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KIR2DL5 genotyping kit and KIR2DL5 genotyping method

A genotyping method and genotyping technology, applied in the field of KIR2DL5 genotyping kits, can solve problems such as false detection, missed detection, and inability to realize high-resolution typing of KIR2DL5 gene

Pending Publication Date: 2021-04-30
SHENZHEN TISSUEBANK PRECISION MEDICINE CO LTD +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high polymorphism of the KIR gene and the randomness of the distribution of polymorphic sites, the above-mentioned multiple detection methods have the risk of missed detection and false detection, and cannot achieve high-resolution typing of the KIR2DL5 gene

Method used

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  • KIR2DL5 genotyping kit and KIR2DL5 genotyping method
  • KIR2DL5 genotyping kit and KIR2DL5 genotyping method
  • KIR2DL5 genotyping kit and KIR2DL5 genotyping method

Examples

Experimental program
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Effect test

Embodiment 1

[0090] Randomly select two positive samples that have undergone low-resolution typing of the KIR2DL5 gene, and use the kit and typing method of the present invention to carry out KIR2DL5 genotyping on the samples to verify the typing effect of the present invention. Genomic DNA of the sample is extracted, and PCR amplification is performed on the genomic DNA. For the results of PCR amplification, please refer to figure 1 . figure 1 In the agarose gel electrophoresis pattern, the bands from left to right are DL2000 DNA Marker, the P1 sequence of sample ①, the P1 sequence of sample ②, the P2 sequence of sample ①, the P2 sequence of sample ②, and the P3 sequence of sample ① and the P3 sequence of sample ②. The sequence length of P1 is 374bp, the sequence length of P2 is 5084bp, and the sequence length of P3 is 1382bp. The results in the figure are consistent with the actual, indicating that the above sequences have been successfully amplified.

[0091] Sequence the purified se...

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Abstract

The invention discloses a KIR2DL5 genotyping kit and a KIR2DL5 genotyping method. The KIR2DL5 genotyping kit comprises an amplification primer 1, an amplification primer 2, an amplification primer 3, an amplification primer 4, an amplification primer 5, an amplification primer 6, a sequencing primer 1, a sequencing primer 2, a sequencing primer 3, a sequencing primer 4, a sequencing primer 5, a sequencing primer 6, a sequencing primer 7, a sequencing primer 8, a sequencing primer 9, a sequencing primer 10, a sequencing primer 11, a sequencing primer 12, a sequencing primer 13 and a sequencing primer 14. The KIR2DL5 gene is subjected to PCR amplification by virtue of multiple amplification primers, the amplified PCR product is subjected to sequencing amplification by virtue of multiple sequencing primers, and then the sequencing amplification product is sequenced, so that a new allelotype which is difficult to identify by virtue of other methods is found, and high-resolution typing of the KIR2DL5 gene is realized.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a KIR2DL5 genotyping kit and a typing method. Background technique [0002] Natural killer cells (Natural Killer cells, NK cells) in the human body through cytokines and cytotoxicity, resist infection and kill malignant cells, play an important role in innate immunity and acquired immunity. Killer cell Immunoglobulin-like Receptor (KIR) is a group of immunoglobulin-like superfamily members mainly expressed on the surface of NK cells and some T cells, which can specifically recognize human major histocompatibility Sex complex MHC-I molecules (Human Leukocyte Antigen I, HLA-I), mediate and regulate the killing function of NK cells and some effector T cells. More and more evidences from clinical transplantation and animal experiments suggest that the heterogeneous response of NK cells triggered by KIR incompatibility between donor and recipient is beneficial to the prognosis of allogen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2531/113C12Q2535/122C12Q2537/165
Inventor 郑仲征杜可明廖宽镇贺青青李岱阳
Owner SHENZHEN TISSUEBANK PRECISION MEDICINE CO LTD
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