CN112877193B - POCT consumable and detection method - Google Patents
POCT consumable and detection method Download PDFInfo
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- CN112877193B CN112877193B CN202110463483.9A CN202110463483A CN112877193B CN 112877193 B CN112877193 B CN 112877193B CN 202110463483 A CN202110463483 A CN 202110463483A CN 112877193 B CN112877193 B CN 112877193B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
- A61B10/0045—Devices for taking samples of body liquids
- A61B10/0051—Devices for taking samples of body liquids for taking saliva or sputum samples
Abstract
The application discloses POCT consumptive material and detection method, it belongs to medical inspection instrument, molecular diagnosis detecting instrument field, and its design essential lies in: the method comprises the following steps: the device comprises a main pipe module, a piston rod module, a reagent pipe module and a sampling rod module; a supervisor module comprising: the device comprises a vertical main pipe, a transverse main pipe, a first inserting double pipe, a second inserting double pipe, a third inserting double pipe and air holes; the piston rod module includes: a first piston rod and a second piston rod; the reagent tube module comprises: the first reagent tube, the second reagent tube and the third reagent tube are respectively connected with the first inserting double tube, the second inserting double tube and the third inserting double tube, and a first sealing ring is arranged between the reagent tube and the inserting double tube corresponding to the reagent tube for sealing. By adopting the POCT consumable and the detection method, the whole process of nucleic acid extraction and PCR reaction can be totally closed without pollutant leakage after a sample is put in.
Description
Technical Field
The application relates to the field of medical examination and inspection instruments and molecular diagnosis and detection instruments, in particular to a POCT consumable and a detection method.
Background
POCT, point-of-care-testing, also known as on-site rapid multi-parameter biochemical testing; is an important detection means for detecting various epidemics, such as novel coronavirus nucleic acid detection, epidemic virus detection and the like.
As the field of biomedical detection, a POCT consumable is designed, and a biological inhibitor, a lysate and a PCR reaction solution are preset inside the POCT consumable. The consumable can be fully automatic in the whole process of nucleic acid extraction and PCR reaction after being put into a sample, and manual intervention is not needed; the totally closed pollution-free leakage is an extremely important technical requirement.
In contrast, the applicant disclosed a POCT nucleic acid extraction amplification test tube in chinese patent application No. 202011256843. X; the Suzhou New sea biotechnology limited company discloses a POCT detection device and a sample mixing device in a Chinese patent with the application number of 201910609966.8; a portable integrated nucleic acid analysis device is disclosed in Chinese patent with the application number of 201811633096.X of Beijing university of chemical industry.
As known from the above documents, the design of a POCT consumable to achieve the above-mentioned "the whole process of nucleic acid extraction to PCR reaction can be totally closed without leakage of contaminants after the sample is put in" is always the direction of research and development by many manufacturers.
Disclosure of Invention
The present application is directed to provide a POCT consumable aiming at the above-mentioned shortcomings of the prior art.
Another object of the present application is to provide a method for detecting POCT consumables, which overcomes the above-mentioned shortcomings of the prior art.
The technical scheme of the application is as follows:
a POCT consumable comprising: the device comprises a main pipe module, a piston rod module, a reagent pipe module and a sampling rod module;
a supervisor module comprising: the device comprises a vertical main pipe, a transverse main pipe, a first inserting double pipe, a second inserting double pipe, a third inserting double pipe and air holes; the vertical main pipe, the first inserting double pipe, the second inserting double pipe, the third inserting double pipe and the air hole are communicated with the transverse main pipe;
the vertical main pipe and the air holes are respectively arranged at the upper side of the transverse main pipe and are respectively arranged at the left end part and the right end part, and the first inserting double pipe, the second inserting double pipe and the third inserting double pipe are respectively arranged at the lower side of the transverse main pipe in parallel at intervals and are arranged at the middle part;
the piston rod module includes: a first piston rod and a second piston rod; the first piston rod and the second piston rod can be inserted into the transverse main pipe;
the sampling rod module is inserted into the vertical main pipe;
the reagent tube module comprises: the first reagent tube, the second reagent tube and the third reagent tube are respectively connected with the first inserting double tube, the second inserting double tube and the third inserting double tube, and a first sealing ring is arranged between the reagent tube and the inserting double tube corresponding to the reagent tube for sealing.
Further, the sampling stick module includes from last to down in proper order: the sampling device comprises an external holding rod, a first top protruding part, a second top protruding part and a sampling head; further comprising: the connection part of the first top protruding part and the second top protruding part is provided with an annular groove, and the second sealing ring is sleeved in the annular groove; the sampling head adopts sponge;
the vertical main pipe includes: the first vertical main pipe space, the second vertical main pipe space, the third vertical main pipe space, the fourth vertical main pipe space and the bottom liquid outlet hole; the first vertical main pipe space, the second vertical main pipe space, the third vertical main pipe space and the fourth vertical main pipe space are all designed to be large in top opening and small in bottom opening; the first vertical main pipe space, the second vertical main pipe space, the third vertical main pipe space and the fourth vertical main pipe space are sequentially connected from top to bottom, and a bottom liquid outlet hole is formed in the bottom of the fourth vertical main pipe space;
the vertical main pipe is communicated with the transverse main pipe through a liquid outlet hole at the bottom;
the diameter of the second sealing ring is larger than that of the bottom opening of the second vertical main pipe space;
in a natural state, the sum of the lengths of the second top protruding part and the sampling head is larger than the sum of the depths of the third vertical main pipe space and the fourth vertical main pipe space, namely the sampling rod module is inserted into the vertical main pipe, and after the second sealing ring reaches an initial sealing position (the initial sealing position is that the second sealing ring is positioned between the third vertical main pipe spaces), the sampling head sponge can be compressed to extrude saliva.
Further, the first piston rod is identical in structure to a second piston rod, the second piston rod including: the first pipe joint, the second pipe joint, the third pipe joint, the fourth pipe joint, the fifth pipe joint, the first baffle and the end baffle; the first pipe joint, the second pipe joint, the third pipe joint, the fourth pipe joint and the fifth pipe joint are sequentially connected; an end baffle is arranged at the end part of the fifth pipe joint, and a first baffle is arranged in the middle area of the fifth pipe joint; the pipe orifices connected with the adjacent pipe joints are provided with third sealing rings;
the first, second and third insertion double pipes have the same structure and the same distance; the lengths of the second pipe joint, the third pipe joint, the fourth pipe joint and the fifth pipe joint are the same;
the outer diameter of the first baffle is larger than the inner diameter of the transverse main pipe, when the first baffle is in contact with the transverse main pipe, the second pipe joint corresponds to the first inserted double pipe, the third pipe joint corresponds to the second inserted double pipe, and the fourth pipe joint corresponds to the third inserted double pipe.
Furthermore, the first reagent tube is provided with an inhibitor, the second reagent tube is provided with a cracking agent, and the third reagent tube is provided with a PCR reaction solution; each reagent tube comprises: a short matching pipe and a long liquid storage pipe; the first reagent tube, the second reagent tube and the third reagent tube have the same structure, and the solution in the reagent tubes is stored in the long liquid storage tube in advance;
each insertion double tube comprises: a short insertion tube and a long insertion tube; the transverse main pipe is connected with the short inserting pipe and the long inserting pipe of each inserting double pipe through the connecting holes;
the short insertion tube is inserted into the short matching tube, and the long insertion tube is inserted into the long liquid storage tube; .
Further, the diameter of the pipe joint of the second piston rod is smaller than that of the transverse main pipe, and the diameter of the third sealing ring of the pipe orifice is not smaller than (larger than or equal to) that of the transverse main pipe;
the first, second and third reagent tubes are all designed with short matching tubes on the left side and long liquid storage tubes on the right side; correspondingly, the "connecting hole of the short insertion tube and the transverse main tube" is on the left side of the "connecting hole of the long insertion tube and the transverse main tube".
Further, the vertical main pipe, the horizontal main pipe, the first insertion double pipe, the second insertion double pipe, and the third insertion double pipe are integrally formed.
Further, the first reagent tube, the second reagent tube and the third reagent tube have the same structure, and the solution in the reagent tubes is stored in the long liquid storage tube in advance.
A detection method using the POCT consumable comprises the following steps:
s1, collecting saliva: collecting saliva using a sampling wand module;
s2, assembling POCT: inserting the sampling rod module which collects the saliva in the step S1 into the vertical main pipe until the vertical main pipe is sealed by the second sealing ring, wherein the sampling head sponge is compressed to extrude the saliva in the inserting process;
s3, extracting saliva into the transverse main tube;
s4, moving saliva;
s5, extruding saliva into the first reagent tube;
s6, extracting a saliva-inhibitor mixed solution;
s7, moving the saliva-inhibitor mixture;
s8, extruding the saliva-inhibitor mixed solution into the lysate;
s9, extracting the mixed liquid in the second reagent tube;
s10, moving the mixed solution obtained in step S9:
s11, pressing the mixed liquid obtained in the step S9 into the long liquid storage tube in the third reagent tube for the first time;
s12, the mixed solution obtained in step S9 is pressed into the long liquid storage tube in the third reagent tube for the second time:
s12-1, the first piston rod and the second piston rod move rightwards, the first piston rod and the second piston rod are separated, and the third sealing ring on the rightmost side of the first piston rod is arranged on the right side of the third insertion double pipe, so that the area between the first pipe joint of the first piston rod and the first pipe joint of the second piston rod is communicated with the air hole, and the air hole and the third insertion double pipe do not form an air channel;
s12-2, the first piston rod and the second piston rod move to the left side and are symmetrically arranged on two sides of a long insertion tube of a third insertion double tube, then the first piston rod and the second piston rod move to the middle at the same time to perform secondary liquid drainage, and the residual liquid in the transverse main tube is squeezed into a long liquid storage tube of the third reagent tube;
s13, the first piston rod is continuously pushed rightward until the first baffle of the first piston rod contacts the transverse main tube, so that the second tube joint of the first piston rod corresponds to the third reagent tube, i.e. the rightmost third sealing ring of the first piston rod is located on the right side of the connecting hole between the long insertion tube of the third insertion double tube and the transverse main tube; the third sealing ring on the secondary right side of the first piston rod is arranged on the left side of the connecting hole between the short insertion pipe of the third insertion double pipe and the transverse main pipe.
The beneficial effect of this application lies in:
first, the basic scheme of the present application is: the structural design of POCT consumables. The specific design difficulty and the key point are as follows:
1.1 sampling stick module, vertical person in charge, sampling stick module and vertical person in charge complex design.
1.2 the piston rod module, the transverse main pipe and the piston rod module are matched with the transverse main pipe.
1.3 reagent pipe module, main pipe module, reagent pipe module and main pipe module matched design.
1.4 reagent tube module 3, main tube module 1 and piston rod module 2
That is, the core design of the scheme of the present application is as follows: the design that a sampling rod module is matched with a vertical main pipe, the design that a piston rod module is matched with a transverse main pipe, the design that a reagent pipe module is matched with a main pipe module, and the design that the reagent pipe module, the main pipe module and the piston rod module are matched are adopted.
Secondly, the basic scheme of the application also comprises: the PCOT consumable is applied to a detection scheme, namely how to mix saliva with an inhibitor, then mix saliva-inhibitor mixed liquor with lysate, and then mix the saliva-inhibitor-lysate mixed liquor with PCR reaction liquid.
That is, the scheme of this application has solved the demand to POCT consumptive material among the background art: the method comprises the following steps:
1) 50-100ul of saliva was collected using a sampling wand module.
2) 40ul of saliva was extracted and added to 30ul of inhibitor (inhibitor) to form mixture A, and then 70ul of mixture A was heated to 95 ︒ C for 30 minutes.
3) 20ul of the mixed solution A is extracted and added into 20ul of lysate to be mixed into a mixed solution B.
4) 10ul of the mixture was extracted from the mixture B and added to 25ul of the PCR reaction solution to carry out PCR reaction.
The detection scheme is unified with the structural design of the consumable.
Drawings
The present application will be described in further detail with reference to the following examples, which are not intended to limit the scope of the present application.
Fig. 1 is a schematic three-dimensional design diagram of a POCT consumable of the present application.
Fig. 2a is a schematic diagram of explosion effect of each component of the POCT consumable of the present application.
FIG. 2b is a schematic design view of a sample wand module of the present application.
Fig. 2c is a schematic design of the vertical master of the master module of the present application.
Fig. 2d is a schematic design of a piston rod module of the present application.
Fig. 3 is a schematic view of a mid-section of a POCT consumable of the present application.
Fig. 4 is a state diagram at the end of step S2 in the POCT consumable supply detection of the present application.
Fig. 5 is a schematic diagram illustrating the state of step S3 during POCT consumable inspection according to the present application.
Fig. 6 is a schematic diagram illustrating the state of step S4 during POCT consumable inspection according to the present application.
Fig. 7 is a schematic diagram illustrating the state of step S5 during POCT consumable inspection according to the present application.
Fig. 8 is a schematic diagram illustrating the state of step S6 during POCT consumable inspection according to the present application.
Fig. 9 is a schematic diagram illustrating the state of step S7 during POCT consumable inspection according to the present application.
Fig. 10 is a schematic diagram illustrating the state of step S8 during POCT consumable inspection according to the present application.
Fig. 11 is a schematic diagram illustrating the state of step S10 during POCT consumable inspection according to the present application.
Fig. 12 is a schematic diagram illustrating the state of step S11 during POCT consumable inspection according to the present application.
FIG. 13 is a schematic diagram illustrating the state of step S12-1 during the detection of POCT consumables according to the present invention.
FIG. 14 is a schematic diagram illustrating the state of step S12-2 during the detection of POCT consumables according to the present invention.
Fig. 15 is a schematic view of the state after step S13 ends in the POCT consumables detection of the present application.
FIG. 16 is a schematic diagram of steps S5-S6, S8-S9 of the present application.
The reference numerals of fig. 1-16 are illustrated as follows:
a main pipe module 1, a first inserting double pipe 1-1, a second inserting double pipe 1-2, a third inserting double pipe 1-3 and a vertical main pipe 1-4;
short insertion tubes 1-1-1, long insertion tubes 1-1-2;
1-4-1 parts of a first vertical main pipe space, 1-4-2 parts of a second vertical main pipe space, 1-4-3 parts of a third vertical main pipe space, 1-4-4 parts of a fourth vertical main pipe space, 1-4-5 parts of a bottom liquid outlet hole, 1-5 parts of a transverse main pipe and 1-6 parts of an air hole;
the piston rod module 2, a first piston rod 2-1, a second piston rod 2-2, a first pipe joint 2-2-1, a second pipe joint 2-2-2, a third pipe joint 2-2-3, a fourth pipe joint 2-2-4, a fifth pipe joint 2-2-5, a first baffle 2-2-6, an end baffle 2-2-7 and a third sealing ring 2-2-8;
a reagent tube module 3, a first reagent tube 3-1, a second reagent tube 3-2, a third reagent tube 3-3, a short matching tube 3-1-1 and a long liquid storage tube 3-1-2;
the sampling rod module 4, the outer holding rod 4-1, the first top protruding part 4-2, the second sealing ring 4-3, the second top protruding part 4-4 and the sampling head 4-5.
Detailed Description
Embodiment 1, a POCT consumable comprising: the device comprises a main pipe module 1, a piston rod module 2, a reagent pipe module 3 and a sampling rod module 4.
Design of primary pipe module 1
As shown in fig. 2a, the master module 1 includes: 1-4 parts of a vertical main pipe, 1-5 parts of a transverse main pipe, 1-1 parts of a first inserted double pipe, 1-2 parts of a second inserted double pipe, 1-3 parts of a third inserted double pipe and 1-6 parts of air holes.
The bottom of the vertical main pipe 1-4 is provided with an opening, so that the vertical main pipe 1-4 is communicated with the transverse main pipe 1-5.
The vertical main pipe 1-4 and the air holes 1-6 are respectively arranged at the upper side of the transverse main pipe 1-5, and the first insertion double pipe 1-1, the second insertion double pipe 1-2 and the third insertion double pipe 1-3 are respectively arranged at the lower side of the transverse main pipe 1-5 in parallel at intervals.
The vertical main pipe 1-4 is arranged at the left end part of the transverse main pipe, and the air hole 1-6 is arranged at the right end part of the transverse main pipe; the first insertion double pipe 1-1, the second insertion double pipe 1-2 and the third insertion double pipe 1-3 are arranged in the middle of the transverse main pipe 1-5, namely, between the vertical main pipe 1-4 and the air hole 1-6 (the vertical main pipe 1-4 and the air hole 1-6 are arranged above the transverse main pipe, and the first insertion double pipe 1-1, the second insertion double pipe 1-2 and the third insertion double pipe 1-3 are arranged below the transverse main pipe).
Particularly, the first inserting double tube 1-1, the second inserting double tube 1-2 and the third inserting double tube 1-3 have the same structure.
Specifically, the vertical main pipe 1-4, the horizontal main pipe 1-5, the first insertion double pipe 1-1, the second insertion double pipe 1-2, and the third insertion double pipe 1-3 are integrally formed.
Design of sampling rod module 4
As shown in fig. 2b, the sampling rod module 4 includes, from top to bottom: an external holding rod 4-1, a first top protruding part 4-2, a second top protruding part 4-4 and a sampling head 4-5; the external holding rod 4-1, the first top protruding part 4-2 and the second top protruding part 4-4 are made of hard materials, such as: hard plastic; the sampling head 4-5 is arranged at the bottom of the second top projection 4-4 and is made of an elastic component.
The external holding rod 4-1 is in a round rod shape (the external holding rod can be replaced by a cuboid), and the first top protruding part 4-2, the second top protruding part 4-4 and the sampling head 4-5 are all in round rod-shaped structures.
The second sealing ring 4-3 is sleeved between the first top protruding part 4-2 and the second top protruding part 4-4; the sampling heads 4-5 are made of sponge (or made of similar materials).
The sampling wand module 4 is shown inserted into the vertical main tube 1-4.
Thirdly, the matching design of the main pipe module 1 and the sampling rod module 4
In particular, the mating features of the host tube module 1 and the sampling wand module 4 are as follows:
firstly, designing a vertical main pipe 1-4: as shown in fig. 2c, the vertical main tube 1-4 (which is circular in cross-section) comprises: 1-4-1 parts of a first vertical main pipe space, 1-4-2 parts of a second vertical main pipe space, 1-4-3 parts of a third vertical main pipe space, 1-4-4 parts of a fourth vertical main pipe space and 1-4-5 parts of a bottom liquid outlet hole; the first vertical main pipe space 1-4-1, the second vertical main pipe space 1-4-2, the third vertical main pipe space 1-4-3 and the fourth vertical main pipe space 1-4-4 are all designed to have a large top opening and a small bottom opening.
Secondly, the sampling head 4-5 of the sampling rod module 4 adopts a sponge design; the connecting part of the first top protruding part 4-2 and the second top protruding part 4-4 is provided with a circumferential groove, and a second sealing ring 4-3 is sleeved in the circumferential groove, so that the second sealing ring cannot slide on the sampling rod module 4 when sealing is ensured.
Thirdly, the diameter of the second sealing ring is larger than that of the bottom opening of the second vertical main pipe space 1-4-2 according to the relation between the vertical main pipe 1-4 and the sampling rod module 4.
In a natural state, the sum of the lengths of the second top protruding part 4-4 and the sampling head 4-5 is larger than the sum of the depths of the third vertical main pipe space 1-4-3 and the fourth vertical main pipe space 1-4-4, so that the following conditions are ensured: the sampling rod module 4 is inserted into the vertical main pipe 1-4, and after the second sealing ring reaches the initial sealing position, the sampling head sponge is compressed to extrude saliva.
Design of piston rod module 2
As shown in fig. 2d, the piston rod module 2 comprises: a first piston rod 2-1 and a second piston rod 2-2; the first piston rod 2-1 and the second piston rod 2-2 have the same structure.
The second piston rod 2-2 includes: the first pipe joint 2-2-1, the second pipe joint 2-2-2, the third pipe joint 2-2-3, the fourth pipe joint 2-2-4, the fifth pipe joint 2-2-5, the first baffle 2-2-6 and the end baffle 2-2-7.
The first pipe joint 2-2-1, the second pipe joint 2-2-2, the third pipe joint 2-2-3, the fourth pipe joint 2-2-4 and the fifth pipe joint 2-2-5 are connected in sequence; an end baffle 2-2-7 is arranged at the end part of the fifth pipe joint 2-2-5, and a first baffle 2-2-6 is arranged in the middle area of the fifth pipe joint 2-2-5.
And the pipe orifices connected with the adjacent pipe joints are provided with third sealing rings 2-2-8.
Namely, a pipe orifice is arranged between the first pipe joint 2-2-1 and the second pipe joint 2-2-2, and a pipe orifice is arranged between the second pipe joint 2-2-2 and the third pipe joint 2-2-3; a pipe orifice is arranged between the third pipe joint 2-2-3 and the fourth pipe joint 2-2-4; a pipe orifice is arranged between the fourth pipe joint 2-2-4 and the fifth pipe joint 2-2-5; the pipe orifices are all provided with third sealing rings 2-2-8, namely the second piston rod 2-2 comprises 4 third sealing rings 2-2-8.
Particularly, the lengths of the second pipe joint 2-2-2, the third pipe joint 2-2-3, the fourth pipe joint 2-2-4 and the fifth pipe joint 2-2-5 are the same.
Fifthly, the matching design of the main pipe module 1 and the piston rod module 2 is as follows:
design of the first and second piston rods 2-2 (first piston rod 2-1): the length of the second pipe joint is greater than that of the first inserted double pipe; the length of the third pipe joint is greater than that of the second inserted double pipe; the length of the fourth pipe joint is greater than that of the third inserted double pipe; the piston rod must be provided with 5 pipe joints and with 4 third sealing rings.
Secondly, designing the transverse main pipe 1-5: the first, second and third insertion double pipes are spaced apart by the same distance.
Thirdly, the relationship between the vertical main pipe 1-4 and the sampling rod module 4: the outer diameter of the first baffle is larger than the inner diameter of the transverse main pipe, when the first baffle is in contact with the transverse main pipe, the second pipe joint corresponds to the first inserted double pipe, the third pipe joint corresponds to the second inserted double pipe, and the fourth pipe joint corresponds to the third inserted double pipe.
Design of reagent tube module 3
As shown in fig. 2a and 2c, the reagent tube module 3 comprises: a first reagent tube 3-1, a second reagent tube 3-2, and a third reagent tube 3-3; 30ul of inhibitor is preset in the first reagent tube 3-1, 20ul of cracking agent is preset in the second reagent tube 3-2, and 25ul of PCR reaction solution is preset in the third reagent tube 3-3.
The first reagent vessel 3-1 includes: a short matching pipe 3-1-1 and a long liquid storage pipe 3-1-2.
The first insertion double tube 1-1 includes: short insertion tube, long insertion tube.
The first insertion double tube 1-1, the second insertion double tube 1-2 and the third insertion double tube 1-3 are respectively matched with the first reagent tube 3-1, the second reagent tube 3-2 and the third reagent tube 3-3; namely, the first reagent tube 3-1, the second reagent tube 3-2 and the third reagent tube 3-3 are connected with the first insertion double tube 1-1, the second insertion double tube 1-2 and the third insertion double tube 1-3 in a clamping manner.
Specifically, the short insertion tube and the long insertion tube into which the double tubes are inserted are respectively fitted to the short fitting tube and the long solution storage tube of the corresponding reagent tube.
In particular, a first sealing ring seal is arranged between each reagent tube and the corresponding insertion double tube.
Particularly, the first reagent tube 3-1, the second reagent tube 3-2 and the third reagent tube 3-3 have the same structure, and the solution in the reagent tubes is stored in the long liquid storage tube in advance.
Seventhly, the design of the matching of the main pipe module 1 and the reagent pipe module 3 is as follows:
first, the design of the supervisor module 1: the transverse main pipe 1-5 is connected with the short insertion pipe and the long insertion pipe of each insertion double pipe (the first insertion double pipe 1-1, the second insertion double pipe 1-2 and the third insertion double pipe 1-3) through the connecting hole.
Second, the design of the reagent tube module 3: a double-tube design mode of a short matching tube 3-1-1 and a long liquid storage tube 3-1-2.
Thirdly, the relationship between the vertical main pipe 1-4 and the reagent pipe module 3 is as follows: after the short insertion tube and the long insertion tube are inserted into the short matching tube 3-1-1 and the long liquid storage tube 3-1-2, the short matching tube 3-1-1 is communicated with the long liquid storage tube 3-1-2 (the outer diameter of the long liquid storage tube is larger than the inner diameter of the long insertion tube).
Eighthly, the reagent tube module 3, the main tube module 1 and the piston rod module 2 are designed in a matching way as follows:
firstly, the method comprises the following steps: the diameter of the pipe joint of the second piston rod 2-2 (the first piston rod 2-1) is smaller than that of the transverse main pipe, and the diameter of the third sealing ring of the pipe orifice is slightly larger than that of the transverse main pipe.
Secondly, the method comprises the following steps: the first, second and third reagent tubes are all designed with the short matching tube on the left side and the long liquid storage tube on the right side (correspondingly, the connecting hole of the short insertion tube and the transverse main tube is on the left side of the connecting hole of the long insertion tube and the transverse main tube); so that, when saliva (mixed liquid) is squeezed into the long liquid storage tube, air in the long liquid storage tube can flow to the short insertion tube and then to the transverse main tube.
A working method of POCT comprises the following steps:
s1, collecting saliva: saliva is collected using the sampling wand module 4.
And S2, assembling the POCT.
S2-1, the reagent tube module 3 loaded with the solution is installed on the main tube module 1 in a matching way, namely, the first reagent tube 3-1, the second reagent tube 3-2 and the third reagent tube 3-3 are correspondingly installed on the first insertion double tube 1-1, the second insertion double tube 1-2 and the third insertion double tube 1-3.
S2-2, inserting the second piston rod 2-2 to the right side of the transverse main pipe 1-5 until the first baffle plate contacts with the transverse main pipe 1-5; the first piston rod 2-1 is then inserted to the left of the transverse main tube 1-5 so that the first tube section of the first piston rod 2-1 is in contact with the first tube section of the second piston rod 2-2.
And S2-3, inserting the sampling rod module with the saliva collected in the step S1 into the vertical main pipe 1-4 until the second sealing ring 4-3 reaches the initial sealing position, wherein the sampling head sponge is compressed to extrude the saliva in the inserting process.
The results of completion of S2-3 are shown in FIG. 4.
S3, drawing saliva into the transverse main tube: as shown in fig. 5, the first piston rod 2-1 moves leftwards, the second piston rod 2-2 moves rightwards, and 40ul of saliva is measured and flows into the transverse main pipe 1-5 from the connecting hole at the bottom of the vertical main pipe 1-4; meanwhile, the sampling rod module slightly moves downwards due to the action of the negative pressure.
S4, moving saliva: as shown in fig. 6, the first piston rod 2-1 and the second piston rod 2-2 move synchronously to the right (i.e. the moving speed and distance of the two are the same), and 40ul of saliva is transferred to the right to the connecting hole of the long insertion tube of the first insertion double tube 1-1 and the transverse main tube 1-5; (in this process, the short insertion tube and the long insertion tube of the first insertion double tube are both very small in orifice, and the solution does not flow into the insertion tubes by itself under the action of gravity).
S5, salivary ingress inhibitors: as shown in fig. 7, the first piston rod 2-1 is not moved, the second piston rod 2-2 is moved to the left until the first tube sections of the two are contacted, and 40ul of saliva is squeezed into the long insertion tube of the first insertion double tube 1-1 and further into the long storage tube of the first reagent tube 3-1, so that the saliva is mixed with the inhibitor; (Note that saliva does not flow completely into the long reservoir tube, which would fill the long insertion tube (otherwise a vacuum would be created)).
Meanwhile, air in the first long insertion pipe inserted into the double pipes and the first reagent pipe enters a gap between the transverse main pipe and the second pipe section of the first piston rod from the short matching pipe of the first reagent pipe 3-1, so that the sampling rod module is pushed upwards a little, and the sampling rod module and the vertical main pipe are always sealed in the process.
S6, extracting saliva-inhibitor mixture: as shown in FIG. 8, in S6-1, the mixture in the long liquid storage tube of the first reagent tube 3-1 is heated to 95 ℃ and held for 30 minutes or more.
S6-2, the first piston rod 2-1 is fixed, the second piston rod 2-2 moves rightwards, and 20ul of liquid is extracted from the long liquid storage tube of the first reagent tube 3-1 to the transverse main tube.
S7, mobilization of saliva-inhibitor mixture: as shown in fig. 9, the first piston rod 2-1 and the second piston rod 2-2 are moved rightward in synchronization (i.e., the rate and distance of movement of the two are the same), and 20ul of the saliva-inhibitor mixed liquid in the transverse main tube in step S8 is transferred rightward to the connecting hole of the long insertion tube of the second insertion double tube and the transverse main tube.
S8, the saliva-inhibitor mixture goes to lysis: as shown in fig. 10, the first piston rod 2-1 is fixed, the second piston rod 2-2 moves leftwards until the first tube joints of the two piston rods are contacted, and 20ul of liquid is squeezed into the long insertion tube of the second insertion double tube and then flows into the long liquid storage tube of the second reagent tube, so as to be mixed with the lysate; (Note that the saliva-inhibitor mixture does not flow completely into the long reservoir, which would fill the long insertion tube (otherwise a vacuum would be created)).
At the same time, air in the second reagent tube 3-2 will enter the gap between the transverse main tube and the second tube section of the second piston rod from the short mating tube of the second reagent tube.
S9, extracting the mixture in the second reagent tube: the first piston rod 2-1 is fixed, the second piston rod 2-2 moves rightwards, and 10ul of liquid in the long liquid storage tube of the second reagent tube 3-2 is extracted to the transverse main tube through the long insertion tube of the second insertion double tube.
S10, as shown in FIG. 11, the mixed liquid obtained in step S9 is moved: the first piston rod 2-1 and the second piston rod 2-2 are moved rightward in synchronization (i.e., the rate and distance of movement of the two are the same), and 10ul of the mixed liquid in the transverse main pipe in step S9 is transferred rightward to the connecting hole between the long insertion pipe of the third insertion double pipe and the transverse main pipe.
S11, as shown in fig. 12, the mixed solution obtained in step S9 is first pushed into the long storage tube in the third reagent tube: the first piston rod 2-1 is fixed, the second piston rod 2-2 moves leftwards until the first pipe joints of the first piston rod and the second piston rod are contacted, 10ul of liquid is squeezed into the long insertion pipe of the third insertion double pipe and then flows into the long liquid storage pipe of the third reagent pipe, and then is mixed with the PCR reaction liquid.
At the same time, air in the third reagent tube 3-3 will enter the gap between the transverse main tube and the third tube section of the second piston rod from the short mating tube of the third reagent tube.
S12, the mixed solution obtained in the step S9 is pressed into the long liquid storage tube in the third reagent tube for the second time.
S12-1, as shown in fig. 13, the first piston rod and the second piston rod 2-2 both move rightwards, the first piston rod and the second piston rod are separated, and the rightmost third sealing ring of the first piston rod is located on the right side of the third insertion double tube, so that the area between the first tube section of the first piston rod and the first tube section of the second piston rod is communicated with the air hole 1-6, and the air hole 1-6 and the third insertion double tube do not form an air passage.
S12-2, as shown in fig. 14, the first piston rod and the second piston rod 2-2 are both moved to the left side and symmetrically disposed at both sides of the long insertion tube of the third insertion double tube, and then the first piston rod and the second piston rod 2-2 are simultaneously moved to the middle to perform the secondary liquid discharge, so that the remaining liquid in the lateral main tube is pushed into the long liquid storage tube of the third reagent tube, and the purpose of the step S12 is to: it is ensured that after step S11 (i.e., the first drain), a small amount of air is sucked in to ensure that 10. mu.l of the mixture solution is mixed into the PCR solution in the third reagent vessel.
In the process of step S12, the gas holes 1 to 6 and the third insertion double tube do not always form a gas passage.
S13, the first piston rod is further pushed rightward until the first baffle of the first piston rod contacts the transverse main tube, so that the second tube section of the first piston rod corresponds to the third reagent tube, as shown in fig. 15, that is, the rightmost third sealing ring of the first piston rod is located on the right side of the connecting hole between the long insertion tube of the third insertion double tube and the transverse main tube; the third sealing ring on the secondary right side of the first piston rod is arranged on the left side of the connecting hole between the short insertion pipe of the third insertion double pipe and the transverse main pipe.
The difficulty in this process is understood to be:
first, the process of steps S5, S6:
as shown in fig. 16, S5, the first piston rod and the second piston rod are pressed to inevitably press saliva from the lateral main tube into the long liquid storage tube of the first reagent tube, and in this process, the saliva flows from the lateral main tube into the long liquid storage tube of the first reagent tube from the long insertion tube of the first insertion double tube, and when the saliva passes through, air in the long insertion tube of the first insertion double tube is pressed out, and the air flows from the long liquid storage tube of the first reagent tube into the short matching tube of the first reagent tube; due to the effect of air pressure, the long insertion tube does not form vacuum, but is filled with saliva;
s6, when the second piston rod moves to the right side, the mixed liquid is pumped up; the reason why it can be extracted is: the long insertion tube in the first insertion double tube is provided with saliva, and the second piston rod moves towards the right side under the action of air pressure, so that mixed liquid can be continuously extracted from the long liquid storage tube.
Similarly, the long insertion tube of the second insertion double tube is filled with the mixed solution of saliva and the inhibitor when the lysate is extracted.
And secondly, when the mixed solution enters the PCR reaction solution, a double extrusion type mode is adopted.
First, the amount of solution itself is small, on the ul scale.
Secondly, before the second extrusion, air is filled between the first piston rod and the second piston rod, so that the solution in the long insertion tube of the third insertion double tube is not filled and mostly flows into the third reagent tube (which is a technically distinct design from the first reagent tube and the second reagent tube).
The above-mentioned embodiments are merely preferred embodiments of the present application, which are not intended to limit the present application in any way, and it will be understood by those skilled in the art that various changes and modifications can be made without departing from the spirit and scope of the present application.
Claims (4)
1. A POCT consumable, comprising: the device comprises a main pipe module, a piston rod module, a reagent pipe module and a sampling rod module;
a supervisor module comprising: the device comprises a vertical main pipe, a transverse main pipe, a first inserting double pipe, a second inserting double pipe, a third inserting double pipe and air holes; the vertical main pipe, the first inserting double pipe, the second inserting double pipe, the third inserting double pipe and the air hole are communicated with the transverse main pipe;
the vertical main pipe and the air holes are respectively arranged at the upper side of the transverse main pipe and are respectively arranged at the left end part and the right end part, and the first inserting double pipe, the second inserting double pipe and the third inserting double pipe are respectively arranged at the lower side of the transverse main pipe in parallel at intervals and are arranged at the middle part;
the piston rod module includes: a first piston rod and a second piston rod; the first piston rod and the second piston rod can be inserted into the transverse main pipe;
the sampling rod module is inserted into the vertical main pipe;
the reagent tube module comprises: the first reagent tube, the second reagent tube and the third reagent tube are respectively connected with the first inserting double tube, the second inserting double tube and the third inserting double tube, and first sealing rings are arranged between the reagent tubes and the inserting double tubes for sealing;
the sampling stick module includes from last down in proper order: the sampling device comprises an external holding rod, a first top protruding part, a second top protruding part and a sampling head; the sampling head adopts sponge; the sampling wand module further comprises: a second seal ring; a connecting part of the first top protruding part and the second top protruding part is provided with an annular groove, and a second sealing ring is sleeved in the annular groove; the vertical main pipe includes: the first vertical main pipe space, the second vertical main pipe space, the third vertical main pipe space, the fourth vertical main pipe space and the bottom liquid outlet hole; the first vertical main pipe space, the second vertical main pipe space, the third vertical main pipe space and the fourth vertical main pipe space are all designed to be large in top opening and small in bottom opening; the first vertical main pipe space, the second vertical main pipe space, the third vertical main pipe space and the fourth vertical main pipe space are sequentially connected from top to bottom, and a bottom liquid outlet hole is formed in the bottom of the fourth vertical main pipe space; the vertical main pipe is communicated with the transverse main pipe through a liquid outlet hole at the bottom; the diameter of the second sealing ring is larger than that of the bottom opening of the second vertical main pipe space; in a natural state, the sum of the lengths of the second top protruding part and the sampling head is larger than the sum of the depths of the third vertical main pipe space and the fourth vertical main pipe space, namely the sampling rod module is inserted into the vertical main pipe, and after the second sealing ring reaches an initial sealing position, the sampling head sponge can be compressed to extrude saliva;
the first piston rod is identical in structure to a second piston rod, the second piston rod including: the first pipe joint, the second pipe joint, the third pipe joint, the fourth pipe joint, the fifth pipe joint, the first baffle and the end baffle; the first pipe joint, the second pipe joint, the third pipe joint, the fourth pipe joint and the fifth pipe joint are sequentially connected; an end baffle is arranged at the end part of the fifth pipe joint, and a first baffle is arranged in the middle area of the fifth pipe joint; the pipe orifices connected with the adjacent pipe joints are provided with third sealing rings;
the first, second and third insertion double pipes have the same structure and the same distance; the lengths of the second pipe joint, the third pipe joint, the fourth pipe joint and the fifth pipe joint are the same;
the outer diameter of the first baffle is larger than the inner diameter of the transverse main pipe, when the first baffle is contacted with the transverse main pipe, the second pipe joint corresponds to the first inserted double pipe, the third pipe joint corresponds to the second inserted double pipe, and the fourth pipe joint corresponds to the third inserted double pipe;
each reagent tube comprises: a short matching pipe and a long liquid storage pipe; the solution in each reagent tube is stored in the long liquid storage tube in advance; each insertion double tube comprises: a short insertion tube and a long insertion tube; the transverse main pipe is connected with the short inserting pipe and the long inserting pipe of each inserting double pipe through the connecting holes; the short insertion tube is inserted into the short matching tube, and the long insertion tube is inserted into the long liquid storage tube;
after the short insertion tube and the long insertion tube are inserted into the short matching tube and the long liquid storage tube, the short matching tube is communicated with the long liquid storage tube;
the diameter of the pipe joint of the second piston rod is smaller than that of the transverse main pipe, and the diameter of the third sealing ring of the pipe orifice is not smaller than that of the transverse main pipe; the first reagent tube, the second reagent tube and the third reagent tube are all arranged in a mode that the short matching tube is arranged on the left side, and the long liquid storage tube is arranged on the right side, and the connecting hole of the short inserting tube and the transverse main tube is arranged on the left side of the connecting hole of the long inserting tube and the transverse main tube.
2. The POCT consumable of claim 1, wherein the first reagent tube is pre-filled with an inhibitor, the second reagent tube is pre-filled with a lysis agent, and the third reagent tube is pre-filled with a PCR reaction solution.
3. The POCT consumable according to claim 1 or 2, wherein the vertical main tube, the horizontal main tube, the first insertion double tube, the second insertion double tube, and the third insertion double tube are integrally formed.
4. The POCT consumable of claim 1 or 2, wherein the first, second and third reagent tubes are identical in structure.
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| CN202110463483.9A CN112877193B (en) | 2021-04-28 | 2021-04-28 | POCT consumable and detection method |
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| US5961472A (en) * | 1997-09-26 | 1999-10-05 | Baxter International Inc. | Closed, one-handed blood sampling system |
| CN103994901B (en) * | 2014-03-18 | 2016-02-10 | 浙江省海洋水产研究所 | A kind of safety-type sampling tool case |
| GB201615320D0 (en) * | 2016-09-09 | 2016-10-26 | Invitron Ltd | Point of care platform test |
| CN109370896A (en) * | 2018-11-21 | 2019-02-22 | 南京中科拜尔医学技术有限公司 | Cartridge is used in a kind of closed four-way multiple fluorescence PCR detection |
| CN211262857U (en) * | 2019-12-24 | 2020-08-14 | 贵州医科大学附属医院 | Endocrine branch of academic or vocational study secretion sampling device |
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Address after: 215000 rooms 101 and 201, C7 building, biomedical industrial park, 218 Xinghu street, Suzhou Industrial Park, Jiangsu Province Patentee after: Suzhou Yarui Biotechnology Co.,Ltd. Address before: Room 1-2f, C7 / F, bio nanotechnology Park, 218 Xinghu street, Suzhou Industrial Park, Jiangsu Province, 215213 Patentee before: SUZHOU MOLARRAY BIOTECHNOLOGY Co.,Ltd. |
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