CN113846004A - Rotary type nucleic acid amplification product detection device and detection method - Google Patents
Rotary type nucleic acid amplification product detection device and detection method Download PDFInfo
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- CN113846004A CN113846004A CN202111277323.1A CN202111277323A CN113846004A CN 113846004 A CN113846004 A CN 113846004A CN 202111277323 A CN202111277323 A CN 202111277323A CN 113846004 A CN113846004 A CN 113846004A
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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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- 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
Abstract
The invention discloses a rotary nucleic acid amplification product detection device and a detection method, wherein when detection is carried out, a nucleic acid sample and an amplification reagent are added into an amplification part, then the amplification part is connected with a dilution part, and an amplification sample is obtained after reaction is finished; when the diluting part rotates to a preset first angle relative to the amplifying part, the first opening is communicated with the second opening, so that a diluting solution in the diluting part is mixed with the amplified sample after amplification, dilution is completed, and a diluted sample is obtained, wherein the diluting solution is stored in the diluting cavity before the diluting part rotates to the preset first angle; and then, connecting the diluting part with the detecting part, so that the diluting part rotates to a preset second angle relative to the detecting part, at the moment, the sealing plate does not block the third opening any more, the third opening can be communicated with the fifth opening, and the diluted sample flows into the detecting cavity and reacts with the detecting unit to obtain a detection result. In the detection process, the tightness is high, the safety is high, an additional sealing device is not required, and the operation is simple and convenient.
Description
Technical Field
The invention relates to the technical field of nucleic acid amplification detection, in particular to a rotary type nucleic acid amplification product detection device and a detection method.
Background
In recent years, with the rapid development of molecular biology techniques, diagnostic methods based on nucleic acid detection have been widely established and widely applied to laboratory detection of human diseases, isothermal amplification technology is one of them, and isothermal amplification has the advantages of rapidness, high efficiency and specificity compared with other nucleic acid amplification technologies and does not need special equipment, so that it is considered by many scholars as a detection method possibly comparable to PCR once it appears. 2019-when the new coronavirus is in a big outbreak, the PCR molecular diagnosis technology is set as the gold standard for detection, but the requirement on a thermal cycler is high, and the detection in the field and the primary hospitals is not facilitated. Thus, scientists have attempted to use alternative exponential amplification techniques, such as loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification, hyperbranched rolling circle amplification, exponential amplification reactions, and exponential strand displacement amplification, among others. The exponential amplification technology reacts under the constant temperature condition without thermal cycle, thereby not only expanding the application scene of the nucleic acid detection technology, but also exciting the potential of POCT application. The molecular diagnosis has the characteristics of high sensitivity and strong specificity, and if the advantages of quick POCT are further integrated, the molecular diagnosis inevitably occupies an important position in the detection field.
POCT refers to a detection mode which is carried out on a sampling site and utilizes a portable analysis instrument and a matched reagent to quickly obtain a detection result. POCT meaning can be understood from two aspects: spatially, tests performed at the patient's side, i.e. "bedside tests"; temporally, a "point-of-care" test can be performed.
The nucleic acid amplification product produced based on the constant temperature amplification technology can perform single molecule nucleic acid detection at a constant and low temperature within 30 min. The current nucleic acid amplification products in China have low requirements on environment and hardware facilities, and have good application in the aspects of biological protection, water body inspection, food inspection, medical diagnosis, microfluid, veterinarian and the like; however, the current nucleic acid amplification products consider the problems of application scenes and cost, and the colloidal gold test strip is used for terminal detection, so that the integration of amplification and test strip detection is realized, the general scene requirements are met, and the cost is reduced, but the product has the defects when being applied to the nucleic acid detection of the new coronavirus: the amplified product needs to be uncovered by a worker, and the worker puts the product into a corresponding portable instrument, so that false positives of detection can be increased, and a laboratory can be polluted.
Namely, the nucleic acid amplification products in the prior art have the problems of complex operation and low safety.
Disclosure of Invention
The invention aims to provide a rotary type nucleic acid amplification product detection device and a detection method, which are used for solving the problems of complicated operation and low safety of nucleic acid amplification products in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
a rotary nucleic acid amplification product detection device comprises an amplification part, a dilution part and a detection part, wherein two ends of the dilution part are respectively detachably connected with the amplification part and the detection part, and the dilution part can respectively rotate relative to the amplification part and the detection part;
an amplification cavity is formed in the amplification part, a first opening is formed in the cavity wall of the amplification cavity, a dilution cavity is formed in the dilution part, and a second opening is formed in the cavity wall of the dilution cavity; when the dilution part rotates to a preset first angle relative to the amplification part, the first opening is communicated with the second opening;
a detection cavity is formed in the detection part, and a detection unit is arranged in the detection cavity; a fifth opening is formed in the cavity wall of the detection cavity, and a third opening is formed in the cavity wall of the dilution cavity; when the dilution part rotates to a preset second angle relative to the detection part, the third opening is communicated with the fifth opening;
the diluting part is further connected with a sealing plate, and the sealing plate is used for sealing the third opening before the diluting part rotates to the second angle.
Optionally, the sealing plate is disposed outside the dilution cavity and rotatably connected to the dilution part, and a fourth opening is formed in the sealing plate;
when the sealing plate rotates to a preset third angle relative to the dilution part, the third opening is communicated with the fourth opening.
Optionally, be provided with first spacing portion on the shrouding, the detection portion still includes the spacing portion of second, first spacing portion with spacing portion detachably connects of second.
Optionally, the first limiting part comprises a limiting hole formed in the sealing plate, and the second limiting part comprises a limiting block protruding from the detection part; when the diluting part is connected with the detecting part, the limiting block is inserted into the limiting hole.
Optionally, the detection part further includes a surrounding part surrounding the fifth opening, and an installation groove is formed on an inner side of the surrounding part, outside the detection cavity;
when the diluting part is connected with the detecting part, one end of the diluting part and the sealing plate are inserted in the mounting groove.
Optionally, an internal thread is formed on a groove wall of the mounting groove, an external thread is formed on an outer wall of the diluting portion, and the diluting portion is in threaded connection with the mounting groove.
Optionally, the detection portion includes an upper casing and a lower casing, the upper casing and the lower casing are in butt-joint surrounding arrangement to form the detection cavity, and the fifth opening is opened on the wall surface of the upper casing.
Optionally, a baffle portion configured with an avoidance port is arranged between the upper shell and the lower shell, and the baffle portion separates the detection cavity to form a buffer pool and a detection pool; the fifth opening is formed in the cavity wall of the buffer pool;
the detection unit comprises detection test paper, the detection test paper is arranged in the detection pool, and one end of the detection test paper penetrates through the avoidance port and extends into the detection pool.
Optionally, the detection portion further includes a drainage portion, one end of the drainage portion is connected to the fifth opening, and the other end of the drainage portion is connected to the wall surface of the lower casing facing the upper casing.
A detection method applied to the rotary nucleic acid amplification product detection device comprises the following steps:
adding a nucleic acid sample and an amplification reagent into the amplification part, and connecting the amplification part and the dilution part to form a reaction device;
carrying out temperature rise treatment on the amplification part according to a PCR program, and carrying out PCR reaction to obtain an amplification sample;
rotating the diluting part to a preset first angle relative to the amplifying part, wherein the first opening is communicated with the second opening, and the amplified sample is mixed with a diluting solution to obtain a diluted sample;
and connecting the diluting part with the detecting part, enabling the diluting part to rotate to a preset second angle relative to the detecting part, enabling the sealing plate not to block the third opening, enabling the third opening to be communicated with the fifth opening, and enabling the diluted sample to flow into the detecting part to finish detection.
Compared with the prior art, the invention has the following beneficial effects:
when the rotary nucleic acid amplification product detection device provided by the invention is used for detection, a nucleic acid sample and an amplification reagent are added into an amplification part, then the amplification part is connected with a dilution part, and an amplification sample is obtained after the reaction is finished; when the diluting part rotates to a preset first angle relative to the amplifying part, the first opening is communicated with the second opening, so that a diluting solution in the diluting part is mixed with the amplified sample after amplification, dilution is completed, and a diluted sample is obtained, wherein the diluting solution is stored in the diluting cavity before the diluting part rotates to the preset first angle; and then, connecting the diluting part with the detecting part, so that the diluting part rotates to a preset second angle relative to the detecting part, at the moment, the sealing plate does not block the third opening any more, the third opening can be communicated with the fifth opening, and the diluted sample flows into the detecting cavity and reacts with the detecting unit to obtain a detection result. In the detection process, the operation of uncovering and the like which possibly have pollution risks is not needed, the sample is not contacted with the outside in the whole detection process, the safety is high, meanwhile, the rotary type nucleic acid amplification product detection device is high in self sealing performance, a sealing device is not needed to be additionally arranged, the detection of the nucleic acid sample can be completed by assembling all parts and adjusting the angle, and the operation is simple and convenient.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without inventive exercise.
The structure, proportion, size and the like shown in the drawings are only used for matching with the content disclosed in the specification, so that the person skilled in the art can understand and read the description, and the description is not used for limiting the limit condition of the implementation of the invention, so the method has no technical essence, and any structural modification, proportion relation change or size adjustment still falls within the scope of the technical content disclosed by the invention without affecting the effect and the achievable purpose of the invention.
FIG. 1 is a schematic view of the overall structure of a rotary nucleic acid amplification product detection apparatus according to an embodiment of the present invention;
FIG. 2 is a schematic bottom view of an amplification part according to an embodiment of the present invention;
FIG. 3 is a schematic top view of a dilution unit according to an embodiment of the present invention;
FIG. 4 is a first bottom view of the dilution part according to the embodiment of the present invention;
FIG. 5 is a schematic diagram of a first cross-sectional structure of a detecting portion according to an embodiment of the present invention;
FIG. 6 is an enlarged partial schematic view of FIG. 5 at A;
FIG. 7 is a schematic top view of a detecting portion according to an embodiment of the present invention;
FIG. 8 is a second bottom view of the dilution part according to the embodiment of the present invention;
FIG. 9 is a schematic diagram of a second cross-sectional structure of the detecting portion according to the embodiment of the present invention;
fig. 10 is a schematic diagram of a third cross-sectional structure of the detecting portion according to the embodiment of the invention.
Illustration of the drawings: 1. an amplification unit; 2. a dilution unit; 3. a detection unit; 31. a second limiting part; 32. a surrounding portion; 33. an upper housing; 34. a lower housing; 35. a baffle portion; 36. a buffer pool; 37. a detection cell; 38. a drainage part; 41. a first opening; 42. a second opening; 43. a third opening; 44. a fourth opening; 45. a fifth opening; 5. closing the plate; 51. a first limiting part; 6. detecting test paper; 71. a first seal ring; 72. and a second seal ring.
Detailed Description
In order to make the objects, features and advantages of the present invention more obvious and understandable, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the embodiments described below are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention. It should be noted that when one component is referred to as being "connected" to another component, it can be directly connected to the other component or intervening components may also be present.
The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings.
Referring to fig. 1 to 10, fig. 1 is a schematic overall structure diagram of a rotary nucleic acid amplification product detection apparatus according to an embodiment of the present invention, fig. 2 is a schematic bottom structure diagram of an amplification part according to an embodiment of the present invention, fig. 3 is a schematic top structure diagram of a dilution part according to an embodiment of the present invention, fig. 4 is a schematic bottom structure diagram of a dilution part according to an embodiment of the present invention, fig. 5 is a schematic first cross-sectional structure diagram of a detection part according to an embodiment of the present invention, fig. 6 is a schematic partial enlarged view of fig. 5 at a, fig. 7 is a schematic top structure diagram of a detection part according to an embodiment of the present invention, fig. 8 is a schematic bottom structure diagram of a dilution part according to an embodiment of the present invention, fig. 9 is a schematic cross-sectional second structure diagram of a detection part according to an embodiment of the present invention, and fig. 10 is a schematic cross-sectional third structure diagram of a detection part according to an embodiment of the present invention.
Example one
The rotary nucleic acid amplification product detection device provided by the embodiment adopts a constant-temperature nucleic acid amplification technology, can amplify, dilute and detect a nucleic acid product in a normal-temperature environment, and reduces the risk of pollutant diffusion and simplifies the operation steps by improving the detection device.
As shown in FIGS. 1 to 7, the rotary nucleic acid amplification product detection apparatus of the present embodiment includes an amplification part 1, a dilution part 2, and a detection part 3, wherein both ends of the dilution part 2 are detachably connected to the amplification part 1 and the detection part 3, respectively, and the dilution part 2 is rotatable with respect to the amplification part 1 and the detection part 3, respectively. Wherein, the detection part 3 is provided with an observation window which is made of transparent material, the observation window is hermetically connected with the detection part 3, and the detection result of the detection unit can be observed through the observation window.
An amplification cavity is formed in the amplification part 1, a first opening 41 is formed in the cavity wall of the amplification cavity, a dilution cavity is formed in the dilution part 2, and a second opening 42 is formed in the cavity wall of the dilution cavity; when the dilution part 2 is rotated to a predetermined first angle with respect to the amplification part 1, the first opening 41 communicates with the second opening 42. Wherein, there is the diluent in the dilution cavity, and this diluent can carry out the dilution to the amplification sample. Wherein, the first opening 41 is opened on the bottom cavity wall of the amplification cavity, the second opening 42 is opened on the top cavity wall of the dilution cavity, when the dilution part 2 is not at the first angle, the first opening 41 is blocked by the dilution part 2, the second opening 42 is blocked by the amplification part 1, and the respective contents of the amplification part 1 and the dilution part 2 can not contact.
A detection cavity is formed in the detection part 3, and a detection unit is arranged in the detection cavity; a fifth opening 45 is formed in the wall of the detection cavity, and a third opening 43 is formed in the wall of the dilution cavity; when the dilution part 2 is rotated to a preset second angle with respect to the detection part 3, the third opening 43 communicates with the fifth opening 45.
A closing plate 5 is further connected to the dilution part 2, the closing plate 5 being adapted to close off the third opening 43 before the dilution part 2 is rotated to the second angle.
Specifically, when detection is performed, a nucleic acid sample and an amplification reagent are added into the amplification part 1, then the amplification part 1 is connected with the dilution part 2, and an amplification sample is obtained after reaction is finished; when the diluting part 2 rotates to a preset first angle relative to the amplifying part 1 (either the amplifying part 1 or the diluting part 2 is rotated, or the diluting part 2 or the amplifying part 1 is rotated), the first opening 41 is communicated with the second opening 42, so that the diluent in the diluting part 2 is mixed with the amplified sample after amplification, dilution is completed, and a diluted sample is obtained, wherein the diluent is stored in a diluting cavity before the diluting part 2 rotates to the preset first angle; then, the diluting part 2 is connected to the detecting part 3, so that the diluting part 2 rotates to a preset second angle relative to the detecting part 3, at this time, the sealing plate 5 no longer blocks the third opening 43, the third opening 43 can communicate with the fifth opening 45, and the diluted sample flows into the detection cavity and reacts with the detection unit to obtain a detection result. In the detection process, the operation of uncovering and the like which possibly have pollution risks is not needed, the sample is not contacted with the outside in the whole detection process, the safety is high, meanwhile, the rotary type nucleic acid amplification product detection device is high in self sealing performance, a sealing device is not needed to be additionally arranged, the detection of the nucleic acid sample can be completed by assembling all parts and adjusting the angle, and the operation is simple and convenient.
It is necessary to supplement that the side walls of the amplification part 1 and the dilution part 2 are both marked with marks for indicating the relative positions of the first opening 41 and the second opening 42, so that an operator can rotate the amplification part 1 or the dilution part 2 to a proper position to make the first opening communicate with the second opening 41 and the second opening 42.
Further, as shown in fig. 2 to 4, a sealing plate 5 is disposed outside the dilution cavity and rotatably connected to the dilution unit 2, and a fourth opening 44 is formed on the sealing plate 5.
When the closing plate 5 is rotated to a predetermined third angle with respect to the dilution part 2, the third opening 43 communicates with the fourth opening 44.
Illustratively, the diluent is added to the dilution chamber before it is mixed with the amplified sample, and the specific addition process is as follows: placing the side, provided with the sealing plate 5, of the dilution part 2 upward, when the sealing plate 5 rotates to a preset third angle relative to the dilution part 2, the third opening 43 is communicated with the fourth opening 44, at the moment, diluent can be added into the dilution cavity from the third opening 43 and the fourth opening 44 in a mode of injection from top to bottom, and after the diluent is added, the sealing plate 5 is used for plugging the third opening 43 again to seal the dilution cavity; obviously, this step can be performed after the nucleic acid sample is added into the amplification part 1, or before the nucleic acid sample is added into the amplification part 1, after the nucleic acid sample is added into the amplification part 1, the dilution part 2 is connected with the amplification part 1, at this time, the operator shakes the amplification part 1 evenly and heats it according to the PCR program, after obtaining the amplified sample, then the above-mentioned steps are performed to add the diluent into the dilution cavity; and then, the dilution part 2 is rotated to a preset first angle relative to the amplification part 1, so that the dilution liquid is mixed with the amplified sample to obtain the diluted sample, the process reduces the risk of dilution liquid leakage, and the safety of the rotary nucleic acid amplification product detection device is improved.
Further, the closing plate 5 is provided with a first limiting portion 51, the detection portion 3 further includes a second limiting portion 31, and the first limiting portion 51 is detachably connected with the second limiting portion 31. After the dilution part 2 is connected to the detection part 3, the second limiting part 31 is connected to the first limiting part 51, and when the dilution part 2 is rotated to a preset second angle relative to the detection part 3, the dilution part 2 is rotated relative to the sealing plate 5, so that the fourth opening 44 is communicated with the fifth opening 45, and at this time, the diluted sample can enter the detection part 3.
In a specific embodiment, as shown in fig. 2 to 7, the first position-limiting portion 51 includes a position-limiting hole formed on the closing plate 5, and the second position-limiting portion 31 includes a position-limiting block protruding from the detecting portion 3; when the diluting part 2 is connected with the detecting part 3, the limiting block is inserted in the limiting hole. Wherein, the quantity of spacing hole is one, sets up in the inner circle of shrouding 5, and the quantity of stopper is one.
In another alternative embodiment, as shown in fig. 8 to 10, the first position-limiting part 51 includes a position-limiting groove formed on the edge of the closing plate 5, and the second position-limiting part 31 includes a position-limiting block protruding from the detecting part 3; when the diluting part 2 is connected with the detecting part 3, the limiting block is abutted against the groove wall of the limiting groove. Wherein, the quantity of spacing groove is one, and the quantity of stopper is one. It should be noted that, when the limiting groove is formed on the edge of the closing plate 5, the operation difficulty can be reduced, and it is convenient for an operator to connect the first limiting portion 51 with the second limiting portion 31.
It should be understood that, the above mentioned limiting holes and limiting blocks, limiting grooves and limiting blocks are arranged in pairs, the specific number is not limited, and a plurality of groups of limiting holes and limiting blocks or a plurality of groups of limiting grooves and limiting blocks can be added according to the requirement.
Further, outside the detection cavity, the detection part 3 further includes a surrounding part 32 surrounding the fifth opening 45, and an installation groove is formed inside the surrounding part 32.
When the diluting part 2 is connected to the detecting part 3, one end of the diluting part 2 and the sealing plate 5 are inserted into the mounting groove. Wherein, the area of shrouding 5 is less than the tank bottom area of this mounting groove for shrouding 5 can insert in the mounting groove, should play around the portion 32 and increase the leakproofness effect between dilution portion 2 and the detection portion 3.
In a specific embodiment, an internal thread is formed on the wall of the mounting groove, an external thread is formed on the outer wall of the diluting part 2, and the diluting part 2 is screwed with the mounting groove. Illustratively, the sealing plate 5 is embedded in the bottom of the diluting part 2, so that the sealing plate 5 is not rotated when the diluting part 2 is screwed to the wall of the mounting groove, i.e. the inner side of the surrounding part 32. After diluting the rotatory back of going into the certain distance in the mounting groove of calorie of portion 2, first spacing portion 51 is connected with spacing portion 31 of second, and at this moment, shrouding 5 is fixed, and is rotatory for diluting portion 2, and the intercommunication is realized with fourth opening 44 to the final third opening 43, makes and dilutes the sample and flow into in the detection cavity. In the above-described mounting process, the connection between the diluting part 2 and the detecting part 3 and the communication between the third opening 43 and the fourth opening 44 are combined in one step, and the complexity of the operation is reduced. Further, a sealing ring may be added to the inner threaded side of the mounting groove to enhance the sealing property between the detecting part 3 and the diluting part 2.
Preferably, the bottom of mounting groove also can set up a plurality of spacing portion to prevent that the bottom of diluting portion 2 from hugging closely with the bottom of mounting groove, resulting in that the content in diluting portion can't normally flow out.
In another alternative embodiment, as shown in fig. 10, a second sealing ring 72 is disposed at the top end of the surrounding portion 32, when the diluting portion 2 is connected to the detecting portion 3, the volume of the second sealing ring 72 is reduced under the pressure, so that the diluting portion 2 is connected to the detecting portion 3 in a sealing manner, and the relative position between the diluting portion 2 and the detecting portion 3 is fixed, but the diluting portion 2 can still rotate at this time, so that the sealing plate 5 can rotate relative to the diluting portion 2 by the limiting action of the first limiting portion 51 and the second limiting portion 31 on the sealing plate 5, and the third opening 43 is communicated with the fourth opening 44.
In addition to the above embodiment, the detection unit 3 includes the upper case 33 and the lower case 34, the upper case 33 and the lower case 34 are disposed to surround each other to form a detection chamber, and the fifth opening 45 is provided on the wall surface of the upper case 33. Wherein, go up casing 33 and casing 34 down for sealing connection, specifically can select to use connecting methods such as welding, cementing, also can increase the sealing washer in the junction of last casing 33 and casing 34 down simultaneously to increase leakproofness.
Further, a baffle part 35 provided with an avoidance port is arranged between the upper shell 33 and the lower shell 34, and the baffle part 35 separates the detection cavity into a buffer pool 36 and a detection pool 37; the fifth opening 45 opens on the chamber wall of the buffer tank 36.
The detecting unit comprises a detecting test paper 6, the detecting test paper 6 is arranged in the detecting pool 37, and one end of the detecting test paper 6 penetrates through the avoiding port and extends into the detecting pool 37. It should be added that the detecting unit may further include a test tube filled with a detecting liquid, and a one-way valve is disposed between the test tube and the detecting cell 37, so that the diluted sample can enter the detecting cell 37 in a one-way manner after entering the buffer cell 36. Wherein, can should dodge mouthful department and set up the sealing washer, the space outside this sealing washer fills test paper 6, but does not oppress test paper 6, ensures test paper 6's detection function.
Further, the detecting part 3 further includes a drainage part 38, one end of the drainage part 38 is connected to the fifth opening 45, and the other end of the drainage part 38 is connected to the wall surface of the lower casing 34 facing the upper casing 33. Specifically, the drainage portion 38 is made of a water-absorbing material such as sponge or cotton. The drainage part 38 is equivalent to a capacity pool, on one hand, the drainage part enables the sample to be diluted to move directionally through water absorption capacity, so that the diluted sample is prevented from leaking, and on the other hand, the drainage part can regulate and control the speed of the diluted sample reaching the buffer pool 36, so that the phenomenon of sample flushing is prevented; and dilution portion 2 is for encapsulated situation when being connected with detection portion 3, can overcome the pressure differential effect of inside and constantly absorb and dilute the sample, with in its drainage to buffer 36. It should be added that the diluted sample is double buffered by the drainage portion 38 and the buffer pool 36, so as to further prevent the sample washing phenomenon.
Further, as shown in fig. 6, one end of the drainage portion 38 protrudes from the fifth opening 45, and is sleeved with a first sealing ring 71; the first seal ring 71 is fitted to the upper housing 33. In addition, the second stopper portion 31 is provided between the drainage portion 38 and the surrounding portion 32.
It should be added that a sealing ring is provided at the junction of the amplification part 1 and the dilution part 2, and the sealing ring improves the sealing property when the amplification sample and the dilution solution are mixed.
In summary, the rotary nucleic acid amplification product detection device provided by the embodiment has the advantages of simple operation and high safety, and also has the advantages of strong sealing performance, sample flushing prevention, high stability and the like.
Example two
This embodiment describes a detection method applied to the rotary nucleic acid amplification product detection device in the first embodiment to facilitate understanding of the principle of the detection device, the detection method comprising the following steps:
s110, adding a nucleic acid sample and an amplification reagent into an amplification cavity of the amplification part 1, and connecting the amplification part 1 and the dilution part 2 to form a reaction device;
s120, inverting the reaction device, rotating the sealing plate 5, communicating the third opening 43 with the fourth opening 44, and adding a diluent into the diluting part 2; the closing plate 5 is rotated, and the third opening 43 and the fourth opening 44 are staggered;
s130, carrying out temperature rise treatment on the amplification part 1 according to a PCR program, and carrying out PCR reaction to obtain an amplification sample;
s140, rotating the dilution part 2 to a preset first angle relative to the amplification part 1, communicating the first opening 41 with the second opening 42, and mixing the amplified sample with a diluent to obtain a diluted sample;
s150, the diluting part 2 is connected to the detecting part 3, and when the diluting part 2 is rotated to a predetermined second angle with respect to the detecting part 3, the sealing plate 5 does not close the third opening 443, the third opening 43 communicates with the fifth opening 45, and the diluted sample flows into the detecting part 3, thereby completing the detection.
Finally, the diluted sample is brought into contact with the test paper 6 in the detection section 3, and the operator can observe the reaction result through the observation window.
In conclusion, the detection method provided by the embodiment has the advantages of simplicity in operation, high safety, high sealing performance, sample flushing prevention, high stability and the like.
The above-mentioned embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. A rotary nucleic acid amplification product detection device comprising an amplification part (1), a dilution part (2) and a detection part (3), wherein both ends of the dilution part (2) are detachably connected to the amplification part (1) and the detection part (3), respectively, and the dilution part (2) is rotatable relative to the amplification part (1) and the detection part (3), respectively;
an amplification cavity is formed in the amplification part (1), a first opening (41) is formed in the cavity wall of the amplification cavity, a dilution cavity is formed in the dilution part (2), and a second opening (42) is formed in the cavity wall of the dilution cavity; when the dilution part (2) rotates to a preset first angle relative to the amplification part (1), the first opening (41) is communicated with the second opening (42);
a detection cavity is formed in the detection part (3), and a detection unit is arranged in the detection cavity; a fifth opening (45) is formed in the cavity wall of the detection cavity, and a third opening (43) is formed in the cavity wall of the dilution cavity; when the dilution part (2) rotates to a preset second angle relative to the detection part (3), the third opening (43) is communicated with the fifth opening (45);
dilution portion (2) still are connected with shrouding (5), shrouding (5) are used for diluting portion (2) and rotate to before the second angle block third opening (43).
2. The rotary nucleic acid amplification product detection apparatus according to claim 1, wherein the sealing plate (5) is disposed outside the dilution chamber and rotatably connected to the dilution unit (2), and a fourth opening (44) is formed in the sealing plate (5);
when the closing plate (5) is rotated to a preset third angle relative to the dilution part (2), the third opening (43) communicates with the fourth opening (44).
3. The rotary nucleic acid amplification product detection apparatus according to claim 2, wherein the sealing plate (5) is provided with a first stopper portion (51), the detection portion (3) further comprises a second stopper portion (31), and the first stopper portion (51) and the second stopper portion (31) are detachably connected.
4. The rotary nucleic acid amplification product detection apparatus according to claim 3, wherein the first stopper portion (51) comprises a stopper hole provided in the sealing plate (5), and the second stopper portion (31) comprises a stopper protruding from the detection portion (3); when the diluting part (2) is connected with the detecting part (3), the limiting block is inserted into the limiting hole.
5. The rotary nucleic acid amplification product detection apparatus according to claim 3, wherein the detection section (3) further comprises a surrounding section (32) disposed around the fifth opening (45) outside the detection chamber, and an installation groove is formed inside the surrounding section (32);
when the diluting part (2) is connected with the detecting part (3), one end of the diluting part (2) and the sealing plate (5) are inserted into the mounting groove.
6. The rotary nucleic acid amplification product detection apparatus according to claim 5, wherein a groove wall of the installation groove is formed with an internal thread, an outer wall of the dilution part (2) is formed with an external thread, and the dilution part (2) is screwed into the installation groove.
7. The rotary nucleic acid amplification product detection apparatus according to any one of claims 1 to 6, wherein the detection section (3) comprises an upper casing (33) and a lower casing (34), the upper casing (33) and the lower casing (34) are arranged in an abutting-surrounding manner to form the detection chamber, and the fifth opening (45) is provided in a wall surface of the upper casing (33).
8. The rotary nucleic acid amplification product detection apparatus according to claim 7, wherein a baffle portion (35) having an avoidance opening is provided between the upper casing (33) and the lower casing (34), and the baffle portion (35) separates the detection chamber to form a buffer chamber (36) and a detection chamber (37); the fifth opening (45) is formed in the cavity wall of the buffer pool (36);
the detection unit comprises detection test paper (6), the detection test paper (6) is arranged in the detection pool (37), and one end of the detection test paper (6) penetrates through the avoidance port and extends into the detection pool (37).
9. The rotary nucleic acid amplification product detection apparatus according to claim 7, wherein the detection section (3) further comprises a drain section (38), one end of the drain section (38) is connected to the fifth opening (45), and the other end of the drain section (38) is connected to a wall surface of the lower casing (34) facing the upper casing (33).
10. A detection method applied to the rotary nucleic acid amplification product detection apparatus according to any one of claims 1 to 9, comprising:
adding a nucleic acid sample and an amplification reagent into the amplification part, and connecting the amplification part and the dilution part to form a reaction device;
carrying out temperature rise treatment on the amplification part according to a PCR program, and carrying out PCR reaction to obtain an amplification sample;
rotating the diluting part to a preset first angle relative to the amplifying part, wherein the first opening is communicated with the second opening, and the amplified sample is mixed with a diluting solution to obtain a diluted sample;
and connecting the diluting part with the detecting part, enabling the diluting part to rotate to a preset second angle relative to the detecting part, enabling the sealing plate not to block the third opening, enabling the third opening to be communicated with the fifth opening, and enabling the diluted sample to flow into the detecting part to finish detection.
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