CN106526007B - A kind of detection plasma activities compositions, method and the blood plasma card using this method - Google Patents
A kind of detection plasma activities compositions, method and the blood plasma card using this method Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a kind of detection plasma activities compositions, method and the blood plasma cards of application this method, the method for being related to detecting albumen, polypeptide, dissociative DNA, small molecule metabolites, inorganic ions, drug and poisonous substance isoreactivity ingredient in blood plasma, uses that micro blood plasma Preparation equipment separates in real time, the blood plasma in quantitative collection whole blood sample is as sample.The micro blood plasma Preparation equipment haemocyte separating layer is capillary polypropylene class material, and using the haemocyte in the technical principle separating and filtering whole blood of volume exclusion, plasma adsorption layer is cellulose material.The present invention provides it is simple and quick, can quantitative collection blood plasma preparation method, active constituent is cured and keeps stable in blood plasma, and active constituent is detected using chemical method, physical method, chromatography, mass spectrography, biochemical method, bioanalysis, immunization, gene sequencing method.
Description
Technical field
The present invention relates to technical field of medical detection more particularly to the detection methods and equipment of plasma activities ingredient.
Background technique
Conventional blood collection method needs first to puncture vein using syringe in sample collection, extracts several milliliters to tens of milliliters
Peripheral blood is transported to testing laboratory, centrifugation removal haemocyte in heparin tube, then is detected using various methods.Blood sample is adopted
Collection wound is obvious, and blood sampling volume is larger, and the transport time limit is larger to the stability influence of active constituent, needs heparin tube, centrifuge etc.
Extras, it is difficult to implement sample collection in the place other than the special blood sampling mechanism such as health system.In addition, after blood sampling not
The whole blood of haemocyte is centrifuged under room temperature state, unstable active components.By taking homocysteine as an example, in blood
Red blood cell still can largely synthesize homocysteine in vitro, therefore its homotype half after being placed at room temperature for of the blood sample containing red blood cell
Cystine detected value can be higher than actual numerical value.And in practicing, due to blood specimen collection, blood sample transport, centrifugal treating, Samples detection etc.
Link may be by different department liables, accordingly, it is difficult to guarantee timely centrifugal treating.
Dry blood cake (Dried blood spot, DBS) is by whole blood collection in filter paper card on piece, and required blood sample is few, in blood sample
It acquires, transport, relatively conventional blood collection method is led with certain advantage in neonatal screening, the collection of medical jurisprudence material evidence etc. in preservation
Domain is widely used.But DBS needs special punch, it is difficult to accomplish quantitative collection, in addition, it, which is equally existed, to be centrifuged in time
The problem of processing, blood volume are easy to be influenced by hematocrit, not can be removed haemocyte, and erythrocyte hemolysis is to detection activity in whole blood
There may be potential interferences for ingredient, are only used for qualitative or not high to quantitative requirement detection applications.
In view of this, a kind of detection plasma activities compositions, method and the blood plasma card using this method how are designed, to eliminate
Drawbacks described above and deficiency in the prior art are the projects that related technical personnel are urgently to be resolved in the industry.
Summary of the invention
The technical issues of in order to overcome in the prior art, the present invention provides a kind of quick, accurate, micro, sensitive inspections
Survey the blood plasma card of plasma activities compositions, method and application this method.
In order to achieve the above-mentioned object of the invention, the invention discloses a kind of detection plasma activities compositions, method, the detection blood
Active constituent method is starched the following steps are included: step 1 acquires blood, and passes through the real-time washed corpuscles of blood plasma Preparation equipment and blood
Slurry；Step 2 collects and solidifies the blood plasma after separation in real time；Step 3 pair solidify after blood plasma carry out active constituent detection.
Further, in step 2, blood plasma is collected as quantitative collection.
Further, in step 1, by droplet of blood in the haemocyte filter layer of blood plasma Preparation equipment, haemocyte filter layer
Haemocyte is separated by filtration from blood.
Further, in step 2, haemocyte filter layer is torn off, the plasma adsorption layer of blood plasma Preparation equipment adsorbs and consolidates
Change blood plasma.
Further, the active constituent be it is following any one: plasma protein, polypeptide, dissociative DNA, small molecule generation
Thank object, inorganic ions, drug, poisonous substance.
Further, in step 3, the detection method of active constituent be it is following any one: chemical method, physical method, color
Spectrometry, mass spectrography, biochemical method, bioanalysis, immunization, gene sequencing method.
In order to achieve the above-mentioned object of the invention, the invention also discloses a kind of blood plasma card, the blood plasma card includes a haemocyte
Filter layer, a plasma adsorption layer, one first card, one second card, the haemocyte filter layer are fixed on first card
On, for the haemocyte in filtering blood, the plasma adsorption layer is fixed on second card, thin for adsorbing separation blood
Blood plasma after born of the same parents, the haemocyte filter layer are located on the plasma adsorption layer.
Further, first card has a blood sampling window, and the haemocyte filter layer is fixed on the blood sampling
At window.
Further, the haemocyte filter layer is polypropylene based material, has microcellular structure, the microcellular structure
Aperture is less than 1 micron.
Further, the plasma adsorption layer is cellulose material.
Compared with prior art, the technical scheme provided by the invention has the following advantages: first, simplicity is quick, one
Aspect, blood sampling are not limited by environment, time, instrument, and patient can voluntarily acquire anywhere or anytime, and be sent under room temperature
To inspection center, the convenience and patient compliance of sampling is greatly improved, on the other hand, the Trace Blood of 1-2 drop can prepare blood
Slurry easy can rapidly be applied to bioanalysis and clinical examination field convenient for the storage and transport of blood plasma card at room temperature；The
Two, quantitative collection, plasma adsorption layer volume size are fixed, and the quantitative plasma sample of precise acquisition provides for subsequent precise measurement
Material base；Third, real-time washed corpuscles and blood plasma make active constituent in blood plasma be cured and keep stable, reduce and live
The increase or decomposition of property ingredient, it is ensured that the accuracy of active component detection data；4th, it without centrifugation, is automatically separated, passes through blood
Cell separating layer carries out real-time, automatic blood cell separation while realizing blood sampling.
Detailed description of the invention
It can be obtained further by detailed description of the invention below and institute's accompanying drawings about the advantages and spirit of the present invention
Solution.
Fig. 1 is detection plasma activities compositions, method flow chart provided by the present invention；
Fig. 2 is the structural schematic diagram of blood plasma card provided by the present invention；
Fig. 3 is the partial structure diagram of blood plasma card in Fig. 2；
Fig. 4, Fig. 5 are the electron microscope pictures of the haemocyte separating layer of blood plasma card provided by the present invention；
Fig. 6 is whole blood sample and blood plasma card sample homocysteine dynamic changing curve figure at room temperature；
Fig. 7 is half Guang ammonia of homotype in Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) LC-MS/MS detection blood plasma card sample
Sour chromatogram；
Fig. 8 is homocysteine chromatography in Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) detection conventional plasma sample
Figure；
Fig. 9 is homotype in the blood plasma that blood plasma card provided by the present invention obtains and the blood plasma of conventional plasma separation method acquisition
Linear relationship chart between semicystinol concentration.
Specific embodiment
The specific embodiment that the invention will now be described in detail with reference to the accompanying drawings.However, the present invention should be understood as to not office
It is limited to this embodiment described below, and technical concept of the invention can be with other well-known techniques or function and those
The identical other technologies combination of well-known technique is implemented.
In the explanation of following specific embodiments, structure and working method of the invention in order to clearly demonstrate will be by all
Multidirectional word is described, but should by "front", "rear", "left", "right", "outside", "inner", " outside ", " inside ",
The Word Understandings such as " axial direction ", " radial direction " are not construed as word of limitation for convenience of term.
The purpose of the present invention is to provide a kind of detection plasma activities compositions, method and the blood plasma cards of application this method, can
So that the active constituent in blood plasma is cured and keep stable, reduces the increase or degradation of active constituent, especially prevent in blood
Continue synthesis of the homocysteine in vitro blood sample causes its concentration constantly to increase, meanwhile, the detection plasma activities ingredient
The blood plasma card of method and application this method, to realize quick, accurate, micro, sensitive testing goal.
The 1-9 specific embodiment that the present invention will be described in detail with reference to the accompanying drawing.
Fig. 1 is the flow chart of detection plasma activities compositions, method provided by the present invention.As shown, provided by the present invention
Detection plasma activities compositions, method include the following steps:
Step 1 acquires blood, and passes through the real-time washed corpuscles of blood plasma Preparation equipment and blood plasma；
By droplet of blood in the haemocyte filter layer of blood plasma Preparation equipment, haemocyte filter layer filters haemocyte from blood
Separation.
Step 2 collects and solidifies the blood plasma after separation in real time；
Blood plasma is collected as quantitative collection.Tear haemocyte filter layer off, the plasma adsorption layer absorption of blood plasma Preparation equipment is simultaneously
Solidify blood plasma.
Step 3 pair solidify after blood plasma carry out active constituent detection.
The active constituent be it is following any one: plasma protein, polypeptide, dissociative DNA, small molecule metabolites, it is inorganic from
Son, drug, poisonous substance.The detection method of active constituent be it is following any one: chemical method, physical method, chromatography, mass spectrography, life
Change method, bioanalysis, immunization, gene sequencing method.
Fig. 2 is the structural schematic diagram of blood plasma card provided by the present invention, and Fig. 3 is the part-structure signal of blood plasma card in Fig. 2
Figure.As shown, blood plasma card provided by the invention includes upper clamp slice 1, lower card 2, haemocyte separating layer 4, plasma adsorption layer 5,
Wherein, haemocyte separating layer 4 is used for washed corpuscles and blood plasma, is fixed at the blood sampling window 3 of upper clamp slice 1, plasma adsorption
Layer 5 is used to adsorb the blood plasma after haemocyte separating layer 4 separates blood cells, is fixed on lower card 2, meanwhile, 1 He of upper clamp slice
Stick to each other between lower card 2.
Fig. 4, Fig. 5 are the electron microscope pictures of the haemocyte separating layer of blood plasma card provided by the present invention.As shown, should
Haemocyte separating layer 4 is capillary polypropylene class material, and aperture uses the technical principle of volume exclusion to realize certainly less than 1 micron
The dynamic haemocyte and blood plasma being separated by filtration in blood sample, makes haemocyte that can not penetrate haemocyte separating layer 4 and is retained in haemocyte point
Above absciss layer 4；The plasma adsorption layer 5 is cellulose material, and Absorption quantity point is realized by the way of fixed volume size
Blood plasma from after is used for bioanalysis and clinical examination.After the blood plasma card completes sampling, the active constituent in acquired sample,
Especially homocysteine keeps stablizing at room temperature, makes the rapid curing and drying of active constituent.The active constituent includes but not
It is limited to, albumen, polypeptide, dissociative DNA, small molecule metabolites, inorganic ions, drug and poisonous substance in blood plasma.
In use, patient acquires finger tip blood, drop 1-2 drop finger tip blood to window of taking a blood sample by using the hemostix of finger prick
Haemocyte separating layer 4 at 3 is allowed to infiltrate haemocyte separating layer 4 and plasma adsorption layer 5, after placing 3 minutes, tears upper clamp slice 1 off
And it is fixed on the haemocyte separating layer 4 of upper clamp slice 1, it dries 15 minutes, fills in sample message (such as patient last name on lower card 2
The information such as name, date of birth, blood sampling date), blood plasma card is put into aluminum foil sack, saves or transports at room temperature.
The present invention is not especially limited the shape of provided haemocyte separating layer 4 and plasma adsorption layer 5, this field
Those of ordinary skill, which can according to need, makes a change the concrete shape of haemocyte separating layer 4 and plasma adsorption layer 5.
Fig. 6 is whole blood sample and blood plasma card sample homocysteine dynamic changing curve figure at room temperature, with homotype
For cysteine, after illustrating haemocyte and blood plasma separation, the active constituent in blood plasma is more stable.
3 parts of whole blood are acquired from 3 subject's ulnar veins respectively, while acquiring identical 3 subject's finger pricks acquisition
The tip finger blood arrived is dripped on micro blood plasma card.Using the sample collection time as baseline, detection is at room temperature (25 ± 1 DEG C)
The increased percent value of homocysteine after placing 2,6,12,24,48 hours respectively.
Elapsed with standing time, in whole blood (blood 1, blood 2, blood 3) homocysteine level constantly increases and baseline value phase
Than, after 2,6,12,24,48 hours the increased percentage mean value of homocysteine be respectively 4.3%, 11.9%, 32.2%,
49.7%, 108.0%；It is equal in the corresponding increased percentage of time homocysteine in micro blood plasma card (card 1, card 2, card 3)
Value is respectively 0.6%, -6.6%, -2.2%, 13.4%, 1.3%.In whole blood (blood 1, blood 2, blood 3) due to red blood cell in vitro
Still constantly synthesis causes homocysteine constantly to increase, and 48 hourly averages increase 108%；Micro blood plasma card (card 1, card 2, card 3)
The holding in 48 hours of middle homocysteine is stablized, average only to increase 1.3%.
Blood plasma card provided by the present invention, on the one hand, separated in real time while blood collection, effectively prevent blood
Active constituent in slurry is influenced by haemocyte, on the other hand, the blood plasma after separation, quick solidification, enzyme contained therein
Failure, effectively prevents influence of the enzyme to active constituent stability, and therefore, active constituent has higher stability.It is micro-
Amount blood plasma blood sampling is to guarantee the stable effective ways of homocysteine, and the sample of acquisition can obvious homotype half in stable sample
Cystine is horizontal, provides stable, accurate, convenient and fast inspection result for analysis detection.
In addition, acquiring whole blood 10mL from 3 subject's ulnar veins respectively, dripped respectively with dropper in micro blood plasma Preparation equipment
On blood plasma card, each Samples subjects make 100 parts, after placing 3 minutes, tear upper layer haemocyte separating layer off, plasma adsorption in
It on the plasma adsorption layer of lower layer, dries 15 minutes, has marked sample message, micro blood plasma card is put into original aluminum foil sack,
Sealing, is placed at room temperature.It samples, surveys 0 day, 1 day, 2 days, 3 days, 4 days, 5 days, 7 days, 10 days, 14 days, 20 days, 30 days respectively
Determine albumen and polypeptide (biochemical and immunization), nucleic acid (gene magnification+gene sequencing method), small molecule metabolites (liquid phase color
Spectrum-mass spectrography), drug and poisonous substance (liquid chromatography-mass spectrometry), inorganic ions (inductively coupled plasma mass spectrometry method) and 0
It compares, and testing result deviation is more than ± 15% to think unstable.Micro blood plasma Preparation equipment dry plasma collected is in room temperature
Under the stability results of various analytes be shown in Table 1.
The stability of 1 dry plasma of table various analytes at room temperature
Fig. 7 is homocysteine chromatogram in Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) detection blood plasma card sample.
The blood plasma card of the micro blood plasma card acquisition of finger prick is removed, by homocysteine sample treatment program addition internal standard, reduction
Agent, precipitating reagent, treated sample is for LC-MS/MS sample detection.LC-MS/MS testing conditions: API 4000LC-MS/MS instrument
(AB Sciex), the source ESI, cation scanning；MRM scanning analysis, Q1/Q3 ion channel are respectively selected as m/z:136 → 90amu
(homocysteine) and 140 → 94amu (internal standard).LC-30 high performance liquid chromatograph (Shimadzu), C18 chromatographic column (2 ×
100mm), -0.02% aqueous formic acid (10:90) of 0.02% formic acid methanol solution of mobile phase, flow velocity 0.35mL/min, 5 μ of sample introduction
L.LC-MS/MS detects that homocysteine chromatography is shown in Fig. 7, the chromatographic retention of homocysteine in micro blood plasma card
It is 1.93 minutes, detects signal strength 2150cps.Fig. 8 is Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) detection conventional plasma
Homocysteine chromatogram in sample, compared with conventional plasma sample, the chromatographic behavior of the two is consistent, no abnormality seen interference
Peak.Illustrate the detection that the collection of blood plasma equally may be implemented in micro blood plasma card, for homocysteine.
Fig. 9 is homotype in the blood plasma that blood plasma card provided by the present invention obtains and the blood plasma of conventional plasma separation method acquisition
Linear relationship chart between semicystinol concentration.As shown, 26 clinical whole blood samples are taken, it respectively will be complete with micro blood plasma card
Drop of blood collects micro blood plasma automatically and blood plasma, the side LC-MS/MS is collected in the centrifugation of conventional plasma separation method in sample collection area
Method detects homotype semicystinol concentration investigating in two kinds of plasma sample simultaneously, and analyzes the two correlation.It receives in the conventional way
For the homocysteine in blood plasma concentration integrated as x-axis (μm ol/L), micro blood plasma card acquisition plasma homocysteine is y
Axis (μm ol/L) draws linear equation, linear relationship y=0.67x+0.8.Micro blood plasma card acquisition blood plasma and traditional vein are worn
The correlation (r) for piercing blood collection method is 0.988.
Those skilled in the art should know that the detection to active constituent, detection method can be chromatography,
It can also be chemical method, physical method, mass spectrography, biochemical method, bioanalysis, immunization, gene sequencing method, the present invention does not make this to have
Body limits.
In addition, blood plasma card provided by the present invention, the DNA concentration and purity of plasma adsorption layer acquisition are as follows: from micro
The plasma adsorption layer for collecting blood plasma is removed in blood plasma Preparation equipment blood plasma card, is placed in centrifuge tube, to equipped with plasma adsorption layer
Centrifuge tube in be added 500 μ L lysates, 56 DEG C of incubation 1h extract dissociative DNA in plasma adsorption layer.It will be by being incubated for processing
Liquid is added in the hole location of DNA separating kit reagent strip, will place the carrier for extracting reagent strip and is put into 820 nucleic acid of Lab-Aid
In extraction apparatus extraction operation room, dry blood cake sample extraction procedure is selected, instrument is automatically performed extraction.Use Nanodrop 2000
Spectrophotometer carries out light absorption value detection, calculates DNA concentration and purity.Then it is detected using high-throughput gene sequencing method free
DNA carries out thalassemia genetic test, is operated to specifications.The concentration of dissociative DNA is 15.2 μ g/ μ L in blood plasma,
DNA purity is 1.68.
The Plasma volumes of blood plasma card provided by the present invention, the acquisition of plasma adsorption layer are as follows: taking 6 clinical whole bloods
Product are taken a blood sample with blood plasma card provided by the present invention respectively, are weighed the quality of the single plasma adsorption layer in acquisition front and back, are calculated acquisition
Plasma purity.Different Plasma volumes (1 μ L, 2 μ L, 5 μ L, 10 μ of each clinical whole blood sample are measured after whole blood is centrifuged again respectively
L, 50 μ L) plasma purity, using Plasma volumes as x-axis (μ L), plasma purity be y-axis (g), carry out linear regression analysis, return
Equation is y=0.0011x+0.0002, r=0.9999 (n=6), to calculate the Plasma volumes of each plasma adsorption layer acquisition
(table 2).The Plasma volumes of monolithic plasma adsorption layer acquisition are (2.75 ± 0.15) μ L, CV=5.35%, illustrate that blood plasma card is collected
Plasma sample amount it is relative constant.
The Plasma volumes of 2 blood plasma card of table acquisition
Blood plasma card provided by the present invention, homotype semicystinol concentration investigating is as follows in plasma adsorption layer: taking 2 clinical whole bloods
Sample, each sample use 6 micro blood plasma card blood samplings respectively, and LC-MS/MS detects homocysteine result respectively.With
CV≤3.7% (n=6) (table 3) between the different acquisition card of product illustrates that blood plasma card detection homocysteine is reproducible.
Different blood plasma card detections homotype semicystinol concentration investigating (μm ol/L) of table 3
Blood plasma card provided by the present invention, the rate of recovery of each heavy metal element are specific as follows: drop one is bled in micro blood plasma
The sample collection area of card after placing 3 minutes, tears upper layer haemocyte separating layer off, dries 15 minutes, remove plasma adsorption and be placed on
In 96 orifice plates, 10 μ L100% nitric acid is added to be incubated at room temperature 2 hours, then plus 190 μ L water be incubated at room temperature 1 hour, add in 200 μ L
It marks (aqueous solution of gallium element containing 100ppb), direct injected, in icp ms (ICP-MS) test sample
The elements such as heavy metal element lead, arsenic, cadmium, mercury and magnesium, sulphur, manganese, iron, zinc, sodium, potassium, selenium.With national institute of standards and technology
(NIST) reference material is reference sample, lead, arsenic, cadmium, mercury (total) the rate of recovery be respectively 102.1%, 101.5%,
101.8%, 102.9%, the rate of recovery of other elements is between 98.7%-103.5%.
Blood plasma card provided by the present invention, can be used for by detection blood plasma in amino acid, carry out neonatal screening or
Congenital hereditary metabolic disease screening, it is specific as follows: to take the plasma adsorption of blood plasma card to be placed in 96U profiled orifice plate, every hole is added
100 μ L of the working solution of internal standard containing amino acid (amino acid internal standard stoste: extraction working solution is now matched according to 1:110)；With stickiness plastic seal
Set covering microwell plate, it is ensured that 45min, oscillation frequency 650rpm are shaken in upper constant-temperature incubation oscillator after good seal, 40 DEG C of incubations；
Every hole is drawn 75 μ L extract liquors and is transferred in V-type section 96 orifice plate of bottom, and aluminium foil big envelope is used to cover microwell plate；Upper tandem mass spectrometer carries out
Sampled amino acid Concentration Testing.Glycine, alanine, proline, valine, leucine, ornithine, first sulphur in detectable blood plasma
A variety of amino acid such as propylhomoserin, phenylalanine, arginine, citrulling, tyrosine are used for neonatal screening or congenital hereditary generation
Thank to disorder in screening.
Compared with prior art, detection plasma activities compositions, method provided by the present invention and the blood plasma of application this method
Fixture has the advantage that first, simplicity quickly, on the one hand, blood sampling is not limited by environment, time, instrument, and patient can be at any time
Voluntarily acquisition everywhere, and it is passed to inspection center under room temperature, the convenience and patient compliance of sampling is greatly improved, separately
On the one hand, the Trace Blood of 1-2 drop can prepare blood plasma, convenient for the storage and transport of blood plasma card at room temperature, easy can rapidly answer
For bioanalysis and clinical examination field；The second, quantitative collection, plasma adsorption layer volume size are fixed, and precise acquisition is quantitative
Plasma sample, provide material base for subsequent precise measurement；Third, real-time washed corpuscles and blood plasma make living in blood plasma
Property ingredient is cured and keeps stable, reduces the increase or decomposition of active constituent, it is ensured that the accuracy of active component detection data；
4th, it is not necessarily to centrifugation, is automatically separated, realizes that carrying out real-time, automatic blood cell while blood sampling separates by haemocyte separating layer.
It should be noted that blood plasma card provided by the present invention is to illustrate blood plasma Preparation equipment, blood plasma card is only
For one of numerous blood plasma Preparation equipments, also, technical effect possessed by the blood plasma card, blood plasma Preparation equipment equally have
Have.For example, a kind of blood plasma Preparation equipment equally has the haemocyte separating layer of washed corpuscles and the plasma adsorption of adsorbed plasma
Layer, prepares plasma sample with this.In addition, realizing that the real-time separation of haemocyte and blood plasma is only the present invention by haemocyte separating layer
Provided one of the mode realizing haemocyte and blood plasma and separating in real time, the blood plasma Preparation equipment provided by the present invention are simultaneously unlimited
In the real-time separation that it can only realize haemocyte and blood plasma by haemocyte separating layer.
Unless otherwise instructed, the herein presented qualifier similar to " first ", " second " is not meant that suitable to the time
The restriction of sequence, quantity or importance, and be used for the purpose of the technical characteristic and another technology in the technical program
Feature is mutually distinguished.Similarly, the herein presented qualifier similar to " one " does not mean that the restriction to quantity, but describes
The technical characteristic not occurred above.Similarly, what is occurred before number herein is similar to " about ", " approximatively "
Modifier generally comprises this number, and its specific meaning should understand in conjunction with context meaning.Similarly, only have specific
Otherwise the noun of quantity quantifier modification should be regarded as again including plural form comprising singular, herein in the technology
It can include the odd number technical characteristic in scheme, also may include a plurality of technical characteristics.
It is preferred embodiment of the invention described in this specification, above embodiments are only to illustrate the present invention
Technical solution rather than limitation of the present invention.All those skilled in the art pass through logic analysis, reasoning under this invention's idea
Or the limited available technical solution of experiment, it all should be within the scope of the present invention.
Claims (4)
1. a kind of detection plasma activities compositions, method, which is characterized in that the active constituent is homocysteine, the detection
The equipment used in plasma activities compositions, method includes a blood plasma card, and the blood plasma card includes a haemocyte filter layer, a blood plasma
Adsorption layer, one first card, one second card, the haemocyte filter layer are fixed on first card, are used for hemofiltration
Haemocyte in liquid, the plasma adsorption layer are fixed on second card, for the blood plasma after adsorbing separation haemocyte, institute
It states haemocyte filter layer to be located on the plasma adsorption layer, the haemocyte filter layer is capillary polypropylene class material, micropore hole
For diameter less than 1 micron, the plasma adsorption layer is cellulose material；Wherein detection plasma activities compositions, method includes following step
It is rapid:
Step 1 acquires blood, and passes through the real-time washed corpuscles of blood plasma card and blood plasma: by droplet of blood in the haemocyte of blood plasma card
Filter layer is placed 3 minutes, and haemocyte is separated by filtration by haemocyte filter layer from blood；
Step 2 collects and solidifies the blood plasma after separation in real time: tearing haemocyte filter layer, the plasma adsorption layer absorption of blood plasma card off
And solidify blood plasma, it dries 15 minutes；Wherein blood plasma is collected as quantitative collection, and plasma adsorption layer uses the side of fixed volume size
Formula realizes the blood plasma after Absorption quantity separation；
Step 3 pair solidify after blood plasma carry out homocysteine active constituent detection；
Wherein, quickly separation solidifies blood plasma to the detection method in real time, it is ensured that the blood plasma of homocysteine after isolation
Middle stabilization.
2. detection plasma activities compositions, method as described in claim 1, which is characterized in that in step 3, the detection of active constituent
Method be it is following any one: chemical method, physical method, chromatography, mass spectrography, biochemical method, bioanalysis, immunization, gene sequencing
Method.
3. a kind of blood plasma card in the described in any item detection plasma activities compositions, methods of such as claim 1-2, feature
It is, the real-time washed corpuscles of the blood plasma card and blood plasma, including a haemocyte filter layer, a plasma adsorption layer, one first card
Piece, one second card, the haemocyte filter layer is fixed on first card, for the haemocyte in filtering blood；Institute
It states plasma adsorption layer to be fixed on second card, for the blood plasma after adsorbing separation haemocyte；Being collected as blood plasma is quantitative
It collects, plasma adsorption layer realizes the blood plasma after Absorption quantity separation, the haemocyte filtering by the way of fixed volume size
Layer is located on the plasma adsorption layer, and the haemocyte filter layer is polypropylene based material, has microcellular structure, the micropore knot
Less than 1 micron, the blood plasma card, which quickly separates blood in real time and solidifies blood plasma, guarantees homocysteine in vitro in the aperture of structure
Stablize in blood sample.
4. a kind of blood plasma card as claimed in claim 3, which is characterized in that first card has a blood sampling window, described
Haemocyte filter layer is fixed at the blood sampling window.
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| CN201911125625.XA CN111220431A (en) | 2016-10-26 | 2016-10-26 | Method for detecting active components in blood plasma and blood plasma card using same |
| CN201610943036.2A CN106526007B (en) | 2016-10-26 | 2016-10-26 | A kind of detection plasma activities compositions, method and the blood plasma card using this method |
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| CN1714292A (en) * | 2002-11-19 | 2005-12-28 | 积水化学工业株式会社 | Plasma or serum separation membrane and filter apparatus including the plasma or serum separation membrane |
| EP2375249A2 (en) * | 2010-04-09 | 2011-10-12 | F. Hoffmann-La Roche AG | Devices and process for separating plasma from a blood sample |
| CN102553440A (en) * | 2010-12-24 | 2012-07-11 | 成功大学 | Liquid filtering device and filtering method thereof |
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| ES2070942T3 (en) * | 1989-04-07 | 1995-06-16 | Abbott Lab | METHOD AND DEVICE FOR THE SEPARATION OF PLASMA OR WHOLE BLOOD SERUM. |
| AU710228B2 (en) * | 1995-06-07 | 1999-09-16 | Pall Corporation | Microfiltration membranes having high pore density and mixed isotropic and anisotropic structure |
| JP4298555B2 (en) * | 2004-03-26 | 2009-07-22 | 栄研化学株式会社 | Blood separation method and apparatus |
| CA2839281A1 (en) * | 2011-06-16 | 2012-12-20 | Baylor Research Institute | Analysis of total homocysteine and methylmalonic acid in plasma by lc-ms/ms from a plasma separator device (psd) |
| WO2014150900A1 (en) * | 2013-03-15 | 2014-09-25 | Baylor Research Institute | Methods and compositions for enhanced analyte detection from blood |
| CN107430114B (en) * | 2015-03-31 | 2020-07-24 | 豪夫迈·罗氏有限公司 | Blood plasma separating card |
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| CN1714292A (en) * | 2002-11-19 | 2005-12-28 | 积水化学工业株式会社 | Plasma or serum separation membrane and filter apparatus including the plasma or serum separation membrane |
| EP2375249A2 (en) * | 2010-04-09 | 2011-10-12 | F. Hoffmann-La Roche AG | Devices and process for separating plasma from a blood sample |
| CN102553440A (en) * | 2010-12-24 | 2012-07-11 | 成功大学 | Liquid filtering device and filtering method thereof |
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