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Human killer cell immunoglobulin-like receptor (KIR) genotyping inspection primer group and application thereof

A technology for genotyping and detection of primers, used in recombinant DNA technology, microbial assay/inspection, biochemical equipment and methods, etc.

Active Publication Date: 2019-03-19
江苏中济万泰生物医药有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Each individual contains different types and numbers of KIR genes, and the alleles at the same site are rich in polymorphisms, but there is a lack of comprehensive and holistic assay kits for human KIR genotyping detection in the prior art

Method used

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  • Human killer cell immunoglobulin-like receptor (KIR) genotyping inspection primer group and application thereof
  • Human killer cell immunoglobulin-like receptor (KIR) genotyping inspection primer group and application thereof
  • Human killer cell immunoglobulin-like receptor (KIR) genotyping inspection primer group and application thereof

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Embodiment

[0019] A kit for detecting human KIR genotyping, comprising primers of SEQ ID No. 1-32, dNTP-Buffer, cresyl red sodium salt and SYBR GreenⅠ. The detection of each person includes 16 detection wells, in addition to placing a pair of allele primers in each well, it also includes internal reference quality control primers, the sequences of which are shown in SEQ ID No.33 and SEQ ID No.34.

[0020] Storage conditions and expiration date

[0021] All components of the kit should be stored below -18°C, and the validity period is 6 months.

[0022] Sample request:

[0023] 1. The sample required for this kit is whole blood, which needs to be extracted from whole blood DNA, and its DNA sample concentration needs to be determined.

[0024] 2. When extracting DNA from whole blood, it is not allowed to use heparin anticoagulated whole blood samples, and EDTA anticoagulant is recommended.

[0025] 3. The DNA sample concentration should be 40-70ng / μl, and the purity A260 / A280 value shou...

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Abstract

The invention discloses a human killer cell immunoglobulin-like receptor (KIR) genotyping inspection primer group, comprising 16 pairs of primers shown in SEQ ID No.1-32. On the basis of the reactiontheory of polymerase chain reaction (fluorescence PCR) in the invention, the first basic group at the end of primer 3' is complementary to the specific basic group of an allele, specific primer pairsonly amplify alleles matched with the same; the primer design is based on human gene sequence issued by Blood Group Antigen Gene Mutation Database (dbRBC) in DNA sequence database (Genbank) that is established by National Center for Biotechnology Information (NCBI), proper specific mutation point is selected to design primers; a PCR doped dye method uses fluorescent dye to show DNA amplification effect through doping the amplified product of DNA. Melting curve analysis is carried out after amplification, and as a result, whether the target product is amplified specifically can be determined through the melting curve analysis, thereby identifying alleles.

Description

technical field [0001] The invention relates to the technical field of gene diagnosis products, in particular to a human KIR genotyping detection primer set and its application. Background technique [0002] Killer cell immunoglobulin-like receptors (KIR) are a group of receptors expressed on the surface of NK cells and some T cells that specifically recognize human major histocompatibility antigen MHC class I molecules, and play an important role in the effector function of NK cells. Immunomodulatory effect. KIR is encoded by a polymorphic multi-gene family located on human chromosome 19q13.4, and its structure and function are diverse. Each individual contains different types and numbers of KIR genes, and the alleles at the same site are rich in polymorphisms, but the prior art lacks a comprehensive and holistic assay kit for human KIR genotyping detection. Contents of the invention [0003] The technical problem to be solved by the present invention is to overcome the...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 林裕翔张浩钧朱晓洁
Owner 江苏中济万泰生物医药有限公司
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