CN107202899A - A kind of authentication method of good chief of a tribe monkey blood group - Google Patents
A kind of authentication method of good chief of a tribe monkey blood group Download PDFInfo
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- CN107202899A CN107202899A CN201710557796.4A CN201710557796A CN107202899A CN 107202899 A CN107202899 A CN 107202899A CN 201710557796 A CN201710557796 A CN 201710557796A CN 107202899 A CN107202899 A CN 107202899A
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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Abstract
The invention discloses a kind of authentication method of good chief of a tribe monkey blood group, comprise the following steps：(1) collection blood is in the container containing porous cotton, centrifugation；(2) supernatant liquid is drawn, is stored at room temperature, is then stayed overnight for 4 DEG C, then is stored at room temperature, serum is centrifuged to obtain；(3) the O-shaped erythrocyte of people is prepared；(4) serum is mixed with the O-shaped erythrocyte of people, non-specific antibody between Adsorption people's monkey kind；(5) serum between removal people's monkey kind after non-specificity is mixed with protection liquid；(5) step (5) gains are mixed with the A types erythrocyte and Type B erythrocyte of normal person respectively, the blood group of good chief of a tribe monkey is judged according to mixed agglutination phenomenon.This method is simple to operate, and cost is low, damages small to good chief of a tribe monkey, qualification result accuracy rate is high, reproducible.
Description
Technical field
The invention belongs to biomedicine technical field, and in particular to a kind of authentication method of good chief of a tribe monkey blood group.
Background technology
Blood group is one of principal character of human blood, and abo blood group antigen is widely present in human erythrocyte and tissue,
It is topmost erythrocyte blood type system.Good chief of a tribe monkey also has ABO blood group system, but different with the mankind, and the abo blood group of good chief of a tribe monkey resists
Original is all expressed in histoorgan, does not express AB antigens in its peripheral red blood cells, or express very weak.These blood group antibodies
It is same with the AB antigen bindings expressed on transplant organ to cause to be similar to allogeneic organ's shifting that Human Blood Type ABO does not conform to
Plant rejection.
Because the abo blood group antigen of good chief of a tribe monkey is not expressed in peripheral red blood cells, and the fibrin in blood plasma and
Non-specific antibody can all produce interference to partial detection between people-monkey kind, easily produce false positive or false negative result,
So the bracket for blood grouping of good chief of a tribe monkey is all more difficult always.
At present, the bracket for blood grouping method of good chief of a tribe monkey has：(1) traditional test tube method antiglobulin test and pilocarpinum are passed through
The AB Antigen Identifications of saliva after medication, but both approaches be difficult standardization, it is believed that factor is more, easily occur false positive or
False negative, causing blood group to judge can not clearly or misjudgment.(2) histotomy immunohistochemistry staining method, the method
Damage to animal is larger, is normally only used for looking back confirmatory study, it is impossible to meet the requirement that breeding population carries out bracket for blood grouping.
(3) cassette gel method, this method cost is high, and centrifugation needs special centrifugal machine, and special centrifugal machine cannot be used for other test tubes from
The heart, with certain limitation.
The content of the invention
, can be effective the invention provides a kind of authentication method of good chief of a tribe monkey blood group for above-mentioned deficiency of the prior art
Solve cost in the prior art high, with limitation, identify inaccurate, the problem of false positive or false negative easily occur.
To achieve the above object, the technical solution adopted for the present invention to solve the technical problems is：
A kind of authentication method of good chief of a tribe monkey blood group, comprises the following steps：
(1) gather good chief of a tribe's monkey hind leg superficial vein blood to be placed in the container containing porous cotton, shaken with 120-130r/min rotating speed
Swing 10-15min；
(2) blood after aspiration step (1) processing is placed in reddish tone heparin tube, is stored at room temperature 40-60min, is subsequently placed in 4
DEG C overnight, then room temperature place 30-40min, in 3500-6000r/min centrifuge 10-15min, Aspirate supernatant；
(3) the fresh O-shaped blood of normal person is drawn in centrifuge tube, 9-10 times of volume is added, and concentration is 0.9% physiology salt
Water, is slightly shaken up, and 15-20min is centrifuged in 1500-2000r/min, and then abandoning supernatant is cleaned with 0.9% physiological saline
Sediment 3-4 times；
(4) it is 1 by volume with step (3) gains by supernatant obtained by step (2):1 mixing, and in 37 DEG C of incubations
30-40min, centrifuges 8-10min then at 1300-1500r/min, takes supernatant；
(5) supernatant obtained by step (4) and protection liquid are 3-4 by volume:1-2 is mixed；Wherein, protection liquid includes 5%
Bovine serum albumin, 5% sucrose, 5%PVP and the phosphate buffer containing 5% calf serum, phosphate buffering liquid concentration is 5mM,
PH value is 7.4；
(6) it is respectively 1 by volume with the A types erythrocyte and Type B erythrocyte of normal person by step (5) gains:1
Mixing, slightly shakes up and centrifuges 1min after 1000r/min, observes agglutination phenomenon.
If there is agglutination phenomenon after only being mixed with A type erythrocytes, Type B blood is accredited as；If only mixed with Type B erythrocyte
Occur agglutination phenomenon after conjunction, be then accredited as A type blood；If there is agglutination phenomenon under two kinds of mixing conditions, O-shaped blood is accredited as；
If occurring without agglutination phenomenon under two kinds of mixing conditions, AB type blood is accredited as.
Further, centrifugal rotational speed is 120r/min in step (1), and centrifugation time is 15min.
Further, 60min is stored at room temperature in step (2), is subsequently placed in 4 DEG C overnight, then room temperature places 30min, in
3800r/min centrifuges 10min.
Further, 9 times of volumes are added in step (3) into the O-shaped erythrocyte of normal person, concentration is 0.9% physiology
Salt solution, is slightly shaken up, and 15min is centrifuged in 1500r/min.
Further, incubation time is 30min in step (4), and centrifugal speed is 1372r/min, and centrifugation time is
10min。
Further, supernatant obtained by step (4) and protection liquid are 3 by volume:1 mixing.
The authentication method for the good chief of a tribe monkey blood group that the present invention is provided, has the advantages that：
(1) blood of collection is placed in the container containing porous cotton, can be constantly shaken in shaking table, the fiber egg in blood
When white original is transformed into insoluble fibrin, the insoluble fibrin of generation is touched and can adsorbed after porous cotton by porous cotton,
Fibrinogen can effectively be removed, it is to avoid its interference to test result.
(2) blood after porous cotton is handled is stood at room temperature, then stood overnight overnight then at 4 DEG C, finally
Place and be stored at room temperature again, then centrifuge, can effectively remove the impurity in serum, obtain limpid serum.
(3) limpid serum is mixed with O-shaped red blood cell, non-specific antibody between people-monkey kind can be eliminated after adsorption treatment, together
When further remove fibrinogen.
(4), can by non-specific antibody between removal people-monkey kind and the supernatant after fibrinogen through protecting liquid protection
Strengthen the vigor of antibody, then mixed with A types erythrocyte or Type B erythrocyte, can accurately and effectively identify blood group type, no
The problem of false positive or false negative occurs, in addition, this method cost is low, do not have limitation to centrifugation.
Brief description of the drawings
Fig. 1 is single erythrogram under microscope.
Fig. 2 is hemagglutination figure under microscope.
Embodiment
Embodiment 1
A kind of authentication method of good chief of a tribe monkey blood group, comprises the following steps：
(1) 20 good chief of a tribe monkeys are chosen, good chief of a tribe's monkey hind leg superficial vein blood is gathered respectively and is placed in the container containing porous cotton, with
120r/min rotating speed concussion 15min；
(2) the blood 3mL after aspiration step (1) processing is placed in reddish tone heparin tube, is stored at room temperature 60min, is subsequently placed in 4
DEG C overnight, then room temperature place 30min, in 3800r/min centrifuge 10min, Aspirate supernatant；
(3) the fresh O-shaped blood of normal person is drawn in centrifuge tube, 9 times of volumes are added, and concentration is 0.9% physiological saline,
Slightly shake up, 15min centrifuged in 1500r/min, then abandoning supernatant cleans sediment 3 times with 0.9% physiological saline,
Single red blood cell can be observed under the microscope, as shown in Figure 1；
(4) it is 1 by volume with step (3) gains by supernatant obtained by step (2):1 mixing, and in 37 DEG C of incubations
30min, centrifuges 10min then at 1372r/min, takes supernatant；
(5) supernatant obtained by step (4) and protection liquid are 3 by volume:1 mixing；Wherein, protection liquid includes 5% ox blood
Albumin, 5% sucrose, 5%PVP and the phosphate buffer containing 5% calf serum, phosphate buffering liquid concentration are 5mM, pH value
For 7.4；
(6) it is respectively 1 by volume with the A types erythrocyte and Type B erythrocyte of normal person by step (5) gains:1
Mixing, slightly shakes up and centrifuges 1min after 1000r/min, observes agglutination phenomenon, can be red thin in microscopical low power Microscopic observation
Born of the same parents' agglutination phenomenon, if occurring blocking, the grumeleuse of graininess or chilli oil shape under mirror, then shows that aggegation occurs in red blood cell, if under mirror
Red cell distribution is uniform, then shows do not occur aggegation, agglutination phenomenon is as shown in Figure 2.
If there is agglutination phenomenon after only being mixed with A type erythrocytes, Type B blood is accredited as；If only mixed with Type B erythrocyte
Occur agglutination phenomenon after conjunction, be then accredited as A type blood；If there is agglutination phenomenon under two kinds of mixing conditions, O-shaped blood is accredited as；
If occurring without agglutination phenomenon under two kinds of mixing conditions, AB type blood is accredited as.
The blood group that good chief of a tribe monkey is final is judged according to phenomenon is condensed, from result, it is A types there are 8 in 20 good chief of a tribe monkeys
Blood, is Type B blood by 6, and 5 are O-shaped blood, and 1 is AB type blood.
Embodiment 2
A kind of authentication method of good chief of a tribe monkey blood group, comprises the following steps：
(1) 20 good chief of a tribe monkeys are chosen, good chief of a tribe's monkey hind leg superficial vein blood is gathered respectively and is placed in the container containing porous cotton, with
120r/min rotating speed concussion 10min；
(2) the blood 3mL after aspiration step (1) processing is placed in reddish tone heparin tube, is stored at room temperature 40min, is subsequently placed in 4
DEG C overnight, then room temperature place 40min, in 3500r/min centrifuge 15min, Aspirate supernatant；
(3) the fresh O-shaped blood of normal person is drawn in centrifuge tube, 10 times of volumes are added, and concentration is 0.9% physiology salt
Water, is slightly shaken up, and 15min is centrifuged in 1500r/min, and then abandoning supernatant cleans sediment 3 with 0.9% physiological saline
It is secondary；
(4) it is 1 by volume with step (3) gains by supernatant obtained by step (2):1 mixing, and in 37 DEG C of incubations
40min, centrifuges 8min then at 1500r/min, takes supernatant；
(5) supernatant obtained by step (4) and protection liquid are 4 by volume:1 mixing；Wherein, protection liquid includes 5% ox blood
Albumin, 5% sucrose, 5%PVP and the phosphate buffer containing 5% calf serum, phosphate buffering liquid concentration are 5mM, pH value
For 7.4；
(6) it is respectively 1 by volume with the A types erythrocyte and Type B erythrocyte of normal person by step (5) gains:1
Mixing, slightly shakes up and centrifuges 1min after 1000r/min, observes agglutination phenomenon.
If there is agglutination phenomenon after only being mixed with A type erythrocytes, Type B blood is accredited as；If only mixed with Type B erythrocyte
Occur agglutination phenomenon after conjunction, be then accredited as A type blood；If there is agglutination phenomenon under two kinds of mixing conditions, O-shaped blood is accredited as；
If occurring without agglutination phenomenon under two kinds of mixing conditions, AB type blood is accredited as.
The blood group that good chief of a tribe monkey is final is judged according to phenomenon is condensed, from result, it is A types there are 8 in 20 good chief of a tribe monkeys
Blood, is Type B blood by 6, and 5 are O-shaped blood, and 1 is AB type blood.
Embodiment 3
A kind of authentication method of good chief of a tribe monkey blood group, comprises the following steps：
(1) 20 good chief of a tribe monkeys are chosen, good chief of a tribe's monkey hind leg superficial vein blood is gathered respectively and is placed in the container containing porous cotton, with
120r/min rotating speed concussion 15min；
(2) the blood 3mL after aspiration step (1) processing is placed in reddish tone heparin tube, is stored at room temperature 60min, is subsequently placed in 4
DEG C overnight, then room temperature place 30min, in 6000r/min centrifuge 10min, Aspirate supernatant；
(3) the fresh O-shaped blood of normal person is drawn in centrifuge tube, 10 times of volumes are added, and concentration is 0.9% physiology salt
Water, is slightly shaken up, and 15min is centrifuged in 2000r/min, and then abandoning supernatant cleans sediment 3 with 0.9% physiological saline
It is secondary；
(4) it is 1 by volume with step (3) gains by supernatant obtained by step (2):1 mixing, and in 37 DEG C of incubations
40min, centrifuges 10min then at 1400r/min, takes supernatant；
(5) supernatant obtained by step (4) and protection liquid are 2 by volume:1 mixing；Wherein, protection liquid includes 5% ox blood
Albumin, 5% sucrose, 5%PVP and the phosphate buffer containing 5% calf serum, phosphate buffering liquid concentration are 5mM, pH value
For 7.4；
(6) it is respectively 1 by volume with the A types erythrocyte and Type B erythrocyte of normal person by step (5) gains:1
Mixing, slightly shakes up and centrifuges 1min after 1000r/min, observes agglutination phenomenon.
If there is agglutination phenomenon after only being mixed with A type erythrocytes, Type B blood is accredited as；If only mixed with Type B erythrocyte
Occur agglutination phenomenon after conjunction, be then accredited as A type blood；If there is agglutination phenomenon under two kinds of mixing conditions, O-shaped blood is accredited as；
If occurring without agglutination phenomenon under two kinds of mixing conditions, AB type blood is accredited as.
The blood group that good chief of a tribe monkey is final is judged according to phenomenon is condensed, from result, it is A types there are 8 in 20 good chief of a tribe monkeys
Blood, is Type B blood by 6, and 5 are O-shaped blood, and 1 is AB type blood.
Respectively in the blood of other different good chief of a tribe monkeys of above-mentioned 20 of 5 periods collection, according to embodiment 1-3 identification
Method carries out bracket for blood grouping, and its result is consistent, illustrates the authentication method that the present invention is provided, reproducible.
Claims (6)
1. a kind of authentication method of good chief of a tribe monkey blood group, it is characterised in that comprise the following steps：
(1) gather good chief of a tribe's monkey hind leg vein blood to be placed in the container containing porous cotton, 10- is shaken with 120-130r/min rotating speed
15min；
(2) blood after aspiration step (1) processing is placed in reddish tone heparin tube, is stored at room temperature 40-60min, is subsequently placed in 4 DEG C of mistakes
Night, then room temperature place 30-40min, and 10-15min, Aspirate supernatant are centrifuged in 3500-6000r/min；
(3) the fresh O-shaped blood of normal person is drawn in centrifuge tube, 9-10 times of volume is added, and concentration is 0.9% physiological saline,
Slightly shake up, 15-20min is centrifuged in 1500-2000r/min, then abandoning supernatant cleans heavy with 0.9% physiological saline
Starch 3-4 times；
(4) it is 1 by volume with step (3) gains by supernatant obtained by step (2):1 mixing, and it is incubated 30- in 37 DEG C
40min, centrifuges 8-10min then at 1300-1500r/min, takes supernatant；
(5) supernatant obtained by step (4) and protection liquid are 3-4 by volume:1-2 is mixed；Wherein, protection liquid includes 5% ox blood
Albumin, 5% sucrose, 5%PVP and the phosphate buffer containing 5% calf serum, phosphate buffering liquid concentration are 5mM, pH value
For 7.4；
(6) it is respectively 1 by volume with the A types erythrocyte and Type B erythrocyte of normal person by step (5) gains:1 mixes
Close, slightly shake up and centrifuge 1min after 1000r/min, observe agglutination phenomenon.
2. the authentication method of good chief of a tribe monkey blood group according to claim 1, it is characterised in that centrifugal rotational speed is in step (1)
120r/min, centrifugation time is 15min.
3. the authentication method of good chief of a tribe monkey blood group according to claim 1, it is characterised in that step is stored at room temperature in (2)
60min, is subsequently placed in 4 DEG C overnight, then room temperature places 30min, and 10min is centrifuged in 3800r/min.
4. the authentication method of good chief of a tribe monkey blood group according to claim 1, it is characterised in that O-shaped to normal person in step (3)
9 times of volumes are added in erythrocyte, concentration is 0.9% physiological saline, is slightly shaken up, and 15min is centrifuged in 1500r/min.
5. the authentication method of good chief of a tribe monkey blood group according to claim 1, it is characterised in that incubation time is in step (4)
30min, centrifugal speed is 1372r/min, and centrifugation time is 10min.
6. the authentication method of good chief of a tribe monkey blood group according to claim 1, it is characterised in that supernatant obtained by step (4) with
It is 3 by volume to protect liquid:1 mixing.
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| CN201710557796.4A CN107202899A (en) | 2017-07-10 | 2017-07-10 | A kind of authentication method of good chief of a tribe monkey blood group |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104698193A (en) * | 2015-02-14 | 2015-06-10 | 四川普莱美行之生物科技有限公司 | Method for identifying blood group of rhesus monkey |
| 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 |
| CN106726557A (en) * | 2017-01-22 | 2017-05-31 | 杭州聚立医疗用品有限公司 | A kind of blood taking bag and the method that fibrinogen is removed with the blood taking bag |
-
2017
- 2017-07-10 CN CN201710557796.4A patent/CN107202899A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN104698193A (en) * | 2015-02-14 | 2015-06-10 | 四川普莱美行之生物科技有限公司 | Method for identifying blood group of rhesus monkey |
| CN106726557A (en) * | 2017-01-22 | 2017-05-31 | 杭州聚立医疗用品有限公司 | A kind of blood taking bag and the method that fibrinogen is removed with the blood taking bag |
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Application publication date: 20170926 |