Unlock instant, AI-driven research and patent intelligence for your innovation.

A detection primer composition for sideroblastic anemia and its application

A technology of primer composition and erythroblasts, which is applied in the field of molecular biology, can solve the problems of low detection rate and achieve the effects of simple operation, wide coverage and high detection efficiency

Active Publication Date: 2022-03-29
天津见康华美医学诊断技术有限公司
View PDF10 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Patent CN108192880A discloses a human δ-aminolevulinic acid synthase mutant protein and its application. According to the δ-aminolevulinic acid synthase mutant protein, gene chips, monoclonal antibodies and ELISA kits are prepared, which are X-linked genetic The genetic diagnosis of sideroblastic anemia and X-linked erythropoietic porphyria can provide guidance, but the method only targets human δ-aminolevulinic acid synthase mutant protein, and its detection rate is low

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
  • A detection primer composition for sideroblastic anemia and its application
  • A detection primer composition for sideroblastic anemia and its application
  • A detection primer composition for sideroblastic anemia and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Sideroblastic anemia gene, mutation site and primer composition thereof

[0041] (1) Using dbSNP database (http: / / www.ncbi.nlm.nih.gov / snp / ), Thousand Genomes database 1000Genomes (http: / / www.1000genomes.org / ), exome integration database ExAC (http: / / exac.broadinstitute.org / ) and the Human Gene Mutation Database HGMD (http: / / www.hgmd.cf.ac.uk / ac / index.php), and the function prediction tool PolyPhen (http: / / genetics.bwh.harvard.edu / pph2 / ) and SIFT (http: / / sift.jcvi.org / ), and use the following principles to screen for disease-causing mutations: (1) According to the genomic location and type of the mutation, filter Delete the mutations that have no effect on the protein product sequence; (2) Use 1000Genomes data and ExAC data to annotate the proportion of each mutation in the population. If the proportion is less than or equal to 1%, it is not considered a polymorphic site; (3) Search the human gene mutation database to check whether a mutation is recorded in ...

Embodiment 2

[0056] Embodiment 2 A kind of kit for detecting SA and its application method

[0057] 1. Amplification primers in multiplex amplification system

[0058] In the compound amplification system, the forward and reverse amplification primers of each gene and mutation site are shown in Table 4 above. In order to make the amplification efficiency of each gene and mutation site as consistent as possible, by adjusting each pair in the compound system The concentrations of the primers and the final adjusted primer concentrations are shown in Table 4 above.

[0059] 2. DNA extraction: Use a DNA extraction kit to extract whole-genome DNA from peripheral blood or bone marrow samples.

[0060] 3. Carry out multiplex PCR amplification to the extracted DNA according to the primer composition provided in Example 1.

[0061] Wherein, the complex amplification system is shown in Table 5 below.

[0062] Table 5 Compound amplification system

[0063] Element volume AgriSeq T...

Embodiment 3

[0089] Example 3 Repeatability Detection

[0090] A patient diagnosed with SA by clinical symptoms and relevant laboratory tests was selected. The ALAS2 gene c.1355G>A nucleotide mutation was detected by first-generation sequencing, and its amino acid mutation was p.R452H.

[0091] The sample was repeatedly detected three times using the primer composition and kit described in the application, and the specific steps and detection methods were as follows:

[0092] 1. DNA extraction: Use the Tiangen DNA Extraction Kit (Cat. No.: DP318-03) to extract the whole genome DNA of the bone marrow sample.

[0093] 2. Carry out multiplex PCR amplification to the extracted DNA according to the primer composition and kit provided in Examples 1 and 2.

[0094] 3. Prepare the amplified product into a DNA library that can be sequenced by the Ion Torrent sequencing platform. For details, refer to the instruction manual of the Life company kit (name: Ion AmpliSeqTM Library Kit, catalog number: ...

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 a primer composition for detecting sideroblastic anemia gene and application thereof, and relates to the technical field of molecular biology. Using the gene group of SA-related gene mutations provided by the present invention, combined with high-throughput sequencing technology, 342 gene mutation sites of the two genes can be detected at one time, with wide coverage, high detection efficiency, and simple and accurate operation. It solves the technical defects of low detection accuracy and limited detection methods. At the same time, it can realize the auxiliary diagnosis of SA in addition to bone marrow morphology and blood routine examination, especially for the follow-up treatment of the disease. Prediction of direction and effect has special advantages, which can make up for the deficiencies of clinical SA in molecular diagnosis, disease evolution, treatment, prognosis, and medication guidance.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a primer composition for detecting sideroblastic anemia gene and application thereof. Background technique [0002] Sideroblastic anemia (sideroblastic anemia, SA) is a group of rare heterogeneous diseases caused by hemoglobin synthesis disorder and poor iron utilization caused by different etiologies, and its common feature is a large number of ring-shaped sideroblasts in the bone marrow , Ineffective erythroid hematopoiesis, increased tissue iron, and different proportions of hypochromic erythrocytes in peripheral blood. Sideroblastic anemia can be divided into two types: hereditary and acquired, and there are differences in the treatment strategies and outcomes of different types of SA. Hereditary sideroblastic anemia is mostly X-chromosomal sex-linked inheritance, and the degree of anemia varies greatly among patients. Some patients with mild anemia are easily miss...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6869C12N15/11
CPCC12Q1/6883C12Q1/6869C12Q2600/156C12Q2600/16C12Q2535/122
Inventor 李静汝昆蔺亚妮
Owner 天津见康华美医学诊断技术有限公司
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com