A kind of biomarker and application thereof for detecting ventricular septal defect

A technology of detection chamber and chain reaction, which is applied in the field of biomedicine to achieve stable results, convenient material collection and accurate diagnosis

Active Publication Date: 2022-04-01
SHENZHEN CHILDRENS HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But is it relevant as a biomarker for VSD? There is no report on the application of DDIT4 gene in the detection of ventricular septal defect

Method used

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  • A kind of biomarker and application thereof for detecting ventricular septal defect
  • A kind of biomarker and application thereof for detecting ventricular septal defect
  • A kind of biomarker and application thereof for detecting ventricular septal defect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Screening of Rapid Diagnosis VSD Molecular Markers

[0042] 1. Transcriptome sequencing

[0043] (1) Collect samples from the Cardiology Department of Shenzhen Children's Hospital, among which 3 samples were from children (<18 years old) before ASD treatment, 4 samples were from children after ASD treatment, and 4 samples were from children before VSD treatment Samples, 2 samples from children after VSD treatment and 4 samples from healthy volunteers, collected 1ml of peripheral blood from healthy children, children with ASD and children with VSD respectively;

[0044] (2) RNA was extracted from peripheral blood samples according to the Trizol reagent manual (Invitrogen Life Technologies, Carlsbad, CA), and its concentration and purity were determined;

[0045] (4) The extracted mRNA is used for sequencing;

[0046] (5) Preliminary screening of nucleic acid markers: The sequencing results of the blood samples of clinical children before and after VSD and ASD...

Embodiment 2

[0075] Example 2 Verification of Rapid Diagnosis VSD Molecular Markers

[0076] In order to further confirm whether DDIT4 is elevated only in children with VSD or in children with other congenital heart diseases, we detected the level of DDIT4 in the peripheral blood of children with patent ductus atriosus (PDA) and children with ASD. The expression level.

[0077] (1) According to the Trizol reagent manual (Invitrogen Life Technologies, Carlsbad, CA), blood was drawn from 30 healthy groups (control groups), 30 PDA children, 30 ASD children, and 30 VSD children, and RNA was extracted from peripheral blood samples , and measure its concentration and purity;

[0078] (2) Synthesize cDNA using a cDNA synthesis kit;

[0079](3) DDIT4 was detected in these samples by qRT-PCR, normalized in each individual sample using the housekeeping gene GAPDH, and 2 -ΔΔCt Methods quantify relative expression changes.

[0080] The data show that if Figure 7 As shown, compared with the contr...

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PUM

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Abstract

The present invention relates to a molecular marker that can be used for rapid and accurate detection of ventricular septal defect. The molecular marker is DDIT4 gene. The inventor found that the expression level of DDIT4 was only significantly increased in children with VSD. Through quantitative analysis of peripheral blood The content of DDIT4 gene can detect VSD. The present invention has the advantages of convenient material collection, simple operation, strong specificity, fast and accurate results, stable results, etc., and can diagnose VSD in a timely, fast, objective and accurate manner. In particular, VSD can be separated from ASD and PDA through one experiment, which has the advantages of traditional diagnosis. advantages over the method. Therefore, the present invention has great clinical application value for the rapid diagnosis of VSD children, and provides direction for further developing a VSD rapid diagnosis kit.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a biomarker for detecting ventricular septal defect and its application. Background technique [0002] Septal defects are the most common congenital heart disease and cause communication between the left and right sides of the heart (atrium / ventricle). It can be divided into ventricular septal defect (VSD) and atrial septal defect (Atrial septal defect, ASD), which account for 40% and 10% of the incidence of congenital heart disease, respectively. Nearly 50% of these defects will heal naturally within a certain period of time after birth. If the defect is small and has no hemodynamic changes, regular follow-up observation and treatment can be performed, while the other half require medical intervention or surgical repair. Common clinical manifestations of patients with septal defects include growth retardation, decreased activity tolerance, and susceptibility to respiratory diseases, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158
Inventor 李博宁刘琮贾红玲范骏
Owner SHENZHEN CHILDRENS HOSPITAL
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