acid amplification zone.

In some forms, the separator is a film or a thin layer.

In some forms, the sample collector is used to puncture a separator.

In some embodiments, the kit further comprises a piercing member for piercing the reaction vessel to communicate the nucleic acid extraction region and the nucleic acid amplification region.

In some approaches, the sample collector may push a piercing member to pierce the reaction vessel.

In some forms, the piercing member is removably disposed on the sample collector.

In some modes, a placement groove is formed on the puncture piece, and a sample collector is inserted into the placement groove.

In some embodiments, the piercing member has a channel formed thereon for communicating the nucleic acid extraction region and the nucleic acid amplification region.

In some forms, the piercing member has a sharp point formed thereon.

In some embodiments, the reaction vessel comprises an extraction tube and an amplification tube, the nucleic acid extraction zone is disposed within the extraction tube, and the nucleic acid amplification zone is disposed within the amplification tube.

In some aspects, the extraction tube includes a tube body and a separator disposed at one end of the tube body.

In some embodiments, the extraction tube is removably disposed on the amplification tube.

In some embodiments, the extraction tube is inserted within the nucleic acid amplification zone.

In some modes, be formed with the recess on the extraction test tube, be formed with the arch on the amplification test tube, the protruding joint is in the recess.

In some forms, the extraction tube has a bead formed thereon and the groove is formed on the bead.

In some embodiments, the reaction vessel includes a light-permeable plate portion for being disposed between the nucleic acid amplification region and the outside.

In some embodiments, the number of the plate portions is at least three, and three or more plate portions surround a regular polygonal region, and the nucleic acid amplification region is disposed in the regular polygonal region.

In some aspects, the sample collector comprises a rod, and the sampling head and the piston are respectively disposed on the rod.

In some embodiments, the sample collector comprises an end cap configured to cover the nucleic acid extraction zone or configured to cover the extraction tube.

In some forms, the end cap, the piston, and the deployment head are disposed in sequence on the stem.

In some embodiments, a lysis solution can be disposed in the nucleic acid extraction zone, and an amplification reagent and a fluorescence reagent can be disposed in the nucleic acid amplification zone.

Compared with the prior art, the utility model has the advantages that:

(1) the utility model discloses in, the sample collector is used for the sampling, and nucleic acid extraction district is used for carrying out the nucleic acid extraction reaction, and the nucleic acid amplification district is used for carrying out nucleic acid amplification detection reaction, and consequently this a reaction unit for quick nucleic acid detection has the function that sampling, nucleic acid extraction, nucleic acid amplification detection reaction realized concurrently, has simplified the procedure that detects and the equipment of adopting, has reduced medical personnel's working strength.

(2) The utility model discloses in, thereby the piston is used for the slip to set up in nucleic acid extraction district can with the material injection nucleic acid amplification district in the nucleic acid extraction district, insert the nucleic acid extraction district after the sample collector sampling and carry out the nucleic acid extraction reaction earlier in, promote the piston again, inject the material in the nucleic acid extraction district into the nucleic acid amplification district and carry out the nucleic acid amplification detection reaction, consequently sampling, nucleic acid extraction and nucleic acid amplification can go on in order, have guaranteed the stability and the accuracy of detection.

(3) The utility model discloses in, the piston is used for the shutoff in nucleic acid extraction district, nucleic acid extraction reaction is accomplished in sealed nucleic acid extraction district, nucleic acid amplification district is also sealed, nucleic acid amplification detection reaction is accomplished in sealed nucleic acid amplification district, on the one hand, the operating procedure can be reduced, greatly made things convenient for the operator, on the other hand also effectively reduces the oxidation or the volatilization of the reactant in the reaction vessel, consequently avoided the external environment to the interference of detection and this a reaction unit for quick nucleic acid detects the pollution that probably causes, especially can prevent the environmental aerosol pollution that the molecular biology field is difficult to solve.

Drawings

Fig. 1 is a schematic perspective view of a first embodiment of the present invention;

fig. 2 is a schematic cross-sectional structure diagram of a sample collector arranged at a first position according to a first embodiment of the present invention;

fig. 3 is a schematic cross-sectional structure diagram of a sample collector disposed at a second position according to a first embodiment of the present invention;

FIG. 4 is a schematic cross-sectional view of a puncturing member according to an embodiment of the present invention;

FIG. 5 is a schematic cross-sectional view of an extraction tube according to a first embodiment of the present invention;

FIG. 6 is a schematic cross-sectional view of a pierced extraction tube according to a first embodiment of the present invention;

fig. 7 is a schematic diagram of an explosion structure according to a first embodiment of the present invention;

fig. 8 is a schematic perspective view of a second embodiment of the present invention;

fig. 9 is a schematic sectional view of a second embodiment of the present invention;

fig. 10 is a schematic sectional view of a puncturing member according to a second embodiment of the present invention;

fig. 11 is a schematic cross-sectional view of a sample collector of the second embodiment of the present invention disposed at a first position;

fig. 12 is a schematic cross-sectional view of a sample collector of the second embodiment of the present invention disposed at a second position;

in the figure: 1-extraction test tube, 2-amplification test tube, 3-sample collector, 4-piercing part, 11-nucleic acid extraction area, 12-tube, 13-separator, 21-nucleic acid amplification area, 22-bulge, 23-flat plate, 31-rod, 32-sampling head, 33-end cap, 34-piston, 41-tip, 42-diversion groove, 43-placement groove, 121-crimp, 122-groove, 123-slot, 124-countersunk groove, 131-hole, 311-breaking groove and 312-positioning groove.

Detailed Description

To facilitate understanding of those skilled in the art, the present invention will be further described with reference to the accompanying drawings and specific embodiments.

As shown in fig. 1 to 12, a reaction apparatus for rapid nucleic acid detection includes a reaction vessel having a nucleic acid extraction region 11 and a nucleic acid amplification region 21, and a sample collector 3 including a sampling head 32 and a piston 34, the piston 34 being slidably disposed in the nucleic acid extraction region 11 so as to inject substances in the nucleic acid extraction region 11 into the nucleic acid amplification region 21.

Reaction vessel refers to a vessel device used to carry out a reaction of a substance. The substance reaction includes an extraction reaction of nucleic acid and an amplification reaction of nucleic acid. The nucleic acid extraction reaction refers to a reaction for extracting nucleic acid in a substance, and can be realized by at least one of a paramagnetic particle method, an alkaline lysis method, a freeze-thaw lysis method, an enzyme lysis method, a chromatographic column method and an alcohol precipitation method. The nucleic acid amplification detection reaction refers to a reaction for amplifying a nucleic acid sequence to be detected in nucleic acid and a fluorescence detection for the amplified nucleic acid sequence, the amplification reaction can be realized by adopting an isothermal nucleic acid amplification reaction, and the fluorescence detection can be realized by Crispr fluorescence cutting signal detection based on Cas enzyme. The substance may be a sample, a reaction product, a lysate, a stabilizer, or the like.

The reaction vessel has a nucleic acid extraction region 11 and a nucleic acid amplification region 21. The nucleic acid extraction region 11 refers to a chamber for performing a nucleic acid extraction reaction. The nucleic acid amplification region 21 refers to a chamber for performing a nucleic acid amplification reaction. The nucleic acid extraction region 11 and the nucleic acid amplification region 21 are disposed at different positions of the reaction vessel, respectively, so that the nucleic acid extraction reaction and the nucleic acid amplification detection reaction can be separately performed at different positions.

The sample collector 3 refers to a device for collecting a sample. The sample can be oral flora, nasal flora, throat flora, cervical flora, etc., or human or animal tissue, blood, secretion, etc., or plant leaf and rhizome tissue, etc., or microorganism on object surface. In practical use, the sample collector 3 can collect samples at the mouth, nose, throat, cervix and other positions of the patient.

The piston 34 refers to a mechanism that can slide reciprocally in the nucleic acid extracting region 11. The substance in the nucleic acid extraction region 11 may be a product of a nucleic acid extraction reaction. If the piston 34 is slidably disposed in the nucleic acid extraction zone 11, the piston 34 is blocked in the nucleic acid extraction zone 11. When the piston 34 slides within the nucleic acid extracting region 11, the interior of the nucleic acid extracting region 11 is compressed, increasing the pressure within the nucleic acid extracting region 11, so that the substance within the nucleic acid extracting region 11 can be injected into the nucleic acid amplification region 21 by the pressure. Additionally, the sampling head 32 is used to collect samples.

The sample sampler 3 has a first position on the reaction vessel, and the sampling head 32 is located in the nucleic acid extraction zone 11 if the sample sampler 3 is disposed in the first position.

The first position refers to a position where the sample sampler 3 performs the nucleic acid extraction reaction 11 on the reaction vessel. When the sampling head 32 is located in the nucleic acid extraction region 11, the sample on the sampling head 32 is disposed in the nucleic acid extraction region 11 so that the sample can undergo a nucleic acid extraction reaction in the nucleic acid extraction region 11.

When the sample sampler 3 is disposed at the first position, the nucleic acid extracting region 11 and the nucleic acid amplifying region 21 are not communicated.

The nucleic acid extraction region 11 and the nucleic acid amplification region 21 are not communicated, so that substances in the nucleic acid extraction region 11 are prevented from entering the nucleic acid amplification region 21, substances in the nucleic acid amplification region 21 are prevented from entering the nucleic acid extraction region 11, and therefore only nucleic acid extraction reaction can be carried out on the reaction vessel, and nucleic acid amplification detection reaction is prevented.

In addition, the piston 34 may seal the nucleic acid extraction zone 11 if the sample collector 3 is disposed in the first position.

The piston 34 seals the nucleic acid extracting region 11, and the sealed nucleic acid extracting region 11 constitutes a chamber which is not in contact with the outside. On one hand, dust and foreign matters in the outside are prevented from entering the nucleic acid extraction area 11, interference of the dust, the foreign matters and the like on the nucleic acid extraction reaction is avoided, operation steps are reduced, and great convenience is brought to an operator; on the other hand, substances in the nucleic acid extraction region 11 are prevented from leaving the nucleic acid extraction region 11 by means of weathering, volatilization or evaporation, and the like, thereby avoiding aerosol pollution of the reactant or the product to the environment.

The sample collector 3 has a second position on the reaction vessel, and if the sample collector 3 is disposed at the second position, the nucleic acid extracting region 11 and the nucleic acid amplifying region 21 are in communication.

The second position refers to a position where the sample collector 3 performs a nucleic acid amplification detection reaction in the reaction vessel. When the sample extraction region 11 and the nucleic acid amplification region 21 are in communication, the substance in the nucleic acid extraction region 11 can enter the nucleic acid amplification region 21, thereby facilitating the substance to perform a nucleic acid amplification detection reaction in the nucleic acid amplification region 21.

When the sample collector 3 is disposed at the second position, the piston 34 is also slidably disposed in the nucleic acid extraction region 11 so that the substance in the nucleic acid extraction region 11 can be injected into the nucleic acid amplification region 21.

If the piston 34 is slidably disposed in the nucleic acid extracting region 11, the piston 34 can push the substance in the nucleic acid extracting region 11 to inject into the nucleic acid amplification region 21, thereby promoting the substance to rapidly enter into the nucleic acid amplification region 21 and increasing the reaction efficiency. The reaction vessel includes an extraction tube 1 and an amplification tube 2, a nucleic acid extraction region 11 is provided in the extraction tube 1, and a nucleic acid amplification region 21 is provided in the amplification tube 2.

The nucleic acid extraction region 11 has an overall structure in which a chamber having an opening at one end is formed. The piston 34 is fitted in the nucleic acid extracting region 11, and the cross section of the piston 34 is circular, and the cross section of the nucleic acid extracting region 11 is also circular. If the piston 34 slides within the nucleic acid extraction zone 11, the piston 34 can slide between the opening of the nucleic acid extraction zone 11 and the bottom within the nucleic acid extraction zone 11. The nucleic acid amplification region 21 is also constructed in an overall structure having a chamber with an opening at one end, and the extraction tube 1 is sealed at the opening of the amplification tube 2, thereby sealing the nucleic acid amplification region 21. The extraction test tube 1 and the amplification test tube 2 are both made of plastic, and have the advantages of inactive chemical properties and low cost.

Specifically, the extraction test tube 1 is composed of a tube body 12 and a partition 13, the entire structure of the tube body 12 is tubular, the partition 13 is provided at one end of the tube body 12, and the other end of the tube body 13 is open. The tube 12 and the partition 13 enclose the nucleic acid extraction region 11.

When the extraction tube 1 is mounted on the amplification tube 2, the extraction tube 1 is blocked on the nucleic acid amplification region 21. At this time, if the sample collector 3 is set at the second position, the piston 34 is also blocked on the nucleic acid extracting region 11, so that the nucleic acid amplification region 21 is also sealed, and the nucleic acid amplification region 21 forms a chamber which is not in contact with the outside, thereby avoiding external interference and aerosol pollution.

The extraction test tube 1 is inserted in the nucleic acid amplification zone 21. Specifically, the tube body 12 is inserted in the nucleic acid amplification region 21.

The extraction test tube 1 and the amplification test tube 2 are connected together in a splicing mode, so that the connection firmness and the sealing performance of the extraction test tube 1 and the amplification test tube 2 are improved, and the compactness of the structure is also improved. The overall structure of the extraction test tube 1 is barrel-shaped, and the overall structure of the amplification test tube 2 is also barrel-shaped, so that the extraction test tube 1 is adapted in the amplification test tube 2.

The extraction test tube 1 can be detachably arranged on the amplification test tube 2, and the extraction test tube 1 and the amplification test tube 2 are matched to form a reaction container, so that the reaction container with the nucleic acid extraction area 11 and the nucleic acid amplification area 21 can be conveniently produced and manufactured, and substances such as lysate, amplification reaction reagents and the like can be conveniently added into the nucleic acid extraction area 11 and the nucleic acid amplification area 21 in the follow-up process.

Specifically, the extraction tube 1 is formed with a groove 122, the amplification tube 2 is formed with a protrusion 22, and the protrusion 22 is engaged with the groove 122. Connection structure retrencies, and the bare-handed test tube that will draw of convenient to use person is assembled together with amplification test tube 2, has also further improved the firm in connection nature and the leakproofness of drawing test tube 1 and amplification test tube 2. The groove 122 is formed in a ring shape, the groove 122 surrounds the tube body 12, the protrusion 22 is formed in a ring shape, and the protrusion 22 surrounds the amplification cuvette 2.

The extraction tube 1 is formed with a bead 121 and a groove 122 is formed on the bead 121. Due to the thin wall thickness of the bead 121, the bead 121 may be elastically deformed to some extent when the bead is subjected to a force, thereby facilitating the protrusion 22 to be caught in the groove 122 or to be separated from the groove 122.

Specifically, the bead 121 is provided at one end of the tube body 12, and the separator 13 is provided at the other end of the tube body 12.

The bead 121 and the tube body enclose a slot 123, the groove 122 is disposed in the slot 123, and the amplification test tube 2 is further inserted in the slot 123. The overall structure of slot 123 is the annular, and the outer end and the slot 123 looks adaptation of amplification test tube 2, the outer end of amplification test tube 2 are inserted and are established in slot 123, and with draw test tube 1 and insert the structure of establishing in nucleic acid amplification district 21 and constitute the effect of dual connection, further improved the fastness of connection and the leakproofness of drawing test tube 1 and amplification test tube 2.

The amplification tube 2 is transparent. When the fluorescence detection is carried out on the reaction container, the fluorescent reagent with the reaction color is preset in the amplification test tube 2, the fluorescent reagent can be irradiated outwards through the amplification test tube 2, the fluorescence signal can be received by adopting a spectrum detection device or the amplification test tube 2 can be directly observed by naked eyes, and the fluorescence analysis is carried out on the nucleic acid, so that whether the patient suffers from the disease or not is determined. The extraction tube 1 is also transparent. The fluorescent reagent may comprise a fluorescent probe. The spectrum detection device can be any one of a fluorescence analyzer, a constant-temperature fluorescence nucleic acid amplification device, an immunofluorescence analysis device, an ultraviolet analyzer, a full-wavelength spectrum scanner and the like.

The amplification tube 2 is transparent. For example, in the case of performing visible light detection on a reaction vessel, a fluorescent reagent having a reaction color is preliminarily set in the amplification tube 2, and after completion of the reaction, it is directly observed with the naked eye through a color change after the reaction in the amplification tube 2, thereby determining whether or not a specific nucleic acid substance is present in the reaction.

The amplification test tube 2 comprises a light-permeable flat plate part 23, and the flat plate part 23 is arranged between the nucleic acid amplification area 21 and the outside. The flat plate portion 23 has a flat plate body in its entire structure. The flat plate portion 23 is formed on the amplification test tube 2. The fluorescent group in the nucleic acid amplification region 21 can be irradiated outwards through the flat plate part 23, and the light-emitting angle of the fluorescent signal is not changed when the fluorescent signal leaves the flat plate part 23, so that the problem that the signal acquired by the amplification test tube 2 is inaccurate due to the change of the light-emitting angle of the fluorescent signal is solved.

The number of the flat plate portions 23 is at least three, and three or more flat plate portions 23 surround a regular polygonal region in which the nucleic acid amplification region 21 is provided. If the fluorescent group in the nucleic acid amplification region 21 emits light, fluorescent signals can be collected at each position in the circumferential direction of the amplification test tube 2, so that the fluorescent signals can be collected at a plurality of positions, and the signals can be collected conveniently.

The partition 13 is pierceable, and the partition 13 is used for separating the nucleic acid extraction region 11 and the nucleic acid amplification region 21. The partition 13 may be punctured directly by the sample collector 3 or other means may be used to puncture the partition 13. The other component may be a piercing member 4.

The separating element 13 is a flexible film which is as easy to pierce as possible by the piercing element 4 while ensuring the separating effect. A partition 13 is welded to the tube 12, and the partition 13 is provided at the bottom in the nucleic acid extraction region 11.

In this embodiment, a puncture means 4 is further included, and the puncture means 4 is used for puncturing the reaction vessel to communicate the nucleic acid extraction region 11 with the nucleic acid amplification region 21. Specifically, the puncturing member 4 is used to puncture the partition member 13 of the extraction test tube 1, and when the puncturing member 4 punctures the reaction vessel, a hole 131 is formed in the partition member 13, and the hole 131 communicates between the nucleic acid extraction region 11 and the nucleic acid amplification region 21.

In the process of pushing down the sample collector 3, the liquid in the nucleic acid extraction region 11 can be quantitatively injected into the amplification test tube 2 by controlling the sliding stroke of the sample collector 3 on the reaction vessel. For example, by observing the distance between the end cap 33 and the extraction tube 2, the amount of liquid injected can be controlled.

Wherein the partition 13 serves to partition the nucleic acid extraction region 11 and the nucleic acid amplification region 21 such that the nucleic acid extraction reaction is completed solely within the nucleic acid extraction region 11 and the nucleic acid amplification detection reaction is completed solely within the nucleic acid amplification region 21. When the puncture member 4 punctures the partition member 13, a hole 131 is formed in the partition member 13, the hole 131 is used to communicate between the nucleic acid extracting region 11 and the nucleic acid amplification region 21, and the substance in the nucleic acid extracting region 11 can be injected into the nucleic acid amplification region 21 through the hole 131. In practical use, if the sample on the sample collector 3 completes the nucleic acid extraction reaction in the nucleic acid extraction region 11, the puncturing part 4 is used to puncture the separating part 13 and push the piston 34 to slide in the nucleic acid extraction region 11, and the substance in the nucleic acid extraction region 11 is injected into the nucleic acid amplification region 21 to perform the nucleic acid amplification detection reaction, so that the nucleic acid extraction reaction and the nucleic acid amplification detection reaction can be performed in order.

The piercing member 4 is detachably provided on the sample collector 3, so that the sample collector 3 can be used alone or can be assembled with the piercing member 4 for use. If when using sample collection ware 3 alone, can adopt sample collection ware 3 to sample the patient, avoid puncture piece 4 accidental injury patient. When the sample collector 3 and the puncture piece 4 are assembled together for use, the puncture piece 4 can puncture the partition 13 and push the piston 34 to slide in the nucleic acid extracting region 11 by pushing the sample collector 3.

The puncture piece 4 is formed with a placement groove 43, and the placement groove 43 is used for inserting the sample collector 3. The piercing member 4 and the sample collector 3 are connected together in a plug-in manner. The user need not with the help of the device, bare-handed installation.

Specifically, the first position actually refers to a position where the puncture element 4 with the sample collector 3 mounted thereon is located within the nucleic acid extraction zone, at which time the puncture element 4 does not puncture the partition 13, and the sampling head 32 is disposed within the nucleic acid extraction zone 11. The second position actually means a position where the lancet 4 mounted with the sample collector 3 enters the nucleic acid amplification zone 21, at which time the lancet 4 pierces the partition 13, the partition 13 is formed with a hole 131 communicating the nucleic acid extraction zone 11 and the nucleic acid amplification zone 21, and the piston 34 is slidably disposed in the nucleic acid extraction zone 11.

The piercing member 4 is formed with a guide groove 42. The flow guide groove 42 is used for communicating the nucleic acid extraction region 11 and the nucleic acid amplification region 21.

When the piercing member 4 pierces the separator 13, the piercing member 4 can be inserted into the hole 131 and the hole 131 is formed in the separator 13, but since the guide groove 42 is formed in the piercing member 4, the nucleic acid extraction region 11 and the nucleic acid amplification region 21 are communicated with each other via the guide groove 42, so that the piercing member 4 does not block the hole 131, and the substance in the nucleic acid extraction region 11 can be injected into the nucleic acid amplification region 21.

Specifically, the opening of the guide groove 42 is provided on the side of the piercing member 4. If the piercing member 4 is inserted into the hole 131, the opening of the flow guide groove 42 is disposed in the nucleic acid extracting region 11, the opening of the flow guide groove 42 is disposed in the nucleic acid amplifying region 21, and the substance in the nucleic acid extracting region 11 can enter the flow guide groove 42 through the opening of the flow guide groove 42 and then enter the nucleic acid amplifying region 21 through the opening of the flow guide groove 42.

The placing groove 42 and the flow guide groove 42 are integrated. When the partition 13 is actually manufactured, the placement groove 42 and the guide groove 42 are integrally formed on the partition 13.

The puncturing member 4 is formed with a tip 41, the tip 41 is disposed at one end of the puncturing member 4, the tip 41 has a tapered shape in its entire structure, and the tip 41 is used to facilitate puncturing the partitioning member 13.

The sample collector 3 comprises a rod 31, a sampling head 32 is arranged at one end of the rod 31, and a piston 34 is arranged on the rod 31.

The rod 31 has a rod-shaped overall structure, and the rod 31 is used for mounting other components such as a sampling head 32 and a piston 34. If the sampling head 32 is used to sample a patient or the plunger 34 is pushed to slide in the nucleic acid extracting region 11, the user can hold the rod 31 by hand for the convenience of the user.

As shown in fig. 1 to 7, in one embodiment, a reaction device for rapid nucleic acid detection is provided, in which a piston 34 is a sealing ring, and the entire structure of the sealing ring is annular, and the piston 34 is sleeved on a rod 31. The rod 31 is formed with a positioning groove 312, the positioning groove 312 is also annular, the positioning groove 312 surrounds the outside of the rod 31, and the piston 34 is partially disposed in the positioning groove 312. The piston 34 is made of rubber with high elasticity, so that the sealing effect of the piston 34 on the nucleic acid extracting area 11 is improved.

As shown in fig. 8 to 12, in the second embodiment, another reaction device for rapid nucleic acid detection is provided, the piston 34 has a ring-shaped overall structure, and the piston 34 is integrally formed on the rod 31, so that the number of parts is reduced, and the production cost is reduced.

The sampling head 32 is made of a material having water absorbing capacity. The material with water absorption capacity may be cotton. Cotton may be mounted on one end of the rod body 31 using a flocking process. The flocking process refers to the fact that cotton is adsorbed at one end of the rod body 31 by the aid of the electrostatic adsorption principle, glue is preset at one end of the rod body 31, and the sampling head 32 and the rod body 31 are bonded together through the glue.

The sampling head 32 may be inserted into the channel 42. If the piercing member 4 is disposed in the nucleic acid extraction region 11, the sample on the sampling head 32 can react with the substance in the nucleic acid extraction region 11 through the guide groove 42.

The length of the rod body 31 is 100 mm and 300mm, so that the sampling head 32 can be conveniently inserted into the mouth, nose, throat, cervix and other positions of a patient for sampling.

The sample collector 3 comprises an end cover 33, and the end cover 33 is used for covering the nucleic acid extraction area 11.

If the cover is covered on the nucleic acid extraction area 11, the cover is arranged on the opening of the nucleic acid extraction area 11, which plays a role in sealing and protecting the nucleic acid extraction area 11 and also plays a role in limiting the sample collector 3, and limits the sliding stroke of the piston 34 in the nucleic acid extraction area 11.

Specifically, the end cap 33 is integrally formed on the rod 31, so that the number of parts is reduced, and the cost is reduced.

The reaction vessel is formed with a countersunk groove 124, the countersunk groove 124 is provided at the opening of the nucleic acid extraction region 11, and the countersunk groove 124 is provided with an end cap 33.

A countersunk groove 124 is provided at one end of the body 12. The end cap 33 fits within the counter sink 124, improving compactness and sealing.

The end cap 33, the piston 34 and the sampling head 32 are arranged in sequence on the rod 31. If the piston 34 slides in the nucleic acid extracting region 11, the sampling head 32 and the piston 34 are disposed in the nucleic acid extracting region 11, and the end cap 33 is disposed outside the reaction vessel. The breaking groove 311 is arranged on one side of the end cover 33, and the sampling head 32 and the piston 34 are arranged on the other side of the end cover 33, so that the end cover 33, the sampling head 32 and the piston 34 are prevented from being separated from each other when the rod body 31 is broken.

The rod 31 is formed with a breaking groove 311, and the breaking groove 311 is used for facilitating a user to break the rod 31.

The breaking groove 311 is annular in overall structure, and the breaking groove 311 surrounds the outside of the rod body 31. If the sampling of the sample collector 3 is finished, the rod body 31 can be broken off, and then the sample collector 3 is inserted into the reaction vessel, so that the user can conveniently control the piston 34 to slide in the nucleic acid extraction area 11 by pressing the sample collector 3, and the operation of the user is further facilitated.

Specifically, the breaking groove 131 is provided at one end of the end cap. If the rod 31 is broken through the breaking groove 311, the end cap 33 is located at one end of the remaining rod 31, and the user can press the rod 31 with his hand, which further facilitates the operation of the user.

The nucleic acid extraction zone 11 can be provided with a lysis solution, and the nucleic acid amplification zone 21 can be provided with an amplification reagent and a fluorescence reagent. The lysate is used for carrying out nucleic acid extraction reaction on a sample, the amplification reaction reagent is used for carrying out amplification reaction on specific nucleic acid, and the fluorescent reagent has a fluorescent group and is used for carrying out nucleic acid detection on the specific nucleic acid product.

An example of detecting Vibrio parahaemolyticus in an environment using the apparatus for nucleic acid detection is described below. Vibrio Parahaemolyticus (VP) is a halophilic bacterium. Vibrio parahaemolyticus food poisoning is caused by eating food containing the bacteria, and is mainly derived from marine products. In the following examples, the lysis solution used is an alkaline lysis solution consisting of NaOH, Tris-HCl, etc. The kit comprises an amplification reaction reagent and a fluorescent reagent, wherein the detection reaction reagent is a common reaction reagent, can be configured by persons skilled in the relevant fields, and comprises an amplification primer, dNTPs, Bst DNA polymerase, a buffer solution, a fluorescent probe, a proper amount of ultrapure water and the like. This example reagent formulation was performed with reference to documents B.Pang, S.Yao, K.xu, J.Wang, X.Song, Y.mu, C.ZHao, J.Li, A novel visual-mixed-dye for LAMP and its application in the detection of foodborne diseases, Analytical Biochemistry (2019), doi: https:// doi.org/10.1016/j.ab.2019.03.002.

The detection mainly comprises the following steps:

s1, wiping the sample to be detected for 3-5 times by using the sampling head 32 of the sample collector 3;

s2, inserting the sampling head 32 into the nucleic acid extraction area 11 to enable the sample collector 3 to be arranged at a first position, wherein the sampling head 32 is located in the nucleic acid extraction area 11, the nucleic acid extraction area 11 is not communicated with the nucleic acid amplification area 21 of the nucleic acid amplification area, the piston 34 seals the nucleic acid extraction area 11, slightly stirring or washing up and down for 5 times to enable the sample and a lysate in the nucleic acid extraction area 11 to be uniformly mixed, and standing at room temperature for 30 seconds;

s3, continuously inserting the sampling head 32 into the nucleic acid extraction area 11 to enable the sample collector 3 to be arranged at a second position, in the process, inserting the sampling head 32 on the puncture piece 4 in the nucleic acid extraction area 11 and pushing the puncture piece 4 to puncture the separating piece 13, pushing the product in the nucleic acid extraction area 11 to be injected into the nucleic acid amplification area 21 by the piston 34, covering the end cover 33 on the extraction test tube 1, breaking the rod body 31 along the breaking groove 311, shaking the reaction container for 10 seconds to enable the nucleic acid extraction reaction product and the amplification reaction reagent to be uniformly mixed, and heating the reaction container at 65 ℃ for 15-20 minutes;

s4, directly observing by naked eyes, wherein the dark blue is a positive reaction, which indicates that the amplified sample contains vibrio parahaemolyticus; the dark gray color is negative reaction, i.e., no vibrio parahaemolyticus was detected in the amplified sample.

For the produced and subpackaged detection reagent, a user only needs to dip or scrape the sample collector 3 for 3-5 times, slightly and uniformly mix the swab in the reagent for 5-10 seconds, forcibly insert the sample collector 3 into the nucleic acid extraction area 11, puncture the separating element 13 so as to quantitatively release the nucleic acid substance to be detected and the lysate into the reaction solution, and observe the reaction result by naked eyes through heating reaction. The whole process is extremely simple to operate.

An example of the detection of mycoplasma cellularis contamination using the nucleic acid detecting apparatus in combination with a fluorescence detecting apparatus is described below. It is statistical that about 15% -35% of cell cultures worldwide suffer from mycoplasma contamination. Since 2013, published articles such as mycoplasma detection related to cell culture have been formally required in Nature journal. In the following examples, the lysis solution used is an alkaline lysis solution consisting of NaOH, Tris-HCl, etc. The amplification reaction reagent and the fluorescent reagent are common reaction reagents, and are carried out by adopting commercial reagents of a company where the applicant is located, wherein the product number is as follows: KS 201. The activator, the amplification reaction reagent and the fluorescent reagent are separately arranged in the amplification kit, the reagents except the activator in the amplification kit are pre-added into the reaction container, and the lysate and the activator in the amplification kit are pre-added into the nucleic acid extraction area 1 for standby.

During detection, the reaction vessel with the prepared reagent is taken out. The detection mainly comprises the following steps:

s1, dipping the cell sap to be detected by adopting the sampling head 32 of the sample collector 3;

s2, inserting the sampling head 32 into the nucleic acid extraction area 11 to enable the sample collector 3 to be arranged at a first position, wherein the sampling head 32 is located in the nucleic acid extraction area 11, the nucleic acid extraction area 11 is not communicated with the nucleic acid amplification area 21, the piston 34 seals the nucleic acid extraction area 11, slightly stirring or washing up and down for 5 times to enable the sample and the lysate in the nucleic acid extraction area 11 to be uniformly mixed, and standing for 20 seconds at room temperature;

s3, continuously inserting the sampling head 32 into the nucleic acid extraction area 11 to enable the sample collector 3 to be arranged at a second position, in the process, the sampling head 32 is inserted on the puncture piece 4 in the nucleic acid extraction area 11 and pushes the puncture piece 4 to puncture the separating piece 13, the piston 34 pushes the product in the nucleic acid extraction area 11 to be injected into the nucleic acid amplification area 21 in the nucleic acid amplification area, the end cover 33 is covered on the extraction test tube 1, the rod body 31 is broken along the breaking groove 311, the reaction container is shaken for 10 seconds to enable the nucleic acid extraction reaction product and the amplification reaction reagent to be uniformly mixed, the reaction container is placed on a fluorescence detection device to be heated for 15-20 minutes, and the fluorescence detection device reads a fluorescence signal value.

S4, when the fluorescence signal change is read by the fluorescence detection device, the reaction is positive, which indicates that the amplified sample contains mycoplasma contamination; otherwise, no cell contamination was detected.

For the produced and subpackaged detection reagent, a user only needs to dip or scrape a sample collector 3 for 3-5 times, lightly mix the sample collector 3 in the reagent for 5-10 seconds, forcibly insert the sample collector 3 into the reaction test tube 1, puncture the separating part 13 so as to quantitatively release nucleic acid substances and lysate to be detected into the nucleic acid amplification area 21 of the nucleic acid amplification area, and observe a reaction result by naked eyes through heating reaction. The whole process is extremely simple to operate.

The above-mentioned embodiment is only the preferred embodiment of the present invention, and does not limit the protection scope of the present invention according to this, so: all equivalent changes made according to the structure, shape and principle of the utility model should be covered within the protection scope of the utility model.

Claims (12)

1. A reaction device for rapid nucleic acid detection comprises a reaction container and a sample collector (3), and is characterized in that the reaction container is provided with a nucleic acid extraction area (11) and a nucleic acid amplification area (21), the sample collector (3) comprises a sampling head (32) and a piston (34), and the piston (34) is slidably arranged in the nucleic acid extraction area (11) so as to inject substances in the nucleic acid extraction area (11) into the nucleic acid amplification area (21).

2. The reaction device for rapid nucleic acid detection according to claim 1, wherein the sample collector (3) has a first position on the reaction vessel, and when the sample collector (3) is disposed at the first position, the sampling head (32) is located in the nucleic acid extraction region (11), and the nucleic acid extraction region (11) and the nucleic acid amplification region (21) are not communicated.

3. The reaction device for rapid nucleic acid detection according to claim 1 or 2, wherein the sample collector (3) has a second position on the reaction vessel, and when the sample collector (3) is disposed at the second position, the piston (34) is slidably disposed in the nucleic acid extraction region (11), and the nucleic acid extraction region (11) and the nucleic acid amplification region (21) are in communication.

4. The reaction device for rapid nucleic acid detection according to claim 1, wherein the reaction container has a partition (13) that can be punctured, and the partition (13) is used to separate the nucleic acid extraction region (11) and the nucleic acid amplification region (21).

5. The reaction device for rapid nucleic acid detection according to claim 4, wherein the partition (13) is a thin film.

6. The reaction device for rapid nucleic acid detection according to claim 1, 2, 4 or 5, further comprising a puncturing element (4), wherein the puncturing element (4) is used for puncturing the reaction vessel so as to communicate the nucleic acid extracting region (11) and the nucleic acid amplifying region (21).

7. The reaction device for rapid nucleic acid detection according to claim 6, wherein the piercing member (4) is detachably disposed on the sample collector (3).

8. The reaction device for rapid nucleic acid detection according to claim 6, wherein the piercing member (4) is formed with a channel (42), and the channel (42) is used for connecting the nucleic acid extraction region (11) and the nucleic acid amplification region (21).

9. A reaction device for rapid nucleic acid detection according to claim 1 or 2 or 4 or 5 or 7 or 8, wherein the reaction container comprises an extraction tube (1) and an amplification tube (2), the nucleic acid extraction zone (11) is disposed in the extraction tube (1), and the nucleic acid amplification zone (21) is disposed in the amplification tube (2).

10. The reaction device for rapid nucleic acid detection according to claim 9, wherein the extraction tube (1) is detachably disposed on the amplification tube (2).

11. The reaction device for rapid nucleic acid detection according to claim 9, wherein the reaction vessel comprises a light-permeable plate portion (23), and the plate portion (23) is disposed between the nucleic acid amplification region (21) and the outside.

12. The reaction device for rapid nucleic acid detection according to claim 1, wherein the sample collector (3) comprises an end cap (33), and the end cap (33) is used for covering the nucleic acid extraction area (11) or the extraction test tube (1).

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