WO2022250142A1 - 血液採取容器、血漿の分離方法、細胞外遊離核酸の分離方法及び細胞外小胞の分離方法 - Google Patents
血液採取容器、血漿の分離方法、細胞外遊離核酸の分離方法及び細胞外小胞の分離方法 Download PDFInfo
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- WO2022250142A1 WO2022250142A1 PCT/JP2022/021743 JP2022021743W WO2022250142A1 WO 2022250142 A1 WO2022250142 A1 WO 2022250142A1 JP 2022021743 W JP2022021743 W JP 2022021743W WO 2022250142 A1 WO2022250142 A1 WO 2022250142A1
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- collection container
- blood collection
- blood
- plasma
- aqueous solution
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- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 229940086066 potassium hydrogencarbonate Drugs 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- KCTAWXVAICEBSD-UHFFFAOYSA-N prop-2-enoyloxy prop-2-eneperoxoate Chemical compound C=CC(=O)OOOC(=O)C=C KCTAWXVAICEBSD-UHFFFAOYSA-N 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011403 purification operation Methods 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 238000010526 radical polymerization reaction Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 239000005368 silicate glass Substances 0.000 description 1
- 229920002050 silicone resin Polymers 0.000 description 1
- 239000005361 soda-lime glass Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229920003002 synthetic resin Polymers 0.000 description 1
- 239000000057 synthetic resin Substances 0.000 description 1
- KKEYFWRCBNTPAC-UHFFFAOYSA-L terephthalate(2-) Chemical compound [O-]C(=O)C1=CC=C(C([O-])=O)C=C1 KKEYFWRCBNTPAC-UHFFFAOYSA-L 0.000 description 1
- 229920002725 thermoplastic elastomer Polymers 0.000 description 1
- 229920005992 thermoplastic resin Polymers 0.000 description 1
- 229920001187 thermosetting polymer Polymers 0.000 description 1
- 230000009974 thixotropic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- JNXDCMUUZNIWPQ-UHFFFAOYSA-N trioctyl benzene-1,2,4-tricarboxylate Chemical compound CCCCCCCCOC(=O)C1=CC=C(C(=O)OCCCCCCCC)C(C(=O)OCCCCCCCC)=C1 JNXDCMUUZNIWPQ-UHFFFAOYSA-N 0.000 description 1
- WDRCVXGINNJWPH-UHFFFAOYSA-N tris(6-methylheptyl) benzene-1,2,4-tricarboxylate Chemical compound CC(C)CCCCCOC(=O)C1=CC=C(C(=O)OCCCCCC(C)C)C(C(=O)OCCCCCC(C)C)=C1 WDRCVXGINNJWPH-UHFFFAOYSA-N 0.000 description 1
- FJFYFBRNDHRTHL-UHFFFAOYSA-N tris(8-methylnonyl) benzene-1,2,4-tricarboxylate Chemical compound CC(C)CCCCCCCOC(=O)C1=CC=C(C(=O)OCCCCCCCC(C)C)C(C(=O)OCCCCCCCC(C)C)=C1 FJFYFBRNDHRTHL-UHFFFAOYSA-N 0.000 description 1
- 229920006337 unsaturated polyester resin Polymers 0.000 description 1
Images
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
- A61J—CONTAINERS SPECIALLY ADAPTED FOR MEDICAL OR PHARMACEUTICAL PURPOSES; DEVICES OR METHODS SPECIALLY ADAPTED FOR BRINGING PHARMACEUTICAL PRODUCTS INTO PARTICULAR PHYSICAL OR ADMINISTERING FORMS; DEVICES FOR ADMINISTERING FOOD OR MEDICINES ORALLY; BABY COMFORTERS; DEVICES FOR RECEIVING SPITTLE
- A61J1/00—Containers specially adapted for medical or pharmaceutical purposes
- A61J1/05—Containers specially adapted for medical or pharmaceutical purposes for collecting, storing or administering blood, plasma or medical fluids ; Infusion or perfusion containers
-
- 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
- B01L3/50215—Test tubes specially adapted for centrifugation purposes using a float to separate phases
-
- 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
-
- 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
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0631—Purification arrangements, e.g. solid phase extraction [SPE]
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/16—Reagents, handling or storing thereof
-
- 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
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
Definitions
- the present invention relates to a blood collection container.
- the present invention also relates to a method for separating plasma, a method for separating extracellular free nucleic acid, and a method for separating extracellular vesicles using the blood collection container.
- Blood collection containers such as blood collection tubes are widely used in clinical examinations to collect blood. After collecting blood in a blood collection container containing a plasma separation material, the blood can be separated into plasma and blood cells by centrifuging the blood collection container. At this time, the plasma is positioned above the plasma separation material, and the blood cells are positioned below.
- the blood collection container containing the plasma separation material include a blood collection container containing a composition for plasma separation containing a resin, an inorganic powder, etc. (for example, Patent Document 1), and a blood container containing a jig for plasma separation.
- a collection container for example, Patent Document 2 is known.
- Patent Document 3 describes a device for separating extracellular DNA in blood.
- the separated plasma may contain a relatively large amount of leukocytes. If the plasma contains a relatively large number of leukocytes, the leukocytes will be destroyed over time during storage of the sample, and components such as proteins and nucleic acids in the leukocytes will leak into the plasma, resulting in negative test results. may affect
- test results vary greatly depending on the nucleic acid leaked from the leukocytes.
- a blood collection container main body, a plasma separation material accommodated in the blood collection container main body, and an aqueous solution contained in the blood collection container main body are contained in the aqueous solution.
- a blood collection container is provided, wherein the solute comprises an anticoagulant, and the total concentration of the solute in the aqueous solution is 100 mM or more and 450 mM or less, or 1200 mM or more.
- the total concentration of the solute in the aqueous solution is 250 mM or more and 390 mM or less, or 1200 mM or more and 6500 mM or less.
- the solute contained in the aqueous solution contains a second solute other than an anticoagulant, and the second solute contains inorganic salts or sugars.
- the second solute contains an inorganic salt
- the inorganic salt contains a sodium salt or a potassium salt.
- the second solute contains saccharides, and the saccharides contain glucose, sucrose, or trehalose.
- the anticoagulant is EDTA, a metal salt of EDTA, heparin, a metal salt of heparin, or sodium citrate.
- the blood collection container is a blood collection container from which a predetermined amount of blood is collected, and an amount of physiological fluid equivalent to the predetermined amount of blood collected in the blood collection container.
- the osmotic pressure of the mixed liquid is 330 mOsm/L or more and 380 mOsm/L or less. or 440 mOsm/L or more and 1300 mOsm/L or less.
- the plasma separation material has a specific gravity of 1.030 or more and 1.120 or less at 25°C.
- the plasma separation material is a composition for plasma separation.
- the plasma separation composition contains an organic component having fluidity at 25° C. and an inorganic fine powder, the organic component contains a resin, and the The inorganic fine powder includes finely divided silica.
- the finely divided silica contains hydrophilic silica.
- the hydrophilic silica content is 0.01% by weight or more and 2.50% by weight or less in 100% by weight of the plasma separation composition.
- the finely divided silica contains hydrophilic silica and hydrophobic silica.
- the plasma separation composition has a specific gravity of 1.050 or more at 25° C., and the inorganic fine powder has a higher specific gravity than the fine powder silica. Contains fine powder.
- the resin includes petroleum resin, cyclopentadiene resin, polyester resin, or (meth)acrylic resin.
- the blood collection container is used to separate extracellular free nucleic acids or extracellular vesicles in blood.
- a plasma separation method comprising the steps of: collecting blood in the blood collection container described above; and centrifuging the blood collection container containing the blood. .
- a blood collection container comprises a blood collection container main body, a plasma separation material contained within the blood collection container main body, and an aqueous solution contained within the blood collection container main body.
- the solute contained in the aqueous solution contains an anticoagulant, and the total concentration of the solute in the aqueous solution is 100 mM or more and 450 mM or less, or 1200 mM or more. Since the blood collection container according to the present invention has the above configuration, it is possible to suppress contamination of plasma with leukocytes and components of leukocytes.
- FIG. 1 is a front cross-sectional view schematically showing a blood collection container according to one embodiment of the present invention.
- a blood collection container comprises a blood collection container main body, a plasma separation material contained within the blood collection container main body, and an aqueous solution contained within the blood collection container main body.
- the solute contained in the aqueous solution contains an anticoagulant, and the total concentration of the solute in the aqueous solution is 100 mM or more and 450 mM or less, or 1200 mM or more.
- the blood collection container according to the present invention has the above configuration, it is possible to suppress contamination of plasma with leukocytes and components of leukocytes.
- the blood collection container according to the present invention has the above configuration, contamination of blood plasma with red blood cells can be suppressed.
- the separated plasma may contain a relatively large amount of leukocytes. If the plasma contains a relatively large number of leukocytes, components of the leukocytes may leak into the plasma and affect tests using plasma. With conventional blood collection containers, it is difficult to sufficiently suppress contamination of plasma with leukocytes and contamination of components in leukocytes. Although a blood collection container containing a cell stabilizer that stabilizes blood cells is sometimes used in order to suppress the effect on the test results, the cell stabilizer is expensive, and the type and concentration of the agent are limited. Some may be harmful to humans and the environment.
- the blood collection container according to the present invention contamination of leukocytes into plasma and contamination of components in leukocytes can be effectively suppressed.
- the blood and the aqueous solution are mixed and the osmotic pressure of the blood increases. Therefore, the water content in white blood cells and the water content in red blood cells move out of the blood cells, and the specific gravities of white blood cells and red blood cells increase.
- the white blood cells and red blood cells with increased specific gravity move better downward than the plasma separation material having a specific specific gravity. As a result, contamination of plasma with white blood cells and red blood cells can be suppressed.
- the concentration of the solute is within an appropriate range, so the stress on the blood cells is suppressed, and the leakage of the contents from the blood cells mixed in the plasma is suppressed. can be effectively suppressed.
- (meth)acryl means one or both of “acryl” and “methacryl”.
- the blood collection container includes a plasma separation material housed within the blood collection container main body.
- a conventionally known plasma separation material can be used as the plasma separation material.
- the plasma separation material include plasma separation compositions and plasma separation jigs.
- the plasma separation material is preferably the plasma separation composition, because the plasma separation material can be easily produced.
- the specific gravity of the plasma separation material at 25°C may be 1.030 or more, 1.040 or more, 1.050 or more, or 1.060 or more. well, it may exceed 1.060, or it may be 1.070 or more.
- the specific gravity of the plasma separation material at 25°C may be 1.120 or less, 1.100 or less, 1.080 or less, or less than 1.070. well, it may be less than 1.060, it may be less than 1.050, it may be less than 1.040.
- the storage location of the plasma separation material is not particularly limited as long as it is within the blood collection container main body.
- the plasma separation material may be arranged on the bottom of the blood collection container main body, may be arranged on the inner wall surface of the blood collection container main body, or may be arranged on the side wall surface of the blood collection container main body. may be
- the plasma separation composition is a composition that migrates between the plasma layer and the blood cell layer during centrifugation to form a partition wall. Moreover, the composition for plasma separation is used for the purpose of preventing component migration between the plasma layer and the blood cell layer after centrifugation.
- the plasma separation composition preferably has thixotropic properties.
- the plasma separation composition may be housed in the bottom portion of the blood collection container main body, or may be arranged on the side wall surface of the blood collection container main body. From the viewpoint of exhibiting the effects of the present invention more effectively, the plasma separation composition is preferably contained in the bottom portion of the blood collection container main body.
- a conventionally known composition for plasma separation can be used as the composition for plasma separation.
- the composition for plasma separation preferably contains an organic component having fluidity at 25°C and an inorganic fine powder.
- Each of the organic component having fluidity at 25° C. and the inorganic fine powder may be used alone, or two or more thereof may be used in combination.
- the viscosity of the organic component at 25°C is preferably 30 Pa ⁇ s or more, more preferably 50 Pa ⁇ s or more, preferably 200 Pa ⁇ s or less, more preferably 100 Pa ⁇ s or less.
- the viscosity is equal to or higher than the lower limit and equal to or lower than the upper limit, the fluidity of the composition for plasma separation is enhanced, and the strength of partition walls can be enhanced.
- the viscosity of the above organic component at 25°C is measured using an E-type viscometer (for example, "TVE-35" manufactured by Toki Sangyo Co., Ltd.) under the conditions of 25°C and a shear rate of 1.0 sec -1 . .
- the organic component preferably contains the resin, and more preferably contains the resin and the organic compound.
- the organic component is a mixture of the resin and the organic compound, it is sufficient that the mixture (the organic component) has fluidity, and the resin or the organic compound has fluidity. It doesn't have to be.
- the resin may be, for example, a solid resin at 25°C.
- Each of the above resin and the above organic compound may be used alone, or two or more thereof may be used in combination.
- Examples of the above resins include petroleum resins, cyclopentadiene resins, polyester resins, polyurethane resins, (meth)acrylic resins, silicone resins, ⁇ -olefin-fumarate ester copolymers, sebacic acid and 2,2-dimethyl-1. ,3-propanediol and 1,2-propanediol copolymers, polyether polyurethane resins, and polyether polyester resins. Only one kind of the resin may be used, or two or more kinds thereof may be used in combination.
- the resin preferably contains petroleum resin, cyclopentadiene resin, polyester resin, or (meth)acrylic resin.
- Examples of the cyclopentadiene-based resin include polymers of cyclopentadiene-based monomers, copolymers of cyclopentadiene-based monomers and aromatic monomers, and dicyclopentadiene resins.
- the cyclopentadiene-based resin may be hydrogenated.
- the polymer of the cyclopentadiene monomer and the copolymer of the cyclopentadiene monomer and the aromatic monomer may be oligomers.
- cyclopentadiene-based monomers examples include cyclopentadiene, dicyclopentadiene, and alkyl-substituted derivatives of cyclopentadiene.
- Styrene, methylstyrene, indene, methylindene and the like are mentioned as the aromatic monomers.
- polyester resins include polyalkylene terephthalate resins and polyalkylene naphthalate resins.
- polyalkylene terephthalate resin examples include polyethylene terephthalate, polybutylene terephthalate and poly-1,4-cyclohexanedimethylene terephthalate.
- polyurethane resin examples include a reaction product of a polyol compound and an isocyanate compound.
- the (meth)acrylic resin includes a resin obtained by polymerizing at least one (meth)acrylic acid ester monomer, and at least one (meth)acrylic acid ester monomer and at least one and a resin obtained by polymerizing a monomer other than the (meth)acrylic acid ester monomer.
- Examples of the (meth)acrylic acid ester monomers include (meth)acrylic acid alkyl esters having an alkyl group having 1 to 20 carbon atoms, (meth)acrylic acid polyalkylene glycol esters, (meth)acrylic acid esters, acid alkoxyalkyl ester, (meth)acrylic acid hydroxyalkyl ester, (meth)acrylic acid glycidyl ester, (meth)acrylic acid dialkylaminoalkyl ester, (meth)acrylic acid benzyl ester, (meth)acrylic acid phenoxyalkyl ester, ( meth)acrylic acid cyclohexyl ester, (meth)acrylic acid isobornyl ester, and (meth)acrylic acid alkoxysilylalkyl ester, and the like. Only one kind of the (meth)acrylic acid ester monomer may be used, or two or more kinds thereof may be used in combination.
- Examples of the above organic compounds include benzene polycarboxylic acid alkyl ester derivatives.
- the organic compound is preferably a benzene polycarboxylic acid alkyl ester derivative. Therefore, the organic component is preferably a mixture of the resin and the benzene polycarboxylic acid alkyl ester derivative.
- benzene polycarboxylic acid alkyl ester derivatives examples include phthalates, trimellitates, pyromellitic esters, and the like. Only one kind of the benzene polycarboxylic acid alkyl ester derivative may be used, or two or more kinds thereof may be used in combination.
- trimellitate examples include tri-n-octyl trimellitate, triisooctyl trimellitate, and triisodecyl trimellitate.
- Examples of the pyromellitic acid ester include tetraisooctyl pyromellitic acid.
- trimellitate esters include “Monocizer W700” and “Monocizer W-750” manufactured by DIC, and “Sansocizer TOTM” and “Sansocizer TITM” manufactured by Shin Nippon Rika.
- the benzene polycarboxylic acid alkyl ester derivative is preferably a phthalate, a trimellitate, or a pyromellitate, more preferably a trimellitate.
- the content of the organic component in 100% by weight of the composition for plasma separation is preferably 80% by weight or more, more preferably 85% by weight or more, still more preferably 90% by weight or more, and preferably 97% by weight or less. .
- Inorganic fine powder examples include fine powder silica, titanium oxide powder, calcium carbonate powder, zinc oxide powder, alumina powder, glass fine powder, talc powder, kaolin powder, bentonite powder, titania powder, and zirconium powder.
- the inorganic fine powder preferably contains fine powder silica.
- the inorganic fine powder includes finely powdered silica and an inorganic fine powder other than finely powdered silica (second inorganic fine powder). and more preferably.
- the specific gravity of the plasma separation composition at 25° C. is 1.050 or higher
- the inorganic fine powder may not contain the second inorganic fine powder.
- the specific gravity of the composition for plasma separation at 25° C. is less than 1.050
- the inorganic fine powder may contain the second inorganic fine powder.
- Each of the inorganic fine powder, the fine silica powder, and the second inorganic fine powder may be used alone or in combination of two or more.
- the finely divided silica includes natural silica and synthetic silica.
- Synthetic silica includes hydrophilic silica and hydrophobic silica.
- Hydrophilic silica has the effect of imparting thixotropy to the composition for plasma separation and adjusting the specific gravity, for example, by hydrogen bonding between hydroxyl groups on the particle surfaces.
- hydrophobic silica has a smaller effect of imparting thixotropy than hydrophilic silica.
- the finely divided silica preferably contains hydrophilic silica, and includes hydrophilic silica and hydrophobic silica. is more preferable.
- the finely divided silica preferably contains at least hydrophilic silica.
- the second inorganic fine powder is preferably an inorganic fine powder having a higher specific gravity than finely divided silica, and is an inorganic fine powder having a specific gravity of 3 or more, such as zinc oxide powder, titanium oxide powder, and alumina powder. is more preferred.
- the specific gravity of the second inorganic fine powder is preferably 3 or more, more preferably 3.5 or more, and even more preferably 4 or more. The higher the specific gravity of the second inorganic fine powder, the better. When the specific gravity is equal to or higher than the lower limit, the specific gravity of the composition for plasma separation can be effectively increased.
- the average particle size of the inorganic fine powder, the fine silica powder, and the second inorganic fine powder is not particularly limited.
- the average particle size of the inorganic fine powder, the fine silica powder, and the second inorganic fine powder may be 1 nm or more, 10 nm or more, 500 nm or less, or 100 nm or less. There may be.
- the average particle diameters of the inorganic fine powder, the fine silica powder, and the second inorganic fine powder are the average diameters measured on a volume basis, and are the values of the median diameter (D50) at 50%.
- the volume average particle size (D50) can be measured by a laser diffraction/scattering method, an image analysis method, a Coulter method, a centrifugal sedimentation method, or the like.
- the volume average particle size (D50) is preferably obtained by measurement by a laser diffraction/scattering method or an image analysis method.
- the specific surface area of the finely divided silica is not particularly limited.
- the specific surface area of the finely divided silica may be 20 m 2 /g or more, 100 m 2 /g or more, 500 m 2 /g or less, or 300 m 2 /g or less. good too.
- the specific surface area of the finely divided silica is measured by the BET method.
- the content of the hydrophilic silica in 100% by weight of the composition for plasma separation is preferably 0.01% by weight or more, more preferably 0.10% by weight or more, and still more preferably 0.30% by weight or more. is 2.50% by weight or less, more preferably 2.00% by weight or less.
- the content of the hydrophilic silica is equal to or higher than the lower limit and equal to or lower than the upper limit, both the specific gravity and thixotropy of the composition for plasma separation can be maintained within a more suitable range.
- the content of the finely divided silica in 100% by weight of the composition for plasma separation is preferably 0.1% by weight or more, more preferably 0.5% by weight or more, preferably 10% by weight or less, and more preferably 7% by weight. % by weight or less.
- content of the finely divided silica is above the lower limit and below the upper limit, both the specific gravity and thixotropy of the composition for plasma separation can be maintained within a more suitable range.
- the content of the second inorganic fine powder is preferably 0.01% by weight or more, more preferably 0.1% by weight or more, and preferably 10% by weight or less. Preferably, it is 7% by weight or less.
- the content of the second inorganic fine powder is not less than the lower limit and not more than the upper limit, the specific gravity of the plasma separation composition can be effectively increased.
- the content of the inorganic fine powder in 100% by weight of the composition for plasma separation is preferably 0.1% by weight or more, more preferably 0.5% by weight or more, preferably 10% by weight or less, and more preferably 7% by weight. % by weight or less.
- the specific gravity of the plasma separation composition can be effectively increased.
- composition for plasma separation may contain other ingredients than those mentioned above as long as the effects of the present invention are not impaired.
- other components include organic gelling agents, thermoplastic elastomers, polyalkylene glycols, silicone oils, co-solvents, antioxidants, colorants and water.
- organic gelling agents include organic gelling agents, thermoplastic elastomers, polyalkylene glycols, silicone oils, co-solvents, antioxidants, colorants and water.
- the specific gravity of the plasma separation composition at 25°C is preferably 1.030 or more, more preferably 1.040 or more, and preferably 1.120 or less.
- the specific gravity of the composition for plasma separation at 25°C is not less than the above lower limit and not more than the above upper limit, the effect of the present invention can be exhibited more effectively.
- the specific gravity of the composition for plasma separation at 25°C may be 1.050 or more, 1.060 or more, 1.060 or more, or 1.070 or more. may be The specific gravity of the plasma separation composition at 25°C may be 1.100 or less, 1.080 or less, less than 1.070, or less than 1.060. may be less than 1.050, or less than 1.040.
- the specific gravity of the composition for plasma separation at 25° C. was obtained by dropping one drop of the composition for plasma separation sequentially into saline at 25° C. whose specific gravity was adjusted stepwise at intervals of 0.002, and floating and sinking in the saline. measured by
- the viscosity of the plasma separation composition at 25°C is preferably 100 Pa ⁇ s or more, more preferably 150 Pa ⁇ s or more, preferably 500 Pa ⁇ s or less, more preferably 400 Pa ⁇ s or less. When the viscosity is at least the lower limit and at most the upper limit, the effects of the present invention can be exhibited more effectively.
- the viscosity of the composition for plasma separation at 25° C. was measured using an E-type viscometer (eg, “TVE-35” manufactured by Toki Sangyo Co., Ltd.) under conditions of 25° C. and a shear rate of 1.0 sec ⁇ 1 . measured.
- E-type viscometer eg, “TVE-35” manufactured by Toki Sangyo Co., Ltd.
- the jig for plasma separation is a jig that moves between the plasma layer and the blood cell layer during centrifugation to form a partition wall. Moreover, the jig for plasma separation is used for the purpose of preventing component transfer between the plasma layer and the blood cell layer.
- a conventionally known plasma separation jig can be used as the plasma separation jig.
- Examples of the plasma separation jig include a mechanical separator (plasma separation jig) described in WO2010/132783A1 and the like.
- Examples of materials for the plasma separation jig include elastomers.
- the blood collection container includes an aqueous solution contained within the blood collection container body.
- the aqueous solution contains an anticoagulant and water.
- the solute contained in the aqueous solution includes an anticoagulant.
- the aqueous solution preferably contains a second solute other than the anticoagulant. Therefore, the aqueous solution preferably contains an anticoagulant (first solute), a second solute, and water.
- the solute contained in the aqueous solution preferably contains an anticoagulant (first solute) and a second solute.
- the total concentration of the solute in the aqueous solution is 100 mM or more and 450 mM or less, or 1200 mM or more. That is, the total concentration of the solutes in the aqueous solution is 100 mmol/L or more and 450 mmol/L or less, or 1200 mmol/L or more.
- the total concentration of solutes is the concentration of anticoagulants if the aqueous solution contains only anticoagulants as solutes.
- the total concentration of the solute is the sum of the concentration of the anticoagulant and the concentration of the second solute when the aqueous solution contains an anticoagulant and a second solute as solutes.
- the concentration of the anticoagulant is the concentration of the anticoagulant in the aqueous solution when the aqueous solution contains only one type of anticoagulant, and when the aqueous solution contains two or more types of anticoagulant, is the total concentration of anticoagulant in the aqueous solution.
- the concentration of the second solute is the concentration of the second solute in the aqueous solution when the aqueous solution contains only one second solute, and the aqueous solution contains two or more second solutes. If so, it is the sum of the concentrations of the second solutes in the aqueous solution.
- the aqueous solution contains three types of solutes, an anticoagulant (X), a second solute (Y), and a second solute (Z), and their concentrations (mM) are A, B, C, the total concentration of solutes in the aqueous solution is (A+B+C) mM.
- the total concentration of the solute in the aqueous solution is preferably the total concentration of components other than water contained in the aqueous solution.
- the total concentration of the solutes in the aqueous solution may be 100 mM or more and 450 mM or less, or may be 1200 mM or more.
- the total concentration of the solutes in the aqueous solution is 100 mM or more and 450 mM or less
- the total concentration of the solutes in the aqueous solution is preferably 250 mM or more, more preferably 300 mM or more, preferably 390 mM or less, and more preferably 380 mM or less.
- the total concentration of the solutes is equal to or higher than the lower limit and equal to or lower than the upper limit, the effects of the present invention can be exhibited more effectively.
- the total concentration of the solutes in the aqueous solution is 1200 mM or higher
- the total concentration of the solutes in the aqueous solution is preferably 1500 mM or higher, more preferably 2000 mM or higher, preferably 7000 mM or lower, and more preferably 6500 mM or lower.
- the total concentration of the solutes is equal to or higher than the lower limit and equal to or lower than the upper limit, the effects of the present invention can be exhibited more effectively.
- the aqueous solution contains an anticoagulant.
- Conventionally known anticoagulants can be used as the anticoagulant. Only one kind of the anticoagulant may be used, or two or more kinds thereof may be used in combination.
- anticoagulant examples include heparin, heparin metal salts, ethylenediaminetetraacetic acid (EDTA), EDTA metal salts and citric acid.
- the anticoagulant is preferably EDTA, a metal salt of EDTA, heparin, a metal salt of heparin, or sodium citrate.
- the concentration of the anticoagulant in the aqueous solution is preferably 2 mM or more, more preferably 5 mM or more, still more preferably 10 mM or more, preferably 2000 mM or less, more preferably 1000 mM or less, even more preferably 500 mM or less, still more preferably 250 mM. Below, even more preferably 100 mM or less, particularly preferably 50 mM or less.
- concentration of the anticoagulant is equal to or higher than the lower limit and equal to or lower than the upper limit, good anticoagulant performance can be exhibited.
- the second solute is a solute other than the anticoagulant contained in the aqueous solution.
- the osmotic pressure of the blood collected in the blood collection container can be effectively increased without excessively increasing the content of the anticoagulant. Only one kind of the second solute may be used, or two or more kinds thereof may be used in combination.
- the second solute is not particularly limited as long as it is a component other than an anticoagulant.
- examples of the second solute include inorganic salts, sugars, sugar alcohols, propylene glycol and polyethylene glycol (PEG).
- inorganic salts examples include sodium salts such as sodium chloride and sodium hydrogen phosphate, and potassium salts such as potassium chloride and potassium hydrogen carbonate.
- Examples of the above saccharides include monosaccharides, disaccharides and polysaccharides.
- Examples of the monosaccharides include glucose, dihydroxyacetone, fructose and galactose.
- Examples of the disaccharides include sucrose, trehalose, maltose and lactulose.
- Examples of the polysaccharides include dextran, hydroxyethyl starch, methylcellulose, polysucrose, and the like.
- sugar alcohols examples include D-mannitol and D-sorbitol.
- the second solute preferably contains inorganic salts or sugars. In this case, the effects of the present invention can be exhibited more effectively.
- the inorganic salt preferably contains sodium salt or potassium salt, and more preferably contains sodium chloride or potassium chloride. In this case, the effects of the present invention can be exhibited even more effectively.
- the saccharide preferably contains glucose, sucrose, or trehalose. In this case, the effects of the present invention can be exhibited even more effectively.
- the concentration of the inorganic salt in the aqueous solution is preferably 100 mM or more, more preferably 200 mM or more, still more preferably 300 mM or more, and preferably 450 mM or less. It is more preferably 420 mM or less, still more preferably 400 mM or less.
- concentration of the inorganic salt is equal to or higher than the lower limit and equal to or lower than the upper limit, the effects of the present invention can be exhibited even more effectively.
- the concentration of the inorganic salt in the aqueous solution is preferably 400 mM or higher, more preferably 500 mM or higher, still more preferably 1000 mM or higher, and preferably 5000 mM or lower, and more preferably. is 2000 mM or less, more preferably 1500 mM or less.
- concentration of the inorganic salt is equal to or higher than the lower limit and equal to or lower than the upper limit, the effects of the present invention can be exhibited even more effectively.
- the concentration of the inorganic salt is the concentration of the inorganic salt in the aqueous solution when the aqueous solution contains only one type of inorganic salt, and the concentration of the inorganic salt in the aqueous solution when the aqueous solution contains two or more types of inorganic salts. It is the total concentration of salt.
- the concentration of the saccharides in the aqueous solution is preferably 100 mM or more, more preferably 200 mM or more, still more preferably 300 mM or more, preferably 450 mM or less. It is preferably 420 mM or less, more preferably 400 mM or less.
- concentration of the saccharides is above the lower limit and below the upper limit, the effects of the present invention can be exhibited even more effectively.
- the concentration of the saccharides in the aqueous solution is preferably 400 mM or higher, more preferably 500 mM or higher, still more preferably 1000 mM or higher, preferably 5000 mM or lower, and more preferably It is 3000 mM or less, more preferably 2500 mM or less.
- concentration of the saccharides is above the lower limit and below the upper limit, the effects of the present invention can be exhibited even more effectively.
- the concentration of the saccharides is the concentration of the saccharides in the aqueous solution when the aqueous solution contains only one type of saccharides, and the total concentration of the saccharides in the aqueous solution when the aqueous solution contains two or more types of saccharides. is.
- the concentration of the second solute in the aqueous solution is preferably 80 mM or more, more preferably 100 mM or more, still more preferably 300 mM or more, preferably 450 mM. 420 mM or less, more preferably 400 mM or less.
- concentration of the second solute is equal to or higher than the lower limit and equal to or lower than the upper limit, the effects of the present invention can be exhibited even more effectively.
- the concentration of the second solute in the aqueous solution is preferably 400 mM or higher, more preferably 1000 mM or higher, still more preferably 2000 mM or higher, and preferably 7000 mM or lower. It is more preferably 6500 mM or less, still more preferably 6000 mM or less.
- concentration of the second solute is equal to or higher than the lower limit and equal to or lower than the upper limit, the effects of the present invention can be exhibited even more effectively.
- the amount of the aqueous solution contained in the blood collection container main body is appropriately changed depending on the size of the blood collection container main body, the amount of blood to be collected, and the like.
- the amount of the aqueous solution contained in the blood collection container body is preferably 0.1 mL or more, more preferably 0.5 mL or more, still more preferably 0.7 mL or more, preferably 5 mL or less, more preferably 3 mL or less, More preferably, it is 2.5 mL or less.
- the amount of the aqueous solution is equal to or more than the lower limit and equal to or less than the upper limit, the effects of the present invention can be exhibited more effectively without excessive dilution of blood.
- the shape of the blood collection container main body is not particularly limited.
- the blood collection container main body is preferably a bottomed tubular container.
- the material of the blood collection container body is not particularly limited.
- Materials for the blood collection container body include thermoplastic resins such as polyethylene, polypropylene, polystyrene, polyethylene terephthalate, polymethyl methacrylate, and polyacrylonitrile; thermosetting resins such as unsaturated polyester resins, epoxy resins, and epoxy-acrylate resins; Modified natural resins such as cellulose acetate, cellulose propionate, ethyl cellulose and ethyl chitin; silicate glasses such as soda lime glass, phosphosilicate glass and borosilicate glass; and glasses such as quartz glass. Only one kind of material may be used for the blood collection container main body, or two or more kinds may be used in combination.
- the blood collection container preferably has a stopper.
- a conventionally known plug can be used as the plug. It is preferable that the stopper is made of a material and has a shape that can be attached to the opening of the blood collection container main body in an airtight and liquid-tight manner.
- the plug is preferably configured so that a blood collection needle can be pierced therethrough.
- the plug examples include a plug having a shape that fits into the opening of the blood collection container main body, a sheet-like sealing plug, and the like.
- the plug body may be a plug body including a plug body such as a rubber plug and a cap member made of plastic or the like. In this case, it is possible to reduce the risk of blood coming into contact with the human body when the plug is pulled out from the opening of the blood collection container main body after blood collection.
- the plug body examples include synthetic resin, elastomer, rubber, and metal foil.
- the rubber examples include butyl rubber and halogenated butyl rubber. Aluminum foil etc. are mentioned as said metal foil.
- the material of the stopper is preferably butyl rubber.
- the stopper (or the stopper body) is preferably a butyl rubber stopper.
- the blood collection container is a blood collection container from which a predetermined amount of blood is collected.
- the predetermined amount of blood is appropriately changed depending on the size and internal pressure of the blood collection container.
- the predetermined amount of the blood may be 1 mL or more, 2 mL or more, 4 mL or more, 12 mL or less, 11 mL or less, or 10 mL or less. may be
- a physiological saline solution in an amount equal to a predetermined amount of blood to be collected in the blood collection container is collected in the blood collection container to obtain a mixed solution in which the physiological saline solution and the aqueous solution are mixed.
- a mixed solution in which the physiological saline solution and the aqueous solution are mixed For example, in a blood collection container from which 5 mL of blood is collected, 5 mL of physiological saline is collected into the blood collection container and mixed by inversion or the like to mix the physiological saline and the aqueous solution. get the liquid
- the osmotic pressure of the mixture of the physiological saline and the aqueous solution is preferably 330 mOsm/L or more and 380 mOsm/L or less, or 440 mOsm/L or more.
- the specific gravity of white blood cells and red blood cells can be effectively increased, and contamination of blood plasma with white blood cells and red blood cells can be more effectively suppressed. can.
- excessive stress on blood cells can be suppressed, and contamination of components in blood cells can be suppressed more effectively.
- the osmotic pressure of the mixed liquid is preferably 340 mOsm/L or more, more preferably 350 mOsm/L or more, and preferably 375 mOsm/L or less. More preferably, it is 370 mOsm/L or less.
- the osmotic pressure of the mixed solution is equal to or higher than the lower limit and equal to or lower than the upper limit, the specific gravity of leukocytes and erythrocytes can be effectively increased when blood is collected in a blood collection container, and leukocytes can be added to plasma.
- the osmotic pressure of the mixed solution is equal to or higher than the lower limit and equal to or lower than the upper limit, excessive stress on blood cells can be suppressed, and contamination of components in blood cells can be suppressed more effectively.
- the osmotic pressure of the mixed solution is preferably 450 mOsm/L or higher, more preferably 500 mOsm/L or higher, preferably 1300 mOsm/L or lower, and more preferably 1000 mOsm/L. /L or less.
- the osmotic pressure of the mixed solution is equal to or higher than the lower limit and equal to or lower than the upper limit, the specific gravity of leukocytes and erythrocytes can be effectively increased when blood is collected in a blood collection container, and leukocytes can be added to plasma. and red blood cell contamination can be suppressed more effectively.
- the osmotic pressure of the mixed solution is equal to or higher than the lower limit and equal to or lower than the upper limit, excessive stress on blood cells can be suppressed, and contamination of components in blood cells can be suppressed more effectively. .
- the osmotic pressure of the above mixed solution is measured by the freezing point depression method using an osmometer (for example, "OM-6060” manufactured by ARKRAY).
- 3 mL or more of blood is preferably collected from 1 mL of the aqueous solution contained therein, more preferably 4 mL or more, preferably 11 mL or less, and 7 mL or less is collected. more preferably.
- the effects of the present invention can be exhibited more effectively without excessive dilution of blood.
- the blood collection container is preferably a blood collection tube.
- the blood collection container main body is preferably a blood collection tube main body.
- the blood collection container is preferably used for separating plasma from blood.
- the blood collection container is preferably used for isolating extracellular free nucleic acids or extracellular vesicles in blood. preferably.
- the blood collection container can be manufactured as follows.
- An aqueous solution is obtained by dissolving the anticoagulant and the second solute in water.
- the resulting aqueous solution is added into the body of the blood collection container.
- the plasma separation material is accommodated in the blood collection container main body.
- FIG. 1 is a front cross-sectional view schematically showing a blood collection container according to one embodiment of the present invention.
- the blood collection container main body 2 has an opening at one end and a closed bottom at the other end.
- the plasma separation composition 3 is housed in the bottom of the blood collection container body 2 .
- the stopper 5 is inserted into the opening of the blood collection container main body 2 .
- the aqueous solution 4 is placed on the surface of the plasma separation composition 3, and more specifically, placed on the upper surface (one end side surface) of the plasma separation composition 3.
- the aqueous solution 4 is placed on the surface of the plasma separation composition 3 when the blood collection container 1 is in an upright state.
- the anticoagulant and the second solute are contained in the blood collection container main body 2 in a state of being dissolved in water.
- the composition for plasma separation is arranged on the side wall surface of the blood collection container main body, and the aqueous solution is applied to the blood collection container when the blood collection container is placed in an upright state. It may be arranged at the bottom of the container body. Moreover, the jig for plasma separation may be used instead of the composition for plasma separation.
- the internal pressure of the blood collection container is not particularly limited.
- the blood collection container can also be used as a vacuum blood collection tube whose inside is evacuated and then sealed with the stopper. In the case of a vacuum blood collection tube, a fixed amount of blood can be collected easily regardless of the skill of the blood collector.
- the inside of the blood collection container is sterilized according to ISO or JIS standards.
- the blood collection container can be used to separate plasma from blood.
- a plasma separation method according to the present invention comprises the steps of collecting blood in the above-described blood collection container and centrifuging the blood collection container containing the blood.
- the plasma separation method according to the present invention preferably includes a step of mixing the collected blood and the aqueous solution between the step of collecting the blood and the step of centrifuging.
- a method for mixing the collected blood and the aqueous solution inversion mixing and the like can be mentioned.
- the centrifugation conditions in the centrifugation step are not particularly limited as long as the plasma separation material can form partition walls to separate plasma and blood cells.
- Examples of the centrifugation conditions include conditions for centrifugation at 400 G or more and 4000 G or less for 10 minutes or more and 120 minutes or less.
- the method for separating extracellular free nucleic acid according to the present invention includes the steps of collecting blood in the above-described blood collection container, and centrifuging the blood collection container containing the blood to separate plasma from the blood. and separating extracellular free nucleic acids from the separated plasma.
- a method for separating extracellular vesicles according to the present invention includes the steps of collecting blood in the blood collection container described above, and centrifuging the blood collection container containing the blood to separate plasma from the blood. and separating extracellular vesicles from the separated plasma.
- the collected blood and the aqueous solution are mixed between the step of collecting the blood and the step of centrifuging. It is preferable to include steps.
- steps As a method for mixing the collected blood and the aqueous solution, inversion mixing and the like can be mentioned.
- the centrifugation conditions in the centrifugation step are not particularly limited as long as the plasma separation material can form partition walls to separate plasma and blood cells.
- Examples of the centrifugation conditions include conditions for centrifugation at 400 G or more and 4000 G or less for 10 minutes or more and 120 minutes or less.
- the extracellular free nucleic acid can be separated from plasma using a conventionally known method.
- the extracellular free nucleic acid include cell free DNA (cfDNA), cell free RNA (cfRNA), and the like.
- the method for isolating the extracellularly released nucleic acid from the plasma include a method using a commercially available nucleic acid purification kit.
- Extracellular free nucleic acids can be easily separated from plasma by using a commercially available nucleic acid purification kit.
- nucleic acid purification kits include QIAamp Circulating Nucleic Acid Kit (QIAGEN), QIAamp MinElute ccfDNA Kits (QIAGEN) and MagMAX Cell-Free DNA Isolation Kit (Applied biosystems).
- extracellular vesicles can be separated from plasma using a conventionally known method.
- the following materials were prepared for the plasma separation composition.
- (Material of organic component having fluidity at 25°C) (Meth) acrylic resin: 2-Ethylhexyl acrylate and butyl acrylate were subjected to radical polymerization by a solution polymerization method in the presence of an azo polymerization initiator to obtain a (meth)acrylate polymer having fluidity at 25°C.
- Trimellitate ester (benzene polycarboxylic acid alkyl ester derivative, "Monocizer W700" manufactured by DIC)
- Hydrophilic silica fine powder silica, "200CF” manufactured by Nippon Aerosil Co., Ltd.
- Hydrophobic silica fine powder silica, "R974" manufactured by Nippon Aerosil Co., Ltd.
- Titanium oxide powder second inorganic fine powder, "A-100” manufactured by Ishihara Sangyo Co., Ltd.
- Silicone oil (“SF8410” manufactured by Dow Corning Toray Co., Ltd.)
- Organic gelling agent (“Gelol D” manufactured by Shin Nippon Rika Co., Ltd.) 1-methyl-2-pyrrolidone (co-solvent)
- Preparation of plasma separation compositions A and B An organic component, an inorganic fine powder, and other components having flowability at 25° C. were mixed at the mixing ratios shown in Table 1 to prepare compositions A and B for plasma separation.
- Preparation of plasma separation composition C The materials of the organic component having fluidity at 25°C shown in Table 1 were blended, heated and melted at 130°C, and mixed to prepare an organic component having fluidity at 25°C. Next, the organic component, the inorganic fine powder, and the other components, which are fluid at 25°C, were mixed at the mixing ratio shown in Table 1 to prepare a composition C for plasma separation.
- PET bottomed tube polyethylene terephthalate tube with a length of 100 mm and an inner diameter of 14 mm at the opening
- Example 1 An anticoagulant and a second solute were dissolved in water to obtain an aqueous solution.
- Table 2 shows the types and concentrations of the ingredients in the resulting aqueous solution.
- composition C for plasma separation 1.2 g was accommodated in the bottom of the blood collection container main body. Also, 1 mL of the obtained aqueous solution was added onto the surface of the composition C for plasma separation. The inside of the blood collection container was evacuated so that the blood collection volume was 5 mL, and the container was sealed with a butyl rubber stopper. Thus, a blood collection container was produced.
- Examples 2-7 and Comparative Examples 2-4 The type of composition for plasma separation and the composition of the aqueous solution were changed as shown in Tables 2-4.
- Example 5 and Comparative Examples 2 and 3 the inside of the blood collection container was evacuated so that the blood collection volume was 10 mL, and the container was sealed with a butyl rubber plug.
- a blood collection container was produced in the same manner as in Example 1 except for these.
- composition C for plasma separation was accommodated in the bottom of the blood collection container main body. Also, 60 mg of the obtained mixture was applied to the inner wall surface of the blood collection container main body and dried. The inside of the blood collection container was evacuated so that the blood collection volume was 10 mL, and the container was sealed with a butyl rubber plug. Thus, a blood collection container was produced.
- Osmotic Pressure of Mixture 5 mL of physiological saline was collected in the blood collection container obtained in Examples 1-4, 6, and 7. In addition, 10 mL of physiological saline was collected in the blood collection container obtained in Example 5. After the physiological saline was collected, the mixture was mixed by inversion to mix the physiological saline and the aqueous solution contained in the blood collection container to obtain a mixed solution. The osmotic pressure of the obtained mixture was measured by the freezing point depression method using an osmometer ("OM-6060" manufactured by ARKRAY).
- DNA contained in the collected plasma was purified using a cfDNA purification kit ("QIAamp Circulating Nucleic Acid Kit” manufactured by QIAGEN). The DNA purification operation was performed on the day plasma was collected from the blood collection container.
- the DNA concentration in the extract after purification was measured using the Qubit dsDNA HS Assay kit (Invitrogen). Then, the cfDNA concentration (the content of cfDNA contained per 1 mL of plasma) was calculated according to the following formula.
- cfDNA concentration [A] x [B]/[C]
- the average concentration of cfDNA recovered from plasma on the day of blood collection is defined as "cfDNA concentration (on the day of collection)", and the average concentration of cfDNA collected from plasma after storage for 7 days is defined as “cfDNA concentration (stored for 7 days)”. and In addition, the difference between the cfDNA concentration (stored for 7 days) and the cfDNA concentration (on the day of collection) was defined as "increase in cfDNA concentration”.
- the average value of the concentration of cfDNA is the average value of the results obtained using the blood of three persons.
- the recovered amount of cfDNA was determined according to the following criteria. It should be noted that the greater the amount of leukocytes mixed in the plasma and the more unstable the leukocytes in the plasma, the greater the increase in the cfDNA concentration due to the destruction of the leukocytes during storage. Therefore, the smaller the amount of increase in cfDNA concentration, the more suppressed the contamination of leukocytes into plasma and the contamination of components in leukocytes.
- the concentration of anticoagulant means the concentration of EDTA2K, not the concentration of EDTA2K ⁇ 2H 2 O.
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Abstract
Description
上記血液採取容器は、上記血液採取容器本体内に収容された血漿分離材を備える。上記血漿分離材として、従来公知の血漿分離材を用いることができる。上記血漿分離材としては、血漿分離用組成物、及び血漿分離用冶具等が挙げられる。血漿分離材の作製が容易であることから、上記血漿分離材は、上記血漿分離用組成物であることが好ましい。
上記血漿分離用組成物は、遠心分離時に血漿層と血球層との間に移動して隔壁を形成する組成物である。また、上記血漿分離用組成物は、遠心分離後に血漿層と血球層との間の成分移行を防止する目的で用いられる。上記血漿分離用組成物は、チクソトロピー性を有することが好ましい。上記血漿分離用組成物は、上記血液採取容器本体の底部に収容されていてもよく、上記血液採取容器本体の側壁面上に配置されていてもよい。本発明の効果をより一層効果的に発揮する観点から、上記血漿分離用組成物は、上記血液採取容器本体の底部に収容されていることが好ましい。
上記「25℃で流動性を有する」とは、25℃での粘度が500Pa・s以下であることを意味する。
上記無機微粉末としては、微粉末シリカ、酸化チタン粉末、炭酸カルシウム粉末、酸化亜鉛粉末、アルミナ粉末、ガラス微粉末、タルク粉末、カオリン粉末、ベントナイト粉末、チタニア粉末、及びジルコニウム粉末等が挙げられる。
上記血漿分離用組成物は、本発明の効果を損なわない限り、上述した成分以外の他の成分を含んでいてもよい。上記他の成分としては、有機ゲル化剤、熱可塑性エラストマー、ポリアルキレングリコール、シリコーンオイル、補助溶媒、酸化防止剤、着色剤及び水等が挙げられる。上記他の成分はそれぞれ、1種のみが用いられてもよく、2種以上が併用されてもよい。
上記血漿分離用冶具は、遠心分離時に血漿層と血球層との間に移動して隔壁を形成する冶具である。また、上記血漿分離用冶具は、血漿層と血球層との間の成分移行を防止する目的で用いられる。
上記血液採取容器は、上記血液採取容器本体内に収容された水溶液を備える。上記水溶液は、抗凝固剤と、水とを含む。上記水溶液に含まれる溶質は、抗凝固剤を含む。上記水溶液は、抗凝固剤以外の第2の溶質を含むことが好ましい。したがって、上記水溶液は、抗凝固剤(第1の溶質)と、第2の溶質と、水とを含むことが好ましい。上記水溶液に含まれる溶質は、抗凝固剤(第1の溶質)と、第2の溶質とを含むことが好ましい。
上記水溶液は、抗凝固剤を含む。上記抗凝固剤として、従来公知の抗凝固剤を用いることができる。上記抗凝固剤は、1種のみが用いられてもよく、2種以上が併用されてもよい。
上記第2の溶質は、上記水溶液に含まれる抗凝固剤以外の溶質である。上記第2の溶質を用いることにより、抗凝固剤の含有量を過度に多くすることなく、血液採取容器内に採取された血液の浸透圧を効果的に大きくすることができる。上記第2の溶質は、1種のみが用いられてもよく、2種以上が併用されてもよい。
上記血液採取容器本体の形状としては、特に限定されない。上記血液採取容器本体は、有底の管状容器であることが好ましい。
上記血液採取容器は、栓体を備えることが好ましい。上記栓体として、従来公知の栓体を用いることができる。上記栓体は、血液採取容器本体の開口に、気密的かつ液密的に取付けることが可能な素材、形状からなる栓体であることが好ましい。上記栓体は、採血針が刺通され得るように構成されていることが好ましい。
上記血液採取容器は、所定量の血液が採取される血液採取容器である。上記血液の上記所定量は、血液採取容器のサイズ及び内圧等により適宜変更される。上記血液の上記所定量は、1mL以上であってもよく、2mL以上であってもよく、4mL以上であってもよく、12mL以下であってもよく、11mL以下であってもよく、10mL以下であってもよい。
上記血液採取容器を用いて、血液から血漿を分離することができる。本発明に係る血漿の分離方法は、上述した血液採取容器内に血液を採取する工程と、上記血液が採取された上記血液採取容器を遠心分離する工程とを備える。
本発明に係る細胞外遊離核酸の分離方法は、上述した血液採取容器内に血液を採取する工程と、上記血液が採取された上記血液採取容器を遠心分離して、血液から血漿を分離する工程と、分離された上記血漿から、細胞外遊離核酸を分離する工程とを備える。
(メタ)アクリル系樹脂:
アクリル酸-2-エチルヘキシルとアクリル酸ブチルとをアゾ系重合開始剤の存在下で溶液重合法によりラジカル重合させ、25℃で流動性を有する(メタ)アクリル酸エステル系重合体を得た。
石油樹脂(イーストマンケミカル社製「リガライトS5090」)
ジシクロペンタジエン樹脂1(コロン社製「スコレッツSU500」)
ジシクロペンタジエン樹脂2(コロン社製「スコレッツSU90」)
トリメリット酸エステル(ベンゼンポリカルボン酸アルキルエステル誘導体、DIC社製「モノサイザーW700」)
親水性シリカ(微粉末シリカ、日本アエロジル社製「200CF」)
疎水性シリカ(微粉末シリカ、日本アエロジル社製「R974」)
酸化チタン粉末(第2の無機微粉末、石原産業社製「A-100」)
シリコーンオイル(東レダウコーニング社製「SF8410」)
有機ゲル化剤(新日本理化社製「ゲルオールD」)
1-メチル-2-ピロリドン(補助溶媒)
表1に記載の配合割合で、25℃で流動性を有する有機成分と無機微粉末とその他の成分とを混合し、血漿分離用組成物A,Bを作製した。
表1に記載の25℃で流動性を有する有機成分の材料を配合し、130℃で加熱溶解し、混合して、25℃で流動性を有する有機成分を作製した。次いで、表1に記載の配合割合で、25℃で流動性を有する有機成分と無機微粉末とその他の成分とを混合し、血漿分離用組成物Cを作製した。
エチレンジアミン四酢酸二カリウム二水和物(EDTA2K・2H2O)
塩化カリウム(KCl)
塩化ナトリウム(NaCl)
スクロース
グルコース
プロピレングリコール
ポリエチレングリコール(PEG4000)
抗凝固剤及び第2の溶質を水に溶解して水溶液を得た。得られた水溶液の配合成分の種類及び濃度を表2に示す。
血漿分離用組成物の種類、及び水溶液の組成を表2~4のように変更した。また、実施例5及び比較例2,3では、血液採取容器内部を、血液採取量が10mLとなるように減圧し、ブチルゴム栓により密封した。これら以外は、実施例1と同様にして、血液採取容器を作製した。
抗凝固剤32重量部を、水68重量部に溶解して、混合液を得た。1.2gの血漿分離用組成物Cを、血液採取容器本体の底部に収容した。また、得られた混合液60mgを、血液採取容器本体の内壁面に塗布し、乾燥させた。血液採取容器内部を、血液採取量が10mLとなるように減圧し、ブチルゴム栓により密封した。このようにして血液採取容器を作製した。
(1)混合液の浸透圧
実施例1~4,6,7で得られた血液採取容器に生理食塩水5mLを採取した。また、実施例5で得られた血液採取容器に生理食塩水10mLを採取した。生理食塩水を採取した後、転倒混和して、生理食塩水と血液採取容器内に収容された水溶液とを混合し、混合液を得た。得られた混合液の浸透圧を、浸透圧計(アークレイ社製「OM-6060」)を用いて、氷点降下法により測定した。
3名の血液を使用し、それぞれについて以下の操作を行った。実施例1~4,6,7及び比較例4で得られた血液採取容器を各2本用意し、それぞれに血液5mLを採取した。また、実施例5及び比較例1~3で得られた血液採取容器を各2本用意し、それぞれに血液10mLを採取した。血液を採取した後、転倒混和して、血液と血液採取容器内に収容された水溶液とを混合した。次いで、血液採取容器を1500Gで15分間遠心分離した。遠心分離後、血漿分離用組成物により形成された隔壁よりも上方に血漿が位置していた。
[B]:精製後の抽出液の全量(mL)
[C]:DNA精製に使用した血漿の量(mL)
〇〇:cfDNA濃度の増加量が20ng/血漿1mL未満である
○:cfDNA濃度の増加量が20ng/血漿1mL以上100ng/血漿1mL未満である
×:cfDNA濃度の増加量が100ng/血漿1mL以上である
2…血液採取容器本体
3…血漿分離用組成物
4…水溶液
5…栓体
Claims (19)
- 血液採取容器本体と、
前記血液採取容器本体内に収容された血漿分離材と、
前記血液採取容器本体内に収容された水溶液とを備え、
前記水溶液に含まれる溶質が、抗凝固剤を含み、
前記水溶液における前記溶質の合計濃度が、100mM以上450mM以下であるか、又は、1200mM以上である、血液採取容器。 - 前記水溶液における前記溶質の合計濃度が、250mM以上390mM以下であるか、又は、1200mM以上6500mM以下である、請求項1に記載の血液採取容器。
- 前記水溶液に含まれる前記溶質が、抗凝固剤以外の第2の溶質を含み、
前記第2の溶質が、無機塩、又は糖類を含む、請求項1又は2に記載の血液採取容器。 - 前記第2の溶質が、無機塩を含み、
前記無機塩が、ナトリウム塩、又はカリウム塩を含む、請求項3に記載の血液採取容器。 - 前記第2の溶質が、糖類を含み、
前記糖類が、グルコース、スクロース、又はトレハロースを含む、請求項3又は4に記載の血液採取容器。 - 前記抗凝固剤が、EDTA、EDTAの金属塩、ヘパリン、ヘパリンの金属塩又はクエン酸ナトリウムである、請求項1~5のいずれか1項に記載の血液採取容器。
- 所定量の血液が採取される血液採取容器であり、
血液採取容器に採取される血液の所定量と等量の生理食塩水を血液採取容器内に採取して、前記生理食塩水と前記水溶液とが混合された混合液を得たときに、前記混合液の浸透圧が、330mOsm/L以上380mOsm/L以下であるか、又は、440mOsm/L以上1300mOsm/L以下である、請求項1~6のいずれか1項に記載の血液採取容器。 - 前記血漿分離材の25℃での比重が1.030以上1.120以下である、請求項1~7のいずれか1項に記載の血液採取容器。
- 前記血漿分離材が、血漿分離用組成物である、請求項1~8のいずれか1項に記載の血液採取容器。
- 前記血漿分離用組成物が、25℃で流動性を有する有機成分と、無機微粉末とを含み、
前記有機成分が、樹脂を含み、
前記無機微粉末が、微粉末シリカを含む、請求項9に記載の血液採取容器。 - 前記微粉末シリカが、親水性シリカを含む、請求項10に記載の血液採取容器。
- 前記血漿分離用組成物100重量%中、前記親水性シリカの含有量が、0.01重量%以上2.50重量%以下である、請求項11に記載の血液採取容器。
- 前記微粉末シリカが、親水性シリカと疎水性シリカとを含む、請求項10~12のいずれか1項に記載の血液採取容器。
- 前記血漿分離用組成物の25℃での比重が1.050以上であり、
前記無機微粉末が、前記微粉末シリカよりも比重が大きい無機微粉末を含む、請求項10~13のいずれか1項に記載の血液採取容器。 - 前記樹脂が、石油樹脂、シクロペンタジエン系樹脂、ポリエステル樹脂、又は(メタ)アクリル系樹脂を含む、請求項10~14のいずれか1項に記載の血液採取容器。
- 血液中の細胞外遊離核酸又は細胞外小胞を分離するために用いられる、請求項1~15のいずれか1項に記載の血液採取容器。
- 請求項1~15のいずれか1項に記載の血液採取容器内に血液を採取する工程と、
前記血液が採取された前記血液採取容器を遠心分離する工程とを備える、血漿の分離方法。 - 請求項1~16のいずれか1項に記載の血液採取容器内に血液を採取する工程と、
前記血液が採取された前記血液採取容器を遠心分離して、血液から血漿を分離する工程と、
分離された前記血漿から、細胞外遊離核酸を分離する工程とを備える、細胞外遊離核酸の分離方法。 - 請求項1~16のいずれか1項に記載の血液採取容器内に血液を採取する工程と、
前記血液が採取された前記血液採取容器を遠心分離して、血液から血漿を分離する工程と、
分離された前記血漿から、細胞外小胞を分離する工程とを備える、細胞外小胞の分離方法。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2001083144A (ja) * | 1999-09-09 | 2001-03-30 | Asahi Kasei Corp | 血液処理方法および組成物 |
JP2005017189A (ja) * | 2003-06-27 | 2005-01-20 | Srl Inc | 血中インスリン濃度測定用添加剤組成物及びそれを含む採血管 |
WO2010053180A1 (ja) | 2008-11-07 | 2010-05-14 | 日立化成工業株式会社 | 血清又は血漿分離材及びそれを用いた採血管 |
WO2010132783A1 (en) | 2009-05-15 | 2010-11-18 | Becton, Dickinson And Company | Density phase separation device |
WO2016159236A1 (ja) * | 2015-03-31 | 2016-10-06 | 積水メディカル株式会社 | 血清または血漿分離用組成物、並びに血液採取容器 |
WO2020132747A1 (en) | 2018-12-24 | 2020-07-02 | Deltadna Biosciences Inc | Composition and method for segregating extracellular dna in blood |
JP2021500585A (ja) * | 2017-10-19 | 2021-01-07 | ストレック, インコーポレイテッド | 溶血、凝固調節および細胞外小胞の安定化のための組成物 |
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Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2001083144A (ja) * | 1999-09-09 | 2001-03-30 | Asahi Kasei Corp | 血液処理方法および組成物 |
JP2005017189A (ja) * | 2003-06-27 | 2005-01-20 | Srl Inc | 血中インスリン濃度測定用添加剤組成物及びそれを含む採血管 |
WO2010053180A1 (ja) | 2008-11-07 | 2010-05-14 | 日立化成工業株式会社 | 血清又は血漿分離材及びそれを用いた採血管 |
WO2010132783A1 (en) | 2009-05-15 | 2010-11-18 | Becton, Dickinson And Company | Density phase separation device |
WO2016159236A1 (ja) * | 2015-03-31 | 2016-10-06 | 積水メディカル株式会社 | 血清または血漿分離用組成物、並びに血液採取容器 |
JP2021500585A (ja) * | 2017-10-19 | 2021-01-07 | ストレック, インコーポレイテッド | 溶血、凝固調節および細胞外小胞の安定化のための組成物 |
WO2020132747A1 (en) | 2018-12-24 | 2020-07-02 | Deltadna Biosciences Inc | Composition and method for segregating extracellular dna in blood |
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