WO2014065410A1 - Procédé et trousse pour le typage d'adn de gène hla - Google Patents

Procédé et trousse pour le typage d'adn de gène hla Download PDF

Info

Publication number
WO2014065410A1
WO2014065410A1 PCT/JP2013/079007 JP2013079007W WO2014065410A1 WO 2014065410 A1 WO2014065410 A1 WO 2014065410A1 JP 2013079007 W JP2013079007 W JP 2013079007W WO 2014065410 A1 WO2014065410 A1 WO 2014065410A1
Authority
WO
WIPO (PCT)
Prior art keywords
hla
seq
gene
base sequence
sequence shown
Prior art date
Application number
PCT/JP2013/079007
Other languages
English (en)
Japanese (ja)
Inventor
一善 細道
滋樹 光永
猪子　英俊
逸朗 井ノ上
Original Assignee
ジェノダイブファーマ株式会社
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by ジェノダイブファーマ株式会社 filed Critical ジェノダイブファーマ株式会社
Publication of WO2014065410A1 publication Critical patent/WO2014065410A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6881—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the present invention relates to a method and kit for DNA typing of HLA genes using a massively parallel sequencer.
  • HLA Human leukocyte antigen
  • MHC histocompatibility complex
  • HLA class I antigen consists of ⁇ -chain showing high polymorphism and ⁇ 2-microglobulin with little polymorphism
  • HLA class II antigen is ⁇ -chain with high polymorphism and ⁇ with little polymorphism. Consists of chains. ⁇ chain of class I molecule is encoded by HLA-A, HLA-B and HLA-C genes
  • ⁇ chain of class II antigen is HLA-DRB1, HLA-DQB1, HLA-DPB1
  • ⁇ chain is HLA-DRA1 , HLA-DQA1, and HLA-DPA1 gene.
  • exon 2 and exon 3 of the gene encoding the ⁇ chain show high polymorphism in the HLA class I antigen, and exon 2 of the gene encoding the ⁇ chain in the HLA class II antigen is advanced. Show polymorphism.
  • the gene region encoding HLA is located in human chromosome 6 short arm 6p21.3, class I region (HLA-A, HLA-C, HLA-B, etc.) from the telomere side toward the centromere side, Class III region, class II region (HLA-DRA, HLA-DRB1, HLA-DQA1, HLA-DQB1, HLA-DPA1, HLA-DPB1, etc.) are arranged in this order, and many genes are encoded at very high density. The relationship between blood transfusion and transplantation and various diseases has been reported. There is no HLA gene in the class III region, and genes such as complement components and tumor necrosis factor (TNF) are present.
  • TNF tumor necrosis factor
  • HLA-DRB gene region encoding the ⁇ chain of the HLA-DR antigen.
  • pseudogenes such as HLA-DRB6 and HLA-DRB9 are located on the same chromosome in addition to HLA-DRB1.
  • HLA-DRB5 (DR51) gene and pseudogenes such as HLA-DRB6 and HLA-DRB9 are located on the same chromosome in addition to HLA-DRB1.
  • DR3 DR52
  • HLA-DRB2 HLA-DRB9
  • DR4 DR7 and DR9 types
  • pseudogenes such as HLA-DRB4 (DR53) gene
  • HLA-DRB7, HLA-DRB8 and HLA-DRB9 are located on the same chromosome.
  • DR8 type no HLA-DRB gene other than HLA-DRB1 is located on the same chromosome.
  • Each allele exon has a plurality of polymorphic regions, and the base sequence (amino acid sequence) of a polymorphic region is often common to a plurality of alleles. That is, each HLA allele is defined by a combination of multiple polymorphic regions. In the HLA class I antigen, exon 2 or exon 3 having the same base sequence as well as the polymorphic region in the exon may be common to a plurality of alleles.
  • HLA is known to have an extremely large number of alleles due to the presence of advanced polymorphisms, and their notation is also determined. That is, the first zone for discriminating serological HLA type (2 digit level), the second zone for discriminating alleles with amino acid substitution within the same serological HLA type (4 digit level), and no amino acid mutation The third zone (6 digit level) that discriminates alleles in which base substitution is recognized, and the fourth zone (8 digit level) that discriminates alleles with base substitution outside the gene region encoding the HLA molecule (intron). .
  • the conventional method is a DNA typing method using PCR centering on the exon region of each gene, it misses base substitution in the intron region and the promoter region, and as a result, the gene structure has other expression HLA. It was possible to miss the detection of a null allele that is not different from the gene but whose expression is suppressed.
  • An object of the present invention is to provide a highly accurate DNA typing method and kit that eliminates ambiguity derived from phase ambiguity.
  • the present invention provides an HLA DNA typing method including the following steps.
  • Primer sets sequences that specifically anneal to the upstream region and downstream region of each gene of HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DPB1, and HLA-DQB1 in the human genome sequence
  • Preparing a pair of oligonucleotides comprising a base sequence shown in any of Nos. 1 to 12 Preparing a pair of oligonucleotides comprising a base sequence shown in any of Nos. 1 to 12;
  • PCR-amplifying a test sample (DNA) using the primer set (3) determining the base sequence of the PCR amplified product; and (4) performing a homology search with the database.
  • the method of the present invention provides the entire base sequence necessary for DNA typing of the 6-locus of the HLA gene from one molecule, so that the ultimate DNA typing in which phase ambiguity with unknown cis-trans positional relationship is eliminated Is the law. As a result, high-accuracy HLA matching between the transplant candidate and the donor candidate at the time of transplantation is realized. Since the entire nucleotide sequence of the gene including peripheral regions such as the promoter region, exon region, and intron region of the HLA gene is determined, it is possible to detect null alleles or novel alleles that are not expressed at all or whose expression is suppressed.
  • (A) It is a figure which shows the relationship between HLA class I gene structure and molecular structure.
  • (B) It is a figure which shows the structure of the promoter area
  • (A) It is a figure which shows the relationship between HLA class II gene structure and molecular structure.
  • (B) It is a figure which shows the structure of the promoter area
  • Primer set preparation step In the DNA typing method of the present invention, first, upstream of each gene of HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DPB1, and HLA-DQB1 in the human genome sequence.
  • a primer set (a pair of oligonucleotides comprising a base sequence shown in any of SEQ ID NOs: 1 to 12) that specifically anneals to each of the region and the downstream region is prepared.
  • the genomic base sequence of human chromosome 6 (6p21.3) including the region where the HLA gene is present has already been elucidated, and the relationship between the gene structure and the structure of the expression product (HLA molecule) is also known (Fig. 1 and FIG. 2).
  • HLA-A, HLA-B and HLA-C which are called classical HLA class I molecules, contain 7 or 8 exons (FIG. 1 (a)).
  • HLA-A, HLA-B and HLA-C which are called classical HLA class I molecules, contain 7 or 8 exons (FIG. 1 (a)).
  • enhancers and promoter regions There are two types of enhancers and promoter regions, and expression is regulated (FIG. 1 (b)).
  • the conventional DNA typing method particularly performs PCR using primers prepared based on exons 2 and 3. As a result, the phase ambiguity problem has arisen as described above.
  • HLA-DR, HLA-DQ, and HLA-DP which are said to be classical HLA class II molecules, are composed of an ⁇ chain and a ⁇ chain, and each gene contains 5 or 6 exons (FIG. 2).
  • A) the promoter region is located outside exon 1, and expression is regulated (FIG. 2 (b)).
  • PCR using a primer prepared based on exon 2 is performed, and accordingly As mentioned above, there was a phase ambiguity problem.
  • the classical class I molecules HLA-A, HLA-B, HLA-C
  • all of the classical class II molecules HLA-DRB1, HLA-DPB1 and HLA-DQB1 gene regions (not only exons) Intron, 5 'and 3' untranslated regions, including a promoter region)
  • a primer set capable of PCR amplification is prepared, and PCR amplified PCR products are used for next-generation sequencing described below.
  • Uncertainties such as phase ambiguity can be eliminated, and the presence or absence of null allyl can be accurately detected.
  • PCR primer sets listed in Table 1 below are prepared.
  • SEQ ID NOs: 1 and 2 in Table 1 are PCR primer sets that specifically amplify the HLA-A gene, which is an MHC class I ⁇ chain. These primer sets are base sequences present at positions sandwiching the entire region of the HLA-A gene (including promoter, exon and intron) from the upstream side and the downstream side in the human genome base sequence (reference sequence: hg19).
  • SEQ ID NO: 1 is a forward primer having a nucleotide sequence corresponding to the 29,910,189th position to the 29,910,212th position from the telomere side in the HLA region of the human genome sequence.
  • SEQ ID NO: 2 is a reverse primer having a base sequence complementary to a base corresponding to the 29th, 913, 562th to 29th, 913, 586th from the telomere side in the HLA region of the human genome sequence sequence.
  • the expected length of PCR products obtained using these primer sets is about 3,398 bases (bp).
  • SEQ ID NOs: 3 and 4 in Table 1 are PCR primer sets that specifically amplify the HLA-B gene, which is an MHC class I ⁇ chain. These primer sets are base sequences present at positions sandwiching the entire region of the HLA-B gene (including promoter, exon and intron) from the upstream side and the downstream side in the human genome base sequence (reference sequence: hg19).
  • SEQ ID NO: 3 is a forward primer having a base sequence corresponding to the 31,321,366th position to the 31,321,392rd position from the telomere side in the HLA region of the human genome base sequence.
  • SEQ ID NO: 4 is a reverse primer having a base sequence complementary to the bases corresponding to the 31st, 325th, 632rd to 31st, 325th, 661th positions from the telomere side in the HLA region of the human genome sequence sequence.
  • the expected length of PCR products obtained using these primer sets is approximately 4,296 bases (bp).
  • SEQ ID NOs: 5 and 6 in Table 1 are PCR primer sets that specifically amplify the HLA-C gene, which is an MHC class I ⁇ chain. These primer sets are base sequences present at positions sandwiching the entire region of the HLA-C gene (including promoter, exon and intron) from the upstream side and the downstream side in the human genome base sequence (reference sequence: hg19).
  • SEQ ID NO: 5 is a forward primer having a base sequence corresponding to the 31st, 235, 959th to the 31st, 235, 982th from the telomere side in the HLA region of the human genome base sequence.
  • SEQ ID NO: 6 is a reverse primer having a base sequence complementary to the bases corresponding to the 31st, 240th, 373rd to 31st, 240th, 398th from the telomere side in the HLA region of the human genome sequence sequence.
  • the expected length of PCR products obtained using these primer sets is about 4,440 bases (bp).
  • SEQ ID NOs: 7 and 8 in Table 1 are PCR primer sets that specifically amplify the HLA-DRB1 gene, which is an MHC class II ⁇ chain. These primer sets are base sequences present at positions sandwiching the entire region of the HLA-DRB1 gene (including promoter, exon and intron) from the upstream side and the downstream side in the human genome base sequence (reference sequence: hg19).
  • SEQ ID NO: 7 is a forward primer having a nucleotide sequence corresponding to the 32,546,220th position to the 32,546,247th position from the telomere side in the HLA region of the human genome sequence.
  • SEQ ID NO: 8 is a reverse primer having a base sequence complementary to the bases corresponding to the 32,558,090th position to the 32,558,118th position from the telomere side in the HLA region of the human genome sequence sequence.
  • the expected length of PCR products obtained using these primer sets is approximately 11,899 bases (bp).
  • SEQ ID NOs: 9 and 10 in Table 1 are PCR primer sets that specifically amplify the HLA-DPB1 gene, which is an MHC class II ⁇ chain. These primer sets are base sequences present at positions sandwiching the entire region of the HLA-DPB1 gene (including promoter, exon and intron) from the upstream side and the downstream side in the human genome base sequence (reference sequence: hg19).
  • SEQ ID NO: 9 is a forward primer having a base sequence corresponding to the 33,042,795th to 33,042,823th from the telomere side in the HLA region of the human genome base sequence.
  • SEQ ID NO: 10 is a reverse primer having a base sequence complementary to the base corresponding to the 33,056,373rd position to the 33,056,399th position from the telomere side in the HLA region of the human genome sequence sequence.
  • the expected length of the PCR product obtained using these primer sets is approximately 13,605 bases (bp).
  • SEQ ID NOs: 11 and 12 in Table 1 are PCR primer sets that specifically amplify the HLA-DQB1 gene, which is an MHC class II ⁇ chain. These primer sets are base sequences present at positions sandwiching the entire region of the HLA-DQB1 gene (including promoter, exon and intron) from the upstream side and the downstream side in the human genome base sequence (reference sequence: hg19).
  • SEQ ID NO: 11 is a forward primer having a nucleotide sequence corresponding to the 32,627,718th position to the 32,627,743th position from the telomere side in the HLA region of the human genome sequence.
  • SEQ ID NO: 12 is a reverse primer having a base sequence complementary to the base corresponding to the 32,634,812th position to the 32,634,835th position from the telomere side in the HLA region of the human genome sequence sequence.
  • the expected length of PCR products obtained using these primer sets is approximately 7,118 bases (bp).
  • primers can be prepared by a technique usually used in this field.
  • the primer set shown in Table 1 shows the most preferable example.
  • forward In the method of the present invention, forward (Forward) in which the entire region of each gene of HLA can be annealed at a position between the upstream side and the downstream side.
  • a set of a primer and a reverse primer those having 1 to several base substitutions, insertions or deletions in the base sequences shown in SEQ ID NOs: 1 to 12 can be similarly used.
  • the test sample (DNA) is PCR amplified using the primer set prepared in step (1).
  • the PCR amplification reaction is performed according to a normal protocol. Specifically, it is as follows. 1. DNA is extracted from the sample according to the form of the test sample. 2. The extracted DNA is quantified, and a primer solution is appropriately set to prepare a reaction solution. 3. Set the reaction conditions and perform the PCR reaction. Example: Thermal denaturation step (usually 92-97 ° C) Annealing step (usually 55-72 ° C) Elongation step (usually 65-80 ° C) 4). The obtained PCR product is purified and subjected to the next nucleotide sequencing step.
  • step (3) A base sequence determination step. Next, the base sequence of the PCR product (amplified DNA) generated in step (2) is determined. This step is preferably performed by using a so-called next-generation sequence (or ultra-high speed sequence). For the next-generation sequence, see, for example, “Experimental Medicine” Vol. 27, No. 1, 2009 (Yodosha).
  • a DNA library is prepared using a Nextera TM DNA sample preparation kit manufactured by Illumina and sequenced using a genome sequencer MiSeq system (Illumina).
  • the library can be prepared in a short time of about 90 minutes or less. Paired end analysis is preferably used for sequencing the resulting sample.
  • step (3) by comparing the base sequence obtained in step (3) with the data of the base sequence database of known HLA alleles, the allele type of the DNA contained in the test sample (Up to 8 digits) is determined.
  • the primer set described in Table 1 is used as a representative example. It is characterized in that primers are set at positions sandwiching the entire HLA class I and HLA class II gene regions, and the amplified DNA is sequenced over almost the entire region of each gene. Guity (uncertainty) can be eliminated and information on null allyl can be obtained. That is, the present invention also provides a kit for HLA genotyping comprising a primer comprising an oligonucleotide having the base sequence represented by SEQ ID NOs: 1-12.
  • PCR reaction was performed using each HLA gene-specific primer set (see Table 1) using the already extracted genomic DNA as a template.
  • the specific procedure is as follows.
  • (1) PCR amplification was performed using Prime STAR GXL polymerase (TaKaRa) with the following composition.
  • Genomic DNA solution (10 ng) 0.5 ⁇ L 5x PrimeSTAR GXL buffer 2 ⁇ L dNTP solution 0.8 ⁇ L Forward primer (5 pmol) 0.4 ⁇ L Reverse primer (5 pmol) 0.4 ⁇ L PrimeSTAR GXL polymerase 0.4 ⁇ L Sterile water 5.5 ⁇ L Total 10 ⁇ L
  • the base sequence of the PCR product was determined. Specifically, it was performed as follows. A DNA library was prepared from the resulting PCR amplification product using the Nextera TM DNA sample preparation kit (Illumina) and sequenced at 150 bp paired end using the MiSeq system (Illumina). Nextera TM DNA sample preparation kit can prepare DNA library in 90 minutes including reaction time by simultaneously performing DNA cleavage called tagmentation and insertion of tag sequence via Nextera transposome. It has the feature of becoming. Further, by using two types of indexes, a maximum of 96 types (Index 1: 8 types + Index 2: 12 types) of samples can be analyzed simultaneously. Another feature is that, unlike other next-generation sequencer DNA library preparations, the starting DNA requirement is only 50 ng.
  • the insert size of the prepared DNA library is as wide as 200 bp to 1 kb or more, which is a feature and merit of this method.
  • HLA typing is performed by data analysis. This data analysis is the most original point of this method.
  • the sequence reads obtained were mapped by BWA (Burrows-Wheeler Aligner) and SAMTools (Sequence Alignment / Map tool) against the reference sequence of each PCR amplification region to obtain analysis results as diplotypes, and Picard and Genome A base different from the reference sequence is detected by Analysis Toolkit (GATK).
  • BWA Brownrows-Wheeler Aligner
  • SAMTools Sequence Alignment / Map tool
  • the phases are classified based on the SNV only based on the pair information shown in the sequence read, and this phase is divided into 2 based on the SNV of the haplotype that is divided.
  • Two HLA gene sequences are generated.
  • one paired-end sequence lead needs to contain two or more SNVs.
  • the distance between two or more SNVs varies, but the HLA gene is very diverse and the insert size of the DNA library prepared with the Nextera TM DNA sample preparation kit is 200 bp to 1 kb or more.
  • the wide feature makes it possible to specify a phase containing two or more SNVs.
  • the confirmation of the SNV divided into phases and the phase of Indel are determined.
  • Indel detected in the first mapping result includes false positives, but in the second mapping, it is mapped to a base sequence having no SNV, so that Indel can be determined with certainty.
  • the two consensus sequences obtained in the second mapping are the sequences of the two HLA genes, and searching this in the IMGT / HLA database reveals whether the HLA allele is new or known. If the HLA allyl is known, the HLA allyl number can be obtained.
  • PCR primers that specifically amplify each of HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DPB1, and HLA-DQB1 were designed, and PCR was performed under the conditions described above.
  • PCR amplification product was electrophoresed by agarose gel electrophoresis, a single amplification product was obtained at the position of the target molecular weight for each HLA gene (FIG. 4).
  • Table 2 shows the HLA allyl type determined using the method of the present invention and the HLA allyl type determined using the conventional SSO method for some of the samples employed. While the SSO method could only specify up to the 4-digit level, the method (Example) of the present invention could determine the allyl type in detail up to the 6-digit or 8-digit level.
  • the method of the present invention enables 8-digit level HLA typing without phase ambiguity, and efficiently detects base substitutions, insertions and deletions in promoters and introns that cause null alleles. It is considered an excellent tool.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Analytical Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'objet de la présente invention est de fournir un procédé et une trousse pour le typage d'ADN hautement précis, qui permet d'éliminer l'ambiguïté issue d'une ambiguïté de phase. La solution selon l'invention comprend un procédé pour le typage d'ADN de HLA caractérisé en ce qu'il comprend : (1) une étape de préparation d'un ensemble d'amorces (SEQ ID NO: 1-12) spécifiquement pour s'hybrider respectivement à une région en amont et à une région en aval de chacun des gènes HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DPB1 et HLA-DQB1, dans la séquence de base du génome humain ; (2) une étape visant à soumettre un échantillon de sujet (ADN) à une amplification par PCR à l'aide de l'ensemble d'amorces ; (3) une étape de détermination de la séquence de base d'un produit amplifié par PCR ; et (4) une étape de recherche d'homologie dans une base de donnée.
PCT/JP2013/079007 2012-10-26 2013-10-25 Procédé et trousse pour le typage d'adn de gène hla WO2014065410A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2012-237124 2012-10-26
JP2012237124A JP2016010318A (ja) 2012-10-26 2012-10-26 Ｈｌａ遺伝子のｄｎａタイピング方法及びキット

Publications (1)

Publication Number Publication Date
WO2014065410A1 true WO2014065410A1 (fr) 2014-05-01

Family

ID=50544781

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2013/079007 WO2014065410A1 (fr) 2012-10-26 2013-10-25 Procédé et trousse pour le typage d'adn de gène hla

Country Status (2)

Country Link
JP (1) JP2016010318A (fr)
WO (1) WO2014065410A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106434863A (zh) * 2016-06-15 2017-02-22 广州医科大学附属第二医院 一种鉴定hla‑dqb1第2外显子单倍型的方法
EP3006571A4 (fr) * 2013-05-09 2017-02-22 Genodive Pharma Inc. Méthode et kit de typage multiplex de l'adn du gène hla
WO2018147438A1 (fr) * 2017-02-13 2018-08-16 国立大学法人京都大学 Ensemble d'amorces de pcr pour gène hla, et procédé de séquençage utilisant ledit ensemble d'amorces de pcr
EP3626835A1 (fr) 2018-09-18 2020-03-25 Sistemas Genómicos, S.L. Procédé pour identification génotypique des deux allèles d'au moins un locus du gène hla d'un sujet
WO2021191634A1 (fr) * 2020-03-27 2021-09-30 The University Of Birmingham Procédés, compositions et kits de typage hla
CN113755593A (zh) * 2021-09-23 2021-12-07 中南大学 检测HLA-A基因SNP标记rs1136697-G的PCR扩增引物、试剂盒及方法
WO2023060871A1 (fr) * 2021-10-15 2023-04-20 西安浩瑞基因技术有限公司 Amorce d'amplification du gène hla, kit, procédé d'établissement d'une banque de séquençage, et procédé de séquençage

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013011734A1 (fr) * 2011-07-21 2013-01-24 ジェノダイブファーマ株式会社 Procédé et trousse pour le typage d'adn d'un gène hla

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013011734A1 (fr) * 2011-07-21 2013-01-24 ジェノダイブファーマ株式会社 Procédé et trousse pour le typage d'adn d'un gène hla

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
SHIINA T. ET AL.: "Super high resolution for single molecule-sequence-based typing of classical HLA loci at the 8-digit level using next generation sequencers", TISSUE ANTIGENS, vol. 80, 4 August 2012 (2012-08-04), pages 305 - 316 *
SHINGO SUZUKI ET AL.: "Jisedai Sequencer o Mochiita HLA Class I Idenshi no Cho Ko Kaizodo DNA Typing(Super high resolution Single molecule-Sequence Based Typing;SS-SBT)-ho no Kaihatsu", MHC, vol. 19, no. 1, 20 April 2012 (2012-04-20), pages 43 - 54 *
TAKASHI SHIINA ET AL.: "HLA Kenkyu no Rinsho Oyo o Mezashita 8 Keta Level HLA Typing-ho no Kaihatsu", MHC, vol. 19, no. 2, 10 August 2012 (2012-08-10), pages 133 *

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10711306B2 (en) 2013-05-09 2020-07-14 Genodive Pharma Inc. Method and kit for multiplex DNA typing of HLA gene
EP3006571A4 (fr) * 2013-05-09 2017-02-22 Genodive Pharma Inc. Méthode et kit de typage multiplex de l'adn du gène hla
CN106434863B (zh) * 2016-06-15 2021-04-23 广州医科大学附属第二医院 一种鉴定hla-dqb1第2外显子单倍型的方法
CN106434863A (zh) * 2016-06-15 2017-02-22 广州医科大学附属第二医院 一种鉴定hla‑dqb1第2外显子单倍型的方法
JP2018130036A (ja) * 2017-02-13 2018-08-23 国立大学法人京都大学 Ｈｌａ遺伝子のｐｃｒプライマーセット及びそれを用いたシークエンス法
CN110494562A (zh) * 2017-02-13 2019-11-22 国立大学法人京都大学 用于hla基因的pcr引物组和使用其的测序方法
EP3581652A4 (fr) * 2017-02-13 2021-03-10 Kyoto University Ensemble d'amorces de pcr pour gène hla, et procédé de séquençage utilisant ledit ensemble d'amorces de pcr
WO2018147438A1 (fr) * 2017-02-13 2018-08-16 国立大学法人京都大学 Ensemble d'amorces de pcr pour gène hla, et procédé de séquençage utilisant ledit ensemble d'amorces de pcr
EP3626835A1 (fr) 2018-09-18 2020-03-25 Sistemas Genómicos, S.L. Procédé pour identification génotypique des deux allèles d'au moins un locus du gène hla d'un sujet
WO2021191634A1 (fr) * 2020-03-27 2021-09-30 The University Of Birmingham Procédés, compositions et kits de typage hla
CN113755593A (zh) * 2021-09-23 2021-12-07 中南大学 检测HLA-A基因SNP标记rs1136697-G的PCR扩增引物、试剂盒及方法
CN113755593B (zh) * 2021-09-23 2024-02-20 中南大学 检测HLA-A基因SNP标记rs1136697-G的PCR扩增引物、试剂盒及方法
WO2023060871A1 (fr) * 2021-10-15 2023-04-20 西安浩瑞基因技术有限公司 Amorce d'amplification du gène hla, kit, procédé d'établissement d'une banque de séquençage, et procédé de séquençage

Also Published As

Publication number Publication date
JP2016010318A (ja) 2016-01-21

Similar Documents

Publication Publication Date Title
Norman et al. Sequences of 95 human MHC haplotypes reveal extreme coding variation in genes other than highly polymorphic HLA class I and II
JP6809977B2 (ja) Ｈｌａ遺伝子のｄｎａタイピング方法及びキット
Krause-Kyora et al. Ancient DNA study reveals HLA susceptibility locus for leprosy in medieval Europeans
JP6308724B2 (ja) Ｈｌａ遺伝子のマルチプレックスｄｎａタイピング方法及びキット
WO2014065410A1 (fr) Procédé et trousse pour le typage d'adn de gène hla
US9920370B2 (en) Haplotying of HLA loci with ultra-deep shotgun sequencing
Gabriel et al. Rapid high-throughput human leukocyte antigen typing by massively parallel pyrosequencing for high-resolution allele identification
AU2014355369B2 (en) Simple method and kit for DNA profiling of HLA genes by high-throughput massively parallel sequencer
Smith et al. Next generation sequencing to determine HLA class II genotypes in a cohort of hematopoietic cell transplant patients and donors
Castelli et al. HLA-G variability and haplotypes detected by massively parallel sequencing procedures in the geographicaly distinct population samples of Brazil and Cyprus
JP2022002539A (ja) 主要組織適合遺伝子複合体一塩基多型
WO2017135396A1 (fr) Jeu de sondes pour génotypage hla par procédé de capture sans utilisation de pcr, et procédé de typage dans lequel il est utilisé
Schiller et al. A simplified method for screening siblings for HLA identity using short tandem repeat (STR) polymorphisms
Lin et al. Characterization of the novel HLA allele, HLA‐DRB1* 08: 76.
CN109355366A (zh) Hla-b高分辨基因测序试剂盒
Keppen et al. HLA‐C* 04: 01: 01: 78, a novel variant of HLA‐C* 04, detected in a Sumi individual from Nagaland, North‐East India
Suzuki et al. Identification of three novel HLA‐DRA alleles by next‐generation sequencing.
Calvo‐Ferrer et al. Identification of a novel HLA‐C* 07 allele, HLA‐C* 07: 943, in a Spanish individual.
Claeys Benchmark of NGS-based prediction algorithms for
WO2023196925A2 (fr) Méthodes et systèmes de typage d'allèles
Esfahani et al. Characterization and specification of microsatellite markers in the HLA-DRB1 gene region: A revision to major histocompatibility complex database
Tan et al. Identification of Class I HLA Alleles in Anonymized Cell Therapy Specimens through Real-Time PCR with Melt-Curve Analysis
CN115354072A (zh) 一种用于hla-dqa1基因分型的引物组和分析方法
JP2015027283A (ja) カニクイザルｍｈｃ遺伝子のマルチプレックスｄｎａタイピング方法及びプライマーセット
De Santis et al. 16IHIW HLA typing by NGS: Workshop review

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13848260

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 13848260

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: JP