CN1900314A - Fluorescence labelling ABO gene typing method and its reagent kit - Google Patents
Fluorescence labelling ABO gene typing method and its reagent kit Download PDFInfo
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- CN1900314A CN1900314A CN 200610047231 CN200610047231A CN1900314A CN 1900314 A CN1900314 A CN 1900314A CN 200610047231 CN200610047231 CN 200610047231 CN 200610047231 A CN200610047231 A CN 200610047231A CN 1900314 A CN1900314 A CN 1900314A
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Abstract
The present invention relates to molecular biology, medical jurisprudence and genetics, and is especially fluorescence labeling ABO gene typing method and reagent kit for treating criminous biological material evidence. The method includes designing one group of allele sequence specific primers based on the sequence polymorphism of ABO determinative gene, applying NED and TMR fluorescent dye labels on the 5' terminals of two primers, and performing PCR product segment analysis with DNA genetic analyzer to judge the ABO genotype of the sample. Compared with available technology, the present invention has the advantages of greatly raised detection sensitivity, simple operation, high speed, etc.
Description
Technical field
The present invention relates to molecular biology, medical jurisprudence, genetics field, concrete is to the fluorescence labelling ABO gene typing of the biological material evidence of committing a crime method and test kit thereof.
Background technology
ABO blood group system is the human genetic marker system of finding the earliest, after 1900Landerstainer finds ABO blood group system, immediately by in clinic study such as the medical science that promptly is applied to transfuse blood and the practice.Along with going deep into to ABO blood group system research, the very fast quilt of new achievement in research extends to multiple fields such as anthropology, sibship analysis and medical jurisprudence individual recognition, and occupy critical role, be applied topmost genetic marker during forensic is identified since for a long time always.Such as the various blood cakes of biological material evidence, seminal stain, mixed stain, hair, stub, salivary stain, sweat stain, cartilaginous tissue etc. that forensic science is committed a crime, particularly how accurately on the ABO genotype identification to " nonsecreting type " mixed stain, for the detection of criminal case points the direction and play crucial effects in carrying out large-scale suspect's investigation process.But the check of traditional forensic abo blood group mainly is to adopt such as serological methods such as absorption experiment, the experiment of dissociating, mixed agglutination experiment, enzyme linked immunological experiments, and the target of detection also is an erythrocyte membrane surfaces A BH antigenic substance, i.e. glycolipid or glycoprotein.This method can only be judged the phenotype of abo blood group, judgement that the more important thing is its result may be because ambient conditions to the active destruction of antigen, bacterial contamination, individual secretor state, ontogeny state even may be subjected to the influence of disease that human body takes a disease, and causes checking failure or appearance mistake to declare the situation of type.
The eighties, modern molecular biology technique develops rapidly, makes people become possibility from gene level understanding and test organisms feature.Especially at the beginning of the nineties, along with the widespread use of round pcr, legal medical expert scholar has all over the world carried out the research work of a large amount of abo blood group gene types, and has reported a series of method of inspection.Wherein report early and the method for being used widely mainly contain the directly method of order-checking of PCR-RFLP and PCR.
The PCR-RFLP method mainly is according to the characteristic of blood group gene in the different or disappearance of Different Alkali base location Nucleotide, select some of them site design PCR primer, amplification comprises the specific fragment that some restriction endonuclease sites changes, utilize restriction enzyme to cut, and carry out electrophoresis, distinguish ABO allelotrope according to the different characteristic of different endonuclease bamhi length, thereby determine the method for abo blood group.
The key of this method and technology is cut at enzyme, needs specified conditions.When enzyme cut the time not enough, enzymic activity descends or amplified production enzyme can occur when too much and cuts incomplete phenomenon, can cause the erroneous judgement of blood group.Simultaneously, because the detection method that this method mainly uses silver to dye, being about to amplified production utilizes common electrophoresis chamber to separate, carry out the method that cma staining detects, though method is simple, cost is lower, but the just single color of planting of gained result, the site of once-combined amplification can not surpass 4, otherwise can be difficult to distinguish the genotype of sample because the amplified production of different loci is overlapped, adding the gained result detects by an unaided eye, its sensitivity is subjected to certain restriction, and complicated operation, and labour intensity is big.
The PCR directly method of order-checking mainly is to use the base sequence variation that PCR-direct sequence analytical technology is analyzed human A, B, 3 kinds of genes of O.The PCR direct sequencing is according to the characteristic of blood group gene in the different or disappearance of Different Alkali base location Nucleotide, select some of them site design PCR primer, amplification comprises the specific fragment in some feature site, the PCR product is directly checked order, directly determine the method for abo blood group according to the sequence results that obtains.
Though its detection platform that adopts is the fluorescent mark analytical technology, it exists the sequencing technologies operation loaded down with trivial details, technical requirements height, the shortcoming that round of visits is long.
Goal of the invention
The method and the test kit thereof that the purpose of this invention is to provide a kind of fluorescence labelling ABO gene typing that difficult biological material evidence fluorescent mark allele specific amplification method in the criminal case is carried out.
For achieving the above object, technical solution of the present invention is as follows:
The test kit of fluorescence labelling ABO gene typing, the fluorescence labelling ABO gene primer that provides 6 can discern the 6th and the 7th exon base substitution sequence is provided, and the method that ABO gene type in a kind of difficult biological material evidence that uses in the multiple criminal case of this test kit rapid detection is provided.
Cardinal principle of the present invention is: ABO blood group system is the apparent recessive inheritance system of multiple allelomorphos by A, B, the decision of O triallelic, its decision gene is positioned on No. 9 karyomit(e), form by 7 exons, wherein 796,803 of the 7th exon liang of polymorphisms of locating the base substitution formation of position determine allelotrope A or B, and single base deletion of the 6th exon 2 61 positions produces allelotrope O.The PCR-SSP method is exactly according to the sequence difference between A, B, each allelotrope of O, design corresponding sequence specific primers, make its only its corresponding allelotrope of each primer in the PCR circulation anneal and be extended, simultaneously by incomplementarity polynucleotide at the different numbers of the disconnected interpolation in 5 ' end of each primer, to adjust PCR product sheet segment length, according to the difference of the amplified production fragment length that obtains, carry out the genotypic judgement of ABO.Therefore, scientific and reasonable design of primers is to guarantee to use the deciding factor that this method is carried out the success of ABO gene type.
The fluorescence labelling ABO gene typing method comprises the steps:
1) according to the synthetic Auele Specific Primer of ABO gene order, and two primer 5 ' ends use NED or TMR fluorochrome label respectively therein;
2) extraction of DNA: the method for employing Chelex-100 or organic solvent extraction is extracted the DNA extraction liquid in the testing sample;
3) pcr amplification: 0.2-10ng DNA extraction liquid is joined in the PCR test tube that contains primer, then be placed in the PCR instrument and increase: 95 ℃ of sex change 11min, 90-97 ℃ of sex change 30s subsequently, 50-60 ℃ of annealing 30s, 70-75 ℃ is extended 30s, after 25-40 the circulation, 70-75 ℃ is extended 5-60min;
4) amplified production analysis: the product after will increasing places the dna fragmentation analyser to carry out electrophoretic separation.
5) interpretation of result: use the GeneMapper3.2 analysis software, all samples is analyzed.
Discern 3 allelotrope A, B, O, determine that AA, AO, BB, BO, AB, 006 kind of genotypic clip size judging criterion of combining are: fragment length is respectively 110,98,85,74bps.
The test kit of fluorescence labelling ABO gene typing comprises: PCR test tube composition is: every primer 2-6pmol, 1.0 μ l dNTP (dATP, dTTP, dCTP, the mixture of dGTP, every kind of concentration 2mM), 25mM MgCl
21.0-4 μ l, 10 * buffer, 1.0 μ l, distilled water 0-3 μ l, 5U/ μ l TaqGold enzyme 0.2-1.0 μ l, 2.5mM calf serum 0.1-2.0 μ l, positive control dna solution 0.2-10ng.
Described test kit is stored under the lucifuge ,-20 ℃; Multigelation is avoided in packing in advance.
Described 6 primer lengths are 19-29 base, and its base sequence is respectively:
①5’CTGTGGCGCTTCTGATGTCCTCGTGGTGA-3’
②5’-AACGATGTCCTCGTGGTAC-3’
③5’-GACTGCAGGGCGATTTCTACTACA-3’
④5’-GCGTCTACTACCTGGGGGG-3’
⑤5’-NED-CTTGAGGATGTCGATGTTG-3’
⑥5-’TMR-TAGCATCTGGTCGACCATCATG-3’。
The present invention has following advantage:
1, the target of ABO genotype detection is DNA, the membrane antigen material (protein) of the more traditional serologic test of DNA has better tolerance for the influence and the destruction of various ambient conditionss, therefore adopt test kit of the present invention to having higher success rate that trace, outmoded biological material are checked, sensitivity is also higher.
When 2, adopting serological method to carry out the abo blood group check for nonsecreting type individuality other body fluid components except that blood, may be very few and can't check owing to antigenic component wherein, even obtain mistake and declare type.And when adopting abo blood group methods of genotyping of the present invention, but its blood group decision gene of direct survey is not subjected to the influence of individual secretor state, can directly obtain accurate conclusion.
3, be common biological material during forensic is identified to seminal fluid in the sexual crime in the biological material evidence of committing a crime and vaginal secretion mixed stain, usually adopt serological method can only obtain the blended abo blood group, adopt plus-minus method to get rid of analysis again, its complicated operation, cycle are long, can not determine the blood group of its sperm simultaneously exactly; And adopt the method for gene type of the present invention owing at first separated the man in the mixed stain, women composition, can directly determine the blood group of sperm, and easy to operate, accurate, cycle weak point.
4, the present invention combines fluorescent mark and carries out the ABO gene type and make its operation simpler with the PCR-SSP method, round of visits is shorter, is more suitable for Automated inspection in batches, and is ageing also stronger, check sensitivity is improved greatly, strengthened the practicality of its evaluation.
5, because the present invention does not adopt restriction enzyme cutting, therefore do not exist since enzyme cut not exclusively cause the result is declared the influence that type produces.
Description of drawings
Fig. 1 is primer combining site of the present invention and combination synoptic diagram.
Fig. 2 is an ABO gene type collection of illustrative plates of the present invention.
Embodiment
The present invention is described in further detail below in conjunction with specific embodiment, but not as limitation of the present invention.
Embodiment 1
The fluorescence labelling ABO gene typing method comprises the steps:
1. according to the synthetic Auele Specific Primer of ABO gene order, and therein 5., 6. two primers 5 ' ends use NED, TMR fluorochrome label (referring to Fig. 1) respectively.
2.DNA extraction: adopt the Chelex-100 method, get and have 3 * 3mm that the old trace is done blood stain
2Size blood stain gauze shreds to centrifuge tube, adds 5% Chelex-100 200 μ l, the concussion vortex, and 56 ℃ of water-baths are after 2 hours, boiling water bath 10min, the centrifugal 3min of 12000rpm gets supernatant, is DNA extraction liquid.
3.PCR amplification: 0.2ng DNA extraction liquid joined contain in steps in the 1 described primer PCR test tube, then be placed in the PCR instrument and increase: 95 ℃ of sex change 11min, 90 ℃ of sex change 30s subsequently, 50 ℃ of annealing 30s, 72 ℃ are extended 30s, and after 25 circulations, 70 ℃ are extended 5min;
4. amplified production analysis: product 1 μ l+ deionized formamide 9 μ l+ molecular weight internal standards (ABI GeneScan ROX 500) the 0.5 μ l after will increasing, behind the mixing, 95 ℃ of sex change 5 minutes place the dna fragmentation analyser to carry out electrophoretic separation; Wherein: analyser adopts ABI-3100 type or 3130 types, and dye set is provided with F, operational mode: GS36-POP4 or GS36-SNP, analytical parameters: GS500;
5. interpretation of result: use the GeneMapper3.2 analysis software, open a new analysis window, after all samples is added, at first set each fragment length of molecular weight internal standard, execution analysis order again can obtain each sample PCR product analysis result, (referring to Fig. 2).
Wherein: discern 3 allelotrope A, B, O, determine the 6 kinds of genotypic clip size judging criterions of AA, AO, BB, BO, AB, OO such as the table 1 that combine:
Table 1 ABO gene type is declared the type standard
| A | B | O | |
| A | 98/85 | 110/98/85 | 98/85/74 |
| B | 110/85 | 110/98/85/74 | |
| O | 98/74 |
The test kit of described fluorescence labelling ABO gene typing: PCR test tube composition is: primer is 2pmol, 1.0 μ l dNTP (dATP, dTTP, dCTP, the mixture of dGTP, every kind of concentration 12mM), 25mM 1.0 μ l MgCl respectively
2, 1.0 μ l, 10 * buffer, 5U/ μ l 0.2 μ l TaqGold enzyme, 2.5mM0.1 μ l BSA, positive control dna solution 0.2ng.
Adopt NED, TMR fluorochrome label primer 5., 6. 5 ' end.
Described 6 primers are:
①5’CTGTGGCGCTTCTGATGTCCTCGTGGTGA-3’
②5’-AACGATGTCCTCGTGGTAC-3’
③5’-GACTGCAGGGCGATTTCTACTACA-3’
④5’-GCGTCTACTACCTGGGGGG-3’
⑤5’-NED-CTTGAGGATGTCGATGTTG-3’
⑥5-’TMR-TAGCATCTGGTCGACCATCATG-3’。
Embodiment 2
Difference from Example 1 is:
The fluorescence labelling ABO gene typing method comprises the steps:
1. according to the synthetic Auele Specific Primer of ABO gene order, and therein 5., 6. two primers 5 ' ends use NED, TMR fluorochrome label respectively.
2.DNA extraction: adopt the Chelex-100 method, clip has the gauze of 1 square centimeter of mixed stain, shreds to centrifuge tube, add TES damping fluid 900 μ l and suspend, add SDS and Proteinase K final concentration and be respectively 1% and 100ug/ml, fully mixing, 37 ℃ of water-baths 1 hour, carrier of separating is collected parting liquid, the centrifugal 8min of 7000rpm abandons supernatant, and precipitation adds 500 μ l TES and suspends, again with the centrifugal 8min of 7000rpm, abandon supernatant, 4 times repeatedly, be and remove the vagina epithelial cell fully; Wherein: the TES damping fluid: 10mM Tris, 100mM NaCl 5mM EDTA.Chelex 100 solution that will remove add 200 μ l 5% in the epithelial precipitation sperm of vagina suspend, and add Proteinase K, dithiothreitol (DTT) solution final concentration is respectively 100ug/ml, 40mmol/l, and 56 ℃ of water-baths are after 3 hours, boiling water bath 10min; The centrifugal 3min of 12000rpm gets supernatant, is DNA extraction liquid.
3.PCR amplification: the 5ngDNA extracting solution joined contain in steps in the 1 described primer PCR test tube, then be placed in the PCR instrument and increase: 95 ℃ of sex change 11min, 95 ℃ of sex change 30s subsequently, 54 ℃ of annealing 30s, 72 ℃ are extended 30s, and after 30 circulations, 72 ℃ are extended 45min.
4. amplified production analysis: product 1ul+ deionized formamide 9ul+ molecular weight internal standard (the ABI GeneScan ROX500) 0.5ul after will increasing, behind the mixing, 95 ℃ of sex change 5 minutes place the dna fragmentation analyser to carry out electrophoretic separation; Wherein: analyser adopts ABI-3100 type or 3130 types, dyes
The material group is provided with F, operational mode: GS36-POP4 or GS36-SNP, analytical parameters: GS500;
5. interpretation of result: use the GeneMapper3.2 analysis software, open a new analysis window, after all samples is added, at first set each fragment length of molecular weight internal standard, execution analysis order again can obtain each sample PCR product analysis result.
The test kit of fluorescence labelling ABO gene typing:
PCR reagent pipe composition is: primer is 4pmol, 1.0 μ l dNTP (dATP, dTTP, dCTP, the mixture of dGTP, every kind of concentration 12mM), 25mM 2.5 μ l MgCl respectively
2, 1.0 μ l, 10 * buffer, 1.5ul distilled water, 5U/ μ l 0.7 μ l TaqGold enzyme 2.5mg/ml 0.9 μ l BSA, 5ng positive control dna solution.
Embodiment 3
Difference from Example 1 is:
The fluorescence labelling ABO gene typing method comprises the steps:
1. according to the synthetic Auele Specific Primer of ABO gene order, and therein 5., 6. two primers 5 ' ends use NED, TMR fluorochrome label respectively.
2.DNA extraction: adopt the method for organic solvent extraction, stub tip paper outer ring, the wide left and right sides of clip 3mm shreds to centrifuge tube, add TES 350ul, naturally cool to room temperature behind the boiling water bath 10min, add 10%SDS 40ul, 10ug/ul Proteinase K 10ul, 55 ℃ of digestion are spent the night, after phenol, the chloroform extracting, add 3M NaAc (PH5.2) 20ul, no water-cooled ethanol 1ml, the centrifugal 10min of 12500rpm abandons supernatant, precipitation with 70% cold washing with alcohol is once dissolved with 50ul TE behind the airing.
3.PXR amplification: 10ng DNA extraction liquid joined contain in steps in the 1 described primer PCR test tube, then be placed in the PCR instrument and increase: 95 ℃ of sex change 11min, 97 ℃ of sex change 30s subsequently, 60 ℃ of annealing 30s, 75 ℃ are extended 30s, and after 40 circulations, 75 ℃ are extended 60min.
3. amplified production analysis: product 1ul+ deionized formamide 9ul+ molecular weight internal standard (the ABI GeneScan ROX 500) 0.5ul after will increasing, behind the mixing, 95 ℃ of sex change 5 minute hands place the dna fragmentation analyser to carry out electrophoretic separation; Wherein: analyser adopts ABI-3100 type or 3130 types, dyes
The material group is provided with F, operational mode: GS36-POP4 or GS36-SNP, analytical parameters: GS500;
4. interpretation of result: use the GeneMapper3.2 analysis software, open a new analysis window, after all samples is added, at first set each fragment length of molecular weight internal standard, execution analysis order again can obtain each sample PCR product analysis result,
The test kit of fluorescence labelling ABO gene typing:
PCR reagent pipe composition is: every primer 6pmol, 1.0 μ l dNTP (dATP, dTTP, dCTP, the mixture of dGTP, every kind of concentration 12mM), 25mM 4.0 μ l MgCl
2, 1.0 μ l, 10 * buffer, 3ul distilled water, 5U/ μ l 1 μ l TaqGold enzyme, 2.5mg/ml 2 μ l BSA, positive control dna solution 10ng.
ABO
SEQUENCE LISTING
＜110〉Liaoning Prov. Inst. of Criminal Science and Technology
＜120〉a kind of method of fluorescence labelling ABO gene typing and test kit thereof
<130>
<160>6
<170>PatentIn version 3.1
<210>1
<211>29
<212>DNA
＜213〉synthetic
<220>
<221>gene
<222>(1)..(29)
<223>
<400>1
ctgtggcgct tctgatgtcc tcgtggtga 29
<210>2
<211>19
<212>DNA
＜213〉synthetic
<220>
<221>gene
<222>(1)..(19)
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aacgatgtcc tcgtggtac 19
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＜213〉synthetic
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gactgcaggg cgatttctac taca 24
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<221>gene
ABO
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gcgtctacta cctgggggg 19
<210>5
<211>19
<212>DNA
＜213〉synthetic
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<221>gene
<222>(1)..(19)
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cttgaggatg tcgatgttg 19
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<212>DNA
＜213〉synthetic
<220>
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<222>(1)..(22)
<223>
<400>6
tagcatctgg tcgaccatca tg 22
Claims (4)
1, a kind of fluorescence labelling ABO gene typing method is characterized in that comprising the steps:
1) according to the synthetic Auele Specific Primer of ABO gene order, and two primer 5 ' ends use NED or TMR fluorochrome label respectively therein;
2) extraction of DNA: the method for employing Chelex-100 or organic solvent extraction is extracted the DNA extraction liquid in the testing sample;
3) pcr amplification: 0.2-10ng DNA extraction liquid is joined in the PCR test tube that contains primer, then be placed in the PCR instrument and increase: 95 ℃ of sex change 11min, 90-97 ℃ of sex change 30s subsequently, 50-60 ℃ of annealing 30s, 70-75 ℃ is extended 30s, after 25-40 the circulation, 70-75 ℃ is extended 5-60min;
4) amplified production analysis: the product after will increasing places the dna fragmentation analyser to carry out electrophoretic separation.
5) interpretation of result: use GeneMapper 3.2 analysis software, all samples is analyzed.
2, fluorescence labelling ABO gene typing method according to claim 1, it is characterized in that: discern 3 allelotrope A, B, O, determine that AA, AO, BB, BO, AB, 006 kind of genotypic clip size judging criterion of combining are: fragment length is respectively 110,98,85,74bps.
3. the test kit of a fluorescence labelling ABO gene typing according to claim 1, it is characterized in that: PCR test tube composition is: every primer 2-6pmol, 1.0 μ l dNTP (dATP, dTTP, dCTP, the mixture of dGTP, every kind of concentration 2mM), 25mM MgCl
21.0-4 μ l, 10 * buffer1.0 μ l, distilled water 0-3 μ l, 5U/l TaqGold enzyme 0.2-1.0 μ l, 2.5mM calf serum 0.1-2.0 μ l, positive control dna solution 0.2-10ng.
4. the test kit of fluorescence labelling ABO gene typing according to claim 3, it is characterized in that: described 6 primer lengths are 19-29 base, and its base sequence is respectively:
①5’-CTGTGGCGCTTCTGATGTCCTCGTGGTGA-3’
②5’-AACGATGTCCTCGTGGTAC-3’
③5’-GACTGCAGGGCGATTTCTACTACA-3’
④5’-GCGTCTACTACCTGGGGGG-3’
⑤5’-NED-CTTGAGGATGTCGATGTTG-3’
⑥5-’TMR-TAGCATCTGGTCGACCATCATG-3’。
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| CN 200610047231 CN1900314A (en) | 2006-07-19 | 2006-07-19 | Fluorescence labelling ABO gene typing method and its reagent kit |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921834A (en) * | 2010-05-17 | 2010-12-22 | 浙江省血液中心 | Polymerase chain reaction-sequence based typing (PCR-SBT) method for ABO blood type genotyping and reagent |
| CN104342491A (en) * | 2014-01-16 | 2015-02-11 | 肖文昕 | Novel non-invasive blood type gene detection kit |
| CN106048059A (en) * | 2016-08-10 | 2016-10-26 | 青岛大学附属医院 | SNP sites of A variation blood type for triggering acute hemolytic transfusion reaction |
| CN106834483A (en) * | 2017-02-27 | 2017-06-13 | 公安部物证鉴定中心 | Detect the template of the allelic ladder of the method and abo blood group locus of abo blood group genotype |
| 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 |
| 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 |
-
2006
- 2006-07-19 CN CN 200610047231 patent/CN1900314A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921834A (en) * | 2010-05-17 | 2010-12-22 | 浙江省血液中心 | Polymerase chain reaction-sequence based typing (PCR-SBT) method for ABO blood type genotyping and reagent |
| CN101921834B (en) * | 2010-05-17 | 2012-12-19 | 浙江省血液中心 | Polymerase chain reaction-sequence based typing (PCR-SBT) method for ABO blood type genotyping and reagent |
| CN104342491A (en) * | 2014-01-16 | 2015-02-11 | 肖文昕 | Novel non-invasive blood type gene detection kit |
| CN106048059A (en) * | 2016-08-10 | 2016-10-26 | 青岛大学附属医院 | SNP sites of A variation blood type for triggering acute hemolytic transfusion reaction |
| CN106048059B (en) * | 2016-08-10 | 2019-04-09 | 青岛大学附属医院 | A kind of SNP site for the A anomaly blood group causing acute hemolytic transfusion reaction |
| CN106834483A (en) * | 2017-02-27 | 2017-06-13 | 公安部物证鉴定中心 | Detect the template of the allelic ladder of the method and abo blood group locus of abo blood group genotype |
| CN106834483B (en) * | 2017-02-27 | 2020-05-12 | 公安部物证鉴定中心 | Method for detecting ABO blood group genotype and template of allelic typing standard of ABO blood group locus |
| 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 |
| CN108642163B (en) * | 2018-05-16 | 2021-10-22 | 江苏中济万泰生物医药有限公司 | Human erythrocyte ABO blood group genotyping primer set and application |
| 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 |
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