CN116751786A - Rh blood group D antigen RHD486C > A allele and detection primer thereof - Google Patents
Rh blood group D antigen RHD486C > A allele and detection primer thereof Download PDFInfo
- Publication number
- CN116751786A CN116751786A CN202310555826.3A CN202310555826A CN116751786A CN 116751786 A CN116751786 A CN 116751786A CN 202310555826 A CN202310555826 A CN 202310555826A CN 116751786 A CN116751786 A CN 116751786A
- Authority
- CN
- China
- Prior art keywords
- rhd486c
- seq
- allele
- sequence
- blood group
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 108700028369 Alleles Proteins 0.000 title claims abstract description 24
- 238000001514 detection method Methods 0.000 title abstract description 15
- 101000580024 Homo sapiens Blood group Rh(D) polypeptide Proteins 0.000 title abstract description 11
- 102100027544 Blood group Rh(D) polypeptide Human genes 0.000 title abstract description 9
- 101150108975 Rhd gene Proteins 0.000 claims abstract description 17
- 230000035772 mutation Effects 0.000 claims abstract description 15
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 4
- 238000011144 upstream manufacturing Methods 0.000 claims description 6
- 210000004369 blood Anatomy 0.000 abstract description 17
- 239000008280 blood Substances 0.000 abstract description 17
- 239000000427 antigen Substances 0.000 abstract description 13
- 108091007433 antigens Proteins 0.000 abstract description 13
- 102000036639 antigens Human genes 0.000 abstract description 13
- 238000000034 method Methods 0.000 abstract description 11
- 108090000623 proteins and genes Proteins 0.000 abstract description 11
- 230000000405 serological effect Effects 0.000 abstract description 7
- 230000007812 deficiency Effects 0.000 abstract description 3
- 206010064571 Gene mutation Diseases 0.000 abstract description 2
- 230000004544 DNA amplification Effects 0.000 abstract 1
- 238000003556 assay Methods 0.000 abstract 1
- 238000006243 chemical reaction Methods 0.000 description 16
- 108020004414 DNA Proteins 0.000 description 12
- 238000012408 PCR amplification Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 7
- 238000012163 sequencing technique Methods 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 239000007795 chemical reaction product Substances 0.000 description 5
- 210000003743 erythrocyte Anatomy 0.000 description 5
- 239000008188 pellet Substances 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 239000006166 lysate Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000007400 DNA extraction Methods 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 108010006785 Taq Polymerase Proteins 0.000 description 2
- 239000011543 agarose gel Substances 0.000 description 2
- 238000000246 agarose gel electrophoresis Methods 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 210000000265 leukocyte Anatomy 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 238000011895 specific detection Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000007399 DNA isolation Methods 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- 208000003441 Transfusion reaction Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 238000009582 blood typing Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000003617 erythrocyte membrane Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000007480 sanger sequencing Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000009589 serological test Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses an Rh blood group antigen RHD486C > A allele, the nucleotide sequence of a wild RHD gene is shown as SEQ ID NO. 1, the sequence of a mutant RHD gene is shown as SEQ ID NO. 2, the mutation site is at 209 th site of the sequence shown as SEQ ID NO. 1, and the mutation site is changed into A from base C. The inventors designed primer sequences for gene mutation, and specifically detected the regioselectivity of the mutant genes for RHD486C > A alleles by gene amplification methods. The primer sequence is positive only for RHD486C > A allele detection results, and is negative for wild type or other site mutation detection results. Specific assays can be performed for the RHD486C > A allele to determine Rh blood group D antigen genotype. The method can make up for the deficiency of serological detection technology, and has important clinical practical significance.
Description
Technical Field
The invention belongs to the technical field of molecular biology, and particularly relates to an Rh blood group antigen RHD486C & gtA allele and a detection primer thereof.
Background
Rh blood group is the most complex and polymorphic system of human erythrocyte blood group system, and is also the main erythrocyte blood group causing clinical transfusion reaction and severe neonatal hemolysis. It has now been found that over 50 Rh blood group antigens, of which RhD antigens are strongly immunogenic, are produced by encoding the RhD gene and are the focus of blood group studies. Clinically, rh blood group antigens are classified into two major classes of RhD positive and RhD negative according to whether D antigens are detected on the surface of erythrocyte membranes.
Currently, conventional methods for detecting Rh blood group D antigens employ serological methods, such as serological saline, indirect anti-human globulin tests, and absorption and diffusion tests. However, serological methods have certain limitations for the following reasons: the outcome of certain individuals' erythrocytes in serological typing is difficult to determine due to disease or other factors; serological results of chronic long-term transfusion patients sometimes exhibit the phenomenon of "mixed vision"; when red blood cells of a sample cannot be obtained or the red blood cell sample is insufficient, such as fetal blood typing, forensic identification remnants, etc., serological tests cannot obtain correct results.
The detection of RhD blood group by immune serology mainly depends on the specificity of anti-D antibodies and antigen expression. Along with the development of molecular biotechnology, rhD mutants are increased year by year, and the antigen expression amount and the gene mutation sites of the RhD mutants are different. The detection of the gene mutants mainly determines the Rh blood group D antigen genotype by a molecular biological method, can make up for the deficiency of serology technology, and has important clinical practical significance. The inventor researches and discovers that 1 case of Rh blood group RHD-N162K mutant carries RHD486C & gtA alleles, no related report and corresponding detection method exist in the prior art, and therefore specific detection is needed to be carried out on the RHD486C & gtA alleles to determine the genotype of the Rh blood group D antigen.
Disclosure of Invention
The invention aims to solve the defects of the prior art, provides an Rh blood group antigen RHD486C & gtA allele, and realizes Rh blood group genotype determination by specifically detecting the RHD486C & gtA allele.
The invention also aims at providing a specific primer and a detection method for detecting RHD486C & gtA alleles, which are realized by utilizing the change of specific sequence sites of different alleles of human RhD blood types to design a specific primer sequence for polymerase chain reaction.
Technical proposal
The nucleotide sequence of the wild RHD gene is shown as SEQ ID NO. 1, the mutant RHD gene is shown as SEQ ID NO. 2, the mutation site is at 209 th site of the sequence shown as SEQ ID NO. 1, and the mutation site is changed into A from base C.
A specific primer for detecting RHD486C & gtA alleles comprises an upstream primer and a downstream primer, wherein the sequence of the upstream primer is shown as SEQ ID NO. 3, and the sequence of the downstream primer is shown as SEQ ID NO. 4.
An upstream primer: 5'-CCTGAGGATGGTCATCAGTAATATCTTCAAA-3' (SEQ ID NO: 3),
a downstream primer: 5'-CAACCTCCCAAGTAGCTGGGA-3' (SEQ ID NO: 4).
A method of detecting the RHD486C > a allele comprising the steps of:
(1) Extracting DNA in a sample to be detected;
(2) Taking the DNA as a template, and carrying out PCR amplification reaction by adopting the specific primer to obtain a PCR reaction product;
(3) Sequencing the PCR amplification reaction product;
(4) Comparing the sequencing result with the sequence of the RHD wild-type gene to determine whether there is 486C-A mutation.
The invention has the beneficial effects that:
the invention discloses an Rh blood group antigen RHD486C & gtA allele, and designs a primer to realize the specific detection thereof, and the Rh blood group antigen genotype can be determined according to the detection result. Compared with the prior art, the invention adopts a molecular biology method to detect the gene level, and can detect the existence of the mutant gene doped in the gene library with high sensitivity and high precision. Due to the difference of RHD genes among different nations, the gene is designed specifically for RHD486C & gtA alleles newly found in Chinese according to the molecular background of the RHD genes of Chinese on the basis of related researches. The invention not only can make up the deficiency of serological detection technology, but also has important clinical practical significance and wide scientific research application value.
Drawings
FIG. 1 is a gel electrophoresis of RHD486C > A allele detection in example 2;
FIG. 2 is a sequencing map of RHD486C > A allele mutations in example 2.
Detailed Description
The technical scheme of the invention is clearly and completely described below with reference to the accompanying drawings and the specific embodiments.
Example 1
An Rh blood group antigen RHD486C > a allele having a g.25290791c > a site mutation at position 25290791 of chromosome 1. The gene is numbered NC_000001.11 (25272393-25330445) in NCBl reference database GRCh38.p14, and the nucleotide sequence of partial DNA containing wild type at the site in the database is listed for reference, and is shown in SEQ ID NO:1, the sequence of the corresponding mutant RHD gene is shown as SEQ ID NO:2, wherein the mutation site is mutated from a base C to A at 209 th position of the SEQ ID NO:1 sequence.
SEQ ID NO:1
GAATGAGTGAGAGGCATCCTTCCTTCTCAGTCGTCCTGGCTCTCCCTCTCTCCCCCAGTATTCGGCTGGCCACCATGAGTGCTTTGTCGGTGCTGATCTCAGTGGATGCTGTCTTGGGGAAGGTCAACTTGGCGCAGTTGGTGGTGATGGTGCTGGTGGAGGTGACAGCTTTAGGCAACCTGAGGATGGTCATCAGTAATATCTTCAACGTGAGTCATGGTGCTGGGAGGAGGGACCTGGGAGAAAAGGGCCAAAAGCTCCATTTGGTGGGGTTTCCAGGGTTTTGAAAAATAAAGACAACCTGTAATCCCAGCTACTTGGGAGGTTGAGGAGGG
SEQ ID NO:2
GAATGAGTGAGAGGCATCCTTCCTTCTCAGTCGTCCTGGCTCTCCCTCTCTCCCCCAGTATTCGGCTGGCCACCATGAGTGCTTTGTCGGTGCTGATCTCAGTGGATGCTGTCTTGGGGAAGGTCAACTTGGCGCAGTTGGTGGTGATGGTGCTGGTGGAGGTGACAGCTTTAGGCAACCTGAGGATGGTCATCAGTAATATCTTCAAAGTGAGTCATGGTGCTGGGAGGAGGGACCTGGGAGAAAAGGGCCAAAAGCTCCATTTGGTGGGGTTTCCAGGGTTTTGAAAAATAAAGACAACCTGTAATCCCAGCTACTTGGGAGGTTGAGGAGGG
A method of detecting the RHD486C > a allele comprising the steps of:
(1) Extracting DNA in a sample to be detected;
(2) Designing a specific primer by taking the DNA as a template to carry out PCR amplification reaction to obtain a PCR reaction product;
1) Primer design: according to RHD gene (sequence number: NC_ 000001.11) recorded by GenBank of National Center for Biological Information (NCBI), designing primers by Oligo 6.0 primer software, and finally determining 1 pair of specific oligonucleotide primer sequences, wherein the upstream primer sequences are shown in SEQ ID NO. 3 in Table 1; the sequence of the downstream primer is shown as SEQ ID NO. 4, and the length of the amplified product fragment is 150bp;
TABLE 1 RHD 4816C > A allele detection primer sequences and reaction specificity
Note that: * The position of the base sequence of the exon referred to by the oligonucleotide primer sequence refers to the entire arrangement sequence of 10 exons starting from the ATG initiation codon; the position of the base sequence of the intron referred to means the individual arrangement order of each intron.
2) Reaction system for PCR amplification:
the total volume of the reaction system is 50 mu L; wherein the PCR 5 Xbuffer solution comprises 10. Mu.L, DNA template 5.0. Mu.L, taq polymerase 1.0. Mu.L of 1U/. Mu.L, mgCl 2 The final concentration is 2.0mmol/L, the final concentration of dNTP is 200nmol/L, and the final concentrations of the specific forward primer and the specific reverse primer are 200nmol/L; adding sterilized double distilled water to the total volume of the reaction system of 50 mu L;
3) Reaction procedure for PCR amplification: the reaction system is reacted in a PCR instrument, and the reaction conditions are as follows: pre-denaturing at 95 ℃ for 5min, then sequentially denaturing at 95 ℃ for 30s, then annealing at 59 ℃ for 40s, and extending at 72 ℃ for 1min for 35 cycles;
(3) Sequencing the PCR amplification reaction product;
(4) Comparing the sequencing result with the sequence of the RHD wild-type gene to determine whether there is 486C-A mutation.
Example 2
Instrument: veriti 96 PCR instrument, BIO-RAD Gel Doc xr+ Gel imager (burle, usa), gel electrophoresis instrument (beijing hexa).
Reagent: QIAamp DNA extraction kit (Qiagen, germany); DNA Isolation Kit extraction kit (PELFREEZ company, beijing); PCR buffer, dNTP, taq enzyme (ABI Co., USA); primers were synthesized by Shanghai Biotechnology Co.
RHD486C > A allele detection was performed by the method of example 1, comprising the steps of:
(1) The method for extracting the DNA in the sample to be detected comprises the following specific steps:
1) One sterile 2.0mL centrifuge tube was taken and 1mL of cell lysate was added.
2) Gently shaking the EDTA anticoagulated whole blood sample until thoroughly mixed; then, 500. Mu.L of the blood sample was pipetted into the above centrifuge tube containing the cell lysate, and the centrifuge tube was gently poured and mixed for 5-6 times.
3) Incubate for 10 minutes at room temperature (during which the centrifuge tube is inverted 2-3 times for homogenization).
4) Centrifuge at 12000rpm for 5 minutes at room temperature.
5) The supernatant was slowly removed as clean as possible with a pipette, taking care not to aspirate the white material at the interface of the two phases.
6) Vortex shaker (Votex) was used to mix vigorously until the white blood cells were resuspended (10-15 seconds).
7) To the resuspended cell solution was added 300. Mu.L of nuclear lysate. The solution was aspirated and aspirated with a pipette tip 5-6 times to lyse leukocytes. At this point the solution should become very viscous. If the cell pellet is visible after mixing, the solution is incubated at 37℃until the pellet dissipates. If the cell pellet is still visible after 1 hour of incubation, 100. Mu.L of additional nuclear lysate is added and incubation at 37℃is repeated.
8) 100. Mu.L of protein pellet was added to the nuclear lysate and vigorously shaken with a vortex for 10-20 seconds.
9) Centrifuge at 12000rpm for 5 minutes at room temperature.
10 The supernatant was transferred to a correspondingly numbered 2.0mL centrifuge tube to which 300 μl of room temperature isopropyl alcohol had been added.
11 Gently invert to mix the solution until a white linear DNA precipitate forms.
12 12000rpm for 1 minute at room temperature.
13 The supernatant was discarded, 70% ethanol at room temperature was added in an equal volume to the sample size, and the tube was gently inverted several times.
14 The ethanol solution was slowly removed as clean as possible with a pipette. And (5) baking the centrifuge tube at 50 ℃ for 5-10 minutes, and volatilizing the residual ethanol liquid as much as possible.
15 50-100. Mu.L of DNA solution was added to the centrifuge tube and gently mixed.
16 DNA extraction was evaluated by 1% agarose gel electrophoresis, and the content was determined by Nanodrop nucleic acid, quantified to 20 ng/. Mu.L and stored at-20 ℃.
(2) Taking the DNA as a template, and adopting a specific primer (shown as SEQ ID NO:3 and SEQ ID NO: 4) to carry out PCR amplification reaction to obtain a PCR reaction product;
reaction system for PCR amplification:
the total volume of the reaction system is 50 mu L; wherein the PCR 5 Xbuffer solution comprises 10. Mu.L, DNA template 5.0. Mu.L, taq polymerase 1.0. Mu.L of 1U/. Mu.L, mgCl 2 The final concentration is 2.0mmol/L, the final concentration of dNTP is 200nmol/L, and the final concentrations of the specific forward primer and the specific reverse primer are 200nmol/L; adding sterilized double distilled water to the total volume of the reaction system of 50 mu L;
3) Reaction procedure for PCR amplification: the reaction system is reacted in a PCR instrument, and the reaction conditions are as follows: pre-denaturing at 95 ℃ for 5min, then sequentially denaturing at 95 ℃ for 30s, then annealing at 59 ℃ for 40s, and extending at 72 ℃ for 1min for 35 cycles;
(3) The amplified product obtained in the above step was subjected to electrophoresis using 1.5% agarose gel to detect whether it had the fragment of interest. And observing the result by a gel imager and photographing, wherein the PCR product shows a single band after electrophoresis, and has no impurity band, so that the PCR product is single and has no non-specific amplification. If the band position is at a position of appropriate size, it is the fragment of interest. As shown in fig. 1, M:50bp gradient molecular weight marker, 1: RHD486C > A mutant samples, 2: blank, 3: wild type control.
(4) Purification of amplification products: the PCR product after agarose gel electrophoresis was purified and recovered using a Agarose Gel DNAPurification Kit kit from Takara, and was prepared for sequencing.
(5) Sanger sequencing and outcome determination: the purified PCR products were sequenced on an ABI3730 type fully automatic DNA sequencer. The sequencing results were aligned with the RHD wild-type reference sequence (NCBI Reference Sequence: NC-000001.11) and reported according to the actual mutation. The mutation diagram of the detected gene is shown in figure 2, and the arrow in the diagram shows that the RHD gene shows the mutation of g.25290791C > A.
Claims (2)
1. The nucleotide sequence of the wild RHD gene is shown as SEQ ID NO. 1, the mutant RHD gene is shown as SEQ ID NO. 2, the mutation site is at 209 th site of the sequence shown as SEQ ID NO. 1, and the mutation site is changed into A from base C.
2. A specific primer for detecting RHD486C > A allele comprises an upstream primer and a downstream primer, wherein the sequence of the upstream primer is shown as SEQ ID NO. 3, and the sequence of the downstream primer is shown as SEQ ID NO. 4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310555826.3A CN116751786A (en) | 2023-05-17 | 2023-05-17 | Rh blood group D antigen RHD486C > A allele and detection primer thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310555826.3A CN116751786A (en) | 2023-05-17 | 2023-05-17 | Rh blood group D antigen RHD486C > A allele and detection primer thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116751786A true CN116751786A (en) | 2023-09-15 |
Family
ID=87950437
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310555826.3A Withdrawn CN116751786A (en) | 2023-05-17 | 2023-05-17 | Rh blood group D antigen RHD486C > A allele and detection primer thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116751786A (en) |
-
2023
- 2023-05-17 CN CN202310555826.3A patent/CN116751786A/en not_active Withdrawn
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| AU2004217872A1 (en) | Identification of fetal DNA and fetal cell markers in maternal plasma or serum | |
| CN106834287B (en) | SNP marker for detecting RhD negative phenotype | |
| CN111197091A (en) | Rh blood type DEL type RHD1073T & gtA allele and application thereof | |
| CN106967808B (en) | Primer group for detecting RhD negative blood type and application thereof | |
| CN103937806B (en) | A kind of Rh blood group DEL type RHD838G > A allelotrope and detection method thereof | |
| CN107190071B (en) | It is a kind of for detecting the SNP marker of RhD variation phenotypes | |
| CN111172297B (en) | RhD blood type gene RHD993C & gtT allele and application thereof | |
| CN111304211B (en) | RHD-T268A mutant and detection thereof | |
| CN111218453B (en) | RhD blood type antigen RHD-G353A mutant and detection | |
| CN110029167B (en) | ERBB2-G519V mutant gene and application thereof in breast cancer auxiliary diagnosis | |
| CN111808937A (en) | Fut 1508 dupT allele of Bombay blood group and detection method and application thereof | |
| CN112941205A (en) | RhD blood group gene RHD634G & gtA allele and detection | |
| CN113667673B (en) | Montellike FUT1236delG allele and detection method and application thereof | |
| CN112940099B (en) | RhD-T163P mutant and detection thereof | |
| CN116751786A (en) | Rh blood group D antigen RHD486C > A allele and detection primer thereof | |
| CN103045722B (en) | Detection kit of disease-causing gene CRYGD of crystalline congenital cataract | |
| CN111154766B (en) | RHD-S68R mutant and detection method thereof | |
| Jiao et al. | Gene identification of rare B (A) blood group | |
| JP4889258B2 (en) | Method for determining resistance to the onset of bovine leukemia | |
| Sano et al. | ABO chimerism with a minor allele detected by the peptide nucleic acid-mediated polymerase chain reaction clamping method | |
| CN110055258B (en) | Breast cancer related gene ERBB2 site g.3939700G > A mutant and application thereof | |
| CN111154850A (en) | RHD939G & gtA allele and detection method thereof | |
| CN115011702B (en) | Allele of RhD blood type gene RHD56C & gtG and detection thereof | |
| CN116656828A (en) | Application of gene ERBB2 locus g.3974028A > G mutation in preparation of breast cancer early-stage screening kit | |
| JP2001500001A (en) | Methods and reagents for typing HLA class I genes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20230915 |