Phenylketonuria detection primer group, kit and gene mutation detection method

A technology of phenylketonuria and primer set, which is applied in the field of gene biology, can solve the problems of blind spots, lack of SPR gene detection sites, and insufficient coverage, and achieves complete coverage, high uniformity, and little mutual interference between primers. Effect

Active Publication Date: 2018-03-23
CAPITALBIO GENOMICS +1
View PDF6 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing technology only realizes the point mutation detection system for all exons and regulatory regions of the five genes PAH, GCH1, PTS, QDPR and PCBD1, but it lacks dete...

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
  • Phenylketonuria detection primer group, kit and gene mutation detection method
  • Phenylketonuria detection primer group, kit and gene mutation detection method
  • Phenylketonuria detection primer group, kit and gene mutation detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, the primer group that detects phenylketonuria pathogenic gene

[0023] The inventor designed primers for the phenylketonuria pathogenic genes PAH, PTS, GCH1, QDPR, PCBD1, and SPR based on these 6 genes and their upstream and downstream splicing sites of about 10 bp. After extensive screening and optimization, the primers were optimized 83 pairs of specific primers constitute a multiplex PCR primer set system, and their nucleotide sequences and mixing ratios are shown in Table 1.

[0024] Table 1. Primer sequences and mixing ratios for detecting phenylketonuria pathogenic genes

[0025]

[0026]

[0027]

[0028]

[0029]

Embodiment 2

[0030] Example 2, Construction of sequencing library for detection of pathogenic genes of phenylketonuria

[0031] Using the primer set provided in Example 1, a sequencing library construction method based on the Ion Proton platform is provided for detecting pathogenic gene mutations of phenylketonuria.

[0032] 1. DNA extraction from samples to be tested

[0033] Take 0.1-0.2 mL of peripheral blood samples (or 2-3 pieces of dried blood spots with a diameter of about 6 mm) to extract genomic DNA using MagPure Tissue&Blood DNA LQ Kit, and use Nanodrop2000 to perform purity testing and concentration of the extracted genomic DNA. The initial assessment, followed by verification of the integrity of the extracted genomic DNA by agarose gel electrophoresis,

[0034] 2. Multiplex PCR amplification reaction

[0035] Use the extracted genomic DNA as a template to perform multiple PCR reactions to amplify the target fragment. The 20 μL reaction system is: 200ng of genomic DNA of the s...

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 discloses a phenylketonuria detection primer group, a kit and a gene mutation detection method. The primer group covering six phenylketonuria related genes PAH, PTS, GCH1, QDPR, PCBD1 and SPR and upstream and downstream 10bp splice sites thereof are provided for the first time. The primer group in a multiplex PCR (polymerase chain reaction) reaction system is high in specificity, lowin primer mutual interference, high in uniformity and high in amplicon expression depth. The phenylketonuria detection primer group, the kit and the gene mutation detection method have advantages ofquickness, comprehensiveness, accuracy and simplicity in operation and are applicable to detection of samples of blood, blood spots, leukocytes and the like. Compared with the prior art, the phenylketonuria detection primer group, the kit and the gene mutation detection method have advantages of wide coverage, more detection sites and high accuracy, can be applied to disease prevention in terms ofphenylketonuria gene mutation detection, carrier detection, genetic counseling and the like and can also be applied to disease treatment guidance.

Description

technical field [0001] The invention belongs to the field of gene biology, and in particular relates to a primer set, a kit and a method for detecting a pathogenic gene of phenylketonuria. Background technique [0002] Phenylketonuria (PKU) is a disease of abnormal amino acid metabolism caused by the lack of phenylalanine hydroxylase (phenylalaninehydroxylase, PAH). Children with phenylketonuria are normal at birth, and symptoms usually begin at 3 to 6 months. The clinical manifestations are usually mental retardation, hair color from black to yellow, and more phenylacetic acid is excreted in urine and sweat. Urine smells. There is currently no effective cure for the disease, and the symptoms can only be alleviated and controlled by diet therapy. Therefore, rapid and accurate detection of pathogenic genes, detection of carriers and genetic counseling are the keys to preventing this disease. [0003] Phenylketonuria is an autosomal recessive genetic disease, the cause of wh...

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): C12Q1/6883C12Q1/6806
CPCC12Q1/6806C12Q1/6883C12Q2600/156C12Q2531/113C12Q2537/143C12Q2563/143C12Q2563/149
Inventor 糜庆丰刘海量张小安崔世红刘情刘宇彬王杨周幸芝程国梅张琳琳常慧刘佩
Owner CAPITALBIO GENOMICS
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