CN108753589A - A kind of specimen cup for real-time fluorescence quantitative PCR detection - Google Patents
A kind of specimen cup for real-time fluorescence quantitative PCR detection Download PDFInfo
- Publication number
- CN108753589A CN108753589A CN201810580440.7A CN201810580440A CN108753589A CN 108753589 A CN108753589 A CN 108753589A CN 201810580440 A CN201810580440 A CN 201810580440A CN 108753589 A CN108753589 A CN 108753589A
- Authority
- CN
- China
- Prior art keywords
- sample
- note
- channel
- vessel
- injection unit
- 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.)
- Granted
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 33
- 238000003753 real-time PCR Methods 0.000 title claims abstract description 20
- 238000002347 injection Methods 0.000 claims abstract description 72
- 239000007924 injection Substances 0.000 claims abstract description 72
- 238000005070 sampling Methods 0.000 claims description 21
- 238000007789 sealing Methods 0.000 claims description 13
- 230000003287 optical effect Effects 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 abstract description 13
- 239000007787 solid Substances 0.000 abstract description 6
- 239000000084 colloidal system Substances 0.000 abstract description 3
- 238000010586 diagram Methods 0.000 description 9
- 238000005119 centrifugation Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 230000008676 import Effects 0.000 description 3
- 229920002521 macromolecule Polymers 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000004313 glare Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 208000031295 Animal disease Diseases 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000021393 food security Nutrition 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 238000007731 hot pressing Methods 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 208000007578 phototoxic dermatitis Diseases 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920006255 plastic film Polymers 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/54—Labware with identification means
- B01L3/545—Labware with identification means for laboratory containers
- B01L3/5457—Labware with identification means for laboratory containers for container closures
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
Abstract
The application provides a kind of specimen cup for real-time fluorescence quantitative PCR detection, the note sample channel for injecting sample and the exhaust passage for bubble to be discharged are offered between the sample injection unit and sample vessel portion of the specimen cup, exhaust passage is in the port of sample injection unit higher than note sample channel in the port of sample injection unit, to in centrifugal process, enable and escapes sample vessel since the bubble that note sample or other experimentations generate passes through exhaust passage in sample vessel, and, the specimen cup offers the storage trough for collecting deposit in sample vessel bottom, to make these solids and colloid be detached from light path range, reduce influence of the deposit to fluoroscopic examination accuracy.
Description
Technical field
The application belongs to molecular biology experiment device, more particularly to a kind of sample for real-time fluorescence quantitative PCR detection
Product cup.
Background technology
Real-time fluorescence quantitative PCR (Quantitative Real-time PCR) be one kind in DNA amplification reaction, with glimmering
The method that photosensitiveness chemical substance detects product total amount after each PCR (PCR) cycle.Pcr amplification reaction usually wraps
Multiple cycle is included, cycle includes primary heating dehybridization procedure and primary cooling reproduction process every time.Real-time fluorescence quantitative PCR be
During PCR amplification, PCR processes are measured in real time by detecting the fluorescence signal intensity of sample.Real time fluorescent quantitative
Round pcr is obtained for extensive use in fields such as medical treatment, scientific research and commercial Applications, as clinical disease diagnosis, Animal diseases are examined
It surveys, food security and medicine, agriculture and animal husbandry, bio-related molecules biology quantitative study etc., is widely used in gene expression research, turns
Gene studies, the numerous areas such as curative effect of medication examination, pathogen detection.
Currently, detecting the fluorescence signal intensity of PCR samples usually using Multi-channel optical detecting system, specifically, lead to
The light irradiation PCR samples using specific wavelength are crossed, and use the fluorescence of the optical detections means such as photelectric receiver detection PCR samples
Signal, to carry out quantitative analysis to the specific dna sequence in PCR samples.
The structure of common specimen cup is as shown in Figure 1.The specimen cup includes cup body 001 and is covered on the cup body 001
On sample vessel patch 002, be provided with sealing cover 003 and for connecting sealing cover 003 and institute at the top of the cup body 001
The flexible connecting plate 004 of cup body 001 is stated, the two neighboring side in lower part of the cup body 001 is orthogonal, under the cup body 001
Portion offers the sample vessel 005 for holding sample, and the sample vessel 005 is square, 005 lower part of sample vessel
Two side walls thickness it is uniform and equal, 001 top of the cup body is provided with the sample injection unit for installing sample additional
006, the sample injection unit 006 is connected to the sample vessel 005 by noting sample channel 007.The note sample channel 007 is one
Root truncated cone-shaped channel, the one end being relatively large in diameter are connect with sample injection unit 006, and the smaller one end of diameter connects with sample vessel 005
It connects.
Real-time fluorescence quantitative PCR detection is typically that fluid sample is injected sample vessel, is injected from the side of sample vessel
Light with specific wavelength, then the fluorescence that sample is sent out is received from the other side adjacent thereto, to detect PCR processes.But
There are problems for the specimen cup used using existing technology, for example, as shown in Figure 2：After sample is added in specimen cup, sample
Bubble 008 is usually had in vessel, therefore, be often used centrifuge to be added sample after and cover sealing cover specimen cup carry out from
The heart operates, so as to be full of sample bubble removing side by side in sample vessel.In general, after specimen cup is placed in centrifuge, with horizontal plane
In 45 ° or so of angle, if the rotating speed of centrifuge is inadequate, bubble is difficult to exclude totally, even if starting with centrifugation in experiment
Machine is clean by removal of bubbles, also often bubble occurs in sample vessel in heating and cooling process.The presence of bubble will necessarily
The detection of fluorescence signal is influenced, and the accuracy of experimental result can be reduced.
It is more again as shown in figure 3, real-time fluorescence quantitative PCR detection is in the application that clinical diagnosis, animal test quarantine field
Using blood, tissue fluid, epithelial tissue etc. as detection sample, in the application of food safety monitoring, for example, to meat and breast
In the inspection of product, meat, dairy products etc. can also be used to be used as sample.In the pretreatment process of sample, cell is carried out
Break process, to discharge intracellular nucleic acid, broken cell wall and other intracellular solids can remain among sample,
Usually it is in suspended state, the solid and colloid 009 of this big proportion suspend in the sample or be deposited on sample vessel bottom all
The detection of fluorescence signal can be impacted, reduce the accuracy of experiment.
Invention content
The application provides a kind of specimen cup for real-time fluorescence quantitative PCR detection, is injected in the sample of the specimen cup
The note sample channel for injecting sample and the exhaust passage for bubble to be discharged are offered between portion and sample vessel portion, exhaust is logical
Road is higher than note sample channel in the port of sample injection unit, in centrifugal process so that in sample in the port of sample injection unit
Sample vessel is escaped since the bubble that note sample or other experimentations generate can pass through exhaust passage in product vessel, and
And the specimen cup offers the storage trough for collecting deposit in sample vessel bottom, to make these solids and colloid
It is detached from light path range, reduces influence of the deposit to fluoroscopic examination accuracy.
Specimen cup provided by the present application for real-time fluorescence quantitative PCR detection, including：Cup body 1 and sample vessel patch
2, the cup body 1 includes sample injection unit 11 and sample vessel portion 12, and the sample injection unit 11 is tubular, the sample vessel
Portion 12 is plate, and the sample injection unit 11 is seamlessly transitted with sample vessel portion 12, and sample vessel portion 12 includes channel section
121 and sampling section 122, the plate face of the channel section 121 be rectangle, the plate face of the sampling section 122 is triangle, also, institute
The outer diameter for stating sampling section 122 is gradually reduced, and two adjacent outer walls of the sampling section 122 are orthogonal, the sample vessel patch
Piece 2 is covered on respectively in the both sides plate face in sample vessel portion 12；Sample vessel 1221, institute are offered in sample vessel portion 12
It states sample vessel 1221 to be square, the cavity wall of the sample vessel 1221 and the outer wall of the sampling section 122 are mutually parallel, and make
The thickness for obtaining the two side walls 1222 of 1221 lower part of sample vessel is equal；It is offered in the bottom of the sample vessel 1221
Storage trough 123 for storing deposit；Note sample channel 1211 and exhaust passage 1212, institute are offered in the channel section 121
One end of one end and exhaust passage 1212 for stating note sample channel 1211 is connected to sample injection unit 11 respectively, the note sample channel
1211 other end and the other end of exhaust passage 1212 converge at the top of the sample vessel 1221, and hold with the sample
Chamber 1221 is connected to, and port of the exhaust passage 1212 in sample injection unit 11 is higher than note sample channel 1211 in sample injection unit
Port in 11；It is additionally provided with sealing cover 3 at 1 top of cup body and the flexible connecting plate 4 for connecting sealing cover 3 and cup body 1.
In a kind of achievable mode, the storage trough 123 is set to non-detection optical path area.
Specifically, the storage trough 123 is set to the two side walls extended surface of 1221 bottom end of sample vessel and is formed
Region in, the storage trough 123 is oppositely arranged with the sample vessel 1221.
Optionally, the internal diameter of 123 slot bottom of the storage trough is more than the internal diameter of groove top, and the depth of the storage trough 123 is less than
The 2/3 of 1222 thickness of the side wall；And/or the storage trough 123 is opened in the bottom end of the sample vessel 1221 and is less than 1/5
Place.
In another achievable mode, the bottom end of the sample injection unit 11 is stairstepping, the exhaust passage
1212 port is opened on higher-order ladder, and the port in the note sample channel 1211 is opened on relatively low ladder；The higher-order
The top surface of ladder is inclined-plane, and the inclined-plane is less than the side far from note sample channel close to the side in note sample channel；It is logical in the note sample
1211 top of road forms note sample slot 124, and the note sample slot 124 is in infundibulate, the lowermost end and the note sample of the note sample slot 124
Channel 1211 is connected to.
Optionally, the ladder of 11 bottom end of sample injection unit seamlessly transits, and the top surface of higher-order ladder is to exhaust passage
The curved surface of 1212 side indents, forms diversion trench, and diameter of the diversion trench close to 1212 one end of exhaust passage is more than close to note
The diameter of 1211 one end of sample channel, depth of the diversion trench close to 1212 one end of exhaust passage are more than close to note sample channel 1211
The depth of one end.
In another achievable mode, from close to 11 one end of sample injection unit to close to 1221 one end of sample vessel,
The internal diameter of the exhaust passage 1212 is gradually reduced；From close to 11 one end of sample injection unit to close to 1221 one end of sample vessel,
The internal diameter in the note sample channel 1211 is gradually reduced；Internal diameter of the note sample channel 1211 in 1221 one end port of sample vessel is big
In internal diameter of the exhaust passage 1212 in 1221 one end port of sample vessel, the note sample channel 1211 is in sample injection unit 11
Internal diameter of the internal diameter of one end port more than the exhaust passage 1212 in 11 one end port of sample injection unit.
Optionally, the note sample channel 1211 includes note sample leading portion and note sample back segment, and the note sample leading portion is noted close to sample
Enter portion 11, the note sample back segment is more than the slope of note sample back segment close to sample vessel 1221, the slope of the note sample leading portion；It is described
Exhaust passage 1212 includes exhaust leading portion and exhaust back segment, and the exhaust leading portion is close to sample injection unit 11, the exhaust back segment
Close to sample vessel 1221, the slope of the exhaust leading portion is more than the slope of exhaust back segment.
Further, the note sample channel 1211 converges at the top of the sample vessel 1221 with the exhaust passage 1212
End is less than at 1/5；And/or 1211 side of note sample channel is provided with note sample optical filtering slot 126；And/or the exhaust is led to
1212 side of road is both provided with exhaust optical filtering slot 127.
Compared with specimen cup in the prior art, specimen cup provided by the present application has note sample channel and exhaust passage, and
And sample vessel bottom end is provided with storage trough, thus in carrying out real-time fluorescence quantitative PCR detection process, with centrifugally operated
Progress, the bubble being mingled in sample vessel escapes sample vessel by exhaust passage, reduces the quantity of bubble in sample vessel,
Reduce influence of the bubble to light path, improves the accuracy of real-time fluorescence quantitative PCR detection, meanwhile, it settles down in centrifugally operated
Macromolecular solid content be deposited in the storage trough, to avoid the macromolecular solid content from blocking light path, further increase
The accuracy of real-time fluorescence quantitative PCR detection.
Description of the drawings
It in order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, below will be to institute in embodiment
Attached drawing to be used is needed to be briefly described, it should be apparent that, the accompanying drawings in the following description is only some implementations of the present invention
Example, for those of ordinary skill in the art, without creative efforts, can also obtain according to these attached drawings
Obtain other attached drawings.
Fig. 1 shows a kind of structural schematic diagram of specimen cup in the prior art；
Fig. 2 shows bubble after using Fig. 1 to show specimen cup note sample, there are situation schematic diagrams；
Fig. 3 show using Fig. 1 show specimen cup note sample after macromolecular substances deposition schematic diagram；
Fig. 4 a are shown according to a kind of structural schematic diagram open state schematic diagram of specimen cup provided by the embodiments of the present application；
Fig. 4 b are specimen cup closed state schematic diagram shown in Fig. 4 a；
Fig. 5 shows the dimensional decomposition structure diagram for the specimen cup that one embodiment of the application provides.
Reference sign
1- cup bodies, 11- sample injection units, 12- sample vessels portion, 121- channel sections, 1211- note sample channel, 1212- exhausts
Channel, 122- sampling sections, 1221- sample vessels, 1222- side walls, 123- storage troughs, 124- note sample slot, and 126- notes sample optical filtering slot,
127- is vented optical filtering slot, 2- sample vessel patches, 3- sealing covers, 4- flexibility connecting plates.
Specific implementation mode
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Whole description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Fig. 4 a are a kind of specimen cup open state schematic diagrames shown according to an illustrative examples, and Fig. 4 b are shown in Fig. 4 a
Specimen cup closed state schematic diagram, the specimen cup include：Cup body 1 and sample vessel patch 2.
In the present embodiment, the specimen cup further includes sealing cover 3, and the sealing cover 3 passes through flexible connecting plate 4 with cup body 1
Connection.Optionally, the flexible connecting plate 4 is set to 1 top of cup body.
In the present embodiment, detected sample is sealed specimen cup with the sealing cover 3, is made by after injection cup body 1
It obtains during quantitative PCR detection, the detected sample can be sealed in the cup body 1, to avoid detecting
Detected sample is overflowed from specimen cup in journey or other impurity enter the fortuitous events such as specimen cup pollution detected sample
Occur and reduces the accuracy quantitatively detected.
In a kind of achievable mode, the cup body 1 includes sample injection unit 11 and sample vessel portion 12, the sample
Injection unit 11 is tubular, and sample vessel portion 12 is plate, and the sample injection unit 11 is seamlessly transitted with sample vessel portion 12.
Applicants have discovered that the sample injection unit 11 of tubular can form the sampling injecting space of bigger, convenient for into specimen cup
Sample operation is noted, the risk that detected sample during noting sample spills sample injection unit 11 can also be reduced, can not only be saved
Detected sample can also reduce pollution of the detected sample to experimental situation.
Inner cavity is offered in the sample injection unit 11, the inner cavity is used to temporarily load detected sample after noting sample,
And sealing cover is installed after noting sample, make to form confined space inside specimen cup.
The inner cavity can have any shape, for example, cylindrical, prismatic, truncated cone-shaped or other special-shaped chambers etc., so as to
It is preferred to flow into sample vessel portion 12 in liquid to be detected sample and be sealed to specimen cup convenient for sealing cover.
Correspondingly, the bottom surface of the sample injection unit 11 can be plane, curved surface or it is other convenient for sample injection and
The Special-Shaped Surface of bubble in specimen cup is excluded, for example, can be inclined-plane or cascaded surface etc..
In the present embodiment, incorporated by reference to Fig. 4, sample vessel portion 12 includes channel section 121 and samples section 122, described
Channel section 121 is integrally formed with the sampling section 122.
Note sample channel 1211 and exhaust passage 1212 are offered in the channel section 121, are opened in the sampling section 122
Equipped with sample vessel.
The plate face of the channel section 121 can have any shape, preferably rectangular, on the one hand easy to process, on the other hand
It is stably placed in detecting instrument convenient for specimen cup.
1211 He of note sample channel for injecting detected sample into sample vessel is offered in the channel section 121
Exhaust passage 1212 for bubble in sample vessel to be discharged.
The one end in the note sample channel 1211 and one end of exhaust passage 1212 connect with the inner cavity of sample injection unit 11 respectively
Logical, the other end for noting sample channel 1211 converges at the top of the sample vessel 1221 with the other end of exhaust passage 1212
End, and be connected to the sample vessel 1221 so that note sample channel 1211 is mutually only in channel section 121 with exhaust passage 1212
Vertical, two passes only cross on the top of sample vessel 1221, to detected sample can along the note sample channel 1211 by
Sample injection unit 11 injects sample vessel 1221, and the bubble in sample vessel 1221 can be expelled to sample along exhaust passage 1212
Injection unit 11.
For convenience of description, in the present embodiment, port of the note sample channel 1211 in sample injection unit 11 is referred to as that note sample enters
Mouthful, the port on sample vessel 1221 is sample export, and port of the exhaust passage 1212 on sample vessel is exhaust gas intake port,
Port in sample injection unit 11 is air exit.As shown in figure 4, the exhaust passage 1212 is in sample injection unit 11
Port of the port higher than note sample channel 1211 in sample injection unit 11, i.e., the air exit of exhaust passage is higher than note sample channel
Sample inlet, to avoid the fluid sample of injection sample injection unit to seal the air exit of exhaust passage as possible.
Since quantitative detection is small with sample amount, may there was only tens microlitres or even more than ten microlitres, moreover, note sample channel
Internal diameter is generally also smaller, and therefore, detected sample is after injecting sample injection unit 11, it is difficult to flow directly into sample vessel 1221
In, typically by detected sample injection sample injection unit 11, then detected sample is injected by sample by the operation such as centrifugation and is held
Chamber 1221.In the operating process such as centrifugation, sample vessel can be discharged by exhaust passage 1212 in the bubble in sample vessel
1221 enter in sample injection unit 11, these bubbles may be air in sample vessel, since detected sample injects speed
Bubble etc. that is fast and mixing in the bubble and/or detected sample of generation is spent, if before the implant operations such as centrifugation, exhaust is logical
The air exit in road is by fluid-tight, then detected sample may be difficult to due to the effect of air pressure into sample vessel 1221, and sample
Gas in vessel 1221 is also difficult to be discharged.
In a kind of achievable mode, the bottom surface of the sample injection unit 11 can be inclined surface, and the inclined surface is leaned on
One end of nearly exhaust passage is higher than one end close to note sample channel so that detected sample collects naturally in sample injection unit 11
In in note sample channel side, since sample size is small, the liquid level of detected sample is less than air exit, therefore, test sample to be checked
Product can come together in sample injection unit 11 close to 1211 side of note sample channel, without causing fluid-tight to exhaust passage 1212.
In another achievable mode, the bottom surface of the sample injection unit 11 is stairstepping, the exhaust passage
1212 air exit is opened on higher-order ladder, and the note sample import in the note sample channel 1211 is opened on relatively low ladder, from
And form note sample slot 124 on 1211 top of note sample channel so that detected sample all concentrates in note sample slot 124, both
The risk that detected sample causes exhaust passage fluid-tight can be reduced, can also make detected sample complete by note sample channel 1211
Portion enters sample vessel 1221, reduces the waste of detected sample.
Further, the note sample slot 124 is in infundibulate, the lowermost end of the note sample slot 124 and the note sample channel
1211 connections, to be conducive in detected sample injection sample vessel 1221.
Further, the top surface of the higher-order ladder is inclined-plane, and the inclined-plane is less than remote close to the side in note sample channel
Side from note sample channel.Applicants have discovered that the bubble in sample vessel escapes the mistake of sample vessel along exhaust passage 1212
Cheng Zhong, part detected sample may escape sample vessel together with bubble, and the detected sample of effusion sample vessel can
It is back in note sample slot with the sloped top face using higher-order ladder, and re-injects sample vessel under the influence of centrifugal force
In, detected sample is full of in sample vessel to empty the bubble in sample vessel and can make.
In the present embodiment, the inclination angle on the inclined-plane be 20 °~40 °, preferably 30 °, applicants have discovered that, at this inclination angle
Under, escape sample vessel detected sample can quick backflow to note sample slot in, moreover, it is enough that note sample slot can be made to be formed
Memory space stores detected sample.
In the present embodiment, from close 11 one end of sample injection unit to close to 1221 one end of sample vessel, that is, gone out by exhaust
To exhaust gas intake port, the internal diameter of the exhaust passage 1212 is gradually reduced mouth；From close 11 one end of sample injection unit to close to sample
1221 one end of vessel, that is, by note sample import to note sample outlet, the internal diameter in the note sample channel 1211 is gradually reduced；The note sample
Internal diameter of the channel 1211 in 1221 one end port of sample vessel is more than the exhaust passage 1212 at 1221 one end end of sample vessel
The internal diameter of mouth, that is, the internal diameter for noting sample outlet are more than the internal diameter of exhaust gas intake port；The note sample channel 1211 is in sample injection unit 11 1
Hold internal diameter of the internal diameter of port more than the exhaust passage 1212 in 11 one end port of sample injection unit, that is, note the interior of sample import
Diameter is more than the internal diameter of air exit.To sum up, the internal diameter in note sample channel 1211 is more than the internal diameter of exhaust passage 1212.
Applicants have discovered that detected sample usually has certain viscosity, therefore, the internal diameter in note sample channel 1211 is more than row
The internal diameter in gas channel 1212 can be such that detected sample rapidly injects in sample vessel 1221, further, it is possible to make sample vessel
Sample vessel is successfully discharged in bubble in 1221 along exhaust passage 1212, and can ensure that the structure of channel section 121 is strong
Degree.
Optionally, the note sample channel 1211 includes note sample leading portion and note sample back segment, and the note sample leading portion is noted close to sample
Enter portion 11, the note sample back segment is more than the slope of note sample back segment close to sample vessel 1221, the slope of the note sample leading portion；It is described
Exhaust passage 1212 includes exhaust leading portion and exhaust back segment, and the exhaust leading portion is close to sample injection unit 11, the exhaust back segment
Close to sample vessel 1221, the slope of the exhaust leading portion is more than the slope of exhaust back segment.
Applicants have discovered that when the note sample channel 1211 and exhaust passage 1212 are arranged according to above structure, it is to be detected
Sample note sample leading portion can rapid decrease, into note sample back segment after flow velocity reduce, to make detected sample gently inject
In sample vessel, avoid making to generate excessive bubble in sample vessel since detected sample injection rate is too fast.Moreover, sample
Bubble in product vessel constantly rises with the progress of centrifugation, although exhaust gas intake port crosses with the outlet of note sample, due to note
Sample outlet has hydraulic pressure, and therefore, the bubble of rising is discharged sample vessel along exhaust passage.Hold as bubble is carried over sample
The detected sample of chamber can be flowed back into along the bottom surface of sample injection unit in note sample channel, to complete the note of detected sample
Sample.
Optionally, the angle of the note sample back segment and the exhaust back segment is 30 ° to 45 °, preferably 32 °.
Optionally, the ladder of 11 bottom end of sample injection unit seamlessly transits, and the top surface of higher-order ladder is to exhaust passage
The curved surface of 1212 side indents, forms diversion trench, and diameter of the diversion trench close to 1212 one end of exhaust passage is more than close to note
The diameter of 1211 one end of sample channel, depth of the diversion trench close to 1212 one end of exhaust passage are more than close to note sample channel 1211
The depth of one end.So that the detected sample being discharged by exhaust passage 1212 can be back to note sample channel along the diversion trench
In 1211, and then it is back in sample vessel 1221.
In the present embodiment, note sample outlet is intersected in the top of sample vessel with exhaust gas intake port, is on the one hand convenient for sample
On the other hand the injection of product and the discharge of gas make influence of the sample export with exhaust gas intake port to light path minimum, maximum limit
Degree ground ensures the accuracy of real-time fluorescence quantitative PCR detection.
In a kind of achievable mode, the note sample channel 1211 converges at the sample with the exhaust passage 1212
The top of vessel 1221 is less than at 1/5.The application has found that the note sample channel 1211 is converged at the exhaust passage 1212
Rheme is set, and is hardly impacted to quantitative detection, can ensure the accuracy of quantitative detection.
Further, 1211 side of note sample channel is provided with note sample optical filtering slot 126, note sample is come to mask
The veiling glare that light path is interfered in channel.
Further, it is both provided with exhaust optical filtering slot 127 in 1212 side of the exhaust passage, is come to mask
The veiling glare that light path is interfered of exhaust passage.
In the present embodiment, as shown in figure 4, the plate face of the sampling section 122 is subtriangular, by close to channel section 121
One end to one end far from channel section 121, the outer diameter of the sampling section 122 is gradually reduced, also, under the sampling section 122
Two adjacent outer walls of portion are orthogonal.
In the present embodiment, sample vessel 1221 is offered in the sampling section 122, the sample vessel 1221 is pros
Shape, 1221 two adjacent cavity walls of the sample vessel and the outer wall of the sampling section 122 are mutually parallel so that the sample holds
The thickness of the two side walls 1222 of 1221 lower part of chamber is equal, to carry out real-time fluorescence quantitative PCR detection.
In a kind of achievable mode, as shown in figure 5, the sample vessel 1221 can be formed in the following manner：
The through-hole perpendicular to plate face is offered in sample vessel portion 12, the sample vessel patch 2 is covered on sample appearance respectively
In the both sides plate face of cavity portion 12, sample vessel 1221 is formed with through-hole in sample vessel portion 12, for example, the side of hot pressing can be passed through
Formula pastes plastic film in two plate faces in sample vessel portion and forms sample vessel.
As shown in figure 4, offering the storage trough 123 for storing deposit in the bottom of the sampling section 122.
In a kind of achievable mode, the storage trough 123 is set to non-detection optical path area, to avoid to detection light
The influence on road ensures the accuracy quantitatively detected.
Specifically, the storage trough 123 is set to the two side walls extended surface of 1221 bottom end of sample vessel and is formed
Region in, the storage trough 123 is oppositely arranged with the sample vessel 1221.
The shape of the storage trough 123 can be the shape of any one not Interference Detection light path, such as spherical shape, water-drop-shaped,
Elliposoidal etc., preferably water-drop-shaped, to provide sufficient memory space, Er Qieneng for the macromolecular substances in detected sample
Enough influences reduced to the maximum extent to light path, to ensure the accuracy of real-time fluorescence quantitative PCR detection.
Optionally, the internal diameter of 123 slot bottom of the storage trough is more than the internal diameter of groove top, and the depth of the storage trough 123 is less than
The 2/3 of 1222 thickness of the side wall；And/or the storage trough 123 is opened in the bottom end of the sample vessel 1221 and is less than 1/5
Place.
It is less than at 1/5 in the bottom end of the sample vessel 1221 applicants have discovered that the storage trough 123 of water-drop-shaped is opened in
Storage trough can be avoided to occupy light path, will not be mixed into the interference light from storage trough in light path.
Real-time fluorescence quantitative PCR detection is carried out using specimen cup provided by the present application, can either be discharged in sample vessel and mix
Miscellaneous bubble, and the macromolecular substances in detected sample can be made to fall in the not storage trough of Interference Detection light path, from two
Aspect improves the accuracy of real-time fluorescence quantitative PCR detection.
The application is described in detail above in association with detailed description and exemplary example, but these explanations are simultaneously
It should not be understood as the limitation to the application.It will be appreciated by those skilled in the art that without departing from the application spirit and scope,
A variety of equivalent substitution, modification or improvements can be carried out to technical scheme and embodiments thereof, these each fall within the application
In the range of.The protection domain of the application is determined by the appended claims.
Claims (9)
1. a kind of specimen cup for real-time fluorescence quantitative PCR detection, which is characterized in that the specimen cup includes：Cup body (1) and
Sample vessel patch (2), the cup body (1) include sample injection unit (11) and sample vessel portion (12), the sample injection unit
(11) it is tubular, sample vessel portion (12) is plate, and the sample injection unit (11) and sample vessel portion (12) are smoothed
It crosses, sample vessel portion (12) includes channel section (121) and sampling section (122), and the plate face of the channel section (121) is square
The plate face of shape, the sampling section (122) is triangle, also, the outer diameter of the sampling section (122) is gradually reduced, the sampling
Two adjacent outer walls of section (122) are orthogonal, and the sample vessel patch (2) is covered on the two of sample vessel portion (12) respectively
In lateral plates；
Sample vessel (1221) is offered in the sampling section (122), the sample vessel (1221) is square, the sample
The cavity wall of vessel (1221) and the outer wall of the sampling section (122) are mutually parallel so that sample vessel (1221) lower part
The thickness of two side walls (1222) is equal；
The storage trough (123) for storing deposit is offered in the bottom of the sample vessel (1221)；
Note sample channel (1211) and exhaust passage (1212), the note sample channel (1211) are offered in the channel section (121)
One end and one end of exhaust passage (1212) be connected to respectively with sample injection unit (11), note sample channel (1211) it is another
The other end of end and exhaust passage (1212) converges at the top of the sample vessel (1221), and with the sample vessel
(1221) it is connected to, port of the exhaust passage (1212) in sample injection unit (11) is higher than note sample channel (1211) in sample
Port in injection unit (11)；
It is additionally provided with sealing cover (3) at the top of cup body (1) and the flexible connecting plate for connecting sealing cover (3) and cup body (1)
(4)。
2. specimen cup according to claim 1, which is characterized in that the storage trough (123) is set to non-detection light path area
Domain.
3. specimen cup according to claim 1 or 2, which is characterized in that the storage trough (123) is set to the sample and holds
The two side walls extended surface of chamber (1221) bottom end is formed by region, the storage trough (123) and the sample vessel
(1221) it is oppositely arranged.
4. specimen cup according to any one of claims 1 to 3, which is characterized in that
The internal diameter of storage trough (123) slot bottom is more than the internal diameter of groove top, and the depth of the storage trough (123) is less than the side wall
(1222) the 2/3 of thickness；And/or
The bottom end that the storage trough (123) is opened in the sample vessel (1221) is less than at 1/5.
5. specimen cup according to any one of claims 1 to 4, which is characterized in that the bottom end of the sample injection unit (11)
Port for stairstepping, the exhaust passage (1212) is opened on higher-order ladder, and the port of note sample channel (1211) is opened
On relatively low ladder；
The top surface of the higher-order ladder is inclined-plane, and the inclined-plane is less than one far from note sample channel close to the side in note sample channel
Side；
Note sample slot (124) is formed on note sample channel (1211) top, the note sample slot (124) is in infundibulate, the note sample
The lowermost end of slot (124) is connected to the note sample channel (1211).
6. specimen cup according to any one of claims 1 to 5, which is characterized in that sample injection unit (11) bottom end
Ladder seamlessly transits, and the top surface of higher-order ladder is the curved surface to exhaust passage (1212) side indent, forms diversion trench, described to lead
Diameter of the chute close to exhaust passage (1212) one end is more than the diameter close to note sample channel (1211) one end, and the diversion trench leans on
The depth of nearly exhaust passage (1212) one end is more than the depth close to note sample channel (1211) one end.
7. specimen cup according to any one of claims 1 to 6, which is characterized in that
From close sample injection unit (11) one end to close to sample vessel (1221) one end, the internal diameter of the exhaust passage (1212)
It is gradually reduced；
From close sample injection unit (11) one end to close to sample vessel (1221) one end, the internal diameter of note sample channel (1211)
It is gradually reduced；
Internal diameter of note sample channel (1211) in sample vessel (1221) one end port exists more than the exhaust passage (1212)
The internal diameter of sample vessel (1221) one end port, internal diameter of note sample channel (1211) in sample injection unit (11) one end port
Internal diameter more than the exhaust passage (1212) in sample injection unit (11) one end port.
8. specimen cup according to any one of claims 1 to 7, which is characterized in that
The note sample channel (1211) includes note sample leading portion and note sample back segment, the note sample leading portion close to sample injection unit (11),
The note sample back segment is more than the slope of note sample back segment close to sample vessel (1221), the slope of the note sample leading portion；
The exhaust passage (1212) includes exhaust leading portion and exhaust back segment, the exhaust leading portion close to sample injection unit (11),
The exhaust back segment is more than the slope of exhaust back segment close to sample vessel (1221), the slope of the exhaust leading portion.
9. according to claim 1 to 8 any one of them specimen cup, which is characterized in that
The top that note sample channel (1211) converges at the sample vessel (1221) with the exhaust passage (1212) is less than
At 1/5；And/or
Note sample channel (1211) side is provided with note sample optical filtering slot (126)；And/or
Exhaust passage (1212) side is both provided with exhaust optical filtering slot (127).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810580440.7A CN108753589B (en) | 2018-06-07 | 2018-06-07 | Sample cup for real-time fluorescent quantitative PCR detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810580440.7A CN108753589B (en) | 2018-06-07 | 2018-06-07 | Sample cup for real-time fluorescent quantitative PCR detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108753589A true CN108753589A (en) | 2018-11-06 |
| CN108753589B CN108753589B (en) | 2024-03-19 |
Family
ID=64000368
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810580440.7A Active CN108753589B (en) | 2018-06-07 | 2018-06-07 | Sample cup for real-time fluorescent quantitative PCR detection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108753589B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023178711A1 (en) * | 2022-03-23 | 2023-09-28 | 无锡百泰克生物技术有限公司 | Single-part rapid pcr chip |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6369893B1 (en) * | 1998-05-19 | 2002-04-09 | Cepheid | Multi-channel optical detection system |
| WO2009072332A1 (en) * | 2007-12-04 | 2009-06-11 | Shimadzu Corporation | Reactor and reaction treatment method |
| CN102507445A (en) * | 2011-09-29 | 2012-06-20 | 北京金诺美生物技术有限公司 | Sample cup and multi-channel optical test system containing same |
| CN105695303A (en) * | 2016-03-21 | 2016-06-22 | 北京金诺美生物技术有限公司 | Sample reaction device and reaction method |
| CN208949289U (en) * | 2018-06-07 | 2019-06-07 | 北京金诺美生物技术有限公司 | A kind of specimen cup for real-time fluorescence quantitative PCR detection |
-
2018
- 2018-06-07 CN CN201810580440.7A patent/CN108753589B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6369893B1 (en) * | 1998-05-19 | 2002-04-09 | Cepheid | Multi-channel optical detection system |
| WO2009072332A1 (en) * | 2007-12-04 | 2009-06-11 | Shimadzu Corporation | Reactor and reaction treatment method |
| CN102507445A (en) * | 2011-09-29 | 2012-06-20 | 北京金诺美生物技术有限公司 | Sample cup and multi-channel optical test system containing same |
| CN105695303A (en) * | 2016-03-21 | 2016-06-22 | 北京金诺美生物技术有限公司 | Sample reaction device and reaction method |
| CN208949289U (en) * | 2018-06-07 | 2019-06-07 | 北京金诺美生物技术有限公司 | A kind of specimen cup for real-time fluorescence quantitative PCR detection |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023178711A1 (en) * | 2022-03-23 | 2023-09-28 | 无锡百泰克生物技术有限公司 | Single-part rapid pcr chip |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108753589B (en) | 2024-03-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US9101925B2 (en) | Centrifuge and separation vessel therefore | |
| US8445264B2 (en) | Systems and methods for harvesting target particles of a suspension | |
| CN105229465A (en) | For device, the system and method for sample preparation | |
| CN109564238A (en) | Screening implement packet and method | |
| CN105482989B (en) | A kind of multiple determination micro-fluidic chip and detection method | |
| CN106980018B (en) | A kind of kit and its application using CD45 immunofluorescences joint CEP17 probe identification circulating tumor cells | |
| CN102803467B (en) | Device and method for biological sample purification and enrichment | |
| CN108801746A (en) | A kind of device of separation and enrichment body fluid components | |
| CN208949289U (en) | A kind of specimen cup for real-time fluorescence quantitative PCR detection | |
| CN111057639B (en) | Circulating tumor cell separation and enrichment system and method based on micro-fluidic chip | |
| CN105854966B (en) | A kind of micro-fluidic chip for embedding functionalized nano-fiber film and its application | |
| CN108753589A (en) | A kind of specimen cup for real-time fluorescence quantitative PCR detection | |
| CN108866064B (en) | Aptamer of targeted metastatic human breast cancer cells and application thereof | |
| CN214735781U (en) | Closed nucleic acid detection apparatus | |
| CN114015741B (en) | Non-invasive cell activity analysis method | |
| CN213327596U (en) | Collection container and kit | |
| CN110527624A (en) | A kind of intermittent streaming electrotransfection device | |
| CN209810203U (en) | Micro-fluidic chip with color development detects function | |
| CN108562743B (en) | Modular chamber and application thereof in efficient capture of rare cells in blood | |
| CN108535462A (en) | A kind of blood analyser with autonomous cleaning and sterilizing ability | |
| CN213943199U (en) | Urine sediment test tube | |
| US20220268671A1 (en) | A device for immobilizing rare cells for cytology | |
| US11833514B2 (en) | Biomimetic array device and methods of using same | |
| EP4317400A1 (en) | Centrifugal filtration cartridge and microbial test method | |
| CN215103239U (en) | Novel PBMC cell separating tube |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20181106 Assignee: Zhuhai Qiao Biotechnology Co.,Ltd. Assignor: BEIJING GENOME BIOTECHNOLOGY Co.,Ltd. Contract record no.: X2020990000125 Denomination of invention: Sample cup for real-time fluorescence quantitative PCR (Polymerase Chain Reaction) detection License type: Common License Record date: 20200323 |
|
| EE01 | Entry into force of recordation of patent licensing contract | ||
| CB02 | Change of applicant information |
Address after: 100176 floor 01-5, building 16, yard 25, Jinghai 4th Road, Beijing Economic and Technological Development Zone, Daxing District, Beijing Applicant after: Beijing jinnuomi Technology Co.,Ltd. Address before: 100176 room 407, building 2, No. 19, Yongchang South Road, Yizhuang Economic Development Zone, Daxing District, Beijing Applicant before: BEIJING GENOME BIOTECHNOLOGY CO.,LTD. |
|
| CB02 | Change of applicant information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |