CN110075942A - The biological humor sampling separator and method that solid-liquid is isolated after being centrifuged can be achieved - Google Patents
The biological humor sampling separator and method that solid-liquid is isolated after being centrifuged can be achieved Download PDFInfo
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- CN110075942A CN110075942A CN201910381724.8A CN201910381724A CN110075942A CN 110075942 A CN110075942 A CN 110075942A CN 201910381724 A CN201910381724 A CN 201910381724A CN 110075942 A CN110075942 A CN 110075942A
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- 239000007788 liquid Substances 0.000 title claims abstract description 30
- 238000005070 sampling Methods 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title abstract description 16
- 238000012360 testing method Methods 0.000 claims abstract description 155
- 239000007791 liquid phase Substances 0.000 claims abstract description 37
- 238000005119 centrifugation Methods 0.000 claims abstract description 31
- 239000007787 solid Substances 0.000 claims abstract description 23
- 238000002955 isolation Methods 0.000 claims abstract description 19
- 239000007790 solid phase Substances 0.000 claims abstract description 16
- 229920003023 plastic Polymers 0.000 claims description 16
- 239000004033 plastic Substances 0.000 claims description 16
- 238000000926 separation method Methods 0.000 claims description 16
- 230000000694 effects Effects 0.000 claims description 14
- 229920002457 flexible plastic Polymers 0.000 claims description 12
- 210000001124 body fluid Anatomy 0.000 claims description 8
- 239000010839 body fluid Substances 0.000 claims description 8
- 238000013461 design Methods 0.000 claims description 8
- 238000001746 injection moulding Methods 0.000 claims description 7
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- 239000000463 material Substances 0.000 claims description 4
- 239000012071 phase Substances 0.000 claims description 3
- 230000008569 process Effects 0.000 abstract description 7
- 238000001514 detection method Methods 0.000 abstract description 6
- 239000013060 biological fluid Substances 0.000 abstract description 5
- 230000015572 biosynthetic process Effects 0.000 abstract description 4
- 238000005755 formation reaction Methods 0.000 abstract description 4
- 238000004321 preservation Methods 0.000 abstract description 3
- 239000004615 ingredient Substances 0.000 abstract description 2
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- 239000000523 sample Substances 0.000 description 16
- 230000032258 transport Effects 0.000 description 9
- 238000010586 diagram Methods 0.000 description 8
- 210000004369 blood Anatomy 0.000 description 7
- 239000008280 blood Substances 0.000 description 7
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- 206010028980 Neoplasm Diseases 0.000 description 2
- 208000002151 Pleural effusion Diseases 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 238000007363 ring formation reaction Methods 0.000 description 2
- 210000002700 urine Anatomy 0.000 description 2
- 206010003830 Automatism Diseases 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 206010048612 Hydrothorax Diseases 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
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Classifications
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- 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
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5082—Test tubes per se
-
- 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
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5082—Test tubes per se
- B01L3/50825—Closing or opening means, corks, bungs
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Centrifugal Separators (AREA)
Abstract
The invention discloses the biological humor sampling separators that solid-liquid after a kind of achievable centrifugation is isolated, including test tube, guide rail shell, thimble ring, fixture ring, rotation tooth post, thrust tooth post, push rod and spring, test tube outer diameter is intermediate thin both ends coarse-texture, guide rail shell is the cylindrical structure with cavity, and thimble ring, fixture ring, rotation tooth post, thrust tooth post are successively placed on test tube and are placed in guide rail shell inner cavity.Using the present apparatus; the biological fluid samples containing solid phase particles ingredient can be made to realize the physical isolation between solid phase and liquid phase layer automatically after completing solid liquid phase layering by centrifugation, so that the target detections such as dissociative DNA in liquid phase layer substance be protected not polluted by solid formations such as cells in Sample preservation and transportational process.
Description
Technical field
The biological humor sampling separator and method being isolated the present invention relates to solid-liquid after a kind of achievable centrifugation, belong to
Sample collection field.
Background technique
Dissociative DNA (cfDNA) is the extracellular trip for being present in various body fluid (including blood, urine, Pleural effusions, milk etc.)
From DNA fragmentation.Peripheral blood cfDNA is already used to carry out noninvasive prenatal gene detection (NIPT) at present；Peripheral blood and hydrothorax
CfDNA can be used for the adjoint diagnosis of cancer target medicine；The early screening of cancer based on cfDNA also by extensive concern and
Research.When carrying out various detections using cfDNA, cfDNA is protected not to be degraded in biological fluid extracting, preservation and transportational process and not
It is an important prerequisite by haemocyte contaminating genomic DNA.Currently the only guard method is in vacuum blood collection tube
It adds anticoagulant and DNA and protects mix reagent, can be saved 1-2 weeks under the conditions of 4-30 DEG C, but be higher than 35 DEG C of protecting effects not
Ideal, especially 0 DEG C or less save and transport completely can not because high temperature and freeze thawing can all cause cell to discharge genome
DNA.Therefore, for the blood equal samples of cfDNA detection summer and winter transport safely save difficulty beat, it is at high cost, and
And it is careless slightly will because environment, strenuous vibration factor caused by cfDNA testing result confidence level reduce.
Solve the problems, such as that this basic method is exactly that the sample acquired first carries out Solid-Liquid Separation and (such as manually grasps after centrifugation
Make blood plasma and haemocyte being separated to different vessels) transport of sample is carried out again.But current sample collection method realizes transport
Experimenter of forefathers' work point from needs profession operates and there is the risk of very big maloperation.
Summary of the invention
Technology of the invention solves the problems, such as: in order to overcome the deficiencies of the prior art, providing a kind of solid liquid phase after centrifugation can be achieved
The biological humor sampling separator and method of isolation, the present apparatus is ingenious to utilize spring force and centrifugal force, in sample solid liquid phase
It is automatic to realize solid liquid phase physical isolation after after centrifugation layering, it is not necessarily to artificial lock out operation, is the sample containing cfDNA in pole
The whole-course automation for stablizing transport and subsequent cfDNA extraction under the conditions of the temperature of end provides strong guarantee.
Technical solution of the invention:
The biological humor sampling separator that solid-liquid is isolated after being centrifuged, including test tube, guide rail shell, thimble can be achieved
Ring, fixture ring, rotation tooth post, thrust tooth post, push rod and spring,
Test tube is intermediate thin both ends coarse-texture, and test tube open at one end is socketed on the inner cavity path of test tube connection ring, spring
It is placed in test tube outer wall and the cavity of test tube connection ring formation, guide rail shell one end and the inner cavity bigger diameter end of test tube connection ring are fixed
Connection, push rod one end can enter from the other end of guide rail shell；
Guide rail shell is the cylindrical structure with cavity, on the guide rail outer casing inner wall of test tube connection ring one end, edge
It axially has been arranged circumferentially detent slot, on the guide rail outer casing inner wall of push rod one end, has been arranged circumferentially along axial direction convex
Play guide rail；
Thimble ring, fixture ring, rotation tooth post, thrust tooth post are successively placed on test tube and are placed in guide rail shell inner cavity；
Thimble ring is the cavity structure of hollow out, is connected along axial circumferentially fixed be connected to close to test tube in the thimble of symmetric design
It connects on thimble ring one end of ring one end,
Fixture ring is coaxially socketed on thimble ring, and fixture ring is fixedly connected with guide rail shell, and thimble ring and fixture ring can edges
It is relatively slipping in the circumferential direction；It is provided with the thimble groove with thimble cooperation along axial direction on fixture ring, is symmetrically connected with c-type on fixture ring inner wall
Fixture, it is the annular in opening that C-type clamp, which has original state,；
Rotation tooth post is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with rotation tooth post groove, rotation
Turn tooth post one end and thimble ring other end axial contact,
Thrust tooth post is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with thrust tooth post groove, is pushed away
Power tooth post groove cooperates with the protruding guide rail of guide rail shell always, and thrust tooth post one end is that dislocation is nibbled with the rotation tooth post other end
It closes, the thrust tooth post other end and push rod end thereof contacts.
When work: the present apparatus after body fluid example will be acquired and be put into centrifuge centrifugation, solid phase and liquid phase are being centrifuged
The lower separation of power effect, while under the action of the centrifugal force, thrust tooth post is moved axially along the protruding guide rail of guide rail shell, overcomes bullet
Spring active force, thrust tooth post drive rotation tooth post and thimble ring synchronizing moving, when the tooth tip of rotation tooth post is more than protruding guide rail
When, rotation tooth post rotates in the case where spring force acts on and tooth post top inclined-plane guides, and C-type clamp tool is open state at this time；
When centrifugation is completed, revolving speed decline, centrifugal force is gradually reduced, and spring force can be increasingly greater than centrifugal force, and elastic force pushes thimble ring, makes
Thimble ring pushes rotation tooth post to slide into protruding guide rail and moves backward along protruding guide rail, thimble ring, rotation tooth post and thrust teeth
Column moves backward together, is placed in the thimble of thimble ring in the thimble groove of fixture ring, while thimble acts on the c-type of fixture ring
On fixture, the closure of two C-type clamps tool is realized, by the flow liquid channel enclosed of test tube, realize automatic solid liquid phase physical isolation.
The thimble of original state is less than the distance of subsequent rotation tooth post reverse movement apart from the distance that C-type clamp has.
Test tube is the assembly of different hardness plastic material, including any one in following scheme:
Scheme 1: C-type clamp have upward 5-15mm to C-type clamp have downward 5-15mm range be flexible plastic, shore hardness
A is 50-70；Test tube other parts are rigid plastics, and shore hardness D is 70-80, and test tube connection ring and test tube are that an injection molding is whole
Body；
Scheme 2: the range that test tube bottom to C-type clamp has upward 5-15mm is flexible plastic, and the hard A of shore is 50-70；Test tube
Other parts are rigid plastics, and shore hardness D is 70-80, and test tube connection ring and test tube are that an injection molding is whole；
Scheme 3: test tube inner core is flexible plastic, and shore hardness A is 50-70, the test tube connection of test tube open at one end socket
Ring is rigid plastics, and shore hardness D is 70-80, and test tube connection ring and test tube pass through the outer wall construction clamping of test tube open at one end
And gluing is fastened together.
The C-type clamp tool top distance of test tube entire length 9-13mm, test tube internal diameter 7-11mm, test tube bottom to closure are 3-
8mm, test tube wall thickness are 0.5-2.5mm.
Preferably, the C-type clamp of test tube entire length 11mm, the internal diameter 9mm of test tube, test tube bottom to closure have top distance
For 5mm, C-type clamp have upward 5-15mm to the range test tube wall thickness that C-type clamp has downward 5-15mm be 0.5mm, other region wall thickness
1-2mm, different wall thickness junction gradient thickness.
Thimble circumferential cross-section is L-shaped, and L-type face acts on C-type clamp tool root.
The biological humor sampling separation method that solid-liquid is isolated after being centrifuged can be achieved, specific step is as follows,
Step 1: building collecting and separating device, including test tube, guide rail shell, thimble ring, fixture ring, rotation tooth post, thrust
Tooth post, push rod and spring,
Test tube is intermediate thin both ends coarse-texture, and test tube open at one end is socketed on the inner cavity path of test tube connection ring, spring
It is placed in test tube outer wall and the cavity of test tube connection ring formation, guide rail shell one end and the inner cavity bigger diameter end of test tube connection ring are fixed
Connection, push rod one end can enter from the other end of guide rail shell；
Guide rail shell is the cylindrical structure with cavity, on the guide rail outer casing inner wall of test tube connection ring one end, edge
It axially has been arranged circumferentially detent slot, on the guide rail outer casing inner wall of push rod one end, has been arranged circumferentially along axial direction convex
Play guide rail；
Thimble ring, fixture ring, rotation tooth post, thrust tooth post are successively placed on test tube and are placed in guide rail shell inner cavity；
Thimble ring is the cavity structure of hollow out, is connected along axial circumferentially fixed be connected to close to test tube in the thimble of symmetric design
It connects on thimble ring one end of ring one end,
Fixture ring is coaxially socketed on thimble ring, and fixture ring is fixedly connected with guide rail shell, and thimble ring and fixture ring can edges
It is relatively slipping in the circumferential direction；It is provided with the thimble groove with thimble cooperation along axial direction on fixture ring, is symmetrically connected with c-type on fixture ring inner wall
Fixture, it is the annular in opening that C-type clamp, which has original state,；
Rotation tooth post is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with rotation tooth post groove, rotation
Turn tooth post one end and thimble ring other end axial contact,
Thrust tooth post is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with thrust tooth post groove, is pushed away
Power tooth post groove cooperates with the protruding guide rail of guide rail shell always, and thrust tooth post one end is that dislocation is nibbled with the rotation tooth post other end
It closes, the thrust tooth post other end and push rod end thereof contacts；
Step 2: the present apparatus after body fluid example will be acquired and be put into centrifuge centrifugation, solid phase and liquid phase are being centrifuged
Power effect lower separation when centrifugation, pushes push rod under the action of the centrifugal force, and push rod acts in thrust tooth post, thrust tooth post along
The protruding guide rail of guide rail shell moves axially, and overcomes acting force of the spring, and thrust tooth post drives rotation tooth post and thimble ring synchronous
It is mobile；
Step 3: when the tooth tip for rotating tooth post is more than protruding guide rail, rotation tooth post is on spring force effect and tooth post top
It holds under inclined-plane guidance, it may occur that rotate up to two tooth post flank of tooth occlusions, C-type clamp tool is open state at this time, is not influenced completely
It is centrifugated the effect of solid liquid phase；
Step 4: when centrifugation is completed, revolving speed decline, centrifugal force is gradually reduced, and spring force is increasingly greater than centrifugal force, spring
Elastic force pushes thimble ring, makes thimble ring that rotation tooth post be pushed to slide into protruding guide rail and moves backward along protruding guide rail, in this way,
Under spring-force driven dual, thimble ring, rotation tooth post and thrust tooth post move backward together, and the thimble of thimble ring is made to be placed in fixture ring
In thimble groove, while thimble acts on the C-type clamp tool of fixture ring, the closure of two C-type clamps tool is realized, by the flow liquid of test tube
Channel enclosed realizes automatic solid liquid phase physical isolation；
Step 5: taking out the upper phase isolated, if lower layer's solid phase will also take, can be centrifuged again, or directly uses
Dynamic push rod is forced to the tooth tip of rotation tooth post more than unclamping again after protruding guide rail, thimble ring can be made to exit thimble groove, test tube
Pipeline is open, takes out lower layer's solid phase.
Compared with prior art, the present invention has the following advantages:
(1) apparatus of the present invention are ingenious utilizes spring force and centrifugal force, and having Position Design by test tube size and C-type clamp can
Guarantee the solid liquid phase physical isolation that the various biological fluid samples such as blood are realized by centrifugation, and after being isolated liquid phase part entirely without
Solid formation exists, and avoiding problems the contaminating genomic DNAs that the cfDNA for being present in liquid phase is discharged by cell；Cooperate cfDNA protection
Reagent is, it can be achieved that -80 DEG C~40 DEG C range 2 weeks or more cfDNA stablize and save and transport；
(2) it is all that can only artificially separate liquid phase sample present in cfDNA to take out that current all cfDNA, which are automatically extracted,
Process is extracted in the automation that being added afterwards could start behind instrument sample pond, this efficiency when great amount of samples is extracted is relatively low and easy
Because mutually being polluted between sample caused by human operational error.Apparatus of the present invention realize automatic solid-liquid isolation after centrifugation, can pass through
The devices such as automatic aspirating needle are equipped with to replace artificial separation sampling process, are mentioned for cfDNA automation can be started after sampling centrifugation
It takes process to provide greatly possible, the direct contact of laboratory technician and biological sample is reduced, to laboratory technician and a kind of protection.
Detailed description of the invention
Fig. 1 is complete structure schematic diagram of the present invention；
Fig. 2 is 2 half section structure diagram of test tube scheme 1 of the present invention and scheme；
Fig. 3 is 3 half section structure diagram of test tube scheme of the present invention；
Fig. 4 is 3 structural schematic diagram of test tube scheme of the present invention；
Fig. 5 is thimble ring of the present invention and fixture ring decomposition diagram；
Fig. 6 is thimble ring of the present invention and fixture ring assembling schematic diagram；
Fig. 7 is the construction profile figure of guide rail shell of the present invention；
Fig. 8 is guide rail shell half sectional view of the present invention；
Fig. 9 is guide rail interior of shell view of the present invention；
Figure 10 is present invention rotation tooth post, thrust tooth post and push rod decomposition diagram；
Figure 11 is present clip ring structure schematic diagram；
Figure 12 is structure of the invention monnolithic case figure.
Specific embodiment
The present invention is described in further detail with reference to the accompanying drawing.
The realization that automatic isolation device can be convenient after the acquisition of biological fluid samples designed by the present invention and centrifugation layering
Sample solid liquid phase physical separation (such as haemocyte and blood plasma separation), this physical isolation can effectively block cellular genome
CfDNA in DNA pollution liquid phase layer.Cooperation DNA protective agent may be implemented in the various body fluid samples of all extreme temperature conditions
The stabilization storage and transport of cfDNA ingredient.By taking blood sample as an example, haemocyte and blood plasma object are realized automatically after centrifugal blood layering
Reason isolation can liberate the blood plasma lock out operation of traditional progress that must artificially uncap.This has 3 advantages outstanding, and 1) avoid reality
The direct contact of the person of testing and biological sample are a kind of protections to laboratory technician, while avoiding the mistake that manual operation may cause；
2) cfDNA is present in liquid phase after automatism isolation, it is entirely avoided cfDNA is polluted by cell genomic dna；3) automatic solid-liquid isolation
Prerequisite is provided for the cfDNA whole-course automation extracted.Cooperate cfDNA protection reagent, it can be achieved that -80 DEG C~40 DEG C ranges
2 weeks or more cfDNA, which stablize, to be saved and transports.
The specific scheme is that the biological humor sampling separator that solid-liquid is isolated after a kind of achievable centrifugation, such as Fig. 1, figure
Shown in 12, including test tube 1, guide rail shell 2, thimble ring 3, fixture ring 4, rotation tooth post 5, thrust tooth post 6, push rod 7 and spring 8,
Test tube 1 is intermediate thin both ends coarse-texture, and the intermediate hose position in same inner diameter may be implemented in intermediate thin structure
Wall thickness is smaller, is easy to be had 12 clampings by C-type clamp to realize the physical blocking of solid liquid phase layer；Both ends slightly ensure that test tube both ends are larger
Wall thickness to increase the rigidity of test tube.1 open at one end of test tube is socketed on the inner cavity path of test tube connection ring 11, and spring 8 is set
In the cavity that 1 outer wall of test tube and test tube connection ring 11 are formed, the inner cavity bigger diameter end of 2 one end of guide rail shell and test tube connection ring 11
It is fixedly connected, 7 one end of push rod can enter from the other end of guide rail shell 2；
As shown in Fig. 7, Fig. 8, Fig. 9, guide rail shell 2 is the cylindrical structure with cavity, close to test tube connection ring 11 1
On 2 inner wall of guide rail shell at end, detent geosynclinal concave slot 9 has been arranged circumferentially along axial direction, retaining groove 9 preferably 4, retaining groove
There is constant slope, the detent tongue complementary snap with 4 periphery of fixture ring, it is ensured that after fixture ring 4 is assembled into guide rail shell
It is accurately positioned.On 2 inner wall of guide rail shell of 7 one end of push rod, protruding guide rail 10 has been arranged circumferentially along axial direction；
Thimble ring 3, fixture ring 4, rotation tooth post 5, thrust tooth post 6 are successively placed on test tube 1 and are placed in 2 inner cavity of guide rail shell
In；Protruding guide rail 10 precisely cooperates with the groove on rotation tooth post 5 and 6 outer wall of thrust tooth post, is by centrifugal force and spring force
The key that C-type clamp tool 12 is clamped and unclamped is realized in effect.Thrust tooth post one end is that dislocation is engaged with the rotation tooth post other end, thrust
The tooth post other end can be pushed with push rod end thereof contacts by push rod；6 groove of thrust tooth post is always without departing from protruding guide rail, rotary teeth
For column 5 in the case where being pushed by thrust tooth post 6, rotation 5 groove of tooth post can alternately realize that entering protruding guide rail slides and be detached from protruding guide rail
And it is blocked by protruding guide rail to realize 12 clamping of C-type clamp tool and unclamp.
As shown in Figure 5, Figure 6, thimble ring 3 is the cavity structure of hollow out, in the thimble of symmetric design along axial circumferentially fixed
It is connected on 3 one end of thimble ring of 11 one end of test tube connection ring, 13 circumferential cross-section of thimble is L-shaped, and L-type face acts on c-type
12 root of fixture.
Fixture ring 4 is coaxially socketed on thimble ring 3, and as shown in figure 11, fixture ring 4 is fixedly connected with guide rail shell 2, thimble
Ring 3 and fixture ring 4 can circumferentially opposite slidings；The thimble groove with thimble cooperation, fixture ring 4 are provided with along axial direction on fixture ring 4
C-type clamp tool 12 is symmetrically connected on inner wall, it is the annular in opening that C-type clamp, which has 12 original states, and C-type clamp tool 12 is metal material
Matter does not deform.The design of thimble ring 3, fixture ring 4 and C-type clamp tool is guaranteeing to realize that clamping test tube separates solid liquid phase layer
While function, it is effectively saved space, single unit system is made to keep cramped construction.
As shown in Figure 10, rotation tooth post 5 is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with rotation
5 groove of tooth post, rotation 5 one end of tooth post and 3 other end axial contact of thimble ring, the thimble of original state is apart from C-type clamp tool 12
Distance is less than the distance that subsequent rotation tooth post 5 moves backward.
Thrust tooth post 6 is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with 6 groove of thrust tooth post,
6 groove of thrust tooth post cooperates with the protruding guide rail 10 of guide rail shell 2 always, and 6 one end of thrust tooth post is with rotation 5 other end of tooth post
Dislocation engagement, 6 other end of thrust tooth post and 7 end thereof contacts of push rod.
When work: the present apparatus after body fluid example will be acquired and be put into centrifuge in the centrifugal force of 1200g-2000g
When being centrifuged, solid phase (such as haemocyte) and liquid phase (such as blood plasma) can separate under the action of the centrifugal force, while in the work of centrifugal force
Under, thrust tooth post 6 is moved axially along the protruding guide rail 10 of guide rail shell 2, overcomes 8 active force of spring, and thrust tooth post 6 drives
Tooth post 5 and 3 synchronizing moving of thimble ring are rotated, when the tooth tip for rotating tooth post 5 is more than protruding guide rail 10, rotates tooth post 5 in bullet
Under the effect of 8 elastic force of spring and the guidance of tooth post top inclined-plane, it may occur that rotation, C-type clamp tool 12 is open state at this time, is not influenced completely
Centrifugal separating effect；When centrifugation is completed, revolving speed decline, centrifugal force is gradually reduced, and 8 elastic force of spring can be increasingly greater than centrifugal force, bullet
Power just will push thimble ring 3, make thimble ring 3 that rotation tooth post 5 be pushed to slide into protruding guide rail 10 and reversely move along protruding guide rail 10
It is dynamic, in this way, thimble ring 3, rotation tooth post 5 and thrust tooth post 6 move backward together under the effect of 8 power of spring, make thimble ring 3
Thimble is placed in the thimble groove of fixture ring 4, while thimble acts on the C-type clamp tool 12 of fixture ring 4, realizes two C-type clamps
The flow liquid channel enclosed of test tube 1 is realized automatic solid liquid phase physical isolation by the closure of tool 12.
Test tube 1 is the assembly of different hardness plastic material, including any one in following scheme:
As shown in Fig. 2, scheme 1: having 12 upward 5-15mm (preferably 10mm) to C-type clamp in C-type clamp and have 12 downward 5-15mm
The range of (preferably 10mm) is flexible plastic, and shore hardness A is 50-70；1 other parts of test tube are rigid plastics, shore hardness D
For 70-80, test tube connection ring 11 and test tube 1 are that an injection molding is whole；
Scheme 2: the range that 1 bottom of test tube to C-type clamp has 12 upward 5-15mm (preferably 10mm) is flexible plastic, and shore is hard
A is 50-70；1 other parts of test tube are rigid plastics, and shore hardness D is 70-80, and test tube connection ring 11 and test tube 1 are a note
Modeling is whole；
As shown in Figure 3, Figure 4, scheme 3: 1 inner core of test tube is flexible plastic, and shore hardness A is 50-70, the opening of test tube 1 one
The test tube connection ring 11 that end cap connects is rigid plastics, and shore hardness D is 70-80, and test tube connection ring 11 and test tube 1 pass through test tube 1
Simultaneously gluing is fastened together for the outer wall construction clamping of open at one end.
1 entire length 9-13mm of test tube, 1 internal diameter 7-11mm of test tube, 1 bottom of test tube to closure C-type clamp have 12 tops away from
From for 3-8mm, 1 wall thickness of test tube is 0.5-2mm.
It is preferred that test tube 1 entire length 11mm, the internal diameter 9mm of test tube 1, the C-type clamp of 1 bottom of test tube to closure have 12 tops away from
From for 5mm, the range that C-type clamp has 12 upward 5-15mm (preferably 10mm) to C-type clamp 12 downward 5-15mm (preferably 10mm) of tool is tried
1 wall thickness of pipe is 0.5mm, other regions wall thickness 1-1.5mm, different wall thickness junction carries out gradient thickness processing.
The biological humor sampling separation method that solid-liquid is isolated after being centrifuged is realized using above-mentioned apparatus, the specific steps are as follows:
Step 1: above-mentioned collecting and separating device is constructed；
Step 2: the present apparatus after body fluid example will be acquired and be put into centrifuge centrifugation, solid phase and liquid phase are being centrifuged
Power effect lower separation when centrifugation, pushes push rod 7, push rod 7 acts in thrust tooth post 6, thrust tooth post 6 under the action of the centrifugal force
Moved axially along the protruding guide rail 10 of guide rail shell 2, overcome 8 active force of spring, thrust tooth post 6 drive rotation tooth post 5 and
3 synchronizing moving of thimble ring；
Step 3: when the tooth tip for rotating tooth post 5 is more than protruding guide rail 10, rotation tooth post 5 is in the effect of 8 elastic force of spring and tooth
Under the guidance of top end inclined-plane, it may occur that rotate up to two tooth post flank of tooth occlusions, C-type clamp tool 12 is open state at this time, completely
Do not influence the effect for being centrifugated solid liquid phase；
Step 4: when centrifugation is completed, revolving speed decline, centrifugal force is gradually reduced, and 8 elastic force of spring is increasingly greater than centrifugal force, bullet
8 elastic force of spring pushes thimble ring 3, makes thimble ring 3 that rotation tooth post 5 be pushed to slide into protruding guide rail 10 and reversely move along protruding guide rail 10
It is dynamic, in this way, thimble ring 3, rotation tooth post 5 and thrust tooth post 6 move backward together under the effect of 8 power of spring, make thimble ring 3
Thimble is placed in the thimble groove of fixture ring 4, while thimble acts on the C-type clamp tool 12 of fixture ring 4, realizes two C-type clamps
The flow liquid channel enclosed of test tube 1 is realized automatic solid liquid phase physical isolation by the closure of tool 12；
Step 5: it takes out the upper phase isolated and carries out the Experiment of correlation detection such as dissociative DNA extraction, if lower layer's solid phase
It takes, can be centrifuged again, or the direct tooth tip for firmly pressing push rod 7 to rotation tooth post 5 is loose again more than after protruding guide rail 10
It opens, thimble ring 3 can be made to exit thimble groove, 1 pipeline of test tube is open, takes out lower layer's solid phase.
Using present invention device disclosed above, the biological fluid samples of the object containing solid phase particles can be made by centrifugal solid-liquid phase
Automatically the physical isolation between solid phase and liquid phase layer is realized after layering, to protect liquid phase layer in Sample preservation and transportational process
In the target detections substance such as dissociative DNA do not polluted by solid formations such as cells.Internal test tube 1 can also can be pumped into negative by the present apparatus
Pressure, realizes the negative pressure quantitative collection of sample.The present apparatus can be applied to the bodies such as (but being not limited to) blood, urine, Pleural effusions, milk
The acquisition and separation of solid and liquid of liquid sample.
Apparatus of the present invention are ingenious to utilize 8 elastic force of spring and centrifugal force, has 12 Position Designs by 1 size of test tube and C-type clamp
It can guarantee after realizing solid liquid phase layering isolation by centrifugation, liquid phase part exists entirely without solid formation, and avoiding problems be present in
CfDNA in liquid phase is polluted by cell genomic dna；Cooperate cfDNA protection reagent, it can be achieved that -80 DEG C~40 DEG C ranges 2 weeks with
On cfDNA stablize save and transport；
It is all after can only artificially taking out the separation of liquid phase sample present in cfDNA that current all cfDNA, which are automatically extracted,
Be added instrument sample pond after could start automation extraction process, this great amount of samples extraction when efficiency it is relatively low and also it is easy because
Human operational error causes mutually to pollute between sample.Apparatus of the present invention realize automatic solid-liquid isolation after centrifugation, can be by matching
The devices such as standby automatic aspirating needle artificially separate sampling process to replace, for that can start cfDNA automation extraction after sampling centrifugation
Process provides a kind of greatly possible, the direct contact of reduction laboratory technician and biological sample, to laboratory technician and protection.
The above description is only an embodiment of the present invention, is not intended to limit the scope of the invention, all to utilize this hair
Equivalent structure or equivalent flow shift made by bright specification and accompanying drawing content is applied directly or indirectly in other relevant skills
Art field, is included within the scope of the present invention.
The content that description in the present invention is not described in detail belongs to the well-known technique of professional and technical personnel in the field.
Claims (10)
1. the biological humor sampling separator that solid-liquid is isolated after centrifugation can be realized, which is characterized in that including test tube (1), lead
Rail shell (2), thimble ring (3), fixture ring (4), rotation tooth post (5), thrust tooth post (6), push rod (7) and spring (8),
Test tube (1) is intermediate thin both ends coarse-texture, and test tube (1) open at one end is socketed on the inner cavity path of test tube connection ring (11)
On, in the cavity that spring (8) is placed in test tube (1) outer wall and test tube connection ring (11) are formed, guide rail shell (2) one end and test tube connect
The inner cavity bigger diameter end for connecing ring (11) is fixedly connected, and push rod (7) one end can enter from the other end of guide rail shell (2)；
Guide rail shell (2) is the cylindrical structure with cavity, guide rail shell (2) inner wall close to test tube connection ring (11) one end
On, detent slot (9) are arranged circumferentially along axial direction, on guide rail shell (2) inner wall of push rod (7) one end, along axial week
To being evenly arranged protruding guide rail (10)；
Thimble ring (3), fixture ring (4), rotation tooth post (5), thrust tooth post (6) are successively placed on test tube (1) and are placed in outside guide rail
In shell (2) inner cavity；
Thimble ring (3) is the cavity structure of hollow out, is connected along axial circumferentially fixed be connected to close to test tube in the thimble of symmetric design
It connects on thimble ring (3) one end of ring (11) one end,
Fixture ring (4) is coaxially socketed on thimble ring (3), and fixture ring (4) is fixedly connected with guide rail shell (2), thimble ring (3) with
Fixture ring (4) can circumferentially opposite sliding；The thimble groove with thimble cooperation, fixture ring (4) are provided with along axial direction on fixture ring (4)
C-type clamp tool (12) is symmetrically connected on inner wall, it is the annular in opening that C-type clamp, which has (12) original state,；
Rotating tooth post (5) is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with rotation tooth post (5) groove,
Tooth post (5) one end and thimble ring (3) other end axial contact are rotated,
Thrust tooth post (6) is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with thrust tooth post (6) groove,
Thrust tooth post (6) groove cooperates with the protruding guide rail of guide rail shell (2) always, and thrust tooth post (6) one end and rotation tooth post (5) are another
One end is dislocation engagement, thrust tooth post (6) other end and push rod (7) end thereof contacts.
2. as described in claim 1 can be achieved the biological humor sampling separator that solid-liquid is isolated after being centrifuged, feature exists
When, work: the present apparatus after body fluid example will be acquired be put into centrifuge centrifugation, solid phase and liquid phase and acted in centrifugal force
Lower separation, while under the action of the centrifugal force, thrust tooth post (6) is moved axially along the protruding guide rail of guide rail shell (2), overcomes bullet
Spring (8) active force, thrust tooth post (6) drives rotation tooth post (5) and thimble ring (3) synchronizing moving, when the tooth of rotation tooth post (5)
When point is more than protruding guide rail (10), rotation tooth post (5) occurs to turn in the case where spring (8) elastic force acts on and tooth post top inclined-plane guides
Dynamic, C-type clamp tool (12) is open state at this time；When centrifugation is completed, revolving speed decline, centrifugal force is gradually reduced, spring (8) elastic force meeting
Increasingly greater than centrifugal force, elastic force push thimble ring (3), make thimble ring (3) that rotation tooth post (5) to be pushed to slide into protruding guide rail (10) simultaneously
It is moved backward along protruding guide rail (10), thimble ring (3), rotation tooth post (5) and thrust tooth post (6) move backward together, make to push up
The thimble of needle ring (3) is placed in the thimble groove of fixture ring (4), while thimble acts on the C-type clamp tool (12) of fixture ring (4)
On, it realizes the closure of two C-type clamp tools (12), by the flow liquid channel enclosed of test tube (1), realizes automatic solid liquid phase physical isolation.
3. as described in claim 1 can be achieved the biological humor sampling separator that solid-liquid is isolated after being centrifuged, feature exists
In distance of the thimble of original state apart from C-type clamp tool (12) is less than the distance of subsequent rotation tooth post (5) reverse movement.
4. as described in claim 1 can be achieved the biological humor sampling separator that solid-liquid is isolated after being centrifuged, feature exists
In test tube (1) is the assembly of different hardness plastic material, including any one in following scheme:
Scheme 1: being flexible plastic, shore in the range of the upward 5-15mm of C-type clamp tool (12) to the downward 5-15mm of C-type clamp tool (12)
Hardness A is 50-70；Test tube (1) other parts are rigid plastics, and shore hardness D is 70-80, test tube connection ring (11) and test tube
(1) whole for an injection molding；
Scheme 2: the range of test tube (1) bottom to the upward 5-15mm of C-type clamp tool (12) are flexible plastic, and the hard A of shore is 50-70；
Test tube (1) other parts are rigid plastics, and shore hardness D is 70-80, and test tube connection ring (11) and test tube (1) are an injection molding
It is whole；
Scheme 3: test tube (1) inner core is flexible plastic, and shore hardness A is 50-70, and the test tube of test tube (1) open at one end socket connects
Connecing ring (11) is rigid plastics, and shore hardness D is that 70-80, test tube connection ring (11) and test tube (1) pass through test tube (1) opening one
Simultaneously gluing is fastened together for the outer wall construction clamping at end.
5. as described in claim 1 can be achieved the biological humor sampling separator that solid-liquid is isolated after being centrifuged, feature exists
In the C-type clamp of test tube (1) entire length 9-13mm, test tube (1) internal diameter 7-11mm, test tube (1) bottom to closure have (12) top
Distance is 3-8mm, and test tube (1) wall thickness is 0.5-2.5mm.
6. as described in claim 1 can be achieved the biological humor sampling separator that solid-liquid is isolated after being centrifuged, feature exists
In, it is preferred that the C-type clamp of test tube (1) entire length 11mm, the internal diameter 9mm of test tube (1), test tube (1) bottom to closure have (12)
Top is apart from range test tube (1) wall thickness for 5mm, the upward 5-15mm of C-type clamp tool (12) to the downward 5-15mm of C-type clamp tool (12)
0.5mm, other regions wall thickness 1-2mm, different wall thickness junction gradient thickness.
7. as described in claim 1 can be achieved the biological humor sampling separator that solid-liquid is isolated after being centrifuged, feature exists
In thimble (13) circumferential cross-section is L-shaped, and L-type face acts on C-type clamp tool (12) root.
8. the biological humor sampling separation method that solid-liquid is isolated after centrifugation can be realized, which is characterized in that specific step is as follows,
Step 1: building collecting and separating device, including test tube (1), guide rail shell (2), thimble ring (3), fixture ring (4), rotation
Tooth post (5), thrust tooth post (6), push rod (7) and spring (8),
Test tube (1) is intermediate thin both ends coarse-texture, and test tube (1) open at one end is socketed on the inner cavity path of test tube connection ring (11)
On, in the cavity that spring (8) is placed in test tube (1) outer wall and test tube connection ring (11) are formed, guide rail shell (2) one end and test tube connect
The inner cavity bigger diameter end for connecing ring (11) is fixedly connected, and push rod (7) one end can enter from the other end of guide rail shell (2)；
Guide rail shell (2) is the cylindrical structure with cavity, guide rail shell (2) inner wall close to test tube connection ring (11) one end
On, detent slot (9) are arranged circumferentially along axial direction, on guide rail shell (2) inner wall of push rod (7) one end, along axial week
To being evenly arranged protruding guide rail (10)；
Thimble ring (3), fixture ring (4), rotation tooth post (5), thrust tooth post (6) are successively placed on test tube (1) and are placed in outside guide rail
In shell (2) inner cavity；
Thimble ring (3) is the cavity structure of hollow out, is connected along axial circumferentially fixed be connected to close to test tube in the thimble of symmetric design
It connects on thimble ring (3) one end of ring (11) one end,
Fixture ring (4) is coaxially socketed on thimble ring (3), and fixture ring (4) is fixedly connected with guide rail shell (2), thimble ring (3) with
Fixture ring (4) can circumferentially opposite sliding；The thimble groove with thimble cooperation, fixture ring (4) are provided with along axial direction on fixture ring (4)
C-type clamp tool (12) is symmetrically connected on inner wall, it is the annular in opening that C-type clamp, which has (12) original state,；
Rotating tooth post (5) is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with rotation tooth post (5) groove,
Tooth post (5) one end and thimble ring (3) other end axial contact are rotated,
Thrust tooth post (6) is cavity cylindrical structure, axially circumferential on its external wall to be uniformly provided with thrust tooth post (6) groove,
Thrust tooth post (6) groove cooperates with the protruding guide rail (10) of guide rail shell (2) always, thrust tooth post (6) one end and rotation tooth post
(5) other end is dislocation engagement, thrust tooth post (6) other end and push rod (7) end thereof contacts；
Step 2: the present apparatus after body fluid example will be acquired and be put into centrifuge centrifugation, solid phase and liquid phase in centrifugation masterpiece
With lower separation, when centrifugation, push rod (7) are pushed under the action of the centrifugal force, push rod (7) acts on thrust tooth post (6), thrust tooth post
(6) it is moved axially along the protruding guide rail (10) of guide rail shell (2), overcomes spring (8) active force, thrust tooth post (6) drives rotation
Turn tooth post (5) and thimble ring (3) synchronizing moving；
Step 3: when the tooth tip for rotating tooth post (5) is more than protruding guide rail (10), rotation tooth post (5) is acted in spring (8) elastic force
And under the guidance of tooth post top inclined-plane, it may occur that rotate up to two tooth post flank of tooth occlusions, C-type clamp tool (12) is open shape at this time
State does not influence the effect for being centrifugated solid liquid phase completely；
Step 4: when centrifugation is completed, revolving speed decline, centrifugal force is gradually reduced, and spring (8) elastic force is increasingly greater than centrifugal force, spring
(8) elastic force pushes thimble ring (3), makes thimble ring (3) that rotation tooth post (5) to be pushed to slide into protruding guide rail (10) and along protruding guide rail
(10) it moves backward, in this way, thimble ring (3), rotation tooth post (5) and thrust tooth post (6) are anti-together under the effect of spring (8) power
To movement, it is placed in the thimble of thimble ring (3) in the thimble groove of fixture ring (4), while thimble acts on the C of fixture ring (4)
On type fixture (12), realize that the flow liquid channel enclosed of test tube (1) is realized automatic solid liquid phase by the closure of two C-type clamp tools (12)
Physical isolation；
Step 5: taking out the upper phase isolated, if lower layer's solid phase will also take, can be centrifuged again, or directly with forcing
Dynamic push rod (7) more than unclamping again after protruding guide rail (10), can make thimble ring (3) exit thimble to the tooth tip for rotating tooth post (5)
Groove, test tube (1) pipeline is open, takes out lower layer's solid phase.
9. as claimed in claim 8 can be achieved the biological humor sampling separation method that solid-liquid is isolated after being centrifuged, feature exists
In test tube (1) is the assembly of different hardness plastic material, including any one in following scheme:
Scheme 1: being flexible plastic, shore in the range of the upward 5-15mm of C-type clamp tool (12) to the downward 5-15mm of C-type clamp tool (12)
Hardness A is 50-70；Test tube (1) other parts are rigid plastics, and shore hardness D is 70-80, test tube connection ring (11) and test tube
(1) whole for an injection molding；
Scheme 2: the range of test tube (1) bottom to the upward 5-15mm of C-type clamp tool (12) are flexible plastic, and the hard A of shore is 50-70；
Test tube (1) other parts are rigid plastics, and shore hardness D is 70-80, and test tube connection ring (11) and test tube (1) are an injection molding
It is whole；
Scheme 3: test tube (1) inner core is flexible plastic, and shore hardness A is 50-70, and the test tube of test tube (1) open at one end socket connects
Connecing ring (11) is rigid plastics, and shore hardness D is that 70-80, test tube connection ring (11) and test tube (1) pass through test tube (1) opening one
Simultaneously gluing is fastened together for the outer wall construction clamping at end.
10. as claimed in claim 8 can be achieved the biological humor sampling separation method that solid-liquid is isolated after being centrifuged, feature exists
In the C-type clamp of test tube (1) entire length 9-13mm, test tube (1) internal diameter 7-11mm, test tube (1) bottom to closure have (12) top
Distance is 3-8mm.
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