CN1924583A - ABO blood group inverse reagent - Google Patents
ABO blood group inverse reagent Download PDFInfo
- Publication number
- CN1924583A CN1924583A CN 200610104071 CN200610104071A CN1924583A CN 1924583 A CN1924583 A CN 1924583A CN 200610104071 CN200610104071 CN 200610104071 CN 200610104071 A CN200610104071 A CN 200610104071A CN 1924583 A CN1924583 A CN 1924583A
- Authority
- CN
- China
- Prior art keywords
- reagent
- blood group
- blood
- cell
- red blood
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
This invention relates to one ABO blood reverse shape agent, which is based on the a's liquid and adopts the improved buffer system, that is from single citrate buffer system into citrate and phosphate double buffer system to better keep the system PH value stable, wherein, the invention adds gland purine as nutrient to make the erythrocyte keep bright color during abundant oxygen process; the anti-erosion system not adopting any chemical antigen to avoid impacting the oxygen content to make the red cell dark; adding EDTA sodium salt to prevent blood-stop cell hemolytic phenomenon.
Description
Technical field:
The present invention relates to a kind of detectable, particularly blood test reagent, specifically is the reverse type blood test reagent that is used for abo blood group.
Background technology:
Erythrocyte blood type is the human blood group of finding the earliest, is a kind of genetic marker of the mankind, to anthropology, science of heredity, to transfuse blood, join type, organ transplant and neonatal immunity disease significant.Since Landsteinor (1900) finds abo blood group, Hoktoen has just proposed the importance of blood group evaluation to treatment of blood transfusion as far back as 1907, Ottenberg etc. have further proposed to cooperate for acceptor blood intersection the notion of experiment in 1908, more emphasize the far reaching significance of blood group evaluation to clinical blood transfusion.Because the importance of abo blood group in blood transfusion identifies that correctly abo blood group is the assurance of safe transfusion.The principle that abo blood group is identified is based on antigen-antibody reaction and produces cell agglutination, uses anti-A, anti-B blood grouping reagent to detect red cell antigens and is referred to as apart from typing; Be referred to as reverse type with antibody in A blood group and the B blood group red blood cell detection serum.People's abo blood group is determined that by antibody in red cell antigens and the blood plasma (serum) its positive reverse type result conforms to, and contrast simultaneously also must be set up, and abo blood group just can judge rightly.
The i.e. property sent out hemolytic blood transfusion reaction symptom seriously almost will take place in abo blood group incompatible blood transfusion without exception.Correct typing of blood method and qualified blood grouping reagent are of equal importance.Lay special stress in the British Pharmacopoeia " detects abo blood group and will detect two aspects of antibody in red cell antigens and serum or the blood plasma ".This is that it is positive reverse type that the both detects, and could correctly judge abo blood group by antibody decision in cell surface antigen and the blood because of people's abo blood group.In order to increase the degree of reliability of ABO positive definite form, when accurately carrying out positive definite form, must carry out reverse type test, promptly examined blood group antibody in the serum as reagent with known blood group standard red blood cell.{ Pharmacopoeia of People's Republic of China } version in 2005 is measured for spelling out in the healthy blood donor blood group examination criteria with the anti-A of national drug supervision and management authorities approval, anti-B blood grouping reagent, and abo blood group is identified with positive and negative typing method.In order to improve blood quality, reduce the generation of abo blood group typing mistake and accident, when carrying out positive definite form, also must carry out reverse type.
Carrying out reverse type mensuration need provide a kind of reverse type to measure reagent, this reagent includes the red blood cell of A, B, O blood group, has a surge capability alserver's solution with a kind of, this preserves liquid can be with red blood cell suspension, and effectively preserve, during use this reagent and blood sample are mixed, observe agglutination phenomenon, can draw the positive or negative result.
Existing alserver's solution such as A Shi liquid, it consists of:
Chloromycetin: 0.33 gram
Citric acid list water (C
6H
8O
7H
2O): 0.55 gram
Glucose (C
6H
12O
6):, 20.5 grams
Inosine (inosine): 2.0 grams
Neomycinsulphate (Neomycin sulphate): 0.5 gram
Sodium chloride: 4.2 grams
Rubber rafter acid trisodium (C
6H
5Na
3O
72H
2O): 8.0 grams
Distilled water: to 1000ml
The PH of this preservation liquid is 6.8 ± 0.2, and osmotic pressure is 340 ± 10mOsm/kg.
Blood group reverse type reagent with the preparation of A Shi liquid can be preserved one month at 4 ℃, remove the transportation distribution time, arrive the term of validity of just having only in end user's hand about 2 weeks, also do not consider the transport for long-distance stability of cell membrane and the influence of cell viability, the cost of detection is very high for a lot of users.
For another the MAP liquid of Japan, AS-5 liquid etc., all because preserve the erythrocytic storage life of blood group short (only be January), the erythrocytic face look of blood group deepen turn black undesirable.
Blood group reverse type reagent provided by the invention, not only from prolonging the aspect research of red blood cell storage life, also open development blood group reverse type reagent by aspects such as nutrition system, buffer system, protective system, filtration washing red blood cell, spectrophotometer regulation and control red blood cell concentrations (keep batch and batch between consistance), make red blood cell concentration obtain standardization, and through red blood cell is handled, with the Precerving liquid that contains number of chemical reagent, not only prolong erythrocytic storage life and erythrocytic freshness, also kept the intensity of ABO antigen on the red blood cell simultaneously.We are short for the blood group reverse type reagent holding time, easily problems such as haemolysis, concentration regulation and control take place, and improve from three aspects respectively:
(1) blood group reverse type reagent is detected processing, comprise the removal of the removal of the detection of the various infections sources, non-red blood cell blood constituent, inmature red blood cell and bib.
(2) the high-quality and efficient blood group alserver's solution of development comprises buffer system (regulating and keep suitable pH value), nutrition system (keep red blood cell vigor, color scarlet, regulate erythrocyte sedimentation and suspension complexity etc.), osmotic pressure regulation system (prevent erythrocyte hemolysis, stablize complete, the maintenance antigen active of erythrocyte membrane) and protective system (prevent the reagent red blood cell breed bacteria and go bad).
(3) utilize the printing opacity principle, utilize spectrophotometer accurately to regulate the concentration of blood group reverse type reagent, guarantee standardized concentration.The blood group reverse type reagent of our development has not only that agglutinating reaction is strong, a characteristic of no haemolysis, no prozone phenomenon, also has stronger antijamming capability, can preserve under open state and stable better.
In order to reach the purpose that guarantees blood group reverse type reagent stable in properties, we have successfully developed the preparation method of a kind of standardized alserver's solution system and blood group reverse type reagent, not only prolonged the holding time of blood group red blood cell under 2-8 ℃ of condition, also kept the intensity and the erythrocytic freshness of ABO antigen on the red blood cell simultaneously, haemolysis has not taken place and be easy to sedimentation and suspension.
Summary of the invention:
The invention provides a kind of new abo blood group reverse type reagent, this reagent is to reach goal of the invention by the improvement to the prescription of preserving liquid.
The present invention improves its buffer system respectively on the basis of A Shi liquid, promptly become citrate-phosphate double buffer system from single citrate buffer system, can better keep the stable of preservation system pH value.The increase adenine is a nutritional labeling, makes to keep scarlet face look by red blood cell oxygen level height in the preservation process.Corrosion protection system does not adopt any chemical preservative such as compositions such as Sodium azide, thimerosal, because chemical preservative can influence the red blood cell oxygen level, causes red blood cell face look to deepen blackout.Add the EDTA disodium salt in addition, prevent that the blood group red blood cell from producing haemolysis under the complement inducing action.The present invention has done a large amount of screenings according to prior art, and new preservation formula of liquid is provided, and particularly adds stabilizing agent EDTA disodium, adopts single water glucose, chloromycetin, and, make advantage of the present invention more outstanding by adjusting the ratio of each component.
Blood group reverse type reagent of the present invention is preserved liquid, and it is as follows preferably to fill a prescription:
Citric acid trisodium 10-20 gram
Potassium dihydrogen phosphate (KH
2PO
4) the 0.3-0.5 gram
Sodium hydrogen phosphate (Na
2HPO
4) the 0.5-0.9 gram
Disodium ethylene diamine tetraacetate 0.8-1.2 gram
Sodium chloride (NaCl) 6-10 gram
Single water glucose (Glucose) 15-25 gram
Adenine (Adenine) 0.3-0.5 gram
Inosine (Inosine) 2-3 gram
Chloromycetin (Chloramphenicol) 0.5-0.8 gram
Above component is settled to 2 liters with distilled water.
Most preferred prescription is as follows:
Citric acid trisodium 14.705 grams
Potassium dihydrogen phosphate (KH
2PO
4) 0.408 gram
Sodium hydrogen phosphate (Na
2HPO
4) 0.716 gram
Disodium ethylene diamine tetraacetate 1.0 grams
Sodium chloride (NaCl) 8.182 grams
Single water glucose (Glucose) 19.810 grams
Adenine (Adenine) 0.405 gram
Inosine (Inosine) 2.682 grams
Chloromycetin (Chloramphenicol) 0.66 gram
Above component is settled to 2 liters with distilled water.
Blood group reverse type reagent of the present invention, be to add in the above-mentioned preservation liquid through the red blood cell of accurately having identified A, B, O blood group to allocate, the blood group red blood cell concentration is 2-4%, and with this blood group reverse type reagent examination criteria typing reagent, it is tired and all was not less than 1: 128.The term of validity was not less than 2 months under 2-8 ℃ of condition, and this blood group reverse type reagent detects with the standard typing reagent after diluting 8 times, and blood group red cell agglutination intensity reaches more than the 3+.Wherein said A blood group also can be A
1Blood group.
Blood group reverse type reagent of the present invention, wherein the cytotrophy system adopts carbohydrate (glucose) and nucleotide (adenine) and energy matter thigh glycosides as adjuvant, not only can delay the blood group erythrocyte hemolysis, prevent that antigen intensity from weakening, keep the stable of blood group erythrocyte membrane, guarantee the erythrocytic freshness of blood group (oxygen level): the time carbohydrate can effectively control the erythrocytic heavy speed of blood group, make the blood group red blood cell be easy to suspend, avoid erythrocyte aggregation bonding.
Blood group reverse type reagent of the present invention, wherein buffer system and osmotic pressure maintenance system adopt salt (phosphate, citrate and sodium chloride) to come the pH value of maintenance system at 6.8-6.9.Phosphoric acid-citrate buffer system is compared with single citric acid buffer system on the books, and surge capability is stronger, in the buffering range that helps keeping whole system to be in requirement, has guaranteed to preserve the ionic strength of liquid simultaneously.Preserve liquid and also have osmotic pressure regulating power preferably, utilize salt to regulate osmotic pressure and blood group red blood cell causing property in vivo, prevent the blood group erythrocyte hemolysis.
Blood group reverse type reagent of the present invention, wherein protective system adopts microbiotic (chloromycetin) corrosion protection system, does not adopt chemical preservative.By a certain amount of antibiotic synergy, effectively prevent the pollution of bacterium, controlled the blood group red blood cell and do not had varied bacteria growing, the guarantee reagent germ-free condition.
Blood group reverse type reagent of the present invention, blood group red blood cell generation haemolysis when wherein EDTA disodium salt EDTA can prevent to detect fresh serum by the inhibition complement activation, thus make the blood typing result be more prone to judge.
Blood group reverse type reagent of the present invention, wherein Pei Zhi blood group alserver's solution need pass through aseptic filtration, method is: use stainless steel high pressure filter filter membrane 0.45 μ l, 121 ℃ of following 0.15Mpa high pressure steam sterilizations 30 minutes, and record filtering start time, concluding time and filtration.
Blood group reverse type reagent of the present invention, wherein the erythrocytic concentration adjustment of blood group adopts following method: the packed red cells that the washing of usefulness blood group alserver's solution is 4 times is once abandoned supernatant.Blood group alserver's solution dilute suspension blood group red blood cell is adjusted to 2-4% with the blood group red blood cell concentration.
Alserver's solution of the present invention can be preserved and stable blood group red blood cell for a long time, thereby realizes the standardized production of blood group reverse type reagent, satisfies the requirement of accurately identifying abo blood group better.
The preparation method that the present invention also provides blood group of the present invention instead to decide reagent, this method can reach the purpose that guarantees blood group reverse type reagent stable in properties.
The preparation method of blood group reverse type reagent of the present invention may further comprise the steps:
(1) prepares alserver's solution in proportion;
(2) red blood cell is removed red blood cell and protein ingredient that aggegation has wherein taken place through filtering and carrying out washing treatment;
(3) regulate erythrocytic concentration with preservation liquid.
The raw material blood group red blood cell sample that screening is obtained at first will screen and handle, and blood uses the kit through State Food and Drug Administration approval to detect, and HBsAg, HCV antibody, HIV-1/HIV-2 antibody, the syphilis serology detects should be negative.Alanine aminotransferase (ALT) value should be chosen in the qualified people A that gathers in 48 hours in range of normal value, B, and O blood group erythrocyte blood type red blood cell does not have haemolysis.Identify abo blood group with the anti-A that ratifies through State Food and Drug Administration, anti-B blood grouping reagent, and identify with positive and negative typing method.Select qualified A blood group, B blood group and the O blood group red blood cell of antigenicity, do not use subgroup blood.The blood group red blood cell is through directly antihuman globulin test, and the result should be negative.Filter then, available filter filters or filter paper filtering, and purpose is to remove the red blood cell coagula of leucocyte, blood platelet and aggegation in the raw material blood, prevents that agglutinating particle is mixed in the blood group reverse type reagent, produces false agglutinating reaction; Remove the plasma proteins of acellular composition then by washing, comprise the blood group antibody composition, further remove part leucocyte and blood platelet simultaneously, prevent that cell fragment and content that its back of breaking produces from producing harmful effect to reagent.Use at last through the preservation liquid of aseptic filtration and regulate erythrocytic concentration, be mixed with the normal concentration blood group reverse type reagent that is fit to the typing use.
Concrete compound method is as follows:
Blood group alserver's solution preparation buffer system: adopt phosphoric acid-citrate buffer system, make the pH value of system maintain 6.8-6.9.
Nutrition system: adopt carbohydrate and nucleotide as adjuvant, can guarantee the erythrocytic freshness of blood group, control the erythrocytic settling velocity of blood group effectively, be easy to the blood group red blood cell suspension, avoid the blood group erythrocyte aggregation bonding.
Osmotic pressure regulation system: utilize salt to regulate, make in vivo consistent of its osmotic pressure and blood group red blood cell, prevent that blood group erythrocyte hemolysis screen from stablizing the blood group erythrocyte membrane with a certain amount of macromolecular complex.
Corrosion protection system: adopt microbiotic (chloromycetin), do not adopt chemical preservative.
The blood group alserver's solution aseptic filtration of preparation.
Blood group red blood cell degrease, filtration, washing, hematocrit, preparation; Blood adopts leukocyte depletion filter to remove leucocyte, to prevent the enzyme that leukocytolysis is discharged and the fragment of generation goods is produced harmful effect.
The erythrocytic preparation of hematocrit blood group:
The whole blood slurry of dehematizing, homotype is mixed; A
1Blood group, B blood group, O blood group blood three person-portions or more than, the slurry of dehematizing respectively, the equal proportion homotype is mixed.
Wash centrifugal:
Wash the red born of the same parents of blood group three times with an amount of physiological sodium chloride solution respectively.Preserve liquid washing blood group red blood cell once with an amount of again.Centrifugal 5 minutes of last 1 2000r/min abandons supernatant, and precipitation is hematocrit blood group red blood cell.
Preparation:
With alserver's solution packed red cells being mixed with concentration respectively is 2-4% blood group red cell suspension.The semi-manufacture calibrating
Sterility test:
Undertaken by { Chinese biological goods rules } current edition general rule { biological products sterility test rules } A3.2.3 item.
Specificity:
Material
Treat check reagent (blood group red cell suspension concentration 2-4%).
Anti-A, anti-B blood grouping reagent (monoclonal antibody) reagent through National Drug Administration's approval, batch batch inspection is qualified.
Anti-A
1, anti-AB (mixing of O type 10 person-portion serum) and anti-H.
Normal AB type human serum (mixing of 10 person-portions).
Physiological sodium chloride solution is meant at 22 ± 1 ℃, PH7.0 ± 0.2 o'clock 8.5-9.0g/LNaCL (0.145M-0.154M).
Test (test tube method):
(1) A
1Type red blood cell reagent is arranged by 5 test tubes of every row, and each adds anti-A, anti-A
1, each 0.1ml of anti-B, in each test tube, add 2-4%A to be checked then respectively
1Blood group red blood cell 0.1ml puts 18-22 ℃ and hatched 15 minutes, and centrifugal 1 minute of 1000rpm does contrast with AB type serum and physiological sodium chloride solution, the visual inspection result simultaneously.
(2) Type B red blood cell reagent is arranged by 4 test tubes of every row, each adds anti-A, each 0.1ml of anti-B, in each test tube, add 2-4%B blood group red blood cell 0.1ml to be checked then respectively, putting 18-22 ℃ hatched 15 minutes, centrifugal 1 minute of 1000rpm, do contrast with AB type serum and physiological sodium chloride solution simultaneously, the visual inspection result.
(3) O type red blood cell reagent is arranged by 6 test tubes of every row, each adds anti-H, anti-A, anti-B, anti-AB0.1ml, in each test tube, add 2-4%O blood group red blood cell 0.1ml to be checked then respectively, putting 18-22 ℃ hatched 15 minutes, centrifugal 1 minute of 1000rpm, do contrast with AB type serum and physiological sodium chloride solution simultaneously, the visual inspection result.
The result judges:
A
1Type red blood cell and anti-A and anti-A
1Agglutinating reaction does not take place with anti-B reagent, AB type human serum and physiological sodium chloride solution in reagent generation agglutinating reaction.
Agglutinating reaction does not take place with anti-A reagent, AB type human serum and physiological sodium chloride solution in Type B red blood cell and anti-B reagent generation agglutinating reaction.
Agglutinating reaction takes place in O cell and anti-H, with anti-A, anti-B, anti-AB, AB type human serum, physiological sodium chloride solution agglutinating reaction does not take place.
| Antibody reagent | A 1Blood group red blood cell reagent | B blood group red blood cell reagent | O blood group red blood cell reagent |
| The anti-A of anti-A 1The anti-H AB of the anti-AB of anti-B type human serum physiological sodium chloride solution | + + - / / - - | - / + / / - - | - / - + + - - |
This mini-test "+" ecbatic positive "-" ecbatic feminine gender is not carried out in "/" expression
Antigenicity detects:
Reagent, anti-A, anti-B blood grouping reagent (monoclonal antibody) are used the reagent through National Drug Administration's approval, and batch batch inspection is qualified, and tiring is 1: 128.
Anti-H blood grouping reagent.
Blood group red blood cell to be checked (2-4%).
Test:
Test tube is lined up 2 row, 9 of every row (3 of the every row of O cell), add the 0.2ml physiological sodium chloride solution from every of the 2nd pipe, the internal control product reagent that in first and second test tube, adds 0.2ml respectively, since second pipe anti-A of inner quality standard, anti-B or anti-H reagent are made 2 times of serial dilutions, every diluted sample two rows.(anti-A is used for detecting A as the internal control product to add the blood group red blood cell 0.2ml of 2-4% correspondence to be checked forwards, backwards in Pai every test tube respectively
1The type human red cell), shake up, put 18-22 ℃ 15 minutes, centrifugal 1 minute of 1000rpm substitutes the anti-A of internal control product, anti-B with physiological sodium chloride solution simultaneously or anti-H reagent is negative control visual inspection result.
The result judges:
Judge respectively and treat that check reagent and the generation of corresponding antibodies reagent are not weaker than the high dilution of 3+ aggegation intensity and are not weaker than the high dilution of 1+ aggegation intensity, the measurement result of check reagent is treated in the identical conduct of 2 row test tubes, and should meet following table and require: differ 1 pipe and meet the requirements then to dilute the low terminal point that is judged to as 2 row terminal points, differ 1 pipe and have that 1 row result is undesirable then need to get the double check reagent retrial for the treatment of as 2 row terminal points, the retrial result must not have 1 row degree pipe terminal point undesirable.
The detection of blood group red blood cell concentration:
Get 20ml blood group red cell suspension to be checked, in 1000rpm centrifugal 1 minute, obtain packed red cells after removing supernatant, measure the volume of packed red cells, calculate its shared percent by volume in 20ml blood group red cell suspension.Requiring the concentration of human red cell is 2-4%.
The haemolysis index detects:
The method of employing peroxidase detects the content of haemoglobin in the supernatant, measures under wavelength 510nm, requires hemoglobin concentration not to be higher than 0.29g/L.
Test procedure:
Add reagent according to following table order and volume:
| Reagent | Blank pipe | Standard pipe | Sample tube to be checked |
| 1.0.1g/L haemoglobin standard liquid (ml) is 4. benzidine reagent (ml) 5.1% hydrogen peroxide (ml) of distilled water (ml) sample 3. to be checked (ml) 2. | 0.02 1.00 1.00 | 0.02 1.00 1.00 | 0.02 1.00 1.00 |
| The rearmounted room temperature 5-10min of mixing adds again | |||
| 6.10% acetate (ml) | 10.0 | 10.0 | 10.0 |
| Measure wavelength 510nm, after the zeroing of blank pipe, detect absorption value | |||
The result judges
Free hemoglobin concentration=(measuring pipe absorption value/standard pipe absorption value) X standard items concentration
Free hemoglobin concentration≤0.29g/L is qualified.
Packing
The blood group red cell suspension is carried out packing.
A
1Blood group red blood cell reagent, B blood group red blood cell reagent, O blood group red blood cell reagent, every loading amount is 10ml.
The finished product calibrating
Physical examination
Outward appearance
A
1Blood group red blood cell reagent: the red blood cell of reagent bottom should be cerise, and supernatant is clear and bright, jolting gently, and red blood cell easily suspends, and does not have little polymers foreign matter.
B blood group red blood cell reagent: the red blood cell of reagent bottom should be cerise, and supernatant is clear and bright, shakes gently, and red blood cell easily suspends, and does not have little polymers or foreign matter.
O blood group red blood cell reagent: the red blood cell of reagent bottom should be cerise, and supernatant is clear and bright, jolting gently, and red blood cell easily suspends, and does not have little polymers or foreign matter.
Specificity
Antigenicity detects
Affinity
Material
(1) anti-A, anti-B blood grouping reagent (monoclonal antibody)
(2) (2) require to make 10% blood group cell suspension according to 2.2.7.1 (2).
Test
Will the anti-A of about 0.05ml, anti-B reagent puts on clean porcelain plate or the slide, add the corresponding 10% blood group red cell suspension of equal-volume, mixing immediately, the face diameter requires more than 10mm, and the blood group of dripping red cell suspension writes down the size of naked eyes aggegation piece when visible aggegation time and 3 minutes at once.
The result judges that the aggegation piece must reach 1mm in 3 minutes
2More than, the affinity aggegation time should meet following table and require:
| Red blood cell to be checked | Antibody reagent | The aggegation time | Grumeleuse in the time of 3 minutes |
| A 1 | Anti-A reagent | ≤ 15 seconds | ≥1mm 2 |
| B | Anti-B reagent | ≤ 15 seconds | ≥1mm 2 |
Reagent of the present invention is applicable to that test tube method, micro plate method (robotization blood typing system), gel cards detect the antibody of the ABO system in blood plasma or the serum.
1, test tube method: this product is mixed in 1: 1 ratio with person under inspection's blood plasma or serum, shakes up.In 900g centrifugal 15 seconds, or room temperature left standstill 1 hour.Whether check haemolysis, haemolysis may be positive findings, or bacterial contamination.Jiggle, observe aggegation situation and immediate record result.
2, micro plate method: in micro plate, add the corresponding blood group red blood cell of 25-50ul and examinee's serum or blood plasma respectively with mark according to 1: 1 ratio.And have gentle hands gently detain clap or with micro plate concussion instrument slight and content in the mix aperture fully, in 700g centrifugal 30 seconds.Whether check haemolysis, haemolysis may be positive findings, or bacterial contamination.Gently detain bat or shake instrument Red Blood Cells Suspension button with have gentle hands, observe the aggegation situation and also write down the result with micro plate machinery.
3, gel block-regulations: in the reverse type pipe of cassette typing, add and treat check reagent 50ul (blood group red cell suspension concentration 0.8%).Add person under inspection's human serum or blood plasma 25ul again.Centrifugal in the cassette blood group special centrifugal machine, naked eyes or block the interpretoscope sentence read result automatically.
Explanation to test findings:
With known A
1After blood group, B blood group, O blood group red blood cell reagent and person under inspection's blood plasma or serum mix, as shown in the table: with A
1Aggegation does not take place and is A type blood group with B blood group red blood cell reagent generation aggegation person in blood group and O blood group red blood cell reagent: with A
1The generation aggegation of blood group red blood cell reagent and and B blood group and not aggegation of O blood group red blood cell reagent person be the Type B blood group; With A
1Blood group and the reagent generation aggegation of B blood group red blood cell and and O blood group red blood cell reagent aggegation person is not taken place is O type blood group; With A
1, aggegation person all do not take place and is AB type blood group in B and O blood group red blood cell reagent.
| Blood group | A 1Blood group red blood cell reagent | B blood group red blood cell reagent | O blood group red blood cell reagent |
| A B O AB | - + + - | + - + - | - - - - |
Annotate: "+" has agglutinating reaction, "-" no agglutinating reaction (limitation of this test method)
This test only detects abo blood group as reverse type and uses.
Reagent of the present invention, through clinical trial, proving effect is good.
Hebei province's Blood Center and Inner Mongolia Autonomous Region Blood Center adopt abo blood group reverse type reagent of the present invention (human red cell), lot number: 20050904,20050905,20050906, adopt reverse type reagent Reagent Red Blood Cells (Pooled Cells) with comparison reagent, IMMUCOR.INC produces, lot number: 42221, the anti-A that positive definite form reagent adopts Changchun Boulder Bioisystech Co., Ltd to produce, anti-B blood grouping reagent (monoclonal antibody), lot number: 20050303, equality detects 6184 routine blood donors (762 of ethnic groups) coincidence rate 100%.
Chinese people open dress police force hospital general and adopt abo blood group reverse type reagent of the present invention (human red cell), lot number: 20050904,20050905,20050906, adopt reverse type reagent Reagent Red Blood Cells (Pooled Cells) with comparison reagent, IMMUCOR.INC produces, lot number: 42221, anti-A, anti-B blood grouping reagent (monoclonal antibody) that positive definite form reagent adopts Changchun Boulder Bioisystech Co., Ltd to produce, lot number: 20050303, parallel detection 1680 routine inpatients, coincidence rate 100%.
Chinese send out the Armed Police Force hospital general armed police's health examination personnel are adopted abo blood group reverse type reagent of the present invention (human red cell), lot number: 20050904,20050905,20050906, comparison reagent adopts reverse type reagent (the Reagent Red Blood Cells (PooledCells) that IMMUCOR.INC produces, lot number 42221, anti-A, anti-B blood grouping reagent that positive definite form reagent adopts Changchun Boulder Bioisystech Co., Ltd to produce, lot number: 20050303, parallel detection, detect 1990 examples altogether, coincidence rate 100%.
Blood group reverse type reagent of the present invention is prepared to A respectively
1Blood group red blood cell reagent, B blood group red blood cell reagent, O blood group red blood cell reagent is dressed up every 10ml, its label A
1Blood group red blood cell reagent is (blueness), and B blood group red blood cell reagent is (yellow), and O blood group red blood cell reagent is (redness), abo blood group reverse type reagent (human red cell).
The present invention is with A
1Blood group red blood cell reagent, B blood group red blood cell reagent, three kinds of reagent of O blood group red blood cell reagent lump together packing, and it is blue that their label is respectively, yellow, redness, each 1 of each packing, every 10ml, label is abo blood group reverse type reagent (human red cell) 3 * 10ml, specialize in surveyor's abo blood group reverse type usefulness, this reagent uses authentication code: the accurate word of traditional Chinese medicines through the approval of drug surveilance department.
Embodiment:
Further specify the present invention by the following examples, but not as limitation of the present invention.
Embodiment 1
Preserve the preparation of liquid:
Citric acid trisodium 14.705 grams
Potassium dihydrogen phosphate (KH
2PO
4) 0.408 gram
Sodium hydrogen phosphate (Na
2HPO
4) 0.716 gram
Disodium ethylene diamine tetraacetate 1.0 grams
Sodium chloride (NaCl) 8.182 grams
Single water glucose (Glucose) 19.810 grams
Adenine (Adenine) 0.405 gram
Inosine (Inosine) 2.682 grams
Chloromycetin (Chloramphenicol) 0.66 gram
Above component is settled to 2 liters with distilled water.
Preparation:
Said components is mixed with dissolved in distilled water and constant volume, use stainless steel high pressure filter filter membrane 0.45 μ l, 121 ℃ of following 0.15Mpa high pressure steam sterilizations 30 minutes.
To add through the blood group red blood cell of accurately having identified the A blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
To add through the blood group red blood cell of accurately having identified the B blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
To add through the blood group red blood cell of accurately having identified the O blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
Make A respectively
1Blood group red blood cell reagent, B blood group red blood cell reagent, three kinds of reagent of O blood group red blood cell reagent lump together packing, and three kinds of reagent is each 1 in each packing, every 10ml, label A
1Blood group red blood cell reagent is (blueness), B blood group red blood cell reagent is (yellow), O blood group red blood cell reagent is (redness) abo blood group reverse type reagent (human red cell) 3 * 10ml, specializing in surveyor's abo blood group reverse type uses, this reagent uses authentication code: the accurate word of traditional Chinese medicines through the approval of drug surveilance department.
Embodiment 2
Preserve the preparation of liquid:
Citric acid trisodium 10 grams
Potassium dihydrogen phosphate (KH
2PO
4) 0.3 gram
Sodium hydrogen phosphate (Na
2HPO
4) 0.5 gram
Disodium ethylene diamine tetraacetate 0.8 gram
Sodium chloride (NaCl) 6 grams
Single water glucose (Glucose) 15 grams
Adenine (Adenine) 0.3 gram
Inosine (Inosine) 2 grams
Chloromycetin (Chloramphenicol) 0.5 gram
Above component is settled to 2 liters with distilled water.
Preparation:
Said components is mixed with dissolved in distilled water and constant volume, use stainless steel high pressure filter filter membrane 0.45 μ l, 121 ℃ of following 0.15Mpa high pressure steam sterilizations 30 minutes.
To add through the blood group red blood cell of accurately having identified the A blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
To add through the blood group red blood cell of accurately having identified the B blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
To add through the blood group red blood cell of accurately having identified the O blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
Make A respectively
1Blood group red blood cell reagent, B blood group red blood cell reagent, three kinds of reagent of O blood group red blood cell reagent lump together packing, three kinds of reagent is each 1 in each packing, every 10ml, label are abo blood group reverse type reagent (human red cell) 3 * 10ml, specialize in surveyor's abo blood group reverse type and use, this reagent uses authentication code: the accurate word of traditional Chinese medicines through the approval of drug surveilance department.
Embodiment 3
Preserve the preparation of liquid:
Citric acid trisodium 20 grams
Potassium dihydrogen phosphate (KH
2PO
4) 0.5 gram
Sodium hydrogen phosphate (Na
2HPO
4) 0.9 gram
Disodium ethylene diamine tetraacetate 1.2 grams
Sodium chloride (NaCl) 10 grams
Single water glucose (Glucose) 25 grams
Adenine (Adenine) 0.5 gram
Inosine (Inosine) 3 grams
Chloromycetin (Chloramphenicol) 0.8 gram
Above component is settled to 2 liters with distilled water.
Preparation:
Said components is mixed with dissolved in distilled water and constant volume, use stainless steel high pressure filter filter membrane 0.45 μ l, 121 ℃ of following 0.15Mpa high pressure steam sterilizations 30 minutes.
To add through the blood group red blood cell of accurately having identified the A blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
To add through the blood group red blood cell of accurately having identified the B blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
To add through the blood group red blood cell of accurately having identified the O blood group in the above-mentioned preservation liquid and allocate, the blood group red blood cell concentration is 3%.
Make A respectively
1Blood group red blood cell reagent, B blood group red blood cell reagent, three kinds of reagent of O blood group red blood cell reagent lump together packing, and three kinds of reagent is each 1 in each packing, every 10ml, label A
1Blood group red blood cell reagent is (blueness), B blood group red blood cell reagent is (yellow), and O blood group red blood cell reagent is (redness), abo blood group reverse type reagent (human red cell) 3 * 10ml, specialize in surveyor's abo blood group reverse type usefulness, but this reagent standardization large-scale production.As seen this project has bigger social benefit.
This reagent uses authentication code: the accurate word of traditional Chinese medicines through the approval of drug surveilance department.
Claims (10)
1, a kind of abo blood group reverse type reagent is characterized in that, this reagent is formed by the red blood cell and the cell-preservation liquid mixed preparing of A, B, O blood group.
2, the reverse type reagent of claim 1, wherein the prescription of cell-preservation liquid is composed as follows:
Citric acid trisodium 10-20 gram
Potassium dihydrogen phosphate 0.3-0.5 gram
Sodium hydrogen phosphate 0.5-0.9 gram
Disodium ethylene diamine tetraacetate 0.8-1.2 gram
Sodium chloride 6-10 gram
Single water glucose 15-25 gram
Adenine 0.3-0.5 gram
Inosine 2-3 gram
Chloromycetin 0.5-0.8 gram
Above component is settled to 2 liters with distilled water.
3, the reverse type reagent of claim 1 is characterized in that, the prescription of wherein preserving liquid is composed as follows:
Most preferred prescription is as follows:
Citric acid trisodium 14.705 grams
Potassium dihydrogen phosphate 0.408 gram
Sodium hydrogen phosphate 0.716 gram
Disodium ethylene diamine tetraacetate 1.0 grams
Sodium chloride 8.182 grams
Single water glucose 19.810 grams
Adenine 0.405 gram
Inosine 2.682 grams
Chloromycetin 0.66 gram
Above component is settled to 2 liters with distilled water.
4, the reverse type reagent of claim 1 is characterized in that, wherein the red blood cell of A, B, O blood group obtains through degrease, filtration, washing, process.
5, the reverse type reagent of claim 1 is characterized in that, allocates by adding in the above-mentioned preservation liquid through the blood group red blood cell of accurately having identified A, B, O blood group, and to make red blood cell concentration be 2-4%.
6, the preparation method of the reverse type reagent of claim 1 is characterized in that, the process following steps:
(1) prepares alserver's solution in proportion;
(2) red blood cell is removed red blood cell and protein ingredient that aggegation has wherein taken place through filtering and carrying out washing treatment;
(3) regulate erythrocytic concentration with preservation liquid.
7, the preparation method of claim 6 is characterized in that, the process following steps:
The raw material blood group red blood cell sample that screening is obtained filters, filter or filter paper filtering with filter, remove the plasma proteins of acellular composition then by washing, comprise the blood group antibody composition, further remove part leucocyte and blood platelet simultaneously, prevent that cell fragment and content that its back of breaking produces from producing harmful effect to reagent, using preservation liquid through aseptic filtration that packed red cells is mixed with concentration at last is 2-4% blood group red cell suspension.
8, the preparation method of claim 6 is characterized in that, the preparation alserver's solution uses stainless steel high pressure filter filter membrane 0.45 μ l, 30 minutes step of 121 ℃ of following 0.15Mpa high pressure steam sterilizations through component is mixed with dissolved in distilled water and constant volume.
9, contain the packing box of the reverse type reagent of claim 1, it is characterized in that, this packing box is the A that will make
1Blood group red blood cell reagent, B blood group red blood cell reagent, three kinds of reagent of O blood group red blood cell reagent lump together packing, and three kinds of reagent is each 1 in each packing, every 10ml, label A
1Blood group red blood cell reagent is (blueness), and B blood group red blood cell reagent is (yellow), and O blood group red blood cell reagent is (redness), and abo blood group reverse type reagent 3 * 10ml specializes in surveyor's abo blood group reverse type and uses.
10, the using method of the reverse type reagent of claim 1 is characterized in that, this method comprises known A
1Blood group, B blood group, O blood group red blood cell reagent mix with person under inspection's blood plasma or serum, and its criterion is:
1) and A
1Aggegation does not take place and is A type blood group with B blood group red blood cell reagent generation aggegation person in blood group and O blood group red blood cell reagent;
2) and A
1The generation aggegation of blood group red blood cell reagent and and B blood group and not aggegation of O blood group red blood cell reagent person be the Type B blood group;
3) and A
1Blood group and the reagent generation aggegation of B blood group red blood cell and and O blood group red blood cell reagent aggegation person is not taken place is O type blood group;
4) and A
1, aggegation person all do not take place and is AB type blood group in B and O blood group red blood cell reagent.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610104071 CN1924583A (en) | 2006-08-02 | 2006-08-02 | ABO blood group inverse reagent |
| CN2007101114071A CN101074962B (en) | 2006-08-02 | 2007-06-18 | ABO blood-type anti-grouping reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610104071 CN1924583A (en) | 2006-08-02 | 2006-08-02 | ABO blood group inverse reagent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1924583A true CN1924583A (en) | 2007-03-07 |
Family
ID=37817308
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200610104071 Pending CN1924583A (en) | 2006-08-02 | 2006-08-02 | ABO blood group inverse reagent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1924583A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102466730A (en) * | 2010-11-19 | 2012-05-23 | 南京神州英诺华医疗科技有限公司 | Novel method for matching blood reverse typing reagent |
| CN105445455A (en) * | 2014-08-28 | 2016-03-30 | 山东省肿瘤医院 | Electrophoresis buffer solution for cell in-situ electrophoresis |
| CN107037220A (en) * | 2017-04-01 | 2017-08-11 | 合肥迪安医学检验所有限公司 | The compound method of ABO reverse type red cell suspensions |
| CN109946129A (en) * | 2019-03-12 | 2019-06-28 | 江苏中济万泰生物医药有限公司 | Five classification cellanalyzer quality-control product preparation methods |
| CN111443211A (en) * | 2020-03-04 | 2020-07-24 | 重庆大学 | Automatic detection card and detection method for multi-blood type system |
| CN112684192A (en) * | 2020-12-25 | 2021-04-20 | 天津市宝坻区人民医院 | New application of ammonium bradykinin in ABO blood group reverse typing kit |
| CN114732008A (en) * | 2022-06-13 | 2022-07-12 | 深圳瑞亚力集团有限公司 | Erythrocyte preservation solution, preparation method thereof and erythrocyte suspension |
| CN115428783A (en) * | 2022-09-16 | 2022-12-06 | 天津科技大学 | Erythrocyte membrane fragment freeze-drying protective solution, use method and application |
-
2006
- 2006-08-02 CN CN 200610104071 patent/CN1924583A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102466730A (en) * | 2010-11-19 | 2012-05-23 | 南京神州英诺华医疗科技有限公司 | Novel method for matching blood reverse typing reagent |
| CN105445455A (en) * | 2014-08-28 | 2016-03-30 | 山东省肿瘤医院 | Electrophoresis buffer solution for cell in-situ electrophoresis |
| CN105445455B (en) * | 2014-08-28 | 2017-08-08 | 山东省肿瘤医院 | A kind of electrophoretic buffer of cell in-situ electrophoresis |
| CN107037220A (en) * | 2017-04-01 | 2017-08-11 | 合肥迪安医学检验所有限公司 | The compound method of ABO reverse type red cell suspensions |
| CN109946129A (en) * | 2019-03-12 | 2019-06-28 | 江苏中济万泰生物医药有限公司 | Five classification cellanalyzer quality-control product preparation methods |
| 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 |
| CN112684192A (en) * | 2020-12-25 | 2021-04-20 | 天津市宝坻区人民医院 | New application of ammonium bradykinin in ABO blood group reverse typing kit |
| CN114732008A (en) * | 2022-06-13 | 2022-07-12 | 深圳瑞亚力集团有限公司 | Erythrocyte preservation solution, preparation method thereof and erythrocyte suspension |
| CN115428783A (en) * | 2022-09-16 | 2022-12-06 | 天津科技大学 | Erythrocyte membrane fragment freeze-drying protective solution, use method and application |
| CN115428783B (en) * | 2022-09-16 | 2023-08-04 | 天津科技大学 | Erythrocyte membrane fragment freeze-drying protection liquid, use method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101074962A (en) | ABO blood-type anti-sizing reagent | |
| CN1924583A (en) | ABO blood group inverse reagent | |
| CN1050905C (en) | Pretreating reagent, pretreatment method, assay with pretreated samples, assay kit and method of diagnosing infectious diseases | |
| CN1296171A (en) | Method and system for determining luminous energy effective value transmitted to fluid of luminous energy target | |
| CN1875115A (en) | Method of assyaing glycoprotein | |
| CN1861036A (en) | Preserving a hemoglobin blood substitute with a transparent overwrap | |
| CN1832756A (en) | Method of altering the binding specificity of proteins by oxidation-reduction reactions | |
| CN1681521A (en) | Method for purifying a biological composition | |
| CN1144047C (en) | Method for testing or determining HCV core antigen and the agent for same | |
| CN1166689C (en) | Method for purifying hemoglobin | |
| CN1195869C (en) | Monoclonal antibody, hybridoma, immunoassay method and diagnosis kit | |
| Taylor et al. | The enigmatic roles of Anelloviridae and Redondoviridae in humans | |
| CN1119414A (en) | Stabilised pharmaceutical compositions and methods for preparing same | |
| CN1766618A (en) | ELISA kit for detecting penicillin G and detection method thereof | |
| CN1839319A (en) | Composition containing aldolase for analyzing retinal vascular disease and analyzing method thereof | |
| CN112111456B (en) | Circulating tumor cell separation and enrichment kit | |
| CN1773282A (en) | Autoantibody assay method for immunological mediated I type diabetes diagnosis | |
| CN1967249A (en) | Four conjunction diagnostic kit of antimyocardial antibody | |
| CN1766626A (en) | ELISA kit for detecting estradiol and detection method thereof | |
| CN1266477C (en) | External blood group and blood serum experimental determining method | |
| CN1427258A (en) | Enzyme-linked immunosorbent assay (ELISA) reagent box for assaying low density lipoprotein content in human urine and its preparation method | |
| CN1260569C (en) | Reagent box for external blood group and blood serum experimental determination | |
| CN1427257A (en) | Enzyme-linked immunosorbent assay (ELISA) reagent box for assaying high density lipoprotein content in human urine and its preparation method | |
| CN1490050A (en) | Hepatitis A inactivated vaccine | |
| CN1891037A (en) | Use of sphingomonaspaucimobilis for anguilla culture |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |