WO2013060004A1 - Procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et trousse d'analyse associée - Google Patents

Procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et trousse d'analyse associée Download PDF

Info

Publication number
WO2013060004A1
WO2013060004A1 PCT/CN2011/081359 CN2011081359W WO2013060004A1 WO 2013060004 A1 WO2013060004 A1 WO 2013060004A1 CN 2011081359 W CN2011081359 W CN 2011081359W WO 2013060004 A1 WO2013060004 A1 WO 2013060004A1
Authority
WO
WIPO (PCT)
Prior art keywords
ankylosing spondylitis
base
pcr
dna
reaction
Prior art date
Application number
PCT/CN2011/081359
Other languages
English (en)
Chinese (zh)
Inventor
古洁若
Original Assignee
Gu Jieruo
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 Gu Jieruo filed Critical Gu Jieruo
Priority to PCT/CN2011/081359 priority Critical patent/WO2013060004A1/fr
Priority to CN201180067457.9A priority patent/CN103502451B/zh
Publication of WO2013060004A1 publication Critical patent/WO2013060004A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • 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
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • 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 specific single nucleotide polymorphism associated with detecting ankylosing spondylitis
  • Ankylosing spondylitis is a subtype of spondyloarthropathies (SPA). Its main clinical manifestations are central axis manifestations such as inflammatory low back pain, morning stiffness, and restricted spinal activity, or / and concurrent peripheral arthritis, tendon end, ligament attachment point inflammation, to the advanced stage of the disease, there will be spinal rigidity or deformity.
  • Previous studies have suggested that the role of genetic factors plays a more important role in the pathogenesis of AS, and previous twin-based studies have shown that high consistent incidence in monozygotic and fraternal twins suggests genetic factors. The role in the pathogenesis of AS exceeds 90%.
  • HLA-B27 is an important gene that is also thought to be related to the genetic aggregation of AS family.
  • MHC also known as the major histocompatibility complex
  • HLA-B27 is a tightly linked group of genes encoding major histocompatibility antigens on a chromosome of the vertebrate, associated with immune responses, immune regulation, and transplant rejection. Studies have shown that in monozygotic twins, the consistent probability of HLA-B27 is 63%, and the consistent probability of HLA-B27 in twins twins is 23%.
  • HLA-B27-positive first-degree relatives in AS patients are 6-16 times more likely to have a family history of HLA-B27-positive individuals, suggesting that non-HLA-B27 familial factors are present in the disease. Development also plays an important role. This opens up a whole new way of thinking about the mechanism of the occurrence and development of ankylosing spondylitis. Through the whole-genome SNP chip scanning and analysis, we can find the susceptible SNP associated with ankylosing spondylitis and explore the ankylosing spondylitis. Pathogenic or susceptible genes.
  • Single nucleotide polymorphism mainly refers to DNA sequence polymorphism caused by single nucleotide variation at the genomic level, which is the most common one in human heritable variation. Species, accounting for more than 90% of all known polymorphisms. SNPs are widespread in the human genome, with an average of one in every 500 to 1000 base pairs, with an estimated total of 3 million or more. The polymorphisms exhibited by SNPs involve only a single base variation, which can be caused by a single base transition or transversion, or by the insertion or deletion of a base. In genomic DNA, any base may be mutated, so SNPs may be both within the gene sequence and possibly non-coding sequences outside the gene.
  • cSNP SNP located in the coding region
  • the mutation rate is only 1/5 of the surrounding sequence, but it is of great significance in the study of hereditary diseases. Therefore, the research of cSNP is more concerned.
  • cSNP can be further divided into two types: one is synonymous cSNP (synonymous cSNP), that is, the change of the coding sequence caused by SNP does not affect the amino acid sequence of the translated protein.
  • the mutated base has the same meaning as the unmutated base; the other is a non-synonymous cSNP, which means that the change in the base sequence can change the protein sequence translated by the blueprint, thereby affecting the protein.
  • the function, this change is often the direct cause of changes in biological traits, about half of cSNPs are non-synonymous cSNPs; in addition, SNPs can also be used as genetic markers to help find and identify disease susceptibility genes.
  • Brown et al. performed a full scan and verification of a large sample of AS patients and healthy volunteers based on a self-made intensive SNP chip. It also pointed out that the two genes IL23R and ARTS1 may be new to AS. Sense gene, followed by the research of ARTS1 gene in Korean and Chinese research institutions, the results suggest that ARTS1 is associated with AS; Chinese research institutions have also verified IL-23R, the results No correlation was found between the two, suggesting that there may be differences between different populations. Moreover, although the above-mentioned Brown's research in 2007 used a dense SNP chip, the chip did not perform a full scan of the dense SNP for this genome, but only customized the intensive SNP chip for the location of interest, thus causing the chip.
  • Illumina Human lM-duo chip With the rapid development of chip technology, Illumina Human lM-duo chip, Illumina Human 610-Quad chip and Illumina OmniExpress chip have emerged to provide more dense SNP mark, covering a wider range of areas.
  • Illumina Human lM-duo chip With the rapid development of chip technology, Illumina Human 610-Quad chip and Illumina OmniExpress chip have emerged to provide more dense SNP mark, covering a wider range of areas.
  • GWAS Genome-wide association analysis
  • ankylosing spondylitis is a relatively common disease, the course of the disease is lingering, and it is easy to cause disability, it should be an early diagnosis and treatment, especially for young people aged 16-25.
  • SNPs that are closely related to ankylosing spondylitis, and to design methods and kits for detecting the specificity of ankylosing spondylitis.
  • the present invention is directed to the deficiencies of the currently lacking specificity of detection methods and related kits in the early diagnosis of AS, providing a highly specific examination method and providing a dedicated kit.
  • SNP rs4552569 can be used as a specific SNP for detecting ankylosing spondylitis. It is an important auxiliary diagnostic indicator for ankylosing spondylitis in the early stage, and the present invention has been completed.
  • One of the objects of the present application is to provide a nucleotide fragment of a pathogenicity-related single nucleotide polymorphism (SNP) of ankylosing spondylitis at an rs4552569 site, ankylosing spondylitis (AS)
  • SNP pathogenicity-related single nucleotide polymorphism
  • AS ankylosing spondylitis
  • One of the objects of the present application is to provide a method for early auxiliary diagnosis of ankylosing spondylitis, which is achieved by the following scheme: obtaining a nucleotide sequence from a sample to be tested, and detecting the position at rs4552569 Whether the base on the base is C, and if the base of the site is C, the incidence of ankylosing spondylitis is significantly increased.
  • the sample includes blood, body fluid or tissue.
  • the method includes the following steps:
  • the method of obtaining the nucleotide sequence from the sample to be tested further includes fluorescent quantitative PCR, denaturing high performance liquid chromatography, restriction fragment length polymorphism, ligase detection or DNA fragment sequencing.
  • One of the objects of the present application is to provide a method for detecting an ankylosing spondylitis-specific single nucleotide polymorphism, the method comprising the steps of:
  • PCR amplification primers and single base extension primers of rs4552569 site were designed using Sequenom Genotyping Tools and MassARRAY Assay Design software;
  • DNA from tissues, cells or blood samples was extracted using the TIANamp Blood DNA kit or NucleoSpin Tissue (MN), quantified by spectrophotometry, agarose gel electrophoresis, and the quality of the qualified DNA was adjusted to 5 ( ig / L, -20 ° C storage standby;
  • reaction conditions are 94 °C for 4 minutes; 94 °C20 seconds, 56 °C for 30 seconds, 72 °C for 1 minute, 45 cycles; 72 °C for 3 minutes; 4 °C ⁇ ;
  • the PCR reaction product was treated with 0.5 U SAP (shrimp alkaline phosphatase) to remove free dNTPs from the system.
  • the reaction system was 7 ⁇ , of which the PCR product was 5 ⁇ , and the SAP mixture was 2 ⁇ (SAP 0.5).
  • U, buffer 0.17 ⁇ ) reaction procedure 37 ° C for 20 minutes; 85 ° C for 5 minutes; 40 °C ⁇ ;
  • the total volume of 9 ⁇ reaction system contains 7 ⁇ of the PCR product after SAP treatment, wherein each extension reaction primer mixture is 0.804 ⁇ , iPLEX enzyme is 0.041 ⁇ 1, the extension mixture is 0.2 ⁇ , and the reaction procedure is 94 ° C for 30 seconds; 94 ° C for 5 seconds; °C 5 seconds, 80 °C 5 seconds 5 cycles; return 94 °C 5 seconds, a total of 40 cycles; 72 °C 3 minutes, 4 °C oo;
  • Chip spotting and qualifier detection
  • the purified product was transferred to a 384-well SpectroCHIP (Sequenom) chip for measurement.
  • the SpectroCHIP chip was analyzed by MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight) matrix-assisted laser desorption ionization time-of-flight mass spectrometry. 4.0 software (sequenom) typing and outputting results;
  • kits for detecting a specific single nucleotide polymorphism of ankylosing spondylitis comprising: PCR amplification primer, extension primer, DNA designed for a region near the base of rs4552569 Polymerase, deionized water, dNTP, MgC12, PCR buffer, SAP, clean resin tree.
  • One of the objects of the present application is to provide the above-mentioned ankylosing spondylitis-specific single nucleotide polymorphism (SNP) in the treatment of ankylosing spondylitis, that is, when the base at the rs4552569 site is C, it is risky. , is the application of T in the treatment of ankylosing spondylitis.
  • SNP single nucleotide polymorphism
  • ankylosing spondylitis-specific single nucleotide polymorphism-associated susceptibility or pathogenicity Because the base at the rs4552569 locus is cytosine C.
  • the above applications include the use of susceptibility or pathogenic genes associated with ankylosing spondylitis-specific single nucleotide polymorphism (SNP) in the preparation of a medicament for the treatment of ankylosing spondylitis.
  • SNP single nucleotide polymorphism
  • the application includes the function of susceptibility or pathogenic genes involved in the intervention of rs4552569, including translation, transcription, and protein synthesis processes, preparation of therapeutic drugs, improvement of bone metabolism abnormalities in patients, and in vivo The immune inflammatory response to achieve therapeutic goals.
  • the control population of 4231 healthy volunteers mainly included healthy volunteers without AS symptoms or medical history in three places in China, Guangdong, Anhui and Singapore. These healthy volunteers were also informed. Later agreed to draw blood for the study and signed informed consent.
  • the basic conditions of the case and healthy volunteers were recorded, including: gender, age and place of origin. 4 ml of whole blood was taken, placed in an EDTA tube, and brought back to the laboratory for DNA extraction.
  • rheumatologists also carefully inquired and recorded the situation of each AS patient and healthy volunteers, especially the AS patients also recorded basic conditions (including: gender, age, age of onset, duration) And clinical phenotype (including: waxy fingers, hip involvement, peripheral arthritis, inflammatory low back pain).
  • the composition of the kit is: Cell Lysis Night, Buffer GS, Buffer GB, Buffer Liquid GD, rinse PW, elution buffer TB, proteinase K, adsorption column CB3, collection tube (2 ml), 1.5 mL sterile collection tube.
  • the study used the Illumina HuamHap 610-Quad SNP chip, the Illumina Human lM-duo chip and the Illumina OmniExpress chip to perform a full-genome SNP scan of the sample.
  • the following are three types of Illumina chips;
  • the chip cartridge can detect four samples in parallel on one chip, which significantly increases the amount of sample output information and reduces errors in experimental operations.
  • the chip is widely used in the content of the HumanHap550 chip, with an additional 100,000 genetic markers.
  • Human 610-quad The genome-wide information covered is authoritative for both known and recently reported CNV regions.
  • the SNPs of the Illumina HuamHap 610-Quad SNP chip are evenly distributed on the genome at a density of about 5 kb.
  • the Illumina HuamHap 610-Quad SNP chip was designed for specific studies targeting highly polymorphic CNV regions in the genome (fragment replication regions and genomic regions without SNPs).
  • the chip contains more than one million probe information.
  • the chip contains probes for gene SNPs, tagged SNPs, copy number changes (CNV), and other high-value genomic regions, as well as new sites such as: increased disease-associated SNP sites Flexible selection of high-density SNP sites in some genomic coding regions.
  • the chip provides high sample throughput and comprehensive genomic content.
  • the OmniExpress can hold up to 12 samples.
  • the chip interrogates more than 700,000 variations per sample, with a total of more than 8 million data points on a single chip.
  • the marker on OmniExpress is part of Illumina's HumanOmnil-Quad content, which combines optimized SNP tag combinations from all three phases of the international HapMap program, with the highest data quality and best content, including copy number variation (CNV) ) Comprehensive support for the application.
  • CNV copy number variation
  • the heterozygosity range is statistically analyzed from the mean of the heterozygosity mean (Mean) -3 ⁇ standard deviation (SD) to the mean heterozygosity (Mean) +3 ⁇ standard deviation (SD). Samples not in this interval Will be removed. Not included in further statistics.
  • the estimated SNP for analysis in the three chips is done by the IMPUTE2 software (version number: 2.1.0), which maximizes the chip's coverage of the chromosome.
  • C 6.91 x 10- 5, i.e., the presence of significant correlation ankylosing spondylitis, C is the SNP nucleotide risk type, when T is a protected type.
  • the 3702 healthy volunteers were mainly from the healthy volunteers collected by the Third affiliated Hospital of Sun Yat-sen University and the Cancer Center of Sun Yat-sen University. The population were healthy volunteers without AS symptoms or medical history. These healthy volunteers Also, after being informed, agreed to draw blood for the study and signed an informed consent form.
  • the composition of the kit is: Cell Lysis Night, Buffer GS, Buffer GB, Buffer GD, rinse solution PW, elution buffer TB, proteinase K, adsorption column CB3, collection tube (2 ml), 1.5 mL sterile collection tube.
  • Primer design (including amplification primers and extension primers) was performed using the iPLEX® Gold snp genotyping technique of the Sequenom Massarray® DNA Mass Spectrometry System. Primer information for SNP rs4552569 is as follows:
  • the template DNAlul was added to each well in a 384-well plate, and 4 ul of the above reaction system was added.
  • the 384-well plate was centrifuged at 1000 rpm for 1 minute, and after mixing, amplification was carried out according to the following procedure: 94. C 4 minutes pre-denaturation, 94. C 20 seconds, 56 ° C 30 seconds, 72 ° C 1 minute for a total of 45 cycles, 72 ° C 3 minutes, 4 ° C cooling.
  • the main feature of matrix-assisted laser desorption ionization time-of-flight mass spectrometry is that The target sequence was amplified by PCR, and then the snp sequence-specific extension primer was added, and one base was extended at the SNP site.
  • the prepared sample analyte is co-crystallized with the chip substrate, and the crystal is placed in a vacuum tube of the mass spectrometer, and then excited by a transient nanosecond (1 (T 9 s) strong laser, resulting in energy due to the energy absorbed by the matrix molecules through the radiation.
  • the matrix crystals sublimate the nucleic acid molecules are desorbed and converted into metastable ions, and the generated ions are mostly singly charged ions, which obtain the same kinetic energy in the accelerating electric field, and then A non-electric field drift region is separated according to its mass-to-charge ratio, and flies in a vacuum tube to reach the detector.
  • the ions generated by MALDI are commonly detected by a Time-of-Flight (TOF) detector, and the smaller the ion mass, the smaller the ion mass is. The faster the arrival, the genotype of SNP in the above PCR reaction product can be quickly and conveniently detected by this method.
  • TOF Time-of-Flight
  • the subjects in the study were all AS and healthy controls in Examples 1 and 2, and these AS cases were diagnosed by experienced rheumatologists based on clinical findings and imaging findings. New York revised standard AS patients. For each AS patient, blood was drawn for use in the study and the informed consent was signed after being informed of the purpose of the study. Finally, a total of 3937 AS patients and 7727 healthy volunteers were included in the study.
  • the TIANamp Blood DNA kit is used to extract DNA from human peripheral blood samples.
  • the composition of the kit is: Cell Lysis Night, Buffer GS, Buffer GB, Buffer GD , rinse solution PW, elution buffer TB, proteinase K, adsorption column CB3, collection tube (2 ml) and 1.5 mL sterile collection tube.
  • the TIANamp Blood DNA kit is used to extract DNA from human peripheral blood samples.
  • the composition of the kit is: Cell Lysis Night, Buffer GS, Buffer GB, Buffer GD , rinse solution PW, elution buffer TB, proteinase K, adsorption column CB3, collection tube (2 ml), 1.5 ml sterile collection tube.
  • the template DNAlul was added to each well of the plate to be tested, and 4 ul of the above reaction system was added, and centrifuged at 1000 rpm for 1 minute. After mixing, the amplification was carried out according to the following procedure: 94. C 4 minutes pre-denaturation, 94. C 20 seconds, 56 ° C 30 seconds, 72 ° C 1 minute for a total of 45 cycles, 72 ° C 3 minutes, 4 ° C cooling.
  • the prepared sample analyte is co-crystallized with the chip substrate, and the crystal is placed in a vacuum tube of the grammar instrument, and then excited by a transient nanosecond (10-9 s) strong laser, which can be quickly and conveniently detected by the method.
  • a transient nanosecond (10-9 s) strong laser which can be quickly and conveniently detected by the method.
  • test result When the test result is C, it is a risk type, and when the test result is T, it is a protection type.
  • the kit includes: PCR amplification primers, extension primers, DNA polymerase, deionized water, dNTP, MgC12, PCR buffer, SAP, clean resin designed for the region near rs4552569 base.
  • the amplification primers and extension primers are shown in the following table:
  • the steps of the detection method are as follows:
  • DNA from tissues, cells or blood samples was extracted using the TIANamp Blood DNA kit or NucleoSpin Tissue (MN), quantified by spectrophotometry, agarose gel electrophoresis, and the quality of the qualified DNA was adjusted to 5 ( ig / L, -20 ° C storage standby;
  • each reaction is 5 ⁇ 1, including template DNA 10 g, Hotstar Taq 0.5 U, amplification primers 0.5pmol each, 25mM dNTP ⁇ . ⁇ , reaction conditions 94 °C 4 minutes; 94 ° C20 seconds, 56 °C for 30 seconds, 72 °C for 1 minute, 45 cycles; 72 °C for 3 minutes; 4 °C ⁇ ;
  • the PCR reaction product was treated with 0.5 U SAP (shrimp alkaline phosphatase) to remove the body.
  • Free dNTP in the system 7 ⁇ reaction system, 5 ⁇ PCR product, 2 ⁇ SAP solution (SAP 0.5 U, buffer 0.17 ⁇ ), reaction procedure at 37 °C for 20 minutes; 85 °C for 5 minutes; 40 °C ⁇ ;
  • the total volume of 9 ⁇ reaction system contains 7 ⁇ of the PCR product after SAP treatment, wherein each extension reaction primer mixture is 0.804 ⁇ , iPLEX enzyme is 0.041 ⁇ 1, the extension mixture is 0.2 ⁇ , and the reaction procedure is 94 ° C for 30 seconds; 94 ° C for 5 seconds; °C 5 seconds, 80 °C 5 seconds 5 cycles; return 94 °C 5 seconds, a total of 40 cycles; 72 °C 3 minutes, 4 °C oo;
  • Chip spotting and qualifier detection
  • the purified product was transferred to a 384-well SpectroCHIP (Sequenom) chip for measurement.
  • the SpectroCHIP chip was analyzed by MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight) matrix-assisted laser desorption ionization time-of-flight mass spectrometry. 4.0 software (sequenom) typing and outputting results;

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Genetics & Genomics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Zoology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Rheumatology (AREA)
  • Biophysics (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Epidemiology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Fait l'objet de cette invention un fragment à polymorphisme nucléotidique relatif à la spondylarthrite ankylosante. Parmi les malades atteints de la spondylarthrite ankylosante, le fragment nucléotidique en position rs4552569 représente la fréquence C sensiblement supérieure chez les témoins sains, la base dans cette position représente la fréquence T sensiblement supérieure chez les témoins sains, notamment lorsque la base en position rs4552569 représente C, elle contribue efficacement à diagnostiquer la spondylarthrite ankylosante. Font aussi l'objet de cette invention un procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et une trousse d'analyse associée permettant de contribuer au diagnostic précoce spécifique à la spondylarthrite ankylosante.
PCT/CN2011/081359 2011-10-27 2011-10-27 Procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et trousse d'analyse associée WO2013060004A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/CN2011/081359 WO2013060004A1 (fr) 2011-10-27 2011-10-27 Procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et trousse d'analyse associée
CN201180067457.9A CN103502451B (zh) 2011-10-27 2011-10-27 强直性脊柱炎相关特异性单核苷酸多态性的检测方法及其试剂盒

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2011/081359 WO2013060004A1 (fr) 2011-10-27 2011-10-27 Procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et trousse d'analyse associée

Publications (1)

Publication Number Publication Date
WO2013060004A1 true WO2013060004A1 (fr) 2013-05-02

Family

ID=48167054

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2011/081359 WO2013060004A1 (fr) 2011-10-27 2011-10-27 Procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et trousse d'analyse associée

Country Status (2)

Country Link
CN (1) CN103502451B (fr)
WO (1) WO2013060004A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107299144A (zh) * 2017-08-10 2017-10-27 广州和康医疗技术有限公司 一种用于检测TNF‑a基因SNP位点rs1799724基因型的扩增引物、试剂盒、方法及其应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1710107A (zh) * 2005-07-12 2005-12-21 卫生部北京医院 一种预测强直性脊柱炎易感性的方法及试剂

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1710107A (zh) * 2005-07-12 2005-12-21 卫生部北京医院 一种预测强直性脊柱炎易感性的方法及试剂

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHEN CHAO: "Association of ERAPl, ANTXR2, IL1R2, IL23R, JAK2 and STAT3 Polymorphisms with Ankylosing Spondylitis", CHINESE DOCTORAL DISSERTATIONS FULL-TEXT.-28 DATABASE (MEDICINE AND HEALTH SCIENCES), 15 September 2010 (2010-09-15), pages 24 - 28 *
DATABASE GENBANK 18 August 2010 (2010-08-18), accession no. s253586435 *
DATABASE GENBANK 26 February 2008 (2008-02-26), accession no. s93156442 *

Also Published As

Publication number Publication date
CN103502451B (zh) 2015-06-10
CN103502451A (zh) 2014-01-08

Similar Documents

Publication Publication Date Title
JP7000658B2 (ja) 肝臓病変を評価する方法
WO2013060005A1 (fr) Procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et trousse d'analyse associée
CN110484621B (zh) 一种肝癌早期预警的方法
JP5899527B2 (ja) 第６染色体短腕２１．３３領域の一塩基多型に基づく抗てんかん薬による薬疹リスクの検査方法
CN110699446B (zh) 一种与非综合征型唇腭裂诊断相关的SNP标志物rs3174298及其应用
US9879314B2 (en) Method for detecting HLA-A*31:01 allele
WO2013078690A1 (fr) Prédisposition à la spondylarthrite ankylosante et procédé de détection d'un polymorphisme mononucléotidique, trousse et utilisation associées
WO2016019633A1 (fr) Kit de réactifs diagnostiques applicables à un site de variation correspondant à un polymorphisme génétique permettant une évaluation précoce d'un risque de cancer du sein
JP2008531041A (ja) 乳ガン診断方法及びそのための組成物
WO2013060004A1 (fr) Procédé de détection de polymorphisme nucléotidique relatif à la spondylarthrite ankylosante et trousse d'analyse associée
WO2006085733A1 (fr) Polynucleotide associe au cancer du sein contenant un polymorphisme de nucleotide unique, micro-reseau et kit de diagnostic comprenant la meme chose et procede de diagnostic du cancer du sein associe
CN110029162B (zh) 一种用于检测系统性红斑狼疮易感性位于非编码基因区的snp标志物及其应用
JP6516128B2 (ja) 抗甲状腺薬誘発性無顆粒球症リスクを判定するための検査方法及び判定用キット
WO2015030142A1 (fr) Procédé de détection d'une prédisposition de l'hépatite b à devenir chronique
CN108424959B (zh) 一种用于强直性脊柱炎早期诊断的生物标记物及其在试剂盒中应用
WO2006033384A1 (fr) Oligomere d'adn, marqueur de gene et ensemble d'oligomeres d'adn conçus pour prevoir l'apparition d'un effet secondaire dans le cadre d'une radiotherapie et procede visant a prevoir l'apparition d'un effet secondaire
US20070264648A1 (en) Dna Oligomer, Genetic Marker and Dna Oligomer Set for Prediction of Onset of Side-Effect from Radiation Therapy, and Method for Predicting Onset of Side-Effect
JP2014045744A (ja) 原発性胆汁性肝硬変の発症リスク予測マーカー、プローブ、プライマー及びキット並びに原発性胆汁性肝硬変の発症リスク予測方法
CN115948533A (zh) 一种用于检测mybpc3突变基因的试剂及其应用
CN118166089A (zh) SNP rs11255332在用于预测抗精神病药物所致高泌乳素血症严重程度及易感人群的产品中的应用
CN114752666A (zh) 替格瑞洛相关呼吸困难基因型检测试剂或试剂盒
Class et al. Patent application title: Detection of hepatitis B virus (HBV) DNA and methylated HBV DNA in urine of patients with HBV-associated hepatocellular carcinoma Inventors: Wei Song (Audubon, PA, US) Surbhi Jain (Doylestown, PA, US) Batbold Boldbaatar (Coatesville, PA, US) Sitong Chen (Audubon, PA, US) Assignees: JBS Science Inc.
WO2004101818A1 (fr) Methode de detection d'un gene predisposant a la schizophrenie et utilisation
WO2012063829A1 (fr) Nouveau gène et méthode de détection de l'ostéoporose basée sur un polymorphisme d'un seul nucléotide sur le locus du gène fong
JP2016149988A (ja) 自閉症スペクトラムの検査方法。

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: 11874764

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: 11874764

Country of ref document: EP

Kind code of ref document: A1