Kit for detecting human natural killer cell immunoglobulin-like receptor KIR genotyping

A genotyping and kit technology, applied in the field of biomedical clinical molecular detection, can solve the problems of incomplete KIR gene detection, high sample DNA detection limit, and small sample detection throughput, so as to achieve good practical application value and shorten the experimental time. Time, the effect of increasing the detection flux

Active Publication Date: 2021-03-05
JIANGSU WEIHE BIOTECH
View PDF8 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] In view of the deficiencies in the existing technology, such as small sample detection throughput, non-specific amplification products and false negative results, long experimental process, low accuracy of results, complicated analysis, high min...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kit for detecting human natural killer cell immunoglobulin-like receptor KIR genotyping
  • Kit for detecting human natural killer cell immunoglobulin-like receptor KIR genotyping
  • Kit for detecting human natural killer cell immunoglobulin-like receptor KIR genotyping

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Raw materials and equipment

[0043] 1.1 Preparation of PCR main reaction solution: 0.18mM deoxynucleotide (dNTP), 1.8mM magnesium chloride (MgCl2), 60.3mM potassium chloride (KCl), 18.9mM Tris-hydrochloride (Tris-HCl) , glycerol (glycerol) 0.6% (v / v), dimethyl sulfoxide (DMSO) 5% (v / v) and formamide 2.5% (v / v), prepare 6*PCR reaction mixture 50mL according to the above ratio , Long-term storage at -20°C, temporary storage at 4°C for later use.

[0044] 1.2 Preparation of reaction plate:

[0045] 1.2.1 Preparation of primer-probe mixture: primer 0.54OD / mL, probe 1.05μM, make 6*primer-probe mixture.

[0046] 1.2.2 Plate according to the following table: primers and probes are shown in SEQ ID No.01-48, SEQ ID No.52-53 and SEQ ID No.55-56 in the following table. Internal standard primers and probes are added to each well, and the nucleotides are shown in SEQ ID No.49-51. Primers and probes were synthesized by General Biosystems (Anhui) Co., Ltd.

[0047]

[0048...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides 16 common genes for detecting a human natural killer cell immunoglobulin-like receptor KIR, wherein the 16 common genes comprise 14 KIR functional genes (2DL1, 2DL2, 2DL3, 2DL4,2DL5A/B, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 3DL1, 3DL2, 3DL3 and 3DS1) and 2 pseudo genes (2DP1 and 3DP1), and a combination of a PCR amplification primer group and a Taqman probe of the common variant ofthe 2DS4; and the combination comprises six primer groups. The invention also provides a kit containing the combination of the PCR amplification primer group and the Taqman probe. The invention has the following technical effects that: an ARMS analysis method is combined with a Taqman multiplex fluorescence PCR technology to carry out qualitative typing detection on the KIR gene; furthermore, improvement is carried out on the basis of a traditional ARMS; various defects of an existing detection method are overcome; and the method disclosed by the invention has wide application prospects and clinical reference value.

Description

technical field [0001] The invention relates to a method and a kit for detecting human natural killer cell immunoglobulin-like receptor KIR genotyping, belonging to the field of biomedical clinical molecular detection. Background technique [0002] Natural killer cells (NK) are derived from bone marrow lymphoid stem cells, and their development and maturation depend on the microenvironment of bone marrow and thymus. NK cells are mainly distributed in peripheral blood and spleen, and there are also a small amount in lymph nodes and other tissues. NK cells do not express specific antigen recognition receptors, and are a type of lymphoid cells different from T and B lymphocytes. NK cells can express a variety of surface markers, most of which can also be expressed on the surface of other immune cells. NK cells are non-specific immune cells. They can directly kill some tumors and virus-infected target cells without antigen pre-sensitization, so they play an important role in t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2535/137C12Q2537/143C12Q2531/113C12Q2561/101
Inventor 陈炤源王浩章婷婷王丹娜
Owner JIANGSU WEIHE BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap