CN1657899A - Method and device for drying and preserving liquid sample - Google Patents
Method and device for drying and preserving liquid sample Download PDFInfo
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- CN1657899A CN1657899A CN 200410004387 CN200410004387A CN1657899A CN 1657899 A CN1657899 A CN 1657899A CN 200410004387 CN200410004387 CN 200410004387 CN 200410004387 A CN200410004387 A CN 200410004387A CN 1657899 A CN1657899 A CN 1657899A
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Abstract
This invention discloses dry preserved method for the liquid sample, it includs the liquid sample will be contacts with a average aperture 0.01 to 5 millimeter spatial solid carriers, and has absorbed in the liquid sample solid carrier except the liquid ingredient; After dry solid carrier percentage of voids for the solid carrier original percentage of voids 20%. A device also disclosed.
Description
Technical field
The present invention relates to a kind of method for drying and storing of method for drying and storing, particularly biological sample of the fluid sample based on porosint.The invention still further relates to the device that is used for fluid sample drying, preservation.
Background technology
Biological sample collection in the scientific research, preservation, transportation, reorganization, recovery are the condition precedents of carrying out various experimental science researchs, the various indexs detections of clinical medicine.Liquid biological sample is the biological sample form that the most extensively adopts in the life science, and to reclaim security in its concentration accuracy of back and the biological sample processing overall process be one of biology, medical science, medicine, biological technical field basic problem that need solve for effective active (comprising in conjunction with activity and biologically active), integrality (as cell and other visible component), the sample reorganization of product content how to use the easiest cost-effective method to keep intact.For many years, scientific workers have made effort untiringly for this reason.
At present, the store method of fluid sample mainly is divided into liquid cryopreservation (2-8 degree Celsius); Freezing preservation (-20～-200 degree Celsius); Freezing (-60 degree Celsius are following) kept dry; (2-8 degree Celsius) preserved in low temperature drying; Preserve (10～30 degree or environment temperatures Celsius) with air drying.
There is different shortcomings separately in existing said method: degraded or bacterium mold propagates etc. cause the change of initial liquid properties of samples rapidly in a short time for example to cause the biological fluid ingredient in the normal temperature environment easily; Fluid sample should not be made long preservation in the low temperature environment; The frozen liq sample retention needs low temperature refrigerator to continue to make temperature to remain on below 0 temperature Celsius, and transportation is made troubles to sample; Though the freeze drying liq sample can obtain satisfied sample organic efficiency, its cost is too high and be unsuitable for processing single or the short run fluid sample; Though present air drying sample retention method has solved the kept dry transportation problem of fluid sample, when handling, sample liquids recovery, reorganization and subsequent analysis have a lot of defectives.
Robert Guthrie utilized Schleicher ﹠amp first in 1963; Schuell Bioscience (the S﹠amp of company; S company) 903 filter membranes (back claims Guthrie Card) are collected neonate's blood sample and have been carried out since the PKU examination, and this filter membrane is the kept dry of widespread use biological fluid.But constitute because this absorption of sample matrix is cellulose, go after drying into a kind of fiber/sample entity structure, cause very big difficulty for the reorganization and the recovery of liquid sample, its range of application is very limited when absorbing fluid sample such as blood.Integrate, this is that the sample reorganization is time-consuming with cellulose membrane as the major defect of fluid sample absorption base, generally needs 30 minutes at least to a few hours, and needs heat treated to redissolve to promote sample; Organic efficiency is low, because the plain membrane matrix of the sample fiber of formation is-no gap substantial structure after dry, so the utmost point is difficult to make once more the dry sample aquation; And cellulose membrane causes the low recovery to the absorption and the suction-operated of sample composition, and its optimum recovery rate is lower than 50% at short notice.
In addition, the cell component in the dry artifact blood being reclaimed is the prerequisite of carrying out cytology research.Joseph (USA; Pat.No.:5432,097) having described the leucocyte that utilizes zymetology digest cellulose film (Guthrie Card) method to carry out dry blood in 1993 reclaims.But this method program need be used the cellulose degraded cellulose membrane, and is therefore time-consuming, cost is high.
Therefore, this area needs a kind of easy, quick and fluid sample method for drying and storing that sample recovery rate is high.
Summary of the invention
The invention provides a kind of method for drying and storing of fluid sample, it comprises described fluid sample and a kind of average pore size, and to be 0.01mm contact to the porous solid carrier of 5mm; With from the solid carrier that has absorbed fluid sample, remove liquid component; The porosity of wherein dry back solid carrier is at least 20% of the former porosity of solid carrier.
According to one embodiment of the invention, described porous solid carrier is to be made of one or more materials that are selected from down group: polymeric material, material, metal material and Inorganic Non-metallic Materials through chemical treatment or untreated biogenetic derivation.
According to another embodiment of the present invention, described fluid sample is the biological fluid or the liquid bio reagent of biological fluid, artificial preparation.
The present invention also provides a kind of device that is used for the kept dry fluid sample, and it comprises that a kind of average pore size is 0.01mm to porous solid carrier 5mm, that still keep former porosity at least 20% after load sample and the drying.
According to method and apparatus of the present invention, can realize quick, the easy kept dry of fluid sample, and compare recovery sample fast and efficiently with art methods.
Description of drawings
Describe the present invention in detail below in conjunction with the drawings and specific embodiments.
Fig. 1 shows to utilize method of the present invention to carry out the synoptic diagram of blood sample drying, preservation and recovery, restructuring procedure.Figure 1A-D represents not application of sample porous solid carrier, the porous solid carrier behind the absorption fluid sample (whole blood), the load sample solid carrier after the dehydration and the reconstruct of blood sample before respectively.
Fig. 2 is the electrophoretogram from the genomic DNA of preserving by the inventive method that blood sample reclaims.Among the figure, M is the molecular weight sign; " 1-4 " is respectively the DNA that extracts from the blood sample of 4 parts of kept dry on solid carrier of the present invention.The applied sample amount of each sample is 1 microgram (DNA).
Embodiment
The present inventor is through extensive studies, the porous solid carrier that discovery utilizes macropore not only can fast Absorption, the dry liquid sample, and dry sample easily from the carrier water or other solvent elutions get off, thereby reclaim fast and efficiently, the reconstruct sample.
Therefore, the invention provides a kind of method for drying and storing of fluid sample, it comprises described fluid sample and a kind of average pore size is contacted to the porous solid carrier of about 5mm macropore for about 0.01mm; With from the solid carrier that has absorbed fluid sample, remove liquid component; The porosity of wherein dry back solid carrier is at least 20% of the former porosity of solid carrier.
In the inventive method, the average pore size of used porous solid carrier is preferably about 0.05mm between about 1mm, more preferably arrives between about 0.5mm at about 0.1mm.
In the method for the invention, described porous solid carrier application of sample and dried porosity preferably keep former porosity at least about 30%, more preferably at least about 40%, most preferably at least about 50%.
Porous solid carrier among the present invention can be made by any material, for example is selected from down one or more materials of group: polymeric material, through material, metal material and the Inorganic Non-metallic Materials of the material of chemical treatment or untreated biogenetic derivation.In a preferred embodiment, cellular solid of the present invention is a polymeric material.
Porous solid carrier of the present invention can be single integral material.This material forms a plurality of surperficial perforates through its preparation technology, and perhaps this material is natural has a plurality of surperficial perforates.
Porous solid carrier of the present invention also can be the solid material with a plurality of surperficial perforates that a plurality of unit by different shape form by physics or chemical method stationary arrangement.The unit itself that constitutes this solid material can have porous structure or not have porous structure.The method of this unit of stationary arrangement is well known by persons skilled in the art, for example mechanical connection, bonding, braiding etc.
Porous solid carrier of the present invention has certain rigidity, after avoiding load bearing fluid, because liquid gravity effect or dry run are subsided porous structure, cause the porosity of dry back load sample solid carrier to be lower than 20% of the former porosity of carrier, be unfavorable for the reconstruct of sample.
Polymeric material of the present invention can be non-biocompatible and/or biodegradability, also can be biocompatibility and/or biodegradable.Be in the consideration of environmental protection and for the preservation of living cells, microbiological specimens, preferred polymeric material of the present invention is a biocompatibility and/or biodegradable.
The example of biocompatibility and/or biodegradability polymeric material has: hydroxycarboxylic acid esters, as poly-(3-butyric ester) (PHB), 3-butyric ester and 3-hydroxycaproic acid ester copolymer (PHB-HH), PLA (PLA), poly lactic coglycolic acid (PLGA), polycaprolactone; Poe, poly-acid anhydrides etc.For blood compatibility, the polymeric material with good anticoagulant property for example has: hydrophilic material such as polymethylacrylic acid beta-hydroxy ethyl ester, polyvinyl alcohol (PVA), PMAm and polyvinyl pyrrolidone; Hydrophobic material such as silicon rubber; Macromolecular material such as poly(ether-urethane) with micro phase separation structure surface; With the material on electronegative surface such as the terylene and the spumescence teflon of the level and smooth carbon film of surperficial evaporation.
The example of abiotic degradable materials has: polyvinyl acetate (PVA), tygon, polystyrene, Polyvinylchloride, polyurethane, polycarbonate etc.
Described porous polymer material can obtain easily by art methods, for example by methods such as polymer foamings.The aperture of described porous polymer material also can be controlled according to a conventional method, thereby obtains to be suitable for the concrete porosint of using.
It is known to those skilled in the art that nearly all thermosetting and thermoplastic resin can both make foamed polymer, i.e. porous polymer material.Polymkeric substance through being usually used in preparing porous polymer material has: polystyrene, polyurethane, Polyvinylchloride, tygon, urea-formaldehyde resin, phenolics etc.
The foaming method that is used to make polymkeric substance produce porous structure comprises mechanical foaming, physical blowing, chemical blowing etc.
Mechanical foaming is that the tool stirring means that takes this opportunity is sneaked in the liquid polymeric raw material gas, forms abscess through type-approval process then.The mechanical foaming method can be used for urea-formaldehyde resin, polyvinyl formal, polyvinyl acetate, Polyvinylchloride colloidal sol etc.
The physical blowing method is to utilize the physical state variation of gas-development agent to produce pore in polymeric material.Physical blowing agent comprises pressure gas, volatile liquid and soluble solids.Volumetric expansion produced pore when compressibility gas such as nitrogen, carbon dioxide etc. reduced at pressure.Volatile liquid produces pore through vaporization, for example low aliphatic hydrocarbon, halogenated aliphatic hydrocarbon and lower boiling alcohol, ether, ketone and the aromatic hydrocarbon etc. that boil.Soluble solids is removed the back by dissolving and produces pore, for example water miscible inorganic salts, water miscible polymkeric substance and starch etc. in polymer product.For example monochloro methane and methylene chloride are usually used in making styrofoam, and fluorhydrocarbon is used for making multiple polyfoam, for example foamed polyethylene, styrofoam, soft and hard polyurethane foams, urea-formaldehyde foam and phenol formaldehyde foam etc.
Chemical blowing process is that the chemical foaming agent that adds the polymkeric substance solution produces one or more gas because of chemical change or thermal decomposition at a certain temperature take place, thereby produces pore in polymer product.Chemical foaming agent comprises inorganic foaming agent and organic foaming agent two classes.Inorganic foaming agent mainly comprises hartshorn salt, ammonium bicarbonate and sodium bicarbonate etc.Organic foaming agent mainly comprises nitroso compound, azo-compound, sulfonyl hydrazines and urea derivative etc.
A kind of method of utilizing the inorganic salts physical blowing agent to prepare porous polymer material is for example disclosed in the CN 1117587C Chinese patent.This method comprises polymkeric substance such as PLA is dissolved in solvent such as the chloroform Huo diox that the gas-development agent such as sodium chloride, the potassium chloride etc. that then particle diameter are equivalent to preset aperture add in the polymer solutions.Potpourri is joined in the mould, drying, demoulding immerse a period of time in the deionized water with goods again.Product dried after soaking is obtained porous polymer material.According to the description of the document, can make the porosint that the aperture is the 50-500 micron.
In a preferred embodiment of the invention, the polymeric material that constitutes porous solid carrier is the open celled polymeric foam that makes by foaming, comprises sponge sample polymeric material.
Porous solid carrier among the present invention preferably has the porosity more than 30%, more preferably the porosity more than 40%, 50%, 60%, 70% even 80%.
In a preferred embodiment of the invention, described porous solid carrier is a kind of water absorptivity foamed polymer.The water absorptivity foamed polymer can use the polyvalent alcohol of high-hydrophilic to make according to a conventional method.For example can make the quick water absorbency plastic foam of a kind of polyester-type: 3.5 parts in 100 parts of hexane diacid binaryglycol esters (weight, down with), 46 parts of TDI-80,6 parts of ethoxy monoleates, 1.1 parts of amine catalysts, 0.5 part of foam stabilizer and water by following prescription.
Porous metal material of the present invention comprises powder metallurgy porous material, sintered metal fiber porosint, sintered metal mesh porosint and foam metal material etc.The metal of making foam metal is selected from Al, Cu, Ni, Fe, Mg, Zn, Pb, Sn, Ag, Cr, Mo, Co etc. and alloy thereof.
Powder metallurgy porous material is to be raw material with the metal powder, the porosint that processes such as process shaping, sintering are made.The metal powder of metallurgical porosint of making powder can be used erose powder, also can be with the powder of almost spherical.
The sintered metal fiber porosint is the porosint of making through technologies such as shaping, sintering with metal fibre.
The sintered metal mesh porosint is the integral porous material of preparing by processes such as rolling, sintering (vacuum or atmosphere) or hot pressing with the single or multiple lift wire gauze.Wire gauze can be woven into plain square net (Plain Weave), twill square hole net (Twill Weave), plain weave seat type net (Plain Dutch Weave), twill seat type net (Dutch Twill Weave) etc.
The light porous metal that foam metal refers to that porosity is higher, the aperture is big, be interconnected between hole.The cellular structure that the structure of foam metal is made up of skeleton shape structure, film etc.The common method of making foam metal has motlten metal foaming, metal powder pulp foaming, infiltration powder slurry sintering process and electrocasting etc.
Inorganic Non-metallic Materials of the present invention comprises the porosint of being made by the conventional method of this area by pottery, glass, cement and fire resistive material etc., and wherein the chemical composition of Inorganic Non-metallic Materials comprises silicate, other oxysalt, oxide, nitride, carbon and carbonide, boride, fluoride, chalcogenide compound, silicon, germanium, III-V and II-VI compounds of group etc.
For example, porous ceramics is the stupalith that burns till, has in the body pore passage structure that communicates with each other in a large number and also connect with material surface through high temperature.The method that is formed porous ceramics by stupalith comprises that accumulation, bonding by particles of aggregates form porous ceramics; Utilize after-flame or the volatilization at high temperature of some additive and in ceramic body, stay hole; Utilize thermal decomposition, phase transformation, the segregation of material to form hole; But utilize the porous carrier absorption ceramic slurry of after-flame, afterwards, after-flame carrier material and form pore texture at high temperature.
The porosity of porous solid carrier of the present invention can be measured by the conventional method of this area.For example, the method for measuring porosity indirectly is the method for density measurement, passes through formula
Represent that ε is the porosity of material, ρ
bBe the average density of porosint cumulative volume, ρ
sDensity for solid portion in the material.
Porous solid carrier of the present invention can directly use without any processing.But in order to prevent contaminated samples, preferred water, aqueous detergent solution are cleaned, or with acid or aqueous slkali soaking.For example the PH of used acid solution is below 5.0, and the PH of alkaline solution is more than 8.0.Continue to clean with the solid carrier available water after acid or the alkali treatment, to remove remaining acid or alkali composition.Can after with acid or alkali treatment, not clean according to circumstances yet, solid carrier is still remained in acid or the alkali environment until dehydrating.
According to the difference of concrete application purpose, porous solid carrier of the present invention can be through following one or more method special processings.
Porous solid carrier is carried out heat treated; Autoclaving is handled; The ultraviolet ray irradiation; Radio exposure; Antiseptic kind is handled; The antibiotic class is handled; The antimycoin class is handled; The organic solvent class is handled, and representative examples of organic has ethanol, isopropyl alcohol, acetone, chloroform, phenols, ethers etc.; The scaling agent class is handled, and the example of scaling agent has tween (Tween-20, Tween-100), lauryl sodium sulfate (SDS) etc.; The anticoagulant class is handled, and the example of anticoagulant has heparin, sodium citrate (salt), ethylenediamine tetraacetic acid (EDTA) etc.
For example can be with the protein binding site of protein-contg liquid sealing porous solid carrier surfaces externally and internally, prevent that the absorption of protein molecular in the sample from causing the decline of sample organic efficiency.
Again for example, can on the surfaces externally and internally of porous solid carrier, connect various chemical groups by chemical method and be beneficial to inclusions in the adsorptive liquid sample, for example amino, carboxyl, hydroxyl, alkyl or other chemical group.
Also may by physics or chemical method on carrier, wrap by or connect suitable molecule such as various protein molecular, nucleic acid molecules, sour substrate etc., so as to specificity or non-specific catch corresponding composition contained in the sample.
Fluid sample among the present invention is defined as and anyly contains the potpourri that remains to be preserved material with what liquid condition existed in water-based or non-aqueous solvent.Described sample mixture can be solution, suspending liquid, emulsion or other any liquid forms.
Fluid sample for example of the present invention can be physiology or pathologic biological fluid: any liquid that other biological bodies such as blood, sweat, urine, cerebrospinal fluid, spinal fluid, arthral fluid, vaginal secretion liquid, seminal fluid, blood plasma, serum, amniotic fluid, milk, pleural fluid, peritoneal fluid, bone marrow fluid, saliva, bile, tear are produced under normal physiological state and morbid state.
Fluid sample of the present invention also can be through artificial formulated fluid sample, the liquid that for example contains bacterium, mould, fungi, parasite etc., the various cells of the liquid extract of various biological tissues such as biology are or/and the liquid extract of tissue extract (liquid extracts of the heart, liver, spleen, lung, kidney etc.) and various vegetable cells etc.
Fluid sample of the present invention can also be the liquid reagent that contains solid solute, as various damping fluids and by the liquid of its preparation.Liquid by described liquid reagent preparation for example is the liquid that contains protein, nucleic acid, cell, blood platelet, bacterium, plasmid, virion, parasite, seminal fluid, vaginal fluid etc.
Utilize method of the present invention to carry out in the process of fluid sample kept dry, can add the protective agent that can improve bioactivator tolerance drying and improve hold capacity.Such protective agent is well known by persons skilled in the art.Polyvalent alcohol for example is as carbohydrates such as glucose, maltose, sucrose, xylulose, ribose, mannose, fructose, gossypose, trehaloses; Sugar derivativess such as D-sorbite; Synthetic polymer is as polyglycol, Hydroxyethyl Starch, polyvinylpyrrolidone, polyacrylamide; Polysaccharide such as ficoll and glucosan; Protein; And the combination of above-mentioned substance.
For hematoblastic preservation, protectant example that for example can add has haemocyanin, casein hydrolysate, polyvinyl pyrrolidone and Hydroxyethyl Starch.
Preservation for nucleic acid such as RNA, DNA, oligonucleotides etc. can add SDS, guanidine, Tween etc.
In a preferred embodiment of the invention, described fluid sample is human or animal's the whole blood sample or the sample of blood constituent.
Dehydrating and can carry out under the natural room temperature evaporation conditions behind the fluid sample adding solid carrier also can be heated the removal of quickening the liquid component of fluid sample under (as in the baking ovens of 37 degree Celsius), hot blast (as hair-dryer), the reduced pressure and be formed dry sample.
As mentioned above, the present invention also provides a kind of device that is used for the kept dry fluid sample, and it comprises a kind of porous solid carrier of macropore.Above-mentioned description and optimum condition about the inventive method is equally applicable to device of the present invention.
In device of the present invention, porous solid carrier can be the Any shape that adapts with its holding components, for example sheet, cylindrical, cube shaped, sphere etc.
Be easy to from solid carrier, reclaim according to the sample of absorption dehydration of the present invention by washing, routine operation such as centrifugal, and the reconstruct fluid sample.The fluid sample of reconstruct can be used for subsequent operation and application such as the detection, purification of sample inclusions, the detection and the purifying of for example various albumen, ion, vitamin, antigen, antibody, cell, nucleic acid etc.
According to the inventive method, gain quick return for the sample inclusions, recovery height.For example the reclaimer operation of sample can be finished in several minutes time in the several seconds.According to the sample of kept dry of the present invention, the recovery can reach at least about 70%, and preferably at least 80%, more preferably at least 90%, most preferably at least 95%.
Method and apparatus of the present invention is preferably applicable to the kept dry of various biological cells.The kept dry of the red blood cell in the blood (RBC), leucocyte (WBC), blood platelet etc. for example.Before the absorptive cell sample, preferably solid carrier is handled in advance, as being closed, using organic solvents such as certain density cell fixation agent such as formalin, aldehydes, ethers, alcohols to handle with containing the protein liquid bag.Cell component is fixed and the subsequent dewatering drying, forms the immobilized cell sample that dehydrates.This sample is after adding appropriate solvent, cell can be able to aquation once more, again recover original version, can further be applied to the histochemical stain, immunohistochemical staining, immunofluorescence dyeing, the detection of cell surface molecule, the sorting or the purifying of specific cells of cell, as with magnetic bead or utilize selected by flow cytometry apoptosis cell or classification and Detection etc.
The suitable stabilizing agent of method and apparatus preferred combination of the present invention is applied to dehydrating of range protein molecule, especially various antigen, antibody, various enzymes or vitrifacation is preserved.The advantage of the inventive method is can keep steadily in the long term when above-mentioned molecule is in the state of dehydrating or vitrifacation state it in conjunction with activity and/or enzymatic activity.When adding solvent mentioned component at short notice (in common 5 minutes) rapidly aquation, be evenly distributed in the liquid phase, thereby can carry out effecting reaction with corresponding part or substrate.
Method and apparatus of the present invention also preferably is applied to collection, the drying of blood sample and preserves.Before the blood sample collection, can carry out anti-coagulants and handle or directly sneak into anti-coagulants, for example heparin, EDTA disodium solid carrier.Blood specimen collection directly takes to point puncture blood collecting or sole puncture (baby) blood-sampling method.Allow drop of blood directly splash into porous solid carrier, dehydrate the potpourri that forms dried blood particulate of anti-freezing and solid carrier then.Also can use sample injector that blood is quantitatively transferred on the above-mentioned porous carrier by conventional venipuncture blood sampling.When sample reconstruct, for example add the solvent (being generally pure water or distilled water) that is equivalent to former blood sample volume approximately and can obtain the reconstruct blood sample approaching rapidly with raw sample, can be used for subsequent treatment such as further check and analysis.According to the application purpose difference, also can not use anti-coagulants directly drop of blood not to be gone into and make dried blood sample in the solid carrier with the anti-coagulants processing.
Embodiment
The preparation of cellular solid
Substantially operate according to the method for embodiment 1 among the CN 1117587C.That is, take by weighing 2.0 gram poly 3-hydroxy butyrates, import 20 milliliters of chloroforms.65 ℃ of following water-bath heating 30 minutes, polymkeric substance is dissolved fully.Add the sodium chloride particle of 60 gram pore diameter ranges at the 0.2-0.4 millimeter in this solution, fully stirring makes and mixes.Said mixture is poured in the mould into matched moulds under the pressure of 0.2MPa.At room temperature dry 48 hours.The demoulding then, and the goods of moulding are put into vacuum drying oven 0.01MPa descended dry 48 hours.Then goods are immersed in 200 milliliters and go in the example water, changed with fresh deionized water in per 8 hours.Take out goods after 72 hours.Goods being placed vacuum drying oven under 0.01MPa dry 48 hours, obtain final products.
Utilize embodiment 1 cellular solid kept dry blood sample
Getting thickness from the polylactic acid porous material that obtains by embodiment 1 is that 0.6 centimetre, diameter are 1.1 centimetres right cylinder.Described cylindrical porosint is fixed in the inside surface of centrifuge tube lid.0.5 milliliter of whole blood sample that is added with heparin evenly is carried on the above-mentioned porosint.The above-mentioned porosint that has loaded blood sample is placed the ventilation drying at room temperature, and water cut is lower than 2% in blood sample.The lid and the centrifuge tube driving fit of dry sample will be loaded with, when using.
Parallel therewith, 0.5 milliliter of blood sample in a same source is used to measure content of hemoglobin in addition.
The recovery of dry blood sample
To open by the centrifuge tube that has dry sample that embodiment 2 obtains, in centrifuge tube, add about 0.6 ml deionized water.Build lid, centrifuge tube is put upside down, make water contact about 5 minutes with porosint.Then, lid promptly obtains the blood sample of reconstruct up with the centrifuge tube whipping several times or centrifugal a little.
By the content of hemoglobin of working sample, and with the contrast content of hemoglobin of embodiment relatively, the recovery that calculates blood sample is about 96%.
Utilize sponge sample polymeric material kept dry blood sample
According to method operation similarly to Example 2, but replace polylactic acid porous material with the polymeric sponge of buying from the market (pore diameter range is 0.2-2.0mm).According to the same quadrat method operation of embodiment 3, reclaim blood sample.In the amount of haemoglobin, the recovery of blood sample is about 93%.
Embodiment 5
From dry blood sample, extract genomic DNA
With the dry and preservation of 0.25 milliliter of blood sample according to the method for embodiment 4.In having 1.5 milliliters of centrifuge tubes that carry the blood sample porous carrier, add 1 milliliter of erythrocyte lysate, make it contact 5 minutes with the load sample solid carrier.The jolting test tube, on the Eppendrof hydro-extractor with maximum velocity centrifugation 15 seconds.Supernatant is abandoned in suction.Add 1 milliliter of erythrocyte lysate again, centrifugal behind the jolting test tube, inhale and abandon supernatant.
In test tube, add 0.3 milliliter of DNA and discharge liquid, with leucocyte precipitation mixing and left standstill 5 minutes.Then, add 0.1 milliliter of albumen precipitation liquid, mixing, centrifugal 5 minutes.
The supernatant that will contain genomic DNA is transferred in the test tube, measures DNA concentration.Carry out electrophoresis according to a conventional method, the quality of identification of dna (the results are shown in Figure 2).
With 0.25 milliliter of fresh blood sample in identical source as parallel control.
The average dna yield of 4 experiments is 35 micrograms as a result, and is 36 micrograms from the average yield of new blood.
Embodiment 6
The recovery of serum T 3, T4 and TSH
In 0.2 milliliter of serum adding porous carrier similarly to Example 4, drying is 3 hours under the room temperature.At room temperature preserved 1 month after the sealing.
The above-mentioned solid carrier that has dry blood serum sample is contacted 10 minutes with 0.2 ml distilled water.Supernatant is taken out in centrifugal back.Measure the concentration of T3, T4 and TSH respectively with chemoluminescence method.Blood serum sample to frozen identical source carries out same mensuration simultaneously.
Found that the average recovery rate (with respect to frozen sample) of three experiments of above-mentioned each factor is all near 98%.
Claims (17)
1. the method for drying and storing of a fluid sample, it comprises described fluid sample and a kind of average pore size, and to be 0.01mm contact to the porous solid carrier of 5mm; With from the solid carrier that has absorbed fluid sample, remove liquid component; The porosity of wherein dry back solid carrier is at least 20% of the former porosity of solid carrier.
2. according to the process of claim 1 wherein that the average pore size of described porous solid carrier is that 0.05mm is to 1mm.
3. according to the method for claim 2, the average pore size of wherein said porous solid carrier is that 0.1mm is to 0.5mm.
4. according to the process of claim 1 wherein that the porosity of dry back load sample solid carrier is at least 30% of the former porosity of carrier.
5. according to the process of claim 1 wherein that the porosity of dry back load sample solid carrier is at least 40% of the former porosity of carrier.
6. according to the process of claim 1 wherein that the porosity of dry back load sample solid carrier is at least 50% of the former porosity of carrier.
7. according to the process of claim 1 wherein that described porous solid carrier is to be made of one or more materials that are selected from down group: polymeric material, material, metal material and Inorganic Non-metallic Materials through chemical treatment or undressed biogenetic derivation.
8. according to the method for claim 7, wherein said polymeric material obtains by cross-linking method, foaming, the non-knitting forming method of fiber or fibrage.
9. method according to Claim 8, wherein said polymeric material is the open celled polymeric foam that makes by foaming.
10. according to the method for claim 7, wherein said metal material is foam metal material, powder metallurgy porous material, sintered metal fiber porosint or sintered metal mesh porosint.
11. according to the method for claim 10, wherein said foam metal is made by motlten metal foaming, metal powder pulp foaming, infiltration powder slurry sintering process or electrocasting.
12. according to the method for claim 7, wherein said Inorganic Non-metallic Materials is the porosint of being made by pottery, glass, cement or fire resistive material.
13. according to the process of claim 1 wherein that the described liquid component of removing is undertaken by air-dry from solid carrier.
14. according to the process of claim 1 wherein that the described liquid component of removing under reduced pressure carries out from solid carrier.
15. according to the process of claim 1 wherein that described fluid sample is the biological fluid or the liquid bio reagent of biological fluid, artificial preparation.
16. according to the method for claim 15, wherein said biological fluid is blood or its composition.
17. a device that is used for the kept dry fluid sample, it comprises a kind of porous solid carrier of macropore.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101440364B (en) * | 2008-12-23 | 2012-06-06 | 大连医科大学 | Environment-friendly DNA preservation product and manufacturing method thereof |
| US8237110B2 (en) | 2009-09-25 | 2012-08-07 | Nuctech Company Limited | Ion mobility spectrometer detection method using dopants |
| CN104777012A (en) * | 2015-04-15 | 2015-07-15 | 戚本昊 | Disposable pretreated auto-destruct urine sampler and manufacturing method thereof |
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| CN119756979A (en) * | 2024-12-18 | 2025-04-04 | 浙江清华长三角研究院 | A system, method and application for preserving human body fluids and excrement |
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| WO2011082449A1 (en) * | 2010-01-08 | 2011-07-14 | University Of Tasmania | Porous polymer monoliths, processes for preparation and use thereof |
| CA3067372A1 (en) | 2017-06-13 | 2018-12-20 | Veterinary Diagnostics Institute, Inc. | System and procedure for stabilizing, storing and recovering blood samples |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US4453955A (en) * | 1982-11-16 | 1984-06-12 | Multiform Desiccants, Inc. | Desiccant cartridge for laboratory desiccator |
| ES2268706T3 (en) * | 1995-05-09 | 2007-03-16 | Beckman Coulter, Inc. | DEVICES AND PROCEDURES FOR SEPARATING BLOOD CELL COMPONENTS FROM THE BLOOD LIQUID PORTION. |
| JP2003517057A (en) * | 1999-12-16 | 2003-05-20 | バセル テクノロジー カンパニー ビー．ブイ． | Method and apparatus for forming a supported catalyst system for olefin polymerization |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101440364B (en) * | 2008-12-23 | 2012-06-06 | 大连医科大学 | Environment-friendly DNA preservation product and manufacturing method thereof |
| US8237110B2 (en) | 2009-09-25 | 2012-08-07 | Nuctech Company Limited | Ion mobility spectrometer detection method using dopants |
| CN102033100B (en) * | 2009-09-25 | 2013-03-13 | 同方威视技术股份有限公司 | Detecting system of ion migration spectrometer (IMS) using doping agent and detecting method thereof |
| CN104777012A (en) * | 2015-04-15 | 2015-07-15 | 戚本昊 | Disposable pretreated auto-destruct urine sampler and manufacturing method thereof |
| WO2016165587A1 (en) * | 2015-04-15 | 2016-10-20 | 戚本昊 | Disposable, pre-treated and auto-destructing urine sampler and production method thereof |
| CN105594501A (en) * | 2015-11-19 | 2016-05-25 | 宁夏中青农业科技有限公司 | Urea resin foam particle matrix and processing method thereof |
| CN119756979A (en) * | 2024-12-18 | 2025-04-04 | 浙江清华长三角研究院 | A system, method and application for preserving human body fluids and excrement |
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