CN108624663A - A kind of human erythrocyte's abo blood group antigen multiplex PCR classifying method and kit - Google Patents
A kind of human erythrocyte's abo blood group antigen multiplex PCR classifying method and kit Download PDFInfo
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Abstract
The present invention relates to a kind of human erythrocyte's abo blood group antigen multiplex PCR classifying method and kits, using multiplex PCR, design 4 pairs of specific primers, O, non-O, B and non-B are expanded respectively, using beta actin as internal reference, 2 multi-PRC reaction systems are formed, according to the presence or absence of amplified band, you can the abo blood group of preliminary judgement sample.Multiple PCR method is introduced abo blood group Genotyping by the present invention；Using the SNP site of O allele and B allele, abo blood group Genotyping is realized.The method of the present invention can reduce disposable's PCR reaction times of ABO antigen gene partings, reduce the manpower and reagent cost of detection, improve detection flux, the time required to shortening large sample size detection.
Description
Technical field
The present invention relates to molecular biology and clinical diagnosis technology field, specifically, being a kind of human erythrocyte ABO blood
Type antigen multiplex PCR classifying method and kit.
Background technology
Mankind's ABO genes are located at No. 9 chromosomes, by A, B and O tri- multiple alleles control, including length scale is from 28
~688bp 6 intrones that equal 7 exons and length are not about 19514bp, overall length is about 18~20kb.Its gene
Coded product is glycosyl transferase, and most coded sequences are located on the 6th and the 7th exon, is responsible for urging for encoding glycosyltransferases
Change region.These transferases control the biosynthesis of abo blood group antigen, to determine its blood group.ABO blood group system is mankind's blood
The strongest blood group system of immunogenicity in type system has weight in the clinical medicine such as medicine and organ, stem cell transplantation of transfusing blood
It acts on.In the prior art, there are mainly two types of Human Blood Type ABO detection methods：When classical serological method, second is that gene
Classifying method.Serological method has the characteristics that simple and practical, but there is also certain limitations, it is necessary first to fresh red blood cell
Sample, it is more demanding to sample；Secondly the range of method detection is relatively narrow, can only be carried out shaping to the common blood group in abo blood group,
For some hypotype samples and rare sample, easily there is ABO typing discrepan- cy in Blood grouping, it is difficult to which sizing or sizing are wrong
Accidentally the phenomenon that, and not can determine that its specific genotype.The method of Genotyping compensates for classical serology side to a certain extent
The deficiency of method in this respect.
Chinese patent 201410271048.6 discloses a kind of for detecting drawing for human erythrocyte's abo blood group Genotyping
Object group and kit, the primer sets for abo blood group genetic testing include ABOINTRON2396G primer pairs,
ABOINTRON2396C primer pairs, ABOEXON7272A primer pairs, ABOEXON7272T primer pairs, ABOEXON7429C primers
It is right, ABOEXON7429G primer pairs, ABOEXON7635G primer pairs, ABOEXON7635A primer pairs, ABOEXON622A primers
It is right, ABOEXON7688G primer pairs, ABOEXON7688C primer pairs and internal control primer pair.Chinese patent 201010174588.4 is public
A kind of PCR-SBT methods for abo blood group Genotyping have been opened, have been included the following steps：Prepare human gene group DNA；Expand ABO
The segment of gene extron 1, exon 2-4, exon 5-7；It obtains amplified production and carries out double digestion purifying；By purified product into
Row sequencing PCR reactions；Sequencing product is subjected to sodium acetate-ethanol precipitation purifying, carries out Capillary Electrophoresis order-checking；By acquisition
Sequence is analyzed by software, determines its genotype.Gene point of the above-mentioned technical proposal for human erythrocyte's blood group ABO antigen
Type, single person-portion need more PCR reaction times (>6 times), amplification step is more, is not suitable for analysis and the mirror of great amount of samples
It is fixed.Multiplex PCR is improved on the basis of regular-PCR, is drawn with 2 Duis or 2 pairs or more in same PCR reaction systems
Object expands the round pcr of multiple target fragments for the different zones of multiple DNA profilings or same template, wide at present
General each research field applied to life science.Since abo blood group is clinically particularly significant, occur now a large amount of
The PCR-SSP methods for ABO Genotypings, it is also many for the design of primers of each SNP of not synantigen.Meanwhile it is more
Weight PCR method is also one and is widely used general technology.In currently available technology, the human erythrocyte ABO about the present invention
Blood group antigens multiplex PCR classifying method, yet there are no report.
Invention content
First purpose of the present invention is to be directed to deficiency in the prior art, is provided a kind of for human erythrocyte's ABO blood
Type antigen multiplex PCR classifying method.
Second object of the present invention is to be directed to deficiency in the prior art, is provided a kind of for human erythrocyte's ABO blood
The primer of type antigens genotyping combines.
Third object of the present invention is to be directed to deficiency in the prior art, and it is anti-to provide a kind of human erythrocyte's abo blood group
Former multiplex PCR parting kit.
Fourth object of the present invention is to be directed to deficiency in the prior art, provides multiplex PCR classifying method as described above
Purposes.
To realize above-mentioned first purpose, the technical solution adopted by the present invention is that：
One kind being used for human erythrocyte's abo blood group antigen multiplex PCR classifying method, for A, B, O multiple allele, design
4 pairs of specific primers are formed 2 multi-PRC reaction systems, are expanded using subject's DNA sample as template, according to amplification item
The abo blood group of the presence or absence of band judgement sample；4 pairs of specific primers are respectively following (1) and (2), or the group of (1) and (3)
It closes；
(1) specific primer of O and non-O allele are expanded；
(2) specific primer of A and non-A allele are expanded；
(3) specific primer of B and non-B allele are expanded.
Further, 2 multi-PRC reaction systems are one of following situation：
(1) specific primer of the amplification O allele forms 1 with the specific primer for expanding non-B allele
Multi-PRC reaction system, the specific primer group of the specific primer and amplification B allele of the non-O allele of amplification
At 1 multi-PRC reaction system；Or
(2) specific primer of the amplification O allele forms 1 with the specific primer for expanding non-A allele
Multi-PRC reaction system, the specific primer group of the specific primer and amplification A allele of the non-O allele of amplification
At 1 multi-PRC reaction system.
Further, 4 pairs of specific primer sequences are respectively such as SEQ ID NO:Shown in 1-8.
To realize above-mentioned second purpose, the technical solution adopted by the present invention is that：
A kind of primer for human erythrocyte's abo blood group antigens genotyping combines, including sequence such as SEQ ID NO:1-10
Shown in primer.
To realize above-mentioned third purpose, the technical solution adopted by the present invention is that：
A kind of human erythrocyte's abo blood group antigen multiplex PCR parting kit, the kit include to draw as described above
Object combines.
Further, also comprising operational manual in the kit, the operational manual records the following contents：Into pedestrian
Multiplex PCR is used when class red blood cell abo blood group antigens genotyping；Amplification system is as follows：O, non-B and internal control primer are to forming more than 1
Weight system；Non- O, B and internal reference form 1 multiple system；
Further, the operational manual records the following contents：The multiple system of the O, non-B and internal reference is as follows：DNA 2
Microlitre, each 0.4 microlitre of non-B, internal control primer, 0.15 microlitre of O primers, 10 microlitres of Mix, ddH2O complements to 20 microlitres, the DNA
Concentration is not less than 20ng/ μ L, and the primer concentration is 10 μM；
Further, the operational manual records the following contents：The multiple system of described non-O, B and internal reference is as follows：DNA 2
Microlitre, 0.08 microlitre of non-O primers, 0.5 microlitre of B primers, 0.2 microlitre of internal control primer, 10 microlitres of Mix, ddH2It is micro- that O complements to 20
It rises, the DNA concentration is not less than 20ng/ μ L, and the primer concentration is 10 μM.
Further, the operational manual records the following contents：Amplification program is as follows：95 degree of 10min；Subsequent cyclic program
It is recycled for 95 degree of 20s, 62 degree of 20s, 72 degree of 20s, 10；95 degree of 20s, 60 degree of 20s, 72 degree of 20s, 25 cycles；Last 72 degree,
7min；After amplification, 8-10 microlitres of PCR product, gel electrophoresis, imaging are taken.
Further, the operational manual records the following contents：Imaging results are carried out matching with following table to compare, determines and surveys
The blood group of sample sheet, in table+number represent corresponding positions and be equipped with band, when certain row imaging results phase of test imaging results and following table
Timing determines that subject is the blood group or genotype that the row represents.
To realize above-mentioned 4th purpose, the technical solution adopted by the present invention is that：
Application of any multiplex PCR classifying method in ABO Genotypings as above.
PCR-SSP methods are mostly used in currently available technology and carry out ABO typing, and single person-portion needs more PCR to react
Number (>6 times).Present invention employs multiple PCR methods, and carry out detection site and simplify, and single person-portion need to only expand 2
Preliminary ABO typing can be realized in PCR reactions.
The invention has the advantages that：
1, of the invention that multiple PCR method is introduced into abo blood group Genotyping；Utilize O allele and B allele
SNP site realizes abo blood group Genotyping；The method of the present invention can reduce disposable's PCR reactions of ABO antigen gene partings
Number reduces the manpower and reagent cost of detection, improves detection flux, the time required to shortening large sample size detection.
2, due to ABO encoding genes only 1, when carrying out multiplex PCR, certain difficulty, this hair are brought to design system
It is bright by contrast experiment's screening and optimizing primer sequence and amplification system, have the advantages that high specific, highly sensitive.
3, the present invention improves the specificity of PCR-SSP reactions in design of primers by mispairing.
Description of the drawings
Attached drawing 1 is electrophoresis result (one) of the sample of known ABO Genotypings after the amplification of multiplex PCR system.It swims in figure
Road 1 is Marker (molecular weight 2,000bp, 1,000bp, 750bp, 500bp, 250bp, 100bp)；Swimming lane 2 and 3,4 and 5,6 and 7
The respectively multiple system product of 3 A types (genotype A1O1) sample O and non-B and non-O and B；Swimming lane 8 and 9,10 and 11 points
Not Wei 2 A types (genotype A1O1v) sample O and non-B and non-O and B multiple system product.
Attached drawing 2 is electrophoresis result (two) of the sample of known ABO Genotypings after the amplification of multiplex PCR system.It swims in figure
Road 1 is Marker (molecular weight 2,000bp, 1,000bp, 750bp, 500bp, 250bp, 100bp)；Swimming lane 2 and 3,4 and 5,6 and 7
The respectively multiple system product of 3 A types (genotype AO1) sample O and non-B and non-O and B；Swimming lane 8 and 9,10 and 11,12
It is respectively the multiple system product of 3 A types (genotype AO1v) sample O and non-B and non-O and B with 13；Swimming lane 14 and 15 is distinguished
For the multiple system product of certain A types (genotype A1A1) sample O and non-B and non-O and B.
Attached drawing 3 is electrophoresis result (three) of the sample of known ABO Genotypings after the amplification of multiplex PCR system.It swims in figure
Road 1 is Marker (molecular weight 1,000bp, 750bp, 500bp, 250bp, 100bp)；Swimming lane 2 and 3,4 and 5,6 and 7,8 and 9 points
Not Wei 4 Type B (genotype BB) sample O and non-B and non-O and B multiple system product；Swimming lane 10 and 11,14 and 15 difference
For the multiple system product of 2 AB types (genotype AB) sample O and non-B and non-O and B；Swimming lane 12 and 13 is respectively certain Type B
The multiple system product of (genotype BO1) sample O and non-B and non-O and B.Which part sample has the non-spies of a small amount of about 2000bp
Anisotropic band does not influence parting judgement.
Attached drawing 4 is electrophoresis result (four) of the sample of known ABO Genotypings after the amplification of multiplex PCR system.It swims in figure
Road 1 is Marker (molecular weight 1,000bp, 750bp, 500bp, 250bp, 100bp)；Swimming lane 2 and 3,6 and 7 is respectively 2 Type Bs
The multiple system product of (genotype BO1) sample O and non-B and non-O and B；Swimming lane 4 and 5 is respectively certain AB type (genotype AB)
The multiple system product of sample O and non-B and non-O and B；Swimming lane 8 and 9 is respectively certain Type B (genotype BO1v) sample O and non-B
And the multiple system product of non-O and B；Swimming lane 10 and 11,14 and 15 is respectively 2 O-shaped (genotype O1O1v) sample O and non-B
And the multiple system product of non-O and B；Swimming lane 12 and 13 be respectively certain O-shaped (genotype O1O1) sample O and non-B and non-O and
The multiple system product of B；Swimming lane 16 and 17 is respectively the multiple of certain O-shaped (genotype O1vO1v) sample O and non-B and non-O and B
System product.
Specific implementation mode
The invention will be further elucidated with reference to specific embodiments.It should be understood that these embodiments are merely to illustrate this hair
It is bright rather than limit the scope of the invention.In addition, it should also be understood that, after having read the content of the invention recorded, art technology
Personnel can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited
Fixed range.
The principle of the present invention：
The combination of multiplex PCR and ABO Genotypings can be significantly reduced the amplification needed for single sample detection by the present invention
Number.Concrete thought is as follows：Since A, B, O are multiple allele, wherein A and B are dominant, and O is recessiveness, therefore is directed to A, B, O
Multiple allele designs 4 pairs of specific primers, forms 2 multi-PRC reaction systems, is carried out by template of subject's DNA sample
Amplification, according to the abo blood group of the presence or absence of amplified band judgement sample；4 pairs of specific primers respectively following (1) and (2),
Or the combination of (1) and (3)；
(1) specific primer of O and non-O allele are expanded；
(2) specific primer of A and non-A allele are expanded；
(3) specific primer of B and non-B allele are expanded.
2 multi-PRC reaction systems are one of following situation：
(1) specific primer of the amplification O allele forms 1 with the specific primer for expanding non-B allele
Multi-PRC reaction system, the specific primer group of the specific primer and amplification B allele of the non-O allele of amplification
At 1 multi-PRC reaction system；Or
(2) specific primer of the amplification O allele forms 1 with the specific primer for expanding non-A allele
Multi-PRC reaction system, the specific primer group of the specific primer and amplification A allele of the non-O allele of amplification
At 1 multi-PRC reaction system.
It is expanded using subject's DNA sample as template, amplification program is as follows：95 degree of 10min；Subsequent cyclic program is 95
20s, 62 degree of 20s, 72 degree of 20s, 10 is spent to recycle；95 degree of 20s, 60 degree of 20s, 72 degree of 20s, 25 cycles；Last 72 degree, 7min；Expand
After increasing, 8-10 microlitres of PCR product, gel electrophoresis, imaging, according to the ABO of the presence or absence of amplified band judgement sample are taken
Blood group.
It is illustrated by taking (1) and the combination of (3) as an example in following embodiment.
1 human erythrocyte's abo blood group antigen multiplex PCR classifying method of the present invention of embodiment
(1) method
The multiplex PCR of based on PCR-SSP.
(2) principle
The central genetic mode of abo blood group is：A, B, O are multiple allele, and wherein A and B are dominant, and O is recessiveness.Design
4 pairs of specific primers expand O, non-O (i.e. A and B), B and non-B (i.e. A and O) respectively, using beta-actin as internal reference, form 2
Multi-PRC reaction.According to the presence or absence of amplified band, you can the abo blood group (such as following table) of preliminary judgement sample.
1 amplified band of table and the genotype table of comparisons
(3) amplimer
Note：Italic is specific position, and runic underscore is the base mismatch introduced.
Amplimer is as follows：O and non-B, internal reference form 1 multiple system；Non- O and B, internal reference form 1 multiple system.Expand
Increase enzyme and uses Platinum Green Hot Start PCR Master Mix (2X) (Thermo Fisher
Scientific).20 microlitres of systems.
O and non-B, the multiple system of internal reference are as follows：2 microlitres of DNA, each 0.4 microlitre of non-B, internal control primer, O primers 0.15 are micro-
It rises, 10 microlitres of Mix, ddH2O is supplied.
Non- O and B, the multiple system of internal reference are as follows：2 microlitres of DNA, 0.08 microlitre of non-O primers, 0.5 microlitre of B primers, internal reference
0.2 microlitre of primer, 10 microlitres of Mix, ddH2O is supplied.
Amplification program is as follows：95 degree of 10min；Subsequent cyclic program recycles for 95 degree of 20s, 62 degree of 20s, 72 degree of 20s, 10；
95 degree of 20s, 60 degree of 20s, 72 degree of 20s, 25 cycles；Last 72 degree, 7min.
Experimental result is as follows：Respectively by several samples of known ABO Genotypings, expanded using 2 kinds of multiplex PCR systems,
The result shows that (Fig. 1-Fig. 4), each genotype of detection is consistent with expected results.Abo blood group letter can be realized by this programme
Easy parting.
Multiple PCR method is introduced abo blood group Genotyping by the present invention；Utilize the SNP of O allele and B allele
Abo blood group Genotyping is realized in site；The specificity of PCR-SSP reactions is improved by mispairing in design of primers.
2 human erythrocyte's abo blood group antigen multiplex PCR parting kit of the present invention of embodiment
A kind of human erythrocyte's abo blood group antigen multiplex PCR parting kit, including primer pair as described below；
Operating instruction：
(1) it carries out using multiplex PCR when human erythrocyte's abo blood group antigens genotyping；
(2) amplification system is as follows：O, non-B and internal control primer are to forming 1 multiple system；Non- O, B and internal reference composition are more than 1
Weight system；Amplification enzyme uses Platinum Green Hot Start PCR Master Mix.
(3) the multiple system of O, non-B and internal reference is as follows：2 microlitres of DNA, each 0.4 microlitre of non-B, internal control primer, O primers
0.15 microlitre, 10 microlitres of Mix, ddH2O is supplied.
(3) the multiple system of non-O, B and internal reference is as follows：2 microlitres of DNA, 0.08 microlitre of non-O primers, 0.5 microlitre of B primers,
0.2 microlitre of internal control primer, 10 microlitres of Mix, ddH2O is supplied.
(4) amplification program is as follows：95 degree of 10min；Subsequent cyclic program is 95 degree of 20s, and 62 degree of 20s, 72 degree of 20s, 10 follow
Ring；95 degree of 20s, 60 degree of 20s, 72 degree of 20s, 25 cycles；Last 72 degree, 7min；After amplification, PCR product, gel electricity are taken
Swimming, imaging.
(5) imaging results are carried out matching with following table to compare, determines the blood group of test sample.
It is detected into 50 samples to known ABO Genotypings using the kit of the present embodiment, by testing result
It is compared with known blood group, accuracy 100%.Kit of the present invention is by primer screening and in design of primers simultaneously
The specificity of PCR-SSP reactions is improved by mispairing, when electrophoresis is not in non-specific band, and as a result interpretation is accurate, parting
As a result relatively reliable.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
Member, under the premise of not departing from the method for the present invention, can also make several improvement and supplement, these are improved and supplement also should be regarded as
Protection scope of the present invention.
SEQUENCE LISTING
<110>Shanghai City Blood Center
<120>A kind of human erythrocyte's abo blood group antigen multiplex PCR classifying method and kit
<130> /
<160> 10
<170> PatentIn version 3.3
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Claims (9)
1. one kind being used for human erythrocyte's abo blood group antigen multiplex PCR classifying method, which is characterized in that answer equipotential for A, B, O
Gene designs 4 pairs of specific primers, forms 2 multi-PRC reaction systems, is expanded using subject's DNA sample as template,
According to the abo blood group of the presence or absence of amplified band judgement sample；4 pairs of specific primers are respectively following (1) and (2), or (1)
With the combination of (3)；
(1) specific primer of O and non-O allele are expanded；
(2) specific primer of A and non-A allele are expanded；
(3) specific primer of B and non-B allele are expanded.
2. being used for human erythrocyte's abo blood group antigen multiplex PCR classifying method according to claim 1, which is characterized in that institute
It is one of following situation to state 2 multi-PRC reaction systems：
(1) specific primer of the amplification O allele and the specific primer composition 1 for expanding non-B allele are multiple
The specific primer of PCR reaction systems, the specific primer and amplification B allele of the non-O allele of amplification forms 1
Multi-PRC reaction system；Or
(2) specific primer of the amplification O allele and the specific primer composition 1 for expanding non-A allele are multiple
The specific primer of PCR reaction systems, the specific primer and amplification A allele of the non-O allele of amplification forms 1
Multi-PRC reaction system.
3. being used for human erythrocyte's abo blood group antigen multiplex PCR classifying method according to claim 1, which is characterized in that institute
4 pairs of specific primer sequences are stated respectively such as SEQ ID NO:Shown in 1-8,
4. a kind of primer for human erythrocyte's abo blood group antigens genotyping combines, which is characterized in that include sequence such as SEQ ID
NO:Primer shown in 1-10,
5. a kind of human erythrocyte's abo blood group antigen multiplex PCR parting kit, which is characterized in that the kit includes power
Profit requires the 4 primer combinations.
6. human erythrocyte's abo blood group antigen multiplex PCR parting kit according to claim 5, which is characterized in that described
Also include operational manual in kit, the operational manual records the following contents：Carry out human erythrocyte's abo blood group antigen
Multiplex PCR is used when parting；Amplification system is as follows：O, non-B and internal control primer are to forming 1 multiple system；Non- O, B and internal reference group
At 1 multiple system；
The multiple system of the O, non-B and internal reference is as follows：2 microlitres of DNA, each 0.4 microlitre of non-B, internal control primer, O primers 0.15 are micro-
It rises, 10 microlitres of Mix, ddH2O complements to 20 microlitres, and the DNA concentration is not less than 20ng/ μ L, and the primer concentration is 10 μM；
The multiple system of described non-O, B and internal reference is as follows：2 microlitres of DNA, 0.08 microlitre of non-O primers, 0.5 microlitre of B primers, internal reference
0.2 microlitre of primer, 10 microlitres of Mix, ddH2O complements to 20 microlitres, and the DNA concentration is not less than 20ng/ μ L, and the primer is dense
Degree is 10 μM.
7. human erythrocyte's abo blood group antigen multiplex PCR parting kit according to claim 6, which is characterized in that described
Operational manual records the following contents：Amplification program is as follows：95 degree of 10min；Subsequent cyclic program is 95 degree of 20s, 62 degree of 20s,
72 degree of 20s, 10 cycles；95 degree of 20s, 60 degree of 20s, 72 degree of 20s, 25 cycles；Last 72 degree, 7min；After amplification, PCR is taken
8-10 microlitres of product, gel electrophoresis, imaging.
8. human erythrocyte's abo blood group antigen multiplex PCR parting kit according to claim 7, which is characterized in that described
Operational manual records the following contents：Imaging results are carried out matching with following table to compare, determine the blood group of test sample,
9. application of any multiplex PCR classifying methods of claim 1-3 in ABO Genotypings.
Priority Applications (1)
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| CN108642163A (en) * | 2018-05-16 | 2018-10-12 | 江苏中济万泰生物医药有限公司 | A kind of human erythrocyte's abo blood group genotyping primer group and application |
| CN109182511A (en) * | 2018-11-12 | 2019-01-11 | 青岛市中心血站 | It is a kind of for detecting the SNP site of abo blood group anomaly |
| CN109554448A (en) * | 2018-12-27 | 2019-04-02 | 浙江省血液中心 | A kind of multiplex PCR-SBT the methods of genotyping and reagent of human erythrocyte's blood group system ABO antigen |
| CN112029842A (en) * | 2020-08-31 | 2020-12-04 | 深圳市血液中心(深圳市输血医学研究所) | Kit and method for ABO blood type genotyping based on high-throughput sequencing |
| WO2021135559A1 (en) * | 2019-12-31 | 2021-07-08 | 河南兴龙生物技术有限公司 | Primer group for detecting human red blood cell abo blood type genotyping, kit and application thereof |
| CN114561458A (en) * | 2022-04-20 | 2022-05-31 | 青岛市中心血站 | SNP (Single nucleotide polymorphism) site related to B variant in ABO (anaerobic-baffled oxide) blood group system and application thereof |
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| CN108642163A (en) * | 2018-05-16 | 2018-10-12 | 江苏中济万泰生物医药有限公司 | A kind of human erythrocyte's abo blood group genotyping primer group and application |
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| CN109182511A (en) * | 2018-11-12 | 2019-01-11 | 青岛市中心血站 | It is a kind of for detecting the SNP site of abo blood group anomaly |
| CN109182511B (en) * | 2018-11-12 | 2021-11-02 | 青岛市中心血站 | SNP (Single nucleotide polymorphism) locus for detecting ABO blood type variants |
| CN109554448A (en) * | 2018-12-27 | 2019-04-02 | 浙江省血液中心 | A kind of multiplex PCR-SBT the methods of genotyping and reagent of human erythrocyte's blood group system ABO antigen |
| CN109554448B (en) * | 2018-12-27 | 2019-08-30 | 浙江省血液中心 | A kind of multiplex PCR-SBT the methods of genotyping and reagent of human erythrocyte's blood group system ABO antigen |
| WO2021135559A1 (en) * | 2019-12-31 | 2021-07-08 | 河南兴龙生物技术有限公司 | Primer group for detecting human red blood cell abo blood type genotyping, kit and application thereof |
| CN112029842A (en) * | 2020-08-31 | 2020-12-04 | 深圳市血液中心(深圳市输血医学研究所) | Kit and method for ABO blood type genotyping based on high-throughput sequencing |
| CN112029842B (en) * | 2020-08-31 | 2021-04-27 | 深圳市血液中心(深圳市输血医学研究所) | Kit and method for ABO blood type genotyping based on high-throughput sequencing |
| CN114561458A (en) * | 2022-04-20 | 2022-05-31 | 青岛市中心血站 | SNP (Single nucleotide polymorphism) site related to B variant in ABO (anaerobic-baffled oxide) blood group system and application thereof |
| CN114561458B (en) * | 2022-04-20 | 2023-12-29 | 青岛市中心血站 | SNP locus related to B variant in ABO blood group system and application thereof |
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