CN106290921A - A kind of blood type test card based on microporous membrane, Blood grouping system - Google Patents
A kind of blood type test card based on microporous membrane, Blood grouping system Download PDFInfo
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- CN106290921A CN106290921A CN201610633675.9A CN201610633675A CN106290921A CN 106290921 A CN106290921 A CN 106290921A CN 201610633675 A CN201610633675 A CN 201610633675A CN 106290921 A CN106290921 A CN 106290921A
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- 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
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/80—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells
Abstract
The present invention provides a kind of blood type test card based on microporous membrane, by multiple microtrabeculae pipes be fixed on the microporous membrane in the middle part of described microtrabeculae pipe and form, described microtrabeculae pipe is for accommodating erythrocyte to be measured and the detectable comprising antibody, and the aperture of described microporous membrane is 0.1 micron to 10 microns, and hole density is 105‑108Individual/cm2Needs according to Blood grouping, the aperture selecting described microporous membrane makes not to be settled down to bottom described microtrabeculae pipe with the erythrocyte of antibodies by microporous membrane, is blocked on above described microporous membrane with the erythrocyte after antibodies, judges blood group according to described erythrocytic red locations.The blood type test card of the present invention has long shelf-life, low cost, almost can detect all of blood group and be correlated with the feature of qualification test.
Description
Technical field
The present invention relates to blood group field tests, be specifically related to a kind of blood type test card, Blood grouping system.
Background technology
Blood group is the method to blood classification, it is common that refer to erythrocytic typing, and it is according to being whether erythrocyte surface is deposited
At some heritable antigenic substance.Antigenic substance can be protein, saccharide, glycoprotein or glycolipid.Generally some antigens
From allele or the coded product of the most chain several genes of same gene, one blood group system of these antigen composition
System.Have now been found that the mankind and be that the blood group system that international Blood Transfusion Association recognizes has 30 kinds, such as ABO blood group system, Rh blood
Type system, MNS blood group system, P blood group system etc., and wherein mostly important with ABO blood group system and Rh blood group system.At present
The authentication method of the blood group that clinical laboratory is the most frequently used is blood coagulation tests and Microcolumn gel test (MGT).
ABO blood group system is most important blood group system in all known blood group systemes.Fixed abo blood group antigen
For: A, B, AB, A1, erythrocyte membrane has Staphylococal Protein A and is referred to as red blood cells of type A, and A type also can repartition as A1 and A2 blood subgroup.Sub-at A1
Containing A and A1 antigen on type erythrocyte, and contain only Staphylococal Protein A on A2 type erythrocyte；Erythrocyte membrane has B antigen, and to be referred to as Type B red
Cell, A, B antigen has referred to as AB type erythrocyte, as none, the most referred to as O type erythrocyte.Rh blood group is most polymorphism
Blood group system, the antigen having now been found that has more than 40, but relate to clinical problem mainly have 5 antigens, i.e. D, C, c, E, e and
Its corresponding specific antibody.Antigen peci-order is D > E > c > C > e.Meanwhile, this blood group is most clinical meaning in addition to abo blood group
The blood group system of justice, plays important meaning in the diagnosis of the hemolytic disease of newborn caused in treatment of blood transfusion and tire mother's immunity, treatment
Justice.In Rh blood group system, the antigenicity of D antigen is the strongest, and individual input D antigen positive negative for the Rh of 50%～75% is red carefully
After born of the same parents, immunity produces anti-D, and the importance of D antigen is only second to A and B antigen.Under regular situation, Rh Blood grouping is to donee
Only detection D antigen, needs blood donor to check and determine its weak D antigen further.
Blood ABO and Rh to be detected blood that patient is accepted in blood transfusion or blood constituent the most compatible outside, right
The inspection screening of the antigens such as its internal irregular antibody, altofrequency and low frequency is also very important.This is mainly in view of not
With the blood of people at the different antigen of immunology, it may happen that certain untoward reaction in transfusion procedure, the latest
The property sent out hemolytic blood transfusion reaction.Additionally, for anemia of pregnant woman, irregular antibody can cause hemolytic disease of newborn, affects neonate dirty
The growth of device, and make its intelligent development come to harm, the neonatal life security of severe patient then entail dangers to.Therefore, antibody screening
It is the most necessary and necessary.
So-called irregular antibody refers to not meet the blood group antibody of abo blood group system Landsteiner rule, the most anti-
Blood group antibody beyond A, anti-B.Hypotype in ABO system, the antibody such as the anti-A of anomaly or certain anti-B, the most irregular anti-
Body.It mostly is IgG antibody.In brine media can not coagulation and can only the erythrocyte of sensitization corresponding antigens, it is necessary to by special
Medium (enzyme, antihuman globulin, polybrene etc.), just can make sensitized erythrocyte agglutination occur.
The purpose of antibody screening is to be found to have the irregular antibody of clinical meaning, selects suitable blood targetedly,
It is unlikely to match blindly.The serum of donee is done the Antibody screening test of routine by the most increasing hospital.
Antiglobulin method includes direct antiglobulin test (DAT) and indirect antiglobulin test (IAT).Antibody molecule and
Complement component is globulin, after human immunoglobulin immune animal, stimulates it to produce the antibody for foreign protein.Animal serum warp
The agglutinin that after crossing inactivation complement, absorption removal is not desired to close can occur specific reaction with human immunoglobulin (AHG).AHG can be with knot
The human immunoglobulin reaction being combined on erythrocyte or be free in serum.To containing coated erythrocyte and the system of free globulin
Interior addition AHG, free globulin can neutralize AHG.After the coated erythrocyte of AHG is scrubbed, the Fab fragment of AHG molecule be coated
Human immunoglobulin molecule Fc fragment on erythrocyte combines.So erythrocyte is put up a bridge and coagulation by AHG.The process of coagulation and bag
Being directly proportional by the amount of the globulin on erythrocyte, the most coated erythrocyte then will not coagulation.
Before blood transfusion, patient (donee) and the blood group of blood donors (blood donor) must be checked, and to intersect
Blood mathcing test.Blood transfusion is a kind of to correct the most anti-safety of blood and temporary transient effective method, and cross matching is determined whether permissible
Most important one of the inspection project of blood transfusion, is the work that must carry out before blood transfusion.Bracket for blood grouping and cross matching accurately is
Clinical safety, the important guarantee of effectively blood transfusion.In clinical medicine, in addition to blood transfusion, transplantation immunity, to hemolytic disease of newborn, certainly
The inspection of body immune hemolytic anemia specific antibody, is also required for blood group knowledge and relevant technology.Blood group is not only in blood transfusion
Upper significant, and at aspects such as ethnology, hereditism, prudence, transplantation immunity, disease resistances (or susceptibility) also
There is important using value.
The common method of bracket for blood grouping mainly has:
1. hemagglutination test
Hemagglutination test refers to that in erythrocyte and serum, blood group antigen and antibody occur macroscopic solidifying in liquid medium
Collection reaction, carries out abo blood group with hemagglutination test and identifies and generally comprise three kinds of methods: slide method, test tube method and automatization's method.Slide
The experimental principle of method and test tube method is essentially identical, and the common ground of the two is easy and simple to handle, but, positive definite form phase is only done with slide method
Ratio, carries out positive definite form and the reverse type of abo blood group qualification with test tube method, had positive reverse type to compare, it is to avoid because of blood group antigen
Or the reason of antibody and the blood group mistake that occurs, again because with test tube method carry out abo blood group identify time, erythrocyte to be measured is entered
Row washing dilution (2～the concentration of 5%), decreases the false positive occurred because red blood cell concentration is the denseest, traditional bracket for blood grouping
Although slide method and test tube method are wasted time and energy, sensitivity is poor, operation is difficult to automatization, mass, result are difficult to preserve, but, examination
The qualification of tube method abo blood group because it is easy and simple to handle, result accurately and reliably, be the most still widely used in abo blood group appraisal.
2. micro-column gel agglutination assay
Microcolumn gel technology obtains U.S. FDA approval for 1994, is built upon one on the basis of tradition blood group serology and exempts from
Epidemiology detection technique.Micro-column gel agglutination assay is the hemagglutination test of a kind of improvement, red cell antigens and the reaction of corresponding antibodies during experiment
Carrying out in micro-column gel medium, micro-column gel has the effect of gel molecular sieve, if red cell antigens is tied with corresponding antibodies
Close, the coagulation block of formation by centrifugation after can not be sieved by gel molecular and stay the upper strata of gel media；If erythrocyte is with anti-
Body is not bound with, and is formed without coagulation block after reaction, then centrifugal rear erythrocyte can be at the bottom of by gel molecular sieve precipitate gel layer
Portion, thus reach to identify the purpose of abo blood group.Microcolumn gel test eliminates the operations such as Washed Red Blood Cells, working procedure and knot
The interpretation of fruit is prone to standardization, automatization.This experimental technique also has that required specimen is few, result is accurate, capacity of resisting disturbance is strong, molten
Blood, lipidemia affect less, the advantage such as reproducible, and just because of the above advantage of Microcolumn gel test, the method has become flourishing
The abo blood group authentication method that country is conventional, domestic also have increasing unit to carry out abo blood group qualification with it.
Chinese patent application CN101718794A provides the preparation side of a kind of ABO/RhD blood typing detection reagent card
Method, reagent card has micro-column gel pipe, and gel tube contains Sephacryl gel, and the erythrocyte without coagulation can be complete
By gel particle gap, the erythrocyte of coagulation then can not pass through.Chinese patent CN101701961A provides a kind of abo blood group
The preparation method of sizing detectable card, is that gel tube contains Sephacryl gel equally.
But micro-column gel agglutination assay blood type test card is in addition to antihumanglobulin cards can detect 3 kinds, other each experiments can only make
Detect with dedicated test card.The most optimum micro-column gel agglutination assay blood type test card can only preserve 1 year at 2-25 DEG C.
Summary of the invention
In order to solve the problems referred to above, it is an object of the invention to overcome above-mentioned deficiency, it is provided that a kind of long shelf-life, cost
Low, almost can detect all of blood group and be correlated with the blood type test card of qualification test, Blood grouping system.
It is an object of the invention to provide a kind of blood type test card based on microporous membrane, the microporous membrane in described detection card allows
Single free erythrocyte passes through, and because occurring the erythrocyte of coagulation then cannot pass through with antibodies, can be trapped in film surface shape
Become red red blood cell layer.Microtrabeculae pipe in described detection card adds erythrocyte and antibody, treats erythrocyte and antibody response
Rear gradient centrifugation.If antibody and erythrocyte react and coagulation, then can be trapped within microporous membrane surface and form the red of redness
Cellular layer, it is determined that for positive findings；If antibody does not reacts with erythrocyte, then the erythrocyte that dissociates can be assembled by filter membrane
Bottom microtrabeculae pipe, it is determined that for negative findings.
A further object of the present invention is to provide a kind of Blood grouping system based on microporous membrane.
According to an aspect of the present invention, a kind of blood type test card, by multiple microtrabeculae pipes be fixed in the middle part of described microtrabeculae pipe
Microporous membrane composition, described microtrabeculae pipe for accommodating erythrocyte to be measured and the detectable comprising antibody, described microporous membrane
Aperture is 0.1 micron to 10 microns, and hole density is 105-108Individual/cm2, according to the needs of Blood grouping, select described microporous membrane
Aperture make not to be settled down to bottom described microtrabeculae pipe with the erythrocyte of antibodies by microporous membrane, with antibodies after red
Cell is blocked on above described microporous membrane, judges blood group according to described erythrocytic red locations.
Wherein, described microporous membrane by selected from polyether sulfone, nylon, polypropylene, polyethylene, polyethylene terephthalate,
Merlon, politef, Kynoar, polytetrafluoroethyldispersion dispersion resin, polyurethane, polrvinyl chloride, polyimides, glass
The material of glass fibrous membrane, cellulose mixture and organosilicon membrane is made.
Described microporous membrane is preferably chosen from polyethylene terephthalate, Merlon, polypropylene and polyimides.
Described microporous membrane is prepared by following method:
1) basement membrane imaging: be 6 μm～the polyethylene terephthalate of 50 μm, Merlon, polypropylene with thickness, gather
The film of acid imide material is basement membrane；
2) irradiation: selection energy is 1MeV-500MeV, stream is by force that the heavy ion beam current of 0.1 na-10 microamperes is from one side
Irradiation basement membrane, exposure time is 0.1 second to 9 hours；
3) sensitization: utilize the Burdick lamp of wavelength 200-365nm that the basement membrane after irradiation is irradiated, Burdick lamp power
Being 50 watts-2000 watts, lamp is 0.1 centimetre-50 centimetres with the distance of basement membrane, and sensitization time is 10 seconds-180 minutes；
4) etching: the basement membrane after sensitization is put into highly basic or strong acid solution that concentration is-30 equivalents of 0.1 equivalent
In be etched, etching period is 1-180 minute；
5) dry: through the oven for drying of excess temperature 10 degree to 150 degree, prepare described microporous membrane.
The plurality of microtrabeculae pipe includes detection pipe and Quality Control pipe.
Described microtrabeculae pipe is made up of polypropylene, polyethylene or polrvinyl chloride.
Described detectable is ABO positive definite form, reverse type, Rh blood group, Direct antiglobulin test, indirect anti-human ball egg
White test, cross matching, Rh blood grouping, direct antihuman globulin classification experiments, A2 blood subgroup qualification test, the inspection of neonate blood group
The reagent surveyed.
According to an aspect of the present invention, it is provided that a kind of Blood grouping system, including above-mentioned blood type test card and supporting examination
Agent, described matched reagent includes the antibody for detecting blood group and corresponding antibody diluent, and described antibody diluent contains phosphorus
Acid buffer, bovine serum albumin (BSA) as lubricant, Polyethylene Glycol (PEG), NaN as preservative3, and as steady
Determine the sucrose of agent, BSA.
Wherein, described antibody diluent consists of: Na2HPO4·12H2O 0.715g、KH2PO4 0.408g、NaCl
1.75g, EDTA-2Na 2.0g, BSA 10g, PEG6000 0.5g, sucrose 40g and NaN30.1g, fully dissolves by purified water
To 1000ml, 2～25 DEG C of preservations.
The described antibody for detecting blood group selected from anti-A monoclonal antibody, anti-B monoclonal antibody, anti-D monoclonal antibody,
Anti-human IgG antibodies, anti-C3d antibody titer, anti-human igg+C3d antibody, anti-A1 antibody, anti-C antibody, anti-c antibody, anti-E antibody and
Anti-e antibody.
Blood type test card of the present invention provides the benefit that compared with micro-column gel agglutination assay:
The microtrabeculae pipe of blood type test card the most of the present invention can use the materials such as polypropylene, polyethylene, polrvinyl chloride to make, micro-
Pore membrane can use polyether sulfone, nylon, polypropylene, polyethylene, Merlon, politef, Kynoar, poly terephthalic acid
Glycol ester, polytetrafluoroethyldispersion dispersion resin, polyurethane, polrvinyl chloride, polyimides, glass fibre membrane, cellulose mixture, have
The materials such as machine silicon fiml are made, and these materials have the highest stability, do not have particular/special requirement to storage temperature, are valid up to
More than 10 years.The most optimum micro-column gel agglutination assay blood type test card can only preserve 1 year at 2-25 DEG C.
Not adding gel/bead in blood type test card the most of the present invention, cost is than micro-column gel agglutination assay Blood grouping
Block much lower.
Blood type test card matched reagent the most of the present invention can detect almost all of blood group and be correlated with qualification test, including
ABO positive definite form, reverse type, Rh blood group, Direct antiglobulin test, indirect antihuman globulin test, cross matching, Rh blood group
Typing, direct antihuman globulin classification experiments, A2 blood subgroup qualification test, neonate Blood grouping test etc..And micro-column gel agglutination assay
Blood type test card is in addition to antihumanglobulin cards can detect 3 kinds, and other each experiments can only use dedicated test card to detect.
Blood type test card matched reagent of the present invention greatly facilitates medical operator, improves the work of medical treatment testing agency
Make efficiency.
Microporous membrane the most of the present invention uses track etch method to make, and microporous membrane aperture can be from 0.1 micron to 10 micron, hole
Footpath uniform, controllable, hole density can be by 105-108Individual/cm2Controlled, microporous membrane stability, homogeneity are the best.
Blood type test card matched reagent the most of the present invention can be used for full-automatic blood type analytical instrument, it is achieved detection complete from
Dynamicization.
Matched reagent of the present invention provides the benefit that:
Antibody diluent the most of the present invention uses phosphate buffer, has good buffer capacity, can protect well
Antibody；The NaCl concentration added in buffer is relatively low, compared with the NaCl concentration of normal saline, can reduce antibody anti-with erythrocyte
Seasonable resistance, intensified response sensitivity.
Antibody diluent the most of the present invention adds bovine serum albumin (BSA), Polyethylene Glycol (PEG) as lubricant,
Can protect erythrocyte well so that erythrocyte is the most broken, free erythrocyte, more easily by microporous membrane, improves the spy of reaction
The opposite sex.
Antibody diluent the most of the present invention adds NaN3As preservative, the long bacterium of diluent can be avoided, improve each examination
The stability of agent.
Antibody diluent the most of the present invention addition sucrose, BSA, as stabilizer, can protect various antibody molten well
Stability in liquid.Test result indicate that, with the various antibody reagents of antibody diluent of the present invention preparation at the bar of 2-8 DEG C
Part lower effect duration is no less than 12 months.
Various blood types card matched reagent the most of the present invention all uses independent packaging, it is easy to preserve.It is easy to use, it is possible to
For full-automatic blood type analytical instrument, it is achieved the full-automation of detection.
Accompanying drawing explanation
Fig. 1 is blood type card schematic diagram；
Fig. 2 is blood type card single hole schematic diagram, 1: microtrabeculae pipe；2: foil laminated film seals；3: microporous membrane, microporous membrane allows red
Cell passes through, because occurring the erythrocyte of coagulation then cannot pass through with antibodies；
Fig. 3 is positive findings；
Fig. 4 is negative findings.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the present invention is described in detail.
Material source: following material is purchased by commercially available unless stated otherwise.
Antibody starting material is all purchased from Immucor company, and specifying information is as shown in the table:
Test is all purchased from Shanghai blood biomedical limited liability company with normal erythrocytes, specifying information such as following table institute
Show:
| Article No. | Title | Code | Purposes |
| 5012053 | People's abo blood group reverse type red corpuscle reagent box | 3x10mL | ABO reverse type normal erythrocytes |
| 5012070 | Antibody screening erythrocyte kit | 3x5mL | Irregular antibody screening |
The preparation of [embodiment 1] detection card
Step one, the processing method of a kind of microporous membrane, comprise the following steps:
(1) basement membrane imaging: can be selected for thickness is the polyether sulfone of 6～50 μm, nylon, polypropylene, polyethylene, poly-terephthaldehyde
Acid glycol ester, Merlon, politef, Kynoar, polytetrafluoroethyldispersion dispersion resin, polyurethane, polrvinyl chloride,
A kind of material in polyimides, glass fibre membrane, cellulose mixture, organosilicon membrane is basement membrane；
(2) irradiation: selection energy is 1-500MeV, stream is by force that the heavy ion beam current of 0.1 na-10 microamperes is from an irradiation
Basement membrane, exposure time is 0.1 second to nine hours；
(3) sensitization: utilize the Burdick lamp of wavelength 200-365nm that the basement membrane after irradiation is irradiated, Burdick lamp merit
Rate is 50-2000 watt, and lamp is 0.1-50 centimetre with the distance of basement membrane, and sensitization time is 10 seconds-180 minutes；
(4) etching: the basement membrane after sensitization is put into and carries out in the highly basic or strong acid solution that concentration is 0.1-30 equivalent
Etching, etching period is 1-180 minute；
(5) dry: after the oven for drying of excess temperature 10 degree to 150 degree, produce the microporous membrane reaching requirement.
Step 2, microporous membrane is fixed on detection block each microtrabeculae pipe central authorities.
Step 3, seal with foil laminated film, it is ensured that seal closely knit, completely.
The preparation of [embodiment 2] detection card matched reagent: anti-A reagent, anti-B reagent, anti-D reagent, anti-human igg reagent, anti-
C3d reagent, anti-human igg+C3d reagent, anti-A1 reagent, anti-C reagent, anti-c reagent, anti-E reagent, anti-e reagent
(1) preparation of antibody diluent
Weigh Na2HPO4·12H2O 0.715g、KH2PO4 0.408g、NaCl 1.75g、EDTA-2Na 2.0g、BSA
10g, PEG6000 0.5g, sucrose 40g and NaN30.1g, is fully dissolved to 1000ml, 2～25 DEG C of preservations by purified water.
(2) selection of antibody
Anti-A antibody titer >=128.
Anti-B antibody titer >=128.
Anti-D antibody titer >=64.
Anti-human IgG antibodies titer >=50.
Anti-C3d antibody titer >=50.
Anti-human igg+C3d antibody titer >=50.
Anti-A1 antibody titer >=50.
Anti-C antibody titer >=50.
Anti-c antibody titer >=50.
Anti-E antibody titer >=50.
Anti-e antibody titer >=50.
(3) preparation of antibody reagent
Anti-A monoclonal antibody is joined according to the titer determined antibody diluent is made anti-A reagent.
Anti-B monoclonal antibody is joined according to the titer determined antibody diluent is made anti-B reagent.
Anti-D monoclonal antibody is joined according to the titer determined antibody diluent is made anti-D reagent.
Anti-human IgG antibodies is joined according to the titer determined antibody diluent is made anti-human igg reagent.
Anti-C3d antibody is joined according to the titer determined antibody diluent is made anti-C3d reagent.
Anti-human igg+C3d antibody is joined according to the titer determined antibody diluent is made anti-human igg+C3d reagent.
Anti-A1 antibody is joined according to the titer determined antibody diluent is made anti-A1 reagent.
Anti-C antibody is joined according to the titer determined antibody diluent is made anti-C reagent.
Anti-c antibody is joined according to the titer determined antibody diluent is made anti-c reagent.
Anti-E antibody is joined according to the titer determined antibody diluent is made anti-E reagent.
Anti-e antibody is joined according to the titer determined antibody diluent is made anti-e reagent.
[embodiment 3] positive definite form is tested
(1) by detection card label, every 4 microtrabeculae pipes are 1 person-portion.1-4 microtrabeculae pipe is set to A pipe, B pipe, D pipe,
Ctl. manage.Each detection card can detect 2 person-portions.
(2) in A pipe, add anti-A reagent respectively, B pipe adds anti-B reagent, D pipe adds anti-D reagent, in Ctl. pipe
Add antibody diluent, often pipe 50 μ l.
(3) it is sequentially added in 1-4 microtrabeculae pipe respectively with 4% red cell suspension of normal saline dilution, often pipe 10 μ l.
(4) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(5) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
Blood group result judges, as shown in the table:
Note: "+" it is positive, "-" is negative.
The experiment of [embodiment 4] positive reverse type
(1) by detection card label, each detection card can detect 1 person-portion.1-8 microtrabeculae pipe is set to A pipe, B pipe, D pipe,
Ctl. pipe, Ac pipe, Bc pipe, Oc manage, from barrel.1-4 microtrabeculae pipe detects for positive definite form, and 5-8 microtrabeculae pipe detects for reverse type.
(2) in A pipe, add anti-A reagent respectively, B pipe adds anti-B reagent, D pipe adds anti-D reagent, in Ctl. pipe
Add antibody diluent, often pipe 50 μ l；At Ac pipe, Bc pipe, Oc pipe, in barrel, it is sequentially added into test serum/blood plasma, often
Pipe 50 μ l.
(3) manage at A pipe, B pipe, D pipe, Ctl., be sequentially added in barrel with 4% erythrocyte of normal saline dilution respectively
Suspension, often pipe 10 μ l；Adding the normal erythrocytes of A1 type 2-3% in Ac pipe, the standard adding Type B 2-3% in Bc pipe is red carefully
Born of the same parents, add the normal erythrocytes of O type 2-3%, often pipe 15-20 μ l in Oc pipe.
(4) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(5) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
Blood group result judges, as shown in the table:
Note: "+" it is positive, "-" is negative.
[embodiment 5] directly antihuman globulin experiment
(1) by detection card label, each microtrabeculae pipe can detect 1 person-portion.
(2) in 1-8 microtrabeculae pipe, anti-human igg+C3d reagent, often pipe 50 μ l are added respectively.
(3) it is sequentially added in 1-8 microtrabeculae pipe respectively with 4% red cell suspension of normal saline dilution, often pipe 10 μ l.
(4) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(5) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
[embodiment 6] cross matching is tested
(1) by detection card label, every 2 microtrabeculae pipes can detect 1 person-portion.By 1-2 microtrabeculae pipe respectively labelling " master " and " secondary
Side ".
(2) master hole is initially charged serum or the blood plasma 40 μ l of donee, adds 4% red blood cell suspension of blood donor
10μl。
(3) secondary side opening is initially charged serum or the blood plasma 40 μ l of blood donor, adds 4% red blood cell suspension of donee
10μl。
(4) detection card is erected on table top, the lower left corner of tapping card and the lower right corner, the serum in mixing reaction chamber or blood
Slurry and erythrocyte, mixing be placed on immunity microtrabeculae couveuse in 37 DEG C hatch 15min.
(5) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(6) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
[embodiment 7] irregular antibody examination is tested
(1) by detection card label, every 3 microtrabeculae pipes can detect 1 person-portion.1-3 microtrabeculae pipe is respectively labeled as I, II, III.
(2) corresponding 0.8～1% antibody screening red blood cell suspension is added according to being marked in the reaction chamber of every hole microtrabeculae pipe
Liquid 50 μ l.
(3) in the reaction chamber of every hole microtrabeculae pipe of labelling, it is separately added into the serum/plasma 50 μ l of person to be checked again.
(4) detection card is erected on table top, the lower left corner of tapping card and the lower right corner, the serum in mixing reaction chamber or blood
Slurry and erythrocyte, mixing be placed on immunity microtrabeculae couveuse in 37 DEG C hatch 15min.
(5) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(6) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
[embodiment 8] directly antihuman globulin classification experiments
(1) by detection card label, every 2 microtrabeculae pipes can detect 1 person-portion.1-2 microtrabeculae pipe is respectively labeled as IgG pipe, C3d
Pipe.
(2) in IgG pipe, add anti-human igg reagent, C3d pipe adds anti-C3d reagent, often pipe 50 μ l.
(3) it is sequentially added in each microtrabeculae pipe with 4% red cell suspension of normal saline dilution, often pipe 10 μ l.
(4) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(5) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
[embodiment 9] A2 blood subgroup qualification test
(1) by detection card label, every 2 microtrabeculae pipes can detect 1 person-portion.1-2 microtrabeculae pipe is respectively labeled as A pipe, A1 pipe.
(2) in A pipe, add anti-A reagent, A1 pipe adds anti-A1 reagent, often pipe 50 μ l.
(3) it is sequentially added in each microtrabeculae pipe with 4% red cell suspension of normal saline dilution, often pipe 10 μ l.
(4) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(5) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
Blood group result judges, as shown in the table:
| Blood group | A manages | A1 manages |
| A1 | + | + |
| A2 | + | - |
Note: "+" it is positive, "-" is negative.
[embodiment 10] Rh blood grouping is tested
(1) by detection card label, every 5 microtrabeculae pipes can detect 1 person-portion.1-5 microtrabeculae pipe is respectively labeled as D pipe, C pipe, c
Pipe, E pipe, e pipe.
(2) in D pipe, add anti-D reagent, C pipe adds anti-C reagent, c pipe adds anti-c reagent, adds in E pipe
Enter anti-E reagent, e pipe adds anti-e reagent, often pipe 50 μ l.
(3) it is sequentially added in each microtrabeculae pipe with 4% red cell suspension of normal saline dilution, often pipe 10 μ l.
(4) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(5) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
Blood group result judges, as shown in the table:
Note: "+" it is positive, "-" is negative.
[embodiment 11] neonate blood group test experience
(1) by detection card label, every 5 microtrabeculae pipes can detect 1 person-portion.1-5 microtrabeculae pipe is respectively labeled as A pipe, B pipe, D
Pipe, Ctl. pipe, IgG+C3d pipe.
(2) in A pipe, add anti-A reagent, B pipe adds anti-B reagent, D pipe adds anti-D reagent, Ctl. pipe adds
Antibody diluent, adds anti-human igg+C3d reagent, often pipe 50 μ l in IgG+C3d pipe.
(3) it is sequentially added in each microtrabeculae pipe with 4% red cell suspension of normal saline dilution, often pipe 10 μ l.
(4) card is put into medical centrifuge to be centrifuged, 2 minutes 800rpm, 3 minutes 1500rpm, take out naked eyes result of determination.
(5) result judges
Red cell agglutination is on microporous membrane surface, for positive findings；
Red blood cell condensation is bottom microtrabeculae pipe, for negative findings；
Blood group result judges, as shown in the table:
Note: "+" it is positive, "-" is negative.
Claims (10)
1. a blood type test card, by multiple microtrabeculae pipes be fixed on the microporous membrane in the middle part of described microtrabeculae pipe and form, described microtrabeculae
Pipe is for accommodating erythrocyte to be measured and the detectable comprising antibody, and the aperture of described microporous membrane is 0.1 micron to 10 microns,
Hole density is 105-108Individual/cm2, according to the needs of Blood grouping, select the aperture of described microporous membrane make not with antibodies
Erythrocyte be settled down to, bottom described microtrabeculae pipe, be blocked on described microporous membrane with the erythrocyte after antibodies by microporous membrane
Top, judges blood group according to described erythrocytic red locations.
2. blood type test card as claimed in claim 1, it is characterised in that described microporous membrane by selected from polyether sulfone, nylon, poly-third
Alkene, polyethylene, polyethylene terephthalate, Merlon, politef, Kynoar, politef disperse
The material of resin, polyurethane, polrvinyl chloride, polyimides, glass fibre membrane, cellulose mixture and organosilicon membrane is made.
3. blood type test card as claimed in claim 2, it is characterised in that described microporous membrane is by selected from poly terephthalic acid second two
The material of alcohol ester, Merlon, polypropylene and polyimides is made.
4. blood type test card as claimed in claim 3, it is characterised in that described microporous membrane is prepared by following method:
1) basement membrane imaging: be that 6 μm～the polyethylene terephthalate of 50 μm, Merlon, polypropylene, polyamides are sub-with thickness
The film of amine material is basement membrane；
2) irradiation: selection energy is 1MeV-500MeV, stream is by force that the heavy ion beam current of 0.1 na-10 microamperes is from an irradiation
Basement membrane, exposure time is 0.1 second to 9 hours；
3) sensitization: utilizing the Burdick lamp of wavelength 200-365nm to be irradiated the basement membrane after irradiation, Burdick lamp power is 50
Watts-2000 watts, lamp is 0.1 centimetre-50 centimetres with the distance of basement membrane, and sensitization time is 10 seconds-180 minutes；
4) etching: put into the basement membrane after sensitization to enter in the highly basic or strong acid solution that concentration is-30 equivalents of 0.1 equivalent
Row etching, etching period is 1-180 minute；
5) dry: through the oven for drying of excess temperature 10 degree to 150 degree, prepare described microporous membrane.
5. blood type test card as claimed in claim 1, it is characterised in that the plurality of microtrabeculae pipe includes detection pipe and Quality Control pipe.
6. blood type test card as claimed in claim 1, it is characterised in that described microtrabeculae pipe is by polypropylene, polyethylene or polychlorostyrene second
Alkene is made.
7. blood type test card as claimed in claim 1, it is characterised in that described detectable is ABO positive definite form, reverse type, Rh
Blood group, Direct antiglobulin test, indirect antihuman globulin test, cross matching, Rh blood grouping, direct antihuman globulin
Classification experiments, A2 blood subgroup qualification test, the reagent of neonate Blood grouping.
8. a Blood grouping system, including the blood type test card described in claim 1 and matched reagent, described matched reagent bag
Including the antibody for detecting blood group and corresponding antibody diluent, described antibody diluent contains phosphate buffer, as lubrication
The bovine serum albumin of agent, Polyethylene Glycol, as the NaN of preservative3, and as the sucrose of stabilizer, bovine serum albumin.
9. Blood grouping system as claimed in claim 8, it is characterised in that described antibody diluent consists of: Na2HPO4·
12H2O 0.715g、KH2PO40.408g, NaCl 1.75g, EDTA-2Na 2.0g, BSA 10g, PEG6000 0.5g, sucrose
40g and NaN30.1g, is fully dissolved to 1000ml, 2～25 DEG C of preservations by purified water.
10. Blood grouping system as claimed in claim 8, it is characterised in that the described antibody for detecting blood group is selected from anti-A
Monoclonal antibody, anti-B monoclonal antibody, anti-D monoclonal antibody, anti-human IgG antibodies, anti-C3d antibody titer, anti-human igg+C3d
Antibody, anti-A1 antibody, anti-C antibody, anti-c antibody, anti-E antibody and anti-e antibody.
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|---|---|---|---|
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108680752A (en) * | 2018-04-26 | 2018-10-19 | 合肥迪安医学检验所有限公司 | The novel anti-B titrations test method of serum IgG |
| CN108680753A (en) * | 2018-04-26 | 2018-10-19 | 合肥迪安医学检验所有限公司 | The anti-B titrations test method of serum IgG |
| CN108918878A (en) * | 2018-04-26 | 2018-11-30 | 合肥迪安医学检验所有限公司 | The novel anti-A titration test method of serum IgG |
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0305337A1 (en) * | 1987-08-24 | 1989-03-01 | Stiftung Für Diagnostische Forschung | Method for the detection of antigens and/or antibodies, and a kit for carrying out the method |
| US5552064A (en) * | 1993-02-26 | 1996-09-03 | Ortho Diagnostic Systems, Inc. | Column agglutination assay and device using biphasic centrifugation |
| CN1849514A (en) * | 2003-07-08 | 2006-10-18 | 因韦尔尼斯医药瑞士股份有限公司 | Particle agglutination detection method and device |
| CN101718794A (en) * | 2009-11-25 | 2010-06-02 | 江阴力博医药生物技术有限公司 | Preparation method of ABO/RhD blood typing detection reagent card |
| CN102331504A (en) * | 2011-10-21 | 2012-01-25 | 江苏中盛医学诊断试剂有限公司 | ABO-CDE blood type grouping reagent card and preparation method thereof |
| CN102445550A (en) * | 2010-10-09 | 2012-05-09 | 苏州苏大赛尔免疫生物技术有限公司 | ABO, RhD blood typing reagent card, preparation method thereof and antibody diluting solution adopted in preparation method |
| CN103502819A (en) * | 2011-04-21 | 2014-01-08 | 西门子公司 | Blood typing devices and methods for testing blood type |
| CN103521085A (en) * | 2012-07-03 | 2014-01-22 | 中山国安火炬科技发展有限公司 | Processing method of microporous membrane as well as application of microporous membrane on cigarette filter |
| CN104081209A (en) * | 2011-12-06 | 2014-10-01 | 布鲁塞尔大学 | Method and device for assaying an antigen present on erythrocytes or an antibody binding to an antigen present on erythrocytes |
| CN104714034A (en) * | 2013-12-17 | 2015-06-17 | 天津德祥生物技术有限公司 | Blood type detection method based on membrane structure |
| CN104950115A (en) * | 2014-03-31 | 2015-09-30 | 天津德祥生物技术有限公司 | Reverse typing detecting method for human ABO blood type based on membrane structure |
-
2016
- 2016-08-03 CN CN201610633675.9A patent/CN106290921A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0305337A1 (en) * | 1987-08-24 | 1989-03-01 | Stiftung Für Diagnostische Forschung | Method for the detection of antigens and/or antibodies, and a kit for carrying out the method |
| US5552064A (en) * | 1993-02-26 | 1996-09-03 | Ortho Diagnostic Systems, Inc. | Column agglutination assay and device using biphasic centrifugation |
| CN1849514A (en) * | 2003-07-08 | 2006-10-18 | 因韦尔尼斯医药瑞士股份有限公司 | Particle agglutination detection method and device |
| CN101718794A (en) * | 2009-11-25 | 2010-06-02 | 江阴力博医药生物技术有限公司 | Preparation method of ABO/RhD blood typing detection reagent card |
| CN102445550A (en) * | 2010-10-09 | 2012-05-09 | 苏州苏大赛尔免疫生物技术有限公司 | ABO, RhD blood typing reagent card, preparation method thereof and antibody diluting solution adopted in preparation method |
| CN103502819A (en) * | 2011-04-21 | 2014-01-08 | 西门子公司 | Blood typing devices and methods for testing blood type |
| CN102331504A (en) * | 2011-10-21 | 2012-01-25 | 江苏中盛医学诊断试剂有限公司 | ABO-CDE blood type grouping reagent card and preparation method thereof |
| CN104081209A (en) * | 2011-12-06 | 2014-10-01 | 布鲁塞尔大学 | Method and device for assaying an antigen present on erythrocytes or an antibody binding to an antigen present on erythrocytes |
| CN103521085A (en) * | 2012-07-03 | 2014-01-22 | 中山国安火炬科技发展有限公司 | Processing method of microporous membrane as well as application of microporous membrane on cigarette filter |
| CN104714034A (en) * | 2013-12-17 | 2015-06-17 | 天津德祥生物技术有限公司 | Blood type detection method based on membrane structure |
| CN104950115A (en) * | 2014-03-31 | 2015-09-30 | 天津德祥生物技术有限公司 | Reverse typing detecting method for human ABO blood type based on membrane structure |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019000740A1 (en) * | 2017-06-28 | 2019-01-03 | 苏州长光华医生物医学工程有限公司 | Micro-column gel card agglutination test result identification system and blood grouping analyzer |
| CN108680752A (en) * | 2018-04-26 | 2018-10-19 | 合肥迪安医学检验所有限公司 | The novel anti-B titrations test method of serum IgG |
| CN108680753A (en) * | 2018-04-26 | 2018-10-19 | 合肥迪安医学检验所有限公司 | The anti-B titrations test method of serum IgG |
| CN108918878A (en) * | 2018-04-26 | 2018-11-30 | 合肥迪安医学检验所有限公司 | The novel anti-A titration test method of serum IgG |
| CN108918879A (en) * | 2018-04-26 | 2018-11-30 | 合肥迪安医学检验所有限公司 | The anti-A titration test method of serum IgG |
| CN109342736A (en) * | 2018-10-17 | 2019-02-15 | 深圳市龙华区中心医院 | A kind of microtrabeculae agglutination antihuman globulin detection card |
| CN110286238A (en) * | 2019-07-25 | 2019-09-27 | 深圳市宇诺生物技术有限公司 | ABO/Rh(D/C/E) blood type test card and preparation method thereof |
| CN111443211A (en) * | 2020-03-04 | 2020-07-24 | 重庆大学 | Automatic detection card and detection method for multi-blood type system |
| CN111443211B (en) * | 2020-03-04 | 2024-01-26 | 重庆大学 | Automatic detection card and detection method for multiple blood group system |
| CN111579802A (en) * | 2020-05-29 | 2020-08-25 | 东南大学 | Small device for blood type detection and detection method |
| CN112067827A (en) * | 2020-11-16 | 2020-12-11 | 天津德祥生物技术有限公司 | Antibody diluent and blood type test card comprising same |
| CN113219183A (en) * | 2021-06-16 | 2021-08-06 | 尚建华 | Liquid glue blood type detection card, preparation method and blood type detection system |
| CN113219183B (en) * | 2021-06-16 | 2024-04-19 | 尚建华 | Liquid rubber blood type detection card, preparation method and blood type detection system |
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