CN107653301A - A kind of hermetic multiple fluorescent PCR detection cartridge - Google Patents
A kind of hermetic multiple fluorescent PCR detection cartridge Download PDFInfo
- Publication number
- CN107653301A CN107653301A CN201710960228.9A CN201710960228A CN107653301A CN 107653301 A CN107653301 A CN 107653301A CN 201710960228 A CN201710960228 A CN 201710960228A CN 107653301 A CN107653301 A CN 107653301A
- Authority
- CN
- China
- Prior art keywords
- axis
- suction nozzle
- pcr detection
- base
- filter core
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
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Classifications
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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]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
-
- 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
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
Abstract
The invention discloses a kind of hermetic multiple fluorescent PCR detection cartridge, including upper lid, housing, axis, filter core suction nozzle and base, upper lid, housing and base form to form a closing space, the centre bore that axis upper end passes lid is connected with drive mechanism, in the centre bore of lower end inserted base, it is connected in the middle part of axis by transverse arm with filter core suction nozzle, is provided with spacing collar on the axis between upper lid and transverse arm, spring is provided with the axis between base and transverse arm；Several PCR pipes and several reagent troughs are provided with base；The filter core suction nozzle is connected with aspirator.The hermetic multiple fluorescent PCR detection cartridge of the present invention can be by two steps of paramagnetic particle method nucleic acid extraction and fluorescence quantitative PCR detection, completed in a small closed box body, detected available for the nucleic acid extraction under no molecular diagnostic laboratories environment and fluorescent PCR, have the advantages that it is compact-sized, easily operated, simple in construction, easily assemble, effectively improve the environment fitness of molecule diagnostics classes product.
Description
Technical field
The present invention relates to a kind of extraction of biotinylated nucleic acid molecule and detection cartridge, specially a kind of hermetic multiple fluorescence
PCR detection cartridges.
Background technology
Nucleic acid extraction and detection are an important process of molecule diagnosis, but in the extraction of nucleic acid and detection process,
Aerosol Pollution is also easy to produce, result false positive is occurred, the diagnosis of conventional molecule is needed in specific molecular diagnostic laboratories
Carry out, seriously constrain the application and popularization of this technology.
The content of the invention
The technical problem to be solved in the present invention is to overcome existing routine molecular diagnostics to need to diagnose in fact in specific molecule
Indoor progress is tested, the defects of seriously constraining the application and popularization of this technology, there is provided a kind of enclosed cartridge, can be used for
The research and development of POCT quasi-molecule diagnostic instrments products.
In order to solve the above-mentioned technical problem, the invention provides following technical scheme：
A kind of hermetic multiple fluorescent PCR detection cartridge, including upper lid, housing, axis, filter core suction nozzle and base, on
Lid, housing and base are formed to form a closing space, and the centre bore that axis upper end passes lid is connected with drive mechanism, lower end
In the centre bore of inserted base, it is connected in the middle part of axis by transverse arm with filter core suction nozzle, the junction of axis and upper lid and bottom seat covers
Spacing collar is equipped with, spring is provided with the axis between base and transverse arm；Several PCR pipes and reagent are provided with base
Groove；The filter core suction nozzle is connected with aspirator.
Further, described aspirator is made up of flexible tracheae and syringe.Air-path interface is provided with upper lid,
Lower air-path interface is provided with transverse arm, the flexible tracheae is connected by upper air-path interface and lower air-path interface with filter core suction nozzle.
Preferably, the upper air-path interface both ends use pagoda header structure, are easy to couple and seal with flexible tracheae；
Preferably, the lower air-path interface upper end uses pagoda header structure, is easy to couple and seal with flexible tracheae；Lower end
Using pyramidal structure, and there is sealing ring, can be combined closely with filter core suction nozzle, be not in gas leakage；
Preferably, the axis is using the structure above lower circle, it is preferred to use hard material is made.
Further, the housing is to use cylindrical housings.
Further, reagent trough, PCR pipe, use is transparent, semi-flexible, resistant to elevated temperatures material.
When axis is rotating in the presence of drive mechanism, upper lid and filter core suction nozzle synchronously rotate, meanwhile,
Under axis and spring effect, filter core suction nozzle can also move up and down with axis, so as to complete to different reagent troughs and PCR pipe
Interior liquid is shifted.Because upper lid and filter core suction nozzle synchronously rotate, the space of cartridge can be saved.
Different detection reagents can be added in reagent trough and PCR pipe, rotation central axis, reagent trough are drawn using filter core suction nozzle
Liquid carries out pressure-vaccum mixing to reagent tank liquid, coordinates magnetic material to realize the combination, repeatedly clear of magnetic bead and nucleic acid
Wash, elute, complete nucleic acid extraction and purifying.It can be added to afterwards by extracting with nucleic acid after purification in PCR pipe, addition reagent is simultaneously
Coordinate the hot well of matched fluorescent quantitative PCR detector, available for fluorescence quantitative PCR detection.
The hermetic multiple fluorescent PCR detection cartridge of the present invention can be by paramagnetic particle method nucleic acid extraction and quantitative fluorescent PCR
Two steps are detected, are completed in a small closed box body, available for the nucleic acid under no molecular diagnostic laboratories environment
Extraction and fluorescent PCR detection, have the advantages that it is compact-sized, easily operated, simple in construction, easily assemble, effectively improve molecule and examine
The environment fitness of disconnected class product.
Brief description of the drawings
Accompanying drawing is used for providing a further understanding of the present invention, and a part for constitution instruction, the reality with the present invention
Apply example to be used to explain the present invention together, be not construed as limiting the invention.In the accompanying drawings：
Fig. 1 is the structural representation of the present invention；
Fig. 2 is the part-structure schematic diagram of the present invention.
Embodiment
The preferred embodiments of the present invention are illustrated below in conjunction with accompanying drawing, it will be appreciated that described herein preferred real
Apply example to be merely to illustrate and explain the present invention, be not intended to limit the present invention.
Embodiment
As shown in figure 1, a kind of hermetic multiple fluorescent PCR detection cartridge, including upper lid 1, cylindrical housings 9, axis
13rd, filter core suction nozzle 6 and base 12, upper lid 1, cylindrical housings 9 and base 12, which are formed, forms a closing space, the upper end of axis 13
The centre bore for passing lid 1 is connected with drive mechanism, and in the centre bore of lower end inserted base 12, the middle part of axis 13 passes through transverse arm 5
Be connected with filter core suction nozzle 6, the junction set of axis 13 and upper lid 1 and base 12 is equipped with spacing collar 7, base 12 and transverse arm 5 it
Between axis 13 on be provided with spring 8；6 PCR pipes 10 and reagent trough 11 are provided with base 12；Filter core suction nozzle 6 and flexible air
Pipe 3 connects.Air-path interface 2 is provided with upper lid 1, lower air-path interface 4 is provided with transverse arm 5；Flexible tracheae one end 3 is by upper
Air-path interface 2 and lower air-path interface 4 are connected with filter core suction nozzle 6, and the other end is connected with syringe.
The upper both ends of air-path interface 2 use pagoda header structure, are easy to couple and seal with flexible tracheae；On lower air-path interface 4
End uses pagoda header structure, is easy to couple and seal with flexible tracheae；Lower end uses pyramidal structure, and has sealing ring, Ke Yiyu
Filter core suction nozzle is combined closely, and is not in gas leakage；Axis is using the structure of lower circle top made of hard material, the He of reagent trough 11
PCR pipe 10, it is made of soft exotic material.Upper lid, base, axis, transverse arm, upper air pipe interface, lower air pipe interface and circle
Shape shell is formed using hard transparent plastic injection molding.
The mixed liquor (1000 μ L) of cracking, combination and magnetic bead is placed in reagent trough 1, and adds the μ L of sample to be tested 300, is tried
Place cleaning solution 1 (300 μ L) in agent groove 2, placement cleaning solution 2 (300 μ L) in the hole of reagent trough 3, not liquid feeding in reagent trough 4, reagent
Eluent (100 μ L) is placed in groove 5, mineral oil (1000 μ L) is placed in the hole of reagent trough 6, being placed respectively in PCR pipe to detect
The PCR reaction solutions (40 μ L) of different purpose nucleic acid fragments.It is inserted in spacing collar, transverse arm, spacing collar successively on axis, is led with flexibility
Pipe couples the lower port of upper air pipe interface with the upper port of lower air pipe interface, and filter core suction nozzle is inserted into the lower end of lower air pipe interface
Mouthful, and insert firmly, and do encapsulation process, it is ensured that it is air tight.Cylindrical container is placed on base, the axis worked good first is inserted
Upper lid composition assembly, then the overall loading above circular housing, upper port and the flexible tracheae of syringe of upper gas-tpe fitting
Connection, completes the assembling of whole cartridge.
Body operating process is as follows：
1st, rotation central axis, filter core suction nozzle is directed at reagent trough 1, axis is then pressed downward, by the liquid in suction nozzle patchhole
It is interior, pull syringe, after the liquid in device to hole carries out pressure-vaccum mixing, 10 minutes are stood, the cell cracking in sample to be tested, nucleic acid
It is released in liquid, magnetic bead is combined in the presence of liquid is combined with nucleic acid；
2nd, magnet is placed in the bottom of reagent trough 1, magnetic bead is adsorbed, after absorption, filter core suction nozzle is by liquid in reagent trough 1
Suction out, be transferred in 4 holes, the magnetic bead combined with nucleic acid is retained in reagent trough；
3rd, the cleaning solution 1 in the hole of reagent trough 2 is all added in the hole of reagent trough 1 with filter core suction nozzle, magnet is removed, with filter
Liquid carries out suction and blows mixings in wicking head device to hole, and magnetic bead surfaces are cleaned with reference to except epinucleic other impurities；
4th, after cleaning, magnet is placed on to the bottom of reagent trough 1, magnetic bead is adsorbed, after absorption, filter core suction nozzle will try
Liquid suctions out in agent groove 1, is transferred in reagent trough 2；
5th, the cleaning solution 2 in the hole of reagent trough 3 is all added in reagent trough 1 with filter core suction nozzle, removes magnet, use filter core
Suction nozzle carries out suction and blows mixing, and magnetic bead surfaces are combined except epinucleic other impurities carry out secondary cleaning；
6th, after cleaning, magnet is placed on to the bottom of reagent trough 1, magnetic bead is adsorbed, after absorption, filter core suction nozzle will try
Liquid suctions out in agent groove 1, is transferred in reagent trough 3；
7th, the elution in reagent trough 5 is all added in reagent trough 1 with filter core suction nozzle, removes magnet, with filter core suction nozzle
Liquid carries out suction and blows mixing in device to hole, the Nucleic Acid Elution that magnetic bead surfaces are combined；
8th, after eluting, magnet is placed on to the bottom of reagent trough 1, magnetic bead is adsorbed, after absorption, filter core suction nozzle will try
Liquid suctions out in agent groove 1, is transferred in reagent trough 5, and the liquid in reagent trough 5 contains extraction and nucleic acid after purification, can use
In fluorescence quantitative PCR detection；
9th, using filter core suction nozzle, take the nucleic acid solution of 10 μ L purifying respectively from reagent trough 5, be added to each PCR pipe
Interior (noticing that suction nozzle can not be inserted into the liquid in PCR pipe)；
10 then utilize filter core suction nozzle, take 50 μ L mineral oil respectively from reagent trough 6, be added in each PCR pipe
(noticing that suction nozzle can not be inserted into the liquid in PCR pipe), completes the preparation of PCR reaction systems；
11st, cartridge is positioned in the hot well of matched fluorescent quantitative PCR detector, proceeds by fluorescent quantitation
PCR detection.
This set mechanism can allow suction nozzle in close box body, and the liquid in different reagent troughs and PCR pipe can be shifted.
Cartridge can be used for no molecular diagnostic laboratories environment under nucleic acid extraction and fluorescent PCR detection, have it is compact-sized, be easy to grasp
Make, environment tolerance is high, can detect the advantages that multiple indexs.
Finally it should be noted that：The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention,
Although the present invention is described in detail with reference to the foregoing embodiments, for those skilled in the art, it still may be used
To be modified to the technical scheme described in foregoing embodiments, or equivalent substitution is carried out to which part technical characteristic.
Within the spirit and principles of the invention, any modification, equivalent substitution and improvements made etc., it should be included in the present invention's
Within protection domain.
Claims (7)
1. a kind of hermetic multiple fluorescent PCR detection cartridge, it is characterised in that including upper lid, housing, axis, filter core suction nozzle
And base, upper lid, housing and base form to form a closing space, axis upper end passes the centre bore and drive mechanism of lid
Connect, in the centre bore of lower end inserted base, be connected in the middle part of axis by transverse arm with filter core suction nozzle, axis and upper lid and bottom seat
Junction set is equipped with spacing collar, and spring is provided with the axis between base and transverse arm；Several PCR pipes are provided with base
With several reagent troughs；The filter core suction nozzle is connected with aspirator.
2. hermetic multiple fluorescent PCR detection cartridge as claimed in claim 1, it is characterised in that described aspirator
It is made up of flexible tracheae and syringe.
3. hermetic multiple fluorescent PCR detection cartridge as claimed in claim 2, it is characterised in that be provided with upper lid
Air-path interface, lower air-path interface is provided with transverse arm, the flexible tracheae passes through upper air-path interface and lower air-path interface and filter core
Suction nozzle connects.
4. hermetic multiple fluorescent PCR detection cartridge as claimed in claim 3, it is characterised in that the upper air-path interface
Both ends use pagoda header structure.
5. the hermetic multiple fluorescent PCR detection cartridge as described in claim 3 or 4, it is characterised in that the lower gas circuit connects
Mouth upper end uses pagoda header structure, and lower end uses pyramidal structure.
6. hermetic multiple fluorescent PCR detection cartridge as claimed in claim 1, it is characterised in that under the axis uses
The structure of circle top.
7. hermetic multiple fluorescent PCR detection cartridge as claimed in claim 1, it is characterised in that the housing is with circle
Cylindrical shell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710960228.9A CN107653301A (en) | 2017-10-16 | 2017-10-16 | A kind of hermetic multiple fluorescent PCR detection cartridge |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710960228.9A CN107653301A (en) | 2017-10-16 | 2017-10-16 | A kind of hermetic multiple fluorescent PCR detection cartridge |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107653301A true CN107653301A (en) | 2018-02-02 |
Family
ID=61118670
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710960228.9A Pending CN107653301A (en) | 2017-10-16 | 2017-10-16 | A kind of hermetic multiple fluorescent PCR detection cartridge |
Country Status (1)
| Country | Link |
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| CN (1) | CN107653301A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108828221A (en) * | 2018-04-04 | 2018-11-16 | 美林美邦（厦门）生物科技有限公司 | A kind of sample process being provided with material transfer organization and detection reagent cup box |
| CN110331075A (en) * | 2019-02-02 | 2019-10-15 | 上海思路迪医学检验所有限公司 | Closed sequencing library prepares cartridge |
| CN111378574A (en) * | 2020-03-25 | 2020-07-07 | 苏州新海生物科技股份有限公司 | Nucleic acid extraction and amplification device |
| CN112608813A (en) * | 2020-12-21 | 2021-04-06 | 上海思路迪生物医学科技有限公司 | Multi-degree-of-freedom library preparation card box with external power source and method |
| CN112779117A (en) * | 2019-11-07 | 2021-05-11 | 浙江大学 | Detection card box with isolation effect for nucleic acid detection experiment |
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| CN207435462U (en) * | 2017-10-16 | 2018-06-01 | 南京中科拜尔医学技术有限公司 | A kind of hermetic multiple fluorescent PCR detection cartridge |
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| CN103571740A (en) * | 2013-10-15 | 2014-02-12 | 周国华 | Device integrated with extraction, amplification and detection of nucleic acid |
| CN105802848A (en) * | 2014-11-27 | 2016-07-27 | 江苏奇天基因生物科技有限公司 | Recombinase-mediated isothermal nucleic acid amplification reaction real-time detection apparatus |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108828221A (en) * | 2018-04-04 | 2018-11-16 | 美林美邦（厦门）生物科技有限公司 | A kind of sample process being provided with material transfer organization and detection reagent cup box |
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| CN108828221B (en) * | 2018-04-04 | 2019-11-12 | 美林美邦（厦门）生物科技有限公司 | A kind of sample process being provided with material transfer organization and detection reagent cup box |
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| US20210011046A1 (en) * | 2018-04-04 | 2021-01-14 | Merlin Biomedical (Xiamen) Co., Ltd. | Sample processing and detecting kit with a material transfer structure |
| CN110331075A (en) * | 2019-02-02 | 2019-10-15 | 上海思路迪医学检验所有限公司 | Closed sequencing library prepares cartridge |
| CN112779117A (en) * | 2019-11-07 | 2021-05-11 | 浙江大学 | Detection card box with isolation effect for nucleic acid detection experiment |
| CN112779117B (en) * | 2019-11-07 | 2024-03-12 | 浙江大学 | Detection cartridge with isolation effect for nucleic acid detection experiment |
| CN111378574A (en) * | 2020-03-25 | 2020-07-07 | 苏州新海生物科技股份有限公司 | Nucleic acid extraction and amplification device |
| CN111378574B (en) * | 2020-03-25 | 2023-10-27 | 苏州新海生物科技股份有限公司 | Nucleic acid extraction and amplification device |
| CN112608813A (en) * | 2020-12-21 | 2021-04-06 | 上海思路迪生物医学科技有限公司 | Multi-degree-of-freedom library preparation card box with external power source and method |
| CN112608813B (en) * | 2020-12-21 | 2022-07-01 | 上海思路迪生物医学科技有限公司 | Multi-degree-of-freedom library preparation card box with external power source and method |
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Application publication date: 20180202 |