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Kit for detecting nucleotide-binding oligomerization domain-like receptor 3snp and application thereof

An oligomerization domain and nucleotide technology, which is applied in the field of real-time fluorescent PCR technology of double ligation probes for detection of NLRP3SNP, can solve the problems such as the lack of NLRP3SNP detection kits, and achieve accurate detection and interpretation and high primer efficiency. Effect

Inactive Publication Date: 2016-03-23
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no relevant report on the NLRP3SNP detection kit based on pyrosequencing technology

Method used

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  • Kit for detecting nucleotide-binding oligomerization domain-like receptor 3snp and application thereof
  • Kit for detecting nucleotide-binding oligomerization domain-like receptor 3snp and application thereof
  • Kit for detecting nucleotide-binding oligomerization domain-like receptor 3snp and application thereof

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Embodiment Construction

[0027] In order to make the object, technical solution and advantages of the present invention clearer, preferred embodiments of the present invention are described in detail below.

[0028] In this example, the principle of the double pyrosequencing method is involved: 1) A specific sequencing primer binds to the single-stranded DNA template, and then an enzyme mixture (including DNAPolymerase, ATPSulfurylase, Luciferase and Apyrase) and a substrate mixture are added (including APS and Luciferin). 2) Add 1 dNTP to the reaction system. If it can just pair with the next base of the DNA template, it will be added to the 3' end of the sequencing primer under the action of DNA polymerase, and at the same time release a molecule of Pyrophosphate (PPi). 3) Under the action of ATP sulfurylase, the generated PPi can combine with APS to form ATP; under the catalysis of luciferase, the generated ATP can combine with luciferin to form oxyluciferin, and at the same time generate visible ...

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Abstract

The invention discloses an SNP kit and a method for detecting SNR (Single Nucleotide Polymorphism) of a nucleotide-binding oligomerization domain receptor 3 (NLRP3) by adopting a double pyrophosphoric acid sequencing method. The kit is used for carrying out detection on NLRP3SNP which particularly is rs1539019C / A and rs10754558C / G SNP. The kit comprises upstream and downstream primers, a double reaction system and conditions. The kit and the detection method which are provided by the invention have a parting success rate of 100 percent on the NLRP3SNP and can implement high-flux low-cost high-efficiency accurate detection on the NLRP3SNP.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a real-time fluorescent PCR technique for detecting NLRP3SNP with double junction probes. Background technique [0002] Nucleotide-binding oligomerization domain-like receptor 3 (NLRP3) is mainly expressed in immune system cells such as peripheral blood mononuclear cells and DCs. Immune response, under exogenous stimuli, forms NLRP3 inflammatory complex with linker molecules ASC and caspase1, regulates the mature secretion of plasma IL-1β, IL-18 and other inflammatory factors, plays a role in immune regulation, inflammatory response, apoptosis, etc. important role in the process. [0003] Disease association studies have found that NLRP3 single nucleotide polymorphism rs10754558C→G increases the risk of hypersensitivity diseases (JAllergyClinImmunol2009;124(4):779-785), and reduces type I diabetes (Autoimmunity2010;43(8):583 -589) and HIV risk (JournalofAcquiredImmuneDeficiencySynd...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2537/143C12Q2565/301
Inventor 王海燕蒋建新张安强曾灵顾玮杨策
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA