CN107202898A - Kit for Shanghai irregular antibody - Google Patents
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Abstract
The present invention provides a kind of kit for Shanghai irregular antibody, belongs to blood group antibody detection technique field.The kit of the present invention contains 3 groups of red blood cells that totally 6 parts of independent specifications are packed, and 3 groups of red blood cells are respectively from 3 person-portions, D, C, c, E, e, s, Le in every person-portion red blood cellb、Fya, K, at least two positives of antigen, M, N, S, Lea、JKa、JKb、Fyb、P1At least one positive of antigen, and three person-portion antigenic component it is complementary, the red blood cell that every group of red blood cell is packed by 2 parts of independences and same size is constituted, wherein 1 part red blood cell passes through bromelain ferment treatment, other 1 part of red blood cell is without bromelain ferment treatment, the concentration of 6 parts of red blood cells is identical in kit, can be 1% 5%.Kit of the present invention is applied to the various blood types antibody detection method of prior art, reduces the loss of irregular antibody, reduces Clinical Blood Transfusion Risk.
Description
Technical field
The present invention relates to blood group antibody detection technique field, specifically, it is related to a kind of various blood types antibody that is applied to and examines
The kit for Shanghai irregular antibody of survey method.
Background technology
In order to prevent the generation of all kinds of transfusion reactions or neonatal hemolytic disease, related blood donor and sufferer are carried out irregular
The examination of antibody is very important.The Ministry of Public Health of China《Clinical blood transfusion technical specification》Article 17 is provided：It is all to meet following feelings
Condition must be by《National clinical examination operational procedure》Pertinent regulations make Antibody screening test：During Crossmatching incompatibility；To there is blood transfusion
History, pregnant history need to receive multiple blood donor in a short time.Further for some healthy blood donors, have different occurring blood typing
When view or suspection have irregular antibody, irregular antibody Screening tests can be also carried out.
So-called irregular antibody refers to the antibody beyond anti-A and anti-B, and its approach produced is probably pregnancy, is transfused blood, and is moved
Plant, or injected the material of immunogenicity.Irregular antibody can cause in hemolytic blood transfusion reaction, destruction input blood
Mismatch red blood cell or shorten the life-span of red blood cell, gently then influence therapeutic effect, heavy then threat to life.In addition, for pregnant woman,
Irregular antibody can cause neonatal hemolytic disease, the development of influence neonate internal organs and intelligence development, and then entail dangers to is newborn for severe patient
Youngster's life security.
Country's situation of all-level hospitals, the method for blood station detection blood group irregular antibody have salt water law, polybrene method, protease at present
Method, antihuman globulin method, micro-column gel agglutination assay etc., wherein most widely used is micro-column gel agglutination assay, it is also FDA recommendation method.With
The cardinal principle of upper methodology is that inspected blood plasma reacts with Irregular antibodies screening cell, with find by inspection blood plasma in be
It is no to there is irregular antibody.At present, conventional commercialization Irregular antibodies screening kit is main is matched suitably by 3 spectrotypes
O-shaped red blood cell combine, red blood cell expression D, C, c, E, e, Jka、Jkb、M、N、S、s、Fya、Fyb、Lea、Leb、K、k、P1
Deng antigen, the cell concentration commonly used in micro-column gel agglutination assay is 0.8%~1%, salt water law, polybrene method, protease method, anti-human ball
The conventional cell concentration of protein method is 3%~5%.
Because the raw material of Irregular antibodies screening kit is human red cell, it, which is originated, is limited, therefore different manufacturers are given birth to
The spectrotype generally existing defect of the Irregular antibodies screening red blood cell of production, such as Rh, Kidd, Lewis blood group system are most not
It is homozygote, but heterozygote, the antigen that homozygote gene code is produced is the doubling dosage of heterozygote gene, therefore uses these
The heterozygote red blood cell of blood group system goes to detect irregular antibody, may cause to be directed to these blood group systems due to dosage effect
Weak antibody missing inspection；And Kidd blood group systems, Lewis blood group systems and corresponding antibodies parent on Irregular antibodies screening red blood cell
Poor with power, the intensity of agglutination is weak, it is also possible to make the irregular antibody produced for Kidd blood group systems, Lewis blood group systems
Missing inspection.The antibody produced for blood group systems such as Rh, Kidd, Lewis accounting in all irregular antibodies is larger, and most tool
There is clinical meaning, if missing inspection, more serious anaphylactic type transfusion reaction or delayed transfusion reaction may be caused, jeopardize patient's life
Disease.Therefore, in the urgent need to a kind of detection method for the loss for reducing irregular antibody, to avoid missing inspection, clinical blood transfusion is reduced
Risk.
The content of the invention
Irregular antibody missing inspection can be effectively reduced it is an object of the invention to provide one kind, Clinical Blood Transfusion Risk is reduced, it is accurate
Exactness is high, the strong blood group irregular antibody kit for screening of stability.
In order to realize the object of the invention, the present invention provides a kind of kit for Shanghai irregular antibody, contained
Two red chromatography cell detection systems, a set of red blood cell detection architecture passes through Protease Treatment, another set of red blood cell detection architecture without
Cross ferment treatment.
In the kit of the present invention, the red blood cell of 2-5 person-portions is contained per red chromatography cell detection system, per in person-portion red blood cell
D、C、c、E、e、s、Leb、Fya, K, at least two positives of antigen, M, N, S, Lea、JKa、JKb、Fyb、P1Antigen at least one
It is positive.
Further, per in red chromatography cell detection system, Rh, Lewis, Kidd antigen systems select homozygote.
Contain the red blood cells of 3 person-portions dispensed alone in kit of the present invention per red chromatography cell detection system, and 3 person-portions
Antigenic component is tried one's best complementation.
In kit of the present invention, described protease is bromelain or papain；When selecting bromelain
When, its method for handling red blood cell is：By pH value 7.2-7.4, mass concentration is pressed for 0.1%-0.3% bromelain enzyme solutions
According to packed cell volume volume ratio 2-3:1 adds in red blood cell, and 35-38 DEG C of processing 10-20min, immediately washing removes enzyme solutions.
The bromelain enzyme solutions are the phosphate buffer solutions that bromelain is added to 0.1M pH value 7.2-7.4
In be made.
Preferably, the method for bromelain ferment treatment red blood cell is：By pH value 7.3, mass concentration is 0.2% pineapple egg
White enzyme solutions according to packed cell volume volume ratio 2:1 is added in red blood cell, 37 DEG C of processing 15-20min, and physiological saline is used immediately
Washing, to remove enzyme solutions, then is preserved with alserver's solution.The alserver's solution of the present invention is purchased from the biological section of mikey
Skill limited company.
The influence of the concentration of bromelain and action time, temperature to red blood cell is very big, by lot of experiments, final true
Surely the most suitable initial concentration for being used for the bromelain enzyme solutions of ferment treatment red blood cell is 0.1%~0.3%, and processing red blood cell is most
Suitable condition be 37 DEG C 15~20 minutes, the salt ionic concentration of bromelain enzyme solutions is 0.1mol/L, and pH is 7.3 ± 0.1, pineapple
Protein enzyme solution must be placed in -20 DEG C of preservations after packing, and the same day uses after redissolution, and can not freeze again.
In the kit of the present invention, the concentration of red cell suspension is identical in two red chromatography cell detection systems, is 1%-5%；
Various concentrations red cell suspension is prepared and calculated in the following ways：
The volume of volume=dose volume-raw cell of dilution.
For example, to prepare the red cell suspension 1000ml of 1% concentration, the HCT of raw cell is 50.00%, then needs original
The volume for expecting cell is the HCT50.00%=20ml of (1% × dose volume 1000ml of compound concentration)/raw cell.
The invention provides application of the mentioned reagent box in Shanghai irregular antibody.
The kit detection object of the present invention is blood plasma, during sentence read result, ferment treatment red blood cell and enzyme should be observed simultaneously not
Red thin reaction result is handled, comprehensive analysis is carried out.
Red blood cell is handled to strengthen antigen and the method for antibody response ability on red blood cell present invention also offers a kind of, is
By pH value 7.2-7.4, mass concentration for 0.1%-0.3% bromelain enzyme solutions according to packed cell volume volume ratio 2-3:
1 adds in red blood cell, 35-38 DEG C of processing 10-20min；The solvent of the bromelain enzyme solutions is 0.1M pH value 7.2-7.4
Phosphate buffer solution.The red blood cell handled through this method, its antigen can be significantly increased with antibody response ability in blood plasma,
See the record of the embodiment of the present application 4.
The beneficial effects of the present invention are：
(1) design adds ferment treatment red blood cell on the basis of original Irregular antibodies screening kit, enzyme treated
The red blood cell crossed can be remarkably reinforced Rh blood group systems, Kidd blood group systems, Lewis blood group systems, P blood group systems and phase in blood plasma
The response intensity of antibody is answered, the missing inspection of irregular antibody in blood plasma is reduced.In clinical sample detection, with ferment treatment red blood cell and
Red blood cell is not enzymatically treated while detecting same sample, the missing inspection of weak antibody in blood plasma can not only be reduced, and reduces clinical blood transfusion wind
Danger, moreover it is possible to the substantially distribution of irregular antibody in positive sample, auxiliary operator analysis experiment are tentatively inferred by reacting general layout
As a result.
(2) reaction system and the preservation body in ferment treatment red blood cell later stage of ferment treatment red blood cell are optimized by repetition test
System so that the long-time stability of ferment treatment red blood cell are preferably, basically identical with the red blood cell that is not enzymatically treated.
(3) compound method of traditional various concentrations red cell suspension is that the pressure of respective volume is drawn after sample is centrifuged
Product red blood cell be added in dilution be respective concentration red cell suspension, due between the packed red cells after centrifugation still
Have more liquid, therefore the cell concentration got according to such compound method is generally lower than theoretical concentration, and differences between batches compared with
Greatly.The determination of kit red blood cell concentration of the present invention is calculated using HCT (hematocrit value), using HCT calculating methods come really
Determining the concentration of red blood cell not only makes the concentration of red blood cell and theoretical concentration closer, moreover it is possible to reduces differences between batches, makes kit
Can be more stable.
Embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.Unless otherwise specified, embodiment
In the conventional meanses that are well known to those skilled in the art of used technological means, raw materials used is commercial goods.
The compound method of 0.1-0.3% bromelains enzyme solutions of the present invention is as follows：
1st, the preparation of pH7.3 phosphate buffers (PBS)：
The preparation of 1000mlA liquid
The preparation of 1000ml B liquid
The preparation of 100ml 0.1M pH7.3 phosphate buffer
2nd, the preparation of 1%-0.3% bromelains：1g bromelain dry powder is placed in 100ml pH7.3PBS,
Concussion dissolving 30 minutes at room temperature, then 2000g centrifuge 10 minutes, collect supernatant, face the used time is diluted to pH7.3PBS
0.1%-0.3% bromelain enzyme solutions.
Embodiment 1 selects the wide O-shaped red blood cell of 3 person-portion spectrotypes
Choose the suitable O-shaped red blood cell of 3 person-portion spectrotypes proportioning to combine, the O-shaped red blood cell per person-portion is bifurcated, one
Branch is ferment treatment red blood cell, another Zhi Weiwei is enzymatically treated red blood cell.Three person-portion red blood cells are divided into No. I, No. II, No. III group, three
Group red blood cell follows claimed below：
(1) red cell antigens spectrum should comprising common are clinical meaning blood group system antigen (Rh, MNSs, Lewis,
Kidd、Duffy、Kell、P).Red blood cell expresses D, C, c, E, e, Jka、Jkb、M、N、S、s、Fya、Fyb、Lea、Leb、K、k、P1Deng
Antigen.D, C, c, E, e, s, Le in per person-portion red blood cellb、Fya, K, at least two positives of antigen, M, N, S, Lea、JKa、JKb、
Fyb、P1At least one positive of antigen.
(2) antigen systems for having dosage effect select homozygote (Rh, Lewis, Kidd).
The epitope pattern of (3) 3 groups of red blood cells is tried one's best complementation, and so the irregular antibody for positive reaction has screening and arranged
The effect removed.
(4) because single red blood cell is higher compared with mixing red blood cell sensitiveness, should individually it be dispensed per person-portion red blood cell.
The red blood cell Direct antiglobulin test result of (5) No. I, No. II, No. III groups should be negative.
(6) No. I, No. II, No. III Irregular antibodies screening red blood cell and No. I, No. II, No. III ferment treatment irregular antibody sieve
Examine erythrocyte hemolysis rate all should≤5.0%.
Influence of the ferment treatment red blood cell system experimental condition of embodiment 2 for Antibody screening result of the test
In the present embodiment, quality control test：Using micro-column gel agglutination assay, in the low ion antihumanglobulin cards of BIORAD diamed
In be separately added into the μ l of 1% red cell suspension 50 handled under different condition, viewing test result after centrifugation.Purpose：Detection is not through
With the red blood cell of condition processing, if false positive can be caused.
Specific test：Using micro-column gel agglutination assay, it is separately added into the low ion antihumanglobulin cards of BIORAD diamed
The μ l of 1% red cell suspension 50 handled under different condition, same cell adds 12 holes；In 12 holes for adding same cell
Be separately added into again known specific commercialization blood group antibody (purchase from the anti-D of Shanghai blood, Anti-E, Anti-e,
Anti-C, Anti-c, Anti-M, Anti-N, purchase are from Millipore Anti-Jka, Anti-Jkb, Anti-P1, Anti-
Lea, Anti-Leb) 25 μ l, 37 DEG C are incubated 15 minutes, after centrifugation with reference to spectrotype viewing test result whether occur false negative,
False positive).Purpose：Detect the red blood cell handled through different condition, if false negative, false positive occurs.
Sensitivity test：Using micro-column gel agglutination assay, it is separately added into the low ion antihumanglobulin cards of BIORAD diamed
The μ l of 1% red cell suspension 50 handled through different condition, same cell adds 6 holes；Add 6 Kong Zhongzai of same cell
It is separately added into 1:10,1:100,1:1000,1:4000,1:8000,1:The Anti-D25 μ l of 16000 dilutions, 37 DEG C are incubated 15 points
Result is observed after clock, centrifugation.Purpose：Detect the red cell suspension handled through different condition, the situation of change of sensitivity.
1st, influence of the various concentrations pineapple enzyme solutions to red blood cell irregular antibody screening results
Set the pH value of various concentrations a series of for 7.3 bromelain enzyme solutions (0.05%, 0.1%, 0.3%,
0.5%th, 1%, 1.5%, 2%) handle red blood cell, by observe red blood cell whether haemolysis, with reference to red blood cell quality control test, with
And irregular antibody Screening tests specificity analyzes influence of the pineapple enzyme concentration to result of the test with sensitivity results.
The influence of the bromelain solution concentration of table 1
As a result show that, when the concentration of bromelain is 0.1%~0.3%, red blood cell is red thin without obvious haemolysis
Born of the same parents' quality control test non-false positive, specificity, the sensitivity test result of irregular antibody Screening tests antigen-antibody reaction is best.
When bromelain excessive concentration, erythrocyte membrane structure can be damaged, haemolysis is caused, and false positive can be caused.When pineapple enzyme concentration is too low,
The DeGrain of ferment treatment red blood cell, the susceptibility of irregular antibody Screening tests does not change.
2nd, influence of the different pH value bromelain enzyme solutions to red blood cell irregular antibody screening results
Set the initial concentration of different pH a series of for 0.2% bromelain enzyme solutions (5.5,6,6.5,7,7.2,
7.4th, 7.6,8,8.5) handle red blood cell, by observe red blood cell whether haemolysis, advised with reference to red blood cell quality control test, and not
Then Antibody screening experiment specificity analyzes shadow of the pH value to result of the test of bromelain enzyme solutions with sensitivity test result
Ring.
Influence of the different bromelain pH value of solution of table 2 to red blood cell irregular antibody screening results
As a result show that red blood cell is without haemolysis, red blood cell quality control test when the pH of bromelain is 7.2~7.4
Non-false positive, specificity, the sensitivity test result of irregular antibody Screening tests antigen-antibody reaction is preferable.PH is directly affected
The activity of bromelain, influences the effect of ferment treatment red blood cell, therefore in process of the test, should strictly control pH.
3rd, pineapple enzyme solutions processing red blood cell temperature and the influence of action time
The initial mass concentration of pineapple enzyme solutions is that 0.2%, pH is 7.3, sets series of temperature and time (temperature：25
℃、30℃、35℃、37℃、40℃；Time：5min, 10min, 15min, 20min, 25min, 30min) handle red blood cell,
By observe red blood cell whether haemolysis, it is specific and sensitive with reference to red blood cell quality control test, and irregular antibody Screening tests
Degree result of the test handles the influence of the temperature and action time of red blood cell to result of the test to analyze enzyme solutions.
The pineapple enzyme solutions of table 3 handle red blood cell temperature and the influence of action time
As a result display is 37 DEG C when the temperature of bromelain ferment treatment red blood cell, when processing time is 10~20min, red thin
Born of the same parents are without haemolysis, red blood cell quality control test non-false positive, the specificity of irregular antibody Screening tests antigen-antibody reaction, spirit
Sensitivity result of the test is preferable.Temperature influences the activity of bromelain, the time effects ferment treatment red blood cell of ferment treatment red blood cell
Effect, when enzyme processing time is too short, in irregular antibody Screening tests, DeGrain.It is long between when treated, it can make
Red blood cell is enzymatically treated excessively, and membrane structure is damaged, and nonspecific aggegation is occurred, is caused false positive.
4th, influence of the salt ionic concentration to red blood cell in pineapple enzyme solutions.
Set a series of different phosphate ion concentrations bromelain enzyme solutions (0.05M, 0.1M, 0.2M, 0.3M,
0.4M, 0.5M, 0.6M) handle red blood cell, by observe red blood cell whether haemolysis, with reference to red blood cell quality control test, and not
Regular Antibody screening experiment specificity analyzes bromelain protein enzyme solution salt ionic concentration to experiment with sensitivity test result
As a result influence.
The influence of the bromelain solution salt ionic concentration of table 4
As a result show that red blood cell is red without haemolysis when the phosphate ion concentration of bromelain enzyme solutions is 0.1M
Cell quality control test non-false positive, the specificity of irregular antibody Screening tests antigen-antibody reaction, sensitivity test result compared with
It is good.Salt ionic concentration is related to the osmotic pressure of solution, and when salt ionic concentration is too low, osmotic pressure is relatively low, can expand red blood cell, hair
Raw haemolysis, has a strong impact on the result of the test of bromelain ferment treatment red blood cell.When salt ionic concentration is too high, red blood cell can be made to wrinkle
Contracting, the blood group antigens configuration of Surface of Erythrocytes changes, and influences the effect of ferment treatment red blood cell.
It can be seen that, the experimental condition of ferment treatment red blood cell is very big for the influential effect of ferment treatment red blood cell, only correctly
With each condition, the accuracy of ability guarantee test result plays the advantage of ferment treatment red blood cell.Verified by great amount of samples,
It is final to determine that (solvent is 0.1M pH value 7.2-7.4 phosphoric acid to bromelain enzyme solutions in bromelain ferment treatment red cell body system
Salt buffer solution) optimum concentration be 0.1%~0.3%, the optimum condition of processing red blood cell is 37 DEG C of 15~20 minutes, pineapple
The salt ionic concentration of protein enzyme solution is 0.1mol/L, and pH is 7.3 ± 0.1.
The determination method of red cell suspension concentration in the kit red blood cell detection architecture of the present invention of embodiment 3
20 whole blood samples are selected at random, its HCT (hematocrit value) are measured after fully mixing, according to HCT (red blood cell ratios
Hold) compound concentration be 1% red cell suspension, solvent is physiological saline.Again red cell suspension is determined with o- first benzidine method
Content of hemoglobin, the amount finally according to hemoglobin counter pushes away the real concentration of cell suspension.Result of the test such as table 5：
Table 5 be conventional method and recommendation method of the present invention preparation 1% red cell suspension deviation ratio compared with
It can be seen from upper table experimental data, the red cell suspension actual concentration for 1% concentration prepared according to conventional method is general
All over relatively low, deviation is larger；And more connect with theoretical concentration according to the actual concentration of the red cell suspension of HCT 1% concentration prepared
Closely, deviation is within ± 10%.When distinct methods detect irregular antibody, used Irregular antibodies screening red blood cell
Concentration is inconsistent, therefore should be according to selected methodology come the irregular antibody of the invention from Different Red concentration of cell suspension
Screening kit.The red cell suspension concentration of kit of the present invention is 1%-5%, is the concentration determined according to HCT methods, with
Theoretical concentration is closer, and deviation is small, and detection repeatability and stability are excellent.
Detected in the kit of the present invention of embodiment 4 through bromelain ferment treatment and without the red blood cell of bromelain ferment treatment
The result of positive sample
3 wide person-portion O of the spectrotype of the bromelain optimal treatment condition Processing Example 1 determined according to embodiment 2
Type red blood cell, and the concentration of red cell suspension is determined as 1% in the method for embodiment 3, it is anti-according to the low ions of BIORAD diamed
Human immunoglobulin card specification requires to carry out the detection that detection carries out blood group irregular antibody in following positive sample.
The kit of the present invention of table 6 detects the result of positive sample
Tested simultaneously with ferment treatment red blood cell and the red blood cell not being enzymatically treated, pineapple egg can be observed according to upper table
It is anti-with corresponding antibody that white enzyme ferment treatment red blood cell can be remarkably reinforced Rh blood group systems, Kidd blood group systems, Lewis blood group systems
Should, and when antibody level is relatively low, ferment treatment red blood cell can reduce loss.But ferment treatment red blood cell can also destroy MNSs blood
The antigen of type system, Duffy blood group systems, therefore ferment treatment red blood cell should support the use with the red blood cell that is not enzymatically treated,
The accuracy of testing result can preferably be ensured.Therefore, kit of the present invention uses two red chromatography cell detection systems simultaneously, a set of
The red blood cell of red blood cell detection architecture passes through Protease Treatment, the red blood cell of another set of red blood cell detection architecture without enzyme at
Reason.
In sentence read result, while observing ferment treatment red blood cell and not being enzymatically treated the reaction general layout of red blood cell to analyze knot
Really.If ferment treatment red blood cell result is feminine gender, the red blood cell not being enzymatically treated is the positive, then is probably MNSs systems, Duffy systems
The antibody of system, if ferment treatment red blood cell result is the positive, the red blood cell not being enzymatically treated is feminine gender, then is probably Rh blood groups system
System, Kidd blood group systems, the antibody of Lewis blood group systems.The blood group system belonging to irregular antibody is first substantially locked, further according to
Which irregular antibody is the reaction general layout of positive reaction cell be to speculate.Example：3 cell effect situations of positive findings are cloudy
Property, feminine gender, the positive；It is negative, negative, positive antigen then to correspond to spectrotype and go for 3 cell distribution situations.So in blood plasma
What is existed is probably then the corresponding irregular antibody of the antigen.
The contrast experiment of embodiment 5
1st, the comparison of papain and bromelain in terms of red blood cell effect is handled.
3 wide person-portion O of the spectrotype of the bromelain optimal treatment condition Processing Example 1 determined according to embodiment 2
Type red blood cell, and the concentration of red cell suspension is determined as 1% in the method for embodiment 3, the present invention obtained based on above-mentioned processing
Kit, containing two red chromatography cell detection systems, the red blood cell of a set of red blood cell detection architecture passes through Protease Treatment, another set of
The red blood cell of red blood cell detection architecture is without ferment treatment.Contain the red blood cell of 2-5 person-portions per red chromatography cell detection system, everyone
D, C, c, E, e, s, Le in part red blood cellb、Fya, K, at least two positives of antigen, M, N, S, Lea、JKa、JKb、Fyb、P1Antigen
At least one is positive.Kit outward appearance of the present invention should be clean and tidy, and word, symbol and mark should be clear；No. I in kit, No. II,
No. III Irregular antibodies screening red blood cell and No. I, No. II, No. III ferment treatment Irregular antibodies screening red blood cell should be fresh after mixing
The red red cell suspension without grumeleuse.After stratification, supernatant is clear, without clearly visible haemolysis and foreign matter.Ferment treatment and nothing
The Irregular antibodies screening red blood cell Direct antiglobulin test result of ferment treatment should be negative, and Irregular antibodies screening is red
Cell hemolysis rate all should≤5.0%, Irregular antibodies screening red blood cell concentration sign concentration ± 10% within.
Conventional enzyme has papain, bromelain, ficin, trypsase etc. at present, and the country is used
More is papain and bromelain.Papain and bromelain ferment treatment red blood cell effect are without significance difference
It is different, but ferment treatment red blood cell long-time stability have significant difference, the red blood cell later stage of Papain ferment treatment is red thin when preserving
The easy haemolysis of born of the same parents.Hemolysis rate in terms of detecting two kinds of ferment treatment red blood cell long-time stability using o- first benzidine method.
The comparison of the papain of table 7 and bromelain ferment treatment red blood cell long-time stability hemolysis rate
As shown in Table 7, the red blood cell long-time stability preserved after Papain ferment treatment are bad, and hemolysis rate is apparently higher than spinach
The red blood cell of trailing plants Protease Treatment.Therefore, the application handles red blood cell using bromelain.
2 kits of the present invention are compared with the testing result of commercial kit in the prior art, the results are shown in Table 8.
The kit of the present invention of table 8 and Shanghai blood Irregular antibodies screening kit (control group) contrast test
By upper table, it can be observed significantly reduce the missing inspection of low-level irregular antibody with this kit, two kinds of examinations
The difference of agent box detection is shown in the result for having square frame.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. a kind of kit for Shanghai irregular antibody, it is characterised in that contain two red chromatography cell detection systems, one
The red blood cell of red chromatography cell detection system passes through Protease Treatment, the red blood cell of another set of red blood cell detection architecture without enzyme at
Reason.
2. kit as claimed in claim 1, it is characterised in that contain the red thin of 2-5 person-portions per red chromatography cell detection system
D, C, c, E, e, s, Le in born of the same parents, every person-portion red blood cellb、Fya, K, at least two positives of antigen, M, N, S, Lea、JKa、JKb、
Fyb、P1At least one positive of antigen.
3. kit as claimed in claim 2, it is characterised in that per in red chromatography cell detection system, RH, Lewis, Kidd resist
Original system selects homozygote.
4. kit as claimed in claim 2, it is characterised in that contain 3 people dispensed alone per red chromatography cell detection system
Part red blood cell, and the antigenic component of 3 person-portions tries one's best complementation.
5. kit as claimed in claim 1, it is characterised in that described protease is bromelain or Papain
Enzyme；When from bromelain, its method for handling red blood cell is：By pH value 7.2-7.4, mass concentration is 0.1%-
0.3% bromelain enzyme solutions according to packed cell volume volume ratio 2-3:1 adds in red blood cell, 35-38 DEG C of processing 10-
20min, immediately washing removes enzyme solutions.
6. kit as claimed in claim 5, it is characterised in that the bromelain enzyme solutions are to add bromelain
It is made in 0.1M pH value 7.2-7.4 phosphate buffer solution.
7. the kit as described in claim 1-6 is any, it is characterised in that its method for handling red blood cell is：By pH value
7.3, mass concentration for 0.2% bromelain enzyme solutions according to packed cell volume volume ratio 2:1 adds in red blood cell, 37 DEG C
15-20min is handled, washing immediately removes enzyme solutions.
8. the kit as described in claim 1-6 is any, it is characterised in that red cell suspension in two red chromatography cell detection systems
Concentration it is identical, be 1%-5%；Various concentrations red cell suspension is prepared and calculated in the following ways：
The volume of volume=dose volume-raw cell of dilution.
9. application of any described kits of claim 1-8 in Shanghai irregular antibody.
10. application as claimed in claim 9, it is characterised in that detection object is blood plasma, during sentence read result, should be observed simultaneously
Ferment treatment red blood cell and the untreated red thin reaction result of enzyme, carry out comprehensive analysis.
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| CN201710451473.7A CN107202898A (en) | 2017-06-15 | 2017-06-15 | Kit for Shanghai irregular antibody |
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| CN201710451473.7A CN107202898A (en) | 2017-06-15 | 2017-06-15 | Kit for Shanghai irregular antibody |
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Cited By (5)
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| CN107764985A (en) * | 2017-10-10 | 2018-03-06 | 中国人民解放军第四军医大学 | A kind of serum antibody identifies quick reader and its application method |
| CN108575987A (en) * | 2018-04-26 | 2018-09-28 | 天津德祥生物技术有限公司 | Red blood cell processing method and its application |
| CN109917136A (en) * | 2019-03-13 | 2019-06-21 | 江苏力博医药生物技术股份有限公司 | The preparation method and application of cradin processing antibody screening red blood cell |
| CN110672862A (en) * | 2019-09-29 | 2020-01-10 | 迈克生物股份有限公司 | Blood type detection card and preparation method thereof |
| CN110736832A (en) * | 2019-12-18 | 2020-01-31 | 首都医科大学附属北京友谊医院 | Composition for identifying spectrum cells by antibodies |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107764985A (en) * | 2017-10-10 | 2018-03-06 | 中国人民解放军第四军医大学 | A kind of serum antibody identifies quick reader and its application method |
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| CN108575987A (en) * | 2018-04-26 | 2018-09-28 | 天津德祥生物技术有限公司 | Red blood cell processing method and its application |
| CN109917136A (en) * | 2019-03-13 | 2019-06-21 | 江苏力博医药生物技术股份有限公司 | The preparation method and application of cradin processing antibody screening red blood cell |
| CN110672862A (en) * | 2019-09-29 | 2020-01-10 | 迈克生物股份有限公司 | Blood type detection card and preparation method thereof |
| CN110736832A (en) * | 2019-12-18 | 2020-01-31 | 首都医科大学附属北京友谊医院 | Composition for identifying spectrum cells by antibodies |
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