WO2020099475A2 - Dispositif et procédé pour obtenir une fraction d'échantillon acellulaire - Google Patents

Dispositif et procédé pour obtenir une fraction d'échantillon acellulaire Download PDF

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Publication number
WO2020099475A2
WO2020099475A2 PCT/EP2019/081156 EP2019081156W WO2020099475A2 WO 2020099475 A2 WO2020099475 A2 WO 2020099475A2 EP 2019081156 W EP2019081156 W EP 2019081156W WO 2020099475 A2 WO2020099475 A2 WO 2020099475A2
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WO
WIPO (PCT)
Prior art keywords
tube
cylinder
cover
cell
thread
Prior art date
Application number
PCT/EP2019/081156
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German (de)
English (en)
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WO2020099475A3 (fr
Inventor
Tim Kinitz
Original Assignee
Sarstedt Ag & Co. Kg
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Publication date
Application filed by Sarstedt Ag & Co. Kg filed Critical Sarstedt Ag & Co. Kg
Publication of WO2020099475A2 publication Critical patent/WO2020099475A2/fr
Publication of WO2020099475A3 publication Critical patent/WO2020099475A3/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3693—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/02—Blood transfusion apparatus
    • A61M1/0281—Apparatus for treatment of blood or blood constituents prior to transfusion, e.g. washing, filtering or thawing
    • 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
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5021—Test tubes specially adapted for centrifugation purposes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/49—Blood
    • G01N33/491—Blood by separating the blood components
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2202/00—Special media to be introduced, removed or treated
    • A61M2202/04—Liquids
    • A61M2202/0413—Blood
    • A61M2202/0415—Plasma
    • 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
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/042—Caps; Plugs
    • 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
    • B01L2300/04—Closures and closing means
    • B01L2300/046—Function or devices integrated in the closure
    • 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
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0681—Filter
    • 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
    • B01L2300/14—Means for pressure control
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0478—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure pistons

Definitions

  • the present invention relates to a device and a method for obtaining a cell-free fraction of a cell-containing sample, in particular for obtaining a cell-free plasma fraction from whole blood, and the use of the device as a separation device for the production of a cell-free fraction from a cell-containing sample.
  • the device enables a method which has at most only one centrifugation step, preferably exactly one centrifugation step, in order to separate a cell-free plasma fraction from whole blood.
  • the use of the device allows the method to be carried out, and further preferably a subsequent storage, within a tube which is set up for taking blood, that is to say without transferring a fraction from the one tube which contains a whole blood sample.
  • a whole blood sample taken in a centrifuge tube which can be formed from a blood collection tube, into a cell-containing fraction and a lighter fraction in a first centrifugation step, for example 10 minutes at 2500 ⁇ g.
  • the lighter fraction still contains cells. Therefore, the with the lighter fraction generated in the first centrifugation step is removed, for example removed from the cell-containing fraction by means of a pipette, filled into a further centrifuge tube and in a second centrifugation step, for example 15 min at 15000 ⁇ g, in a cell fraction which forms a pellet and a cell-free lighter plasma fraction Cut.
  • the heavier cell fraction for example removed from the heavier fraction by means of a pipette, and filled into a third vessel.
  • the object of the invention is to provide an alternative device for isolation, preferably also for storage, of a cell-free sample fraction, e.g. from whole blood, and to provide a process for producing a cell-free sample fraction, which is in particular a cell-free plasma fraction.
  • the device is preferably intended to enable a method which contains at most or exactly one centrifugation step and in which the cell-free plasma fraction preferably remains connected to the blood collection tube in which the original whole blood sample was contained.
  • the invention solves the problem with the features of the claims and in particular provides a device that a tube set up for blood collection, also as
  • the tube has at its first end a guide for a piston which is displaceable in the tube and at a second end opposite the first end has a detachably attached cover with an inlet opening arranged thereon.
  • the tube has a preset vacuum and can have a closed bottom, firmly connected to the wall, at the first end and accordingly no piston and
  • a cover which is detachably attached to the second end of the tube is also referred to as the first cover and generally preferably has a connector for a cannula as an inlet opening, the connector being preferably covered by a diaphragm which is pierced by the cannula when the connector is connected.
  • the first cover is preferably attached to the second end of the tube by means of a thread.
  • the device has a cylinder which, with its first end, precedes that of a wall which is impermeable to cells and permeable to serum is closed, can be inserted into the tube and is set up to form a seal between the outer surface of the cylinder and the inner surface of the tube.
  • the cylinder is accessible at its second end opposite the first end for the removal of liquid, e.g. open, optionally provided with a pouring edge, or covered by a pierceable diaphragm and can optionally have a second cover covering its second end.
  • liquid e.g. open, optionally provided with a pouring edge, or covered by a pierceable diaphragm and can optionally have a second cover covering its second end.
  • the cylinder can preferably be driven into the tube by means of a thread.
  • the cylinder is in engagement with the tube by means of a thread.
  • the cylinder can engage the tube directly on the thread, so that the cylinder can be driven to move into the tube by relative rotation against the tube.
  • the thread can be arranged on a second cover or a screw nut and be in engagement with the tube or the cylinder, so that the cylinder can be driven into the tube by rotating the second cover or the screw nut through the second cover or the screw nut.
  • the device with a tube for blood collection is suitable for use as a separating device for producing a cell-free fraction from a cell-containing liquid sample, the tube having a first end and at the opposite second end having an inlet opening arranged on a detachably attached first cover, with an after Pulling the sample into the tube and after removing the first cover insertable cylinder, which is set up between its outer surface and the
  • Inner surface of the tube to form a seal and which is displaceable into the tube with its first end first, the first end of the cylinder with one for cells
  • the cylinder is preferably driven by a screw thread for displacement into the tube, which acts between the tube and the cylinder.
  • a second cover can be arranged to be fixed at the second end of the tube and to cover at least the edge of the cylinder at its second end.
  • the second cover has a diaphragm in the area that covers the second end of the cylinder.
  • the screw thread, through which the cylinder is driven for displacement into the tube is arranged on a second cover and stands with one
  • the second cover is preferably arranged to engage the tube, e.g. by means of a screw thread in order to push the cylinder towards the first end of the tube when the second cover rotates against the tube or against the cylinder.
  • the second cover is further preferably configured to close a gap formed between the tube and the cylinder.
  • the second cover is preferably designed to cover the second end of the cylinder in a liquid-tight manner.
  • the screw thread, through which the cylinder is driven for displacement into the tube is formed in that the cylinder can be directly engaged with the tube by means of a thread, so that the cylinder can be rotated relative to the tube along the Tube can be driven.
  • the screw thread, through which the cylinder is driven for displacement into the tube is formed by a screw nut which is in engagement with the cylinder and the tube.
  • the cylinder is driven along the tube by means of the screw nut which engages the cylinder and the tube.
  • the device is configured by assembling the tube, which has a piston guided at its first end or a preset negative pressure in its internal volume, and a first cover arranged at the second end of the tube, with a separate cylinder and optionally a separate second cover and / or optionally a screw nut.
  • the method for producing a cell-free fraction from a blood sample has the steps of
  • a) centrifuging a tube for drawing blood into which a blood sample has been drawn in the tube having a displaceably guided piston at a first end or a preset negative pressure in its inner volume, and an inlet opening arranged on a detachably attached first cover at its opposite second end has, preferably with exactly 1 centrifugation step,
  • the cylinder is preferably driven for insertion into the tube by means of a screw thread, which acts between the tube and the cylinder.
  • the cylinder is inserted into the tube by rotating a second cover which closes the second end of the cylinder in a liquid-tight manner.
  • the second cover can engage in a thread arranged on the tube.
  • the insertion of the cylinder may be driven by turning a nut which engages the cylinder and the tube, or by rotating the cylinder relative to the tube if the cylinder is directly threadedly engaged with the tube .
  • the inlet opening is preferably formed by a connector for a cannula.
  • the second lid can be arranged to connect to the tube, e.g. a thread on the
  • the second cover can optionally be set up to close the entire cross-sectional opening at the second end of the tube, optionally including the opening at the second end of a cylinder inserted therein, only to close a gap which is formed between the tube and the cylinder inserted therein. or just to close the opening of the cylinder at its second end.
  • the cylinder may have a pouring edge at its second end.
  • the cylinder can be driven along the tube by means of a screw nut.
  • a screw nut grips e.g. on the cylinder and on the tube, once by means of a thread and once on a fixed guide, so that the rotation of the screw nut the
  • a nut can be attached to the cylinder and engaged with a thread on the tube.
  • the cylinder can be threaded directly to the tube, which preferably has a preset vacuum, and has no piston to be brought into engagement, so that the cylinder can be driven by rotating relative to the tube along the tube, in particular into the tube.
  • the tube preferably has an individual identification, e.g. a label.
  • the device can have, as an assembly of the blood collection tube, which optionally has a piston guided at its first end or a preset negative pressure in its internal volume, a first cover arranged at its second end, with a separate cylinder, optionally a screw nut, which is connected to the cylinder and the tube is to be brought into engagement, wherein further optionally the cylinder can be directly engaged with the tube by means of a thread, in particular for a tube which has a preset vacuum and has no piston, and / or optionally with a separate second cover be trained.
  • the device preferably forms a blood collection tube which has at a first end a guide for a piston which is displaceably and liquid-tightly guided in the tube, a first cover being arranged at the opposite second end of the tube, e.g. by means of a screw thread which has an inlet opening, e.g. in the form of a connector for a cannula, e.g. has a bayonet connection.
  • the inlet opening is covered by a diaphragm, which is from a
  • Connector connected cannula is pierced.
  • a blood collection tube is available under the Monovette brand from Sarstedt, Nümbrecht.
  • the blood collection tube can optionally contain a stabilizing composition for cell-free nucleic acids and / or for cells.
  • a stabilizing composition placed in the blood collection tube e.g. when a sample is drawn in, mixed with the latter, has the advantage of stabilizing the content of cell-free nucleic acids contained in the cell-free fraction produced, or of reducing the degradation or the re-synthesis of cell-free nucleic acids. Another advantage of such
  • Stabilizing composition can be the proportion of cells that are in the
  • Inserting the cylinder into the tube can be destroyed and the nucleic acids released can decrease.
  • the sample is whole blood, which is drawn into the tube, for example, immediately after it has been removed from a human or animal body.
  • the stabilizing composition or a solution of the stabilizing composition in water or in the sample, can e.g.
  • mercaptoethanol dithiotreitol (DTT)
  • DTT dithiotreitol
  • glycine an alkylamine and / or polyamine as scavenger for free formaldehyde
  • urotropin and at least one buffer substance that buffers to a pH of 7 or below, preferably urotropin and as Buffer substance citrate, each optionally additionally a buffer substance and / or an anticoagulant, which can be a chelating agent for divalent cations, in particular EDTA and / or citrate, optionally PEG,
  • the stabilizing composition for the cell-free nucleic acids contained in the biological sample and / or for the nucleated cells can be a composition which contains at least one buffer substance which buffers to a pH of 7 or below, at least one anticoagulant, which is preferably a chelating agent. and urotropin, optional PEG, e.g. dissolvable in dry mixture or in aqueous solution or in the sample, or consists of them.
  • the stabilizing composition can consist of at least one
  • Buffer substance the at least one anticoagulant, which is a chelating agent, and urotropin, optionally PEG, in aqueous solution.
  • This preferably contains
  • the buffer substance can buffer the aqueous solution to a pH in the range from 3.5 to 7 and have a buffer capacity which corresponds to a concentration of 10 to 100 mM citrate, 1 to 30 gcw./vol.-% in the solution. Urotropin is dissolved.
  • the at least one buffer substance and the at least one anticoagulant, which is a chelating agent can be formed by citrate buffers.
  • the at least one buffer substance can be, for example, citrate buffer, acetate buffer, MES (2-N-morpholinoethanolsulfonic acid), PIPES (piperazine-N, N'-bis-2-ethanosulfonic acid), MOPS (3-N-morpholinopropanesulfonic acid), phosphate buffer and mixtures of at least two of these may be selected and the at least one anticoagulant that is a chelating agent may be selected from citrate and / or EDTA and mixtures thereof.
  • An aqueous solution of the stabilizing composition also referred to as a stabilizing solution, can be contained in the tube in the tube in a volumetric proportion of, for example, up to 20% of the maximum sample volume for which a tube is set up.
  • the device according to the invention is suitable for use as a separation device for isolation, preferably also for storage, of a cell-free sample fraction, in particular a cell-free plasma fraction from whole blood.
  • the invention provides a method which can be carried out with the device for producing a cell-free sample fraction, which is in particular a cell-free plasma fraction.
  • the method for producing a cell-free fraction which is in particular a plasma fraction, or for separating cell-free plasma from a cell-containing fraction, provides for the tube with a sample contained therein, which is in particular a whole blood sample, and optionally with a stabilizing composition for free nucleic acids and / or for cells is centrifuged in exactly one centrifugation step in order to separate the sample into a heavy fraction, which contains cells, and a light fraction, which has a substantially lower content of cells.
  • the cylinder is then pushed into this tube, ie without removing the light lraction from the tube or without decanting the light lraction into another container.
  • the cylinder Since the cylinder has a seal between its outer surface and the inner surface of the tube and is fluid-tightly slidable along the tube, the slight lraction is caused by inserting the cylinder into the tube through the wall attached to the first end of the cylinder Internal volume of the cylinder is pressed and cells are held back by the wall.
  • the wall can e.g. Have pores that have a maximum diameter of 1 mhi in order to allow only cell-free liquid to pass through, in particular only cell-free serum.
  • the wall covering the first end of the cylinder is preferably against one
  • the sample is divided into a heavy, cell-containing lraction and a light, cell-depleted lraction, in particular by exactly one centrifugation step.
  • the wall can be covered by a grille between the Internal volume of the cylinder and the wall is arranged and covers the clear cross section of the cylinder, be supported.
  • a grid can be connected, for example, to the inside of the cylinder, for example by a clamp connection, or can be formed in one piece with the cylinder.
  • the wall preferably consists of material which does not bind nucleic acids, in particular nucleic acids of the sample, or binds only to a small extent, e.g. made of hydrophobic material.
  • the light fraction obtained by centrifugation since the light fraction obtained by centrifugation has passed through the wall of the cylinder into the cylinder when the cylinder is inserted into the tube, the light fraction through the wall located at the first end of the cylinder is from the cell-containing heavy fraction Cut.
  • the arrangement of the cylinder within the tube has the advantage that the light fraction is still connected to the same tube in which the original sample was contained. This prevents mix-ups of samples that are possible by transferring fractions from one container to another, since a label attached to the tube also remains connected to the cylinder.
  • the method may include the step of removing the cell-free fraction from the cylinder, e.g. by suction or pouring, and to store this fraction in another container and / or to analyze it directly.
  • the cell-free fraction which is preferably the plasma fraction separated from a whole blood sample, can e.g. stored for at least 6 h, for at least 12 h or at least 24 h in the cylinder connected to or inserted into the same tube into which the blood sample was drawn, e.g. at 5 to 25 ° C.
  • the insertion of the cylinder into the tube can be driven by loading the cylinder on its second end towards its first end or against the first end of the tube.
  • the insertion of the cylinder into the tube is driven by rotating a second cover against the tube, the second cover with a Screw thread of the tube is engaged and the second cover is arranged at least in sections, preferably completely, against the second end of the cylinder.
  • the second cover can load the second end of the cylinder in the direction of its first end or against the first end of the tube the second cover can be arranged, for example, against a leading edge at the second end of the cylinder.
  • the insertion of the cylinder into the tube may be driven by turning a nut that is engageable with the tube and the cylinder, or by rotating the cylinder relative to the tube in a thread that the cylinder directly engages the tube with is feasible.
  • Arranged locking device which allows the second cover to be fixed on the tube and / or on the cylinder in the direction of screwing the second cover onto the tube and which blocks in the opposite direction, in particular unscrewing the second cover from the tube and / or the cylinder prevented.
  • the second cover preferably has a pierceable section in an area which is to be arranged above the clear cross section of the cylinder, e.g. a pierceable diaphragm.
  • This area is e.g. pierceable by a hollow needle.
  • the pierceable area is preferably formed by a diaphragm which closes automatically after a hollow needle is pulled out.
  • the seal of the outer surface of the cylinder to the inner surface of the tube can be formed by a sealing edge formed on the cylinder and / or a sealing ring, for example an O-ring.
  • a sealing edge and / or a sealing ring can be arranged on the outer surface of the cylinder, for example at the first end of the cylinder, preferably at a distance from the first end of the cylinder.
  • the arrangement of a sealing edge and / or a sealing ring at a distance from the first end of the cylinder has the advantage that the section of the cylinder between its first end, for example the wall arranged thereon, and the sealing edge or the sealing ring guides the insertion of the cylinder can form in the tube.
  • This distance can be, for example, 1 to 50%, for example 2 to 20% of the length of the cylinder by which the cylinder can be pushed into the tube at the maximum.
  • the cylinder can preferably be attached coaxially to the tube, and the cylinder has a rotationally symmetrical outer surface and the tube has a rotationally symmetrical surface
  • FIG. 2 shows a further embodiment of a device according to the invention
  • FIG. 1 shows a tube 1, which can have a preset negative pressure or at the first end 3 of which a piston (not shown) is displaceably guided.
  • a first cover is provided (not shown), which has an attachment piece for a cannula as an inlet opening.
  • a first cover is e.g. to be releasably attached to the tube by means of a thread 7 which is attached to the outside of the tube.
  • the tube 1 is centrifuged, possibly after removing a protruding plunger which engages a piston, with the filled-in sample, so that the sample is divided into a heavy fraction and a light fraction.
  • the first lid is removed from the tube, e.g. unscrewed, and a cylinder 9 with its first end 10, which is covered by a cell-impermeable wall 11, is inserted into the tube 1, e.g. by exerting a force on the second end 8 of the cylinder 9.
  • the tube 1 and cylinder 9 can be formed without thread 7.
  • the cylinder 9 is preferably driven into the tube 1 by turning along a thread 7 with which the cylinder 9 is directly engaged with the tube 1.
  • FIG. 2 shows the device in the form of a tube 1 for taking blood, a screw nut 5 and a cylinder 9.
  • the screw nut 5 engages in the external thread 7, which is arranged at the second end 4 of the tube 1.
  • the screw nut 5 is connected to the cylinder 9 and is not displaceably or rotatably mounted on the cylinder 9 along the cylinder 9 so that the turning of the screw nut 5 drives the insertion of the cylinder 9 into the tube 1.
  • a seal 16 is arranged on the screw nut 5, which bears against the second end of the tube 1 and / or against the cylinder 9 when the screw nut 5 is fixed on the tube 1
  • the seal is formed by a sealing ring 13, which is fixed to the first end 10 of the cylinder.
  • the cylinder 9 can generally have a plurality of circumferential seals which are distributed over the length of the cylinder 9.
  • the second cover 12 may have a seal (not shown) that bears against the second end of the tube 1 when the second cover 12 is attached to the tube 1.
  • the seal 13 can be formed by at least one circumferential sealing ring.
  • FIG. 3 shows an embodiment in which the insertion of the cylinder 9 is driven in that a second cover 12, which covers the cylinder 9 and which engages in the thread 7 at the second end 4 of the tube 1, is turned against the tube 1 , so that the second cover 12 moves towards the first end 3 of the tube 1 and thereby pushes the cylinder 9 into the tube 1 in the same direction.
  • the second cover 12 has, in the area in which it covers the open second end 8 of the cylinder 9, a diaphragm 15 which can be pierced with a hollow needle to remove cell-free samples. Such a second cover 12 closes the second end 4 of the tube 1 and the open second end 8 of the cylinder 9.
  • Whole blood was drawn into a monovette as a blood collection tube as a sample.
  • the tube optionally contained an aqueous composition
  • Stabilization composition consisted of 15% hexamethylenetetramine, 8%
  • Composition based on the drawn blood volume, be presented in the tube.
  • the tube had a piston guide at a first end and on
  • the whole blood sample or the mixture of whole blood and the stabilizing composition was centrifuged until a heavy, cell-containing and red fraction and above that a yellow light fraction formed.
  • the centrifugation was e.g. run at 2500 to 3000 x g for 10 min.
  • the first lid was removed from the tube and a cylinder, the first end of which was covered by a grid-supported membrane as a cell-impermeable wall, was pushed into the tube with its first end first.
  • the cylinder had a circumferential sealing edge which was attached to its outer surface 1 to 2 mm from its first end and which formed a liquid-tight seal to the inner surface of the tube.
  • the cylinder wall was arranged coaxially and at a distance of approximately 0.2 to 0.5 mm from the inner surface of the tube. During the insertion of the cylinder into the tube, at least a portion of the light fraction was pressed through the membrane and was found inside the cylinder.
  • a second cover was placed on the second end of the tube, which had an internal thread that engaged the external thread on the second end of the tube.
  • the second cover covered the second end of the tube and also the second end of the cylinder, so that when the second cover was screwed onto the tube, the cylinder was pushed into the tube.
  • the second lid had the advantage of preventing sample from escaping from the second end of the tube or cylinder.
  • Periampullary Cancer Direct Quantitative PCR for ALU repeats. Clinical Chemistry, 52 (6), 1062-1069) showed that the filtration removed cells and cell fragments from the samples as well as a second centrifugation step carried out for comparison (15 min, 15,000 x g).

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  • Measurement Of The Respiration, Hearing Ability, Form, And Blood Characteristics Of Living Organisms (AREA)

Abstract

L'invention concerne un tube de prélèvement sanguin comprenant un cylindre qui, par sa première extrémité avant, fermée par une paroi imperméable aux cellules et perméable au sérum sanguin, peut être introduit dans le tube et est conçu pour former un élément d'étanchéité entre la surface extérieure du cylindre et la surface intérieure du tube. Le cylindre est accessible par sa deuxième extrémité opposée à la première extrémité pour le prélèvement de liquide, éventuellement pourvu d'un bord verseur, ou recouvert par une membrane perforable.
PCT/EP2019/081156 2018-11-13 2019-11-13 Dispositif et procédé pour obtenir une fraction d'échantillon acellulaire WO2020099475A2 (fr)

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DE102018219379.7A DE102018219379A1 (de) 2018-11-13 2018-11-13 Vorrichtung und Verfahren zur Gewinnung einer zellfreien Probenfraktion
DE102018219379.7 2018-11-13

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JP3931069B2 (ja) * 2001-08-22 2007-06-13 協和メデックス株式会社 採血用具、これを用いた定量用溶液調製方法、定量用溶液調製器具及びその使用方法並びに定量用溶液を用いた定量方法
AT500459B1 (de) * 2004-01-23 2010-08-15 Greiner Bio One Gmbh Verfahren zum zusammenbau einer kappe mit einem aufnahmebehälter
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JP5730695B2 (ja) * 2011-02-15 2015-06-10 株式会社雅精工 血液分離シリンダ
WO2014064921A1 (fr) * 2012-10-22 2014-05-01 パナソニック株式会社 Dispositif à filtre
EP4122586A1 (fr) * 2015-06-08 2023-01-25 Becton, Dickinson and Company Extracteur de plasma

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