Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Molecular marker for prenatal noninvasive diagnosis of fetuses with cleft lip and palate, neural tube malformation and congenital heart disease and application of molecular marker

A technology for congenital heart disease and neural tube defects, applied in the field of biomedicine, can solve the problem of lack of diagnostic molecular markers and other problems

Active Publication Date: 2020-11-17
SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, many scholars at home and abroad are working on the research of discovering new diagnostic specific markers, but so far, except for neural tube defects and Down syndrome, which can be screened by serum alpha-fetoprotein, there is no effective screening method for other birth defects. diagnostic molecular markers

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
  • Molecular marker for prenatal noninvasive diagnosis of fetuses with cleft lip and palate, neural tube malformation and congenital heart disease and application of molecular marker
  • Molecular marker for prenatal noninvasive diagnosis of fetuses with cleft lip and palate, neural tube malformation and congenital heart disease and application of molecular marker
  • Molecular marker for prenatal noninvasive diagnosis of fetuses with cleft lip and palate, neural tube malformation and congenital heart disease and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Isolation of exosomes from the peripheral blood of mothers with cleft lip and palate and piRNA sequencing.

[0048] 1. Separation of plasma.

[0049] Collect the whole blood sample, put it in an EDTA anticoagulant tube, mix it upside down gently, use a 4°C low-temperature centrifuge, centrifuge at 1600×g for 10 minutes, collect the supernatant (plasma) into a new EP tube, and centrifuge at 16000×g After 10 minutes to remove cell debris, the plasma was divided into several centrifuge tubes.

[0050] 2. Isolation and identification of exosomes.

[0051] Plasma exosomes were isolated by ultracentrifugation. The separation method is as follows: centrifuge at 10,000×g for 1 hour at 4°C, transfer the supernatant to a new ultrahigh-speed centrifuge tube, and centrifuge at 100,000×g for 4 hours at 4°C. Discard the supernatant and resuspend the exosomes with 100 μl cold PBS. The isolated exosome particles were identified using three standard methods (transmission e...

Embodiment 2

[0058] Example 2 The quantitative PCR method was used to verify the differentially expressed piRNAs in the exosomes of pregnant women with cleft lip and palate.

[0059] 1. Design and synthesis of piRNA primers.

[0060] According to the sequence of the target gene, piRNA primers were designed and synthesized by Shanghai Sangon Bioengineering Co., Ltd. The housekeeping gene selects U6. The forward primer of hsa-piR-020492 gene is 5'-GGGGCGGCGGCGGCGGTG-3' (SEQ ID NO.1); the forward primer of hsa-piR-016792 gene is 5'-CCTCCCAAAGTGCTGGGATTACAG-3' (SEQ ID NO.2); The forward primer of hsa-piR-009228 gene is 5'-TAGTTGAACATGGGTCAGTCGGTCC-3' (SEQ ID NO.3); the forward primer of hsa-piR-001311 gene is 5'-ATTGGTGGTTCAGTGGTAGAATTCTCGC-3' (SEQ ID NO.4); The forward primer of hsa-piR-004993 gene is 5'-TCGCGAGTTCAAATCTCGCTGG-3' (SEQ ID NO.5); the forward primer of hsa-piR-016659 gene is 5'-CCCCCCACTGCTAAATTTGACTGG-3' (SEQ ID NO.6); The forward primer of the hsa-piR-016945 gene is 5'-CGGC...

Embodiment 3

[0064] Example 3 The sample size was expanded to verify the piRNAs screened by the above method in the exosomes of pregnant women with cleft lip and palate.

[0065] On the basis of Example 2, continue to expand the number of samples, increase cleft lip and palate pregnant women and healthy control pregnant women to 50 cases each, and further verify the six piRNAs screened above (hsa-piR-001101, hsa-piR-009228, hsa - the possibility of piR-016659, hsa-piR-019912, hsa-piR-020388 and hsa-piR-020496) as molecular markers. The results showed that 4 piRNAs (hsa-piR-009228, hsa-piR-016659, hsa-piR-019912, and hsa-piR-020496) were still differentially expressed after being verified by 50 pairs of samples, so they were included in subsequent studies. The expression of the other 2 piRNAs (hsa-piRNA-001101 and hsa-piR-020388) had no statistical difference, as Figure 5 shown.

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

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a molecular marker for prenatal noninvasive diagnosis of fetuses with cleft lip and palate, neural tube malformation and congenital heart disease and application of the molecular marker. The molecular marker for prenatal noninvasive diagnosis of the fetuses with cleft lip and palate, neural tube malformation and congenital heart disease comprises three piRNAs including hsa-piR-009228, hsa-piR-016659 and hsa-piR-020496. The invention further discloses application of the molecular marker for prenatal noninvasive diagnosis in preparation of products for prenatal screening, early warning, clinical diagnosis and biochemical examination of the fetuses with cleft lip and palate, neural tube malformation and congenital heart disease. The molecular marker and the application have the advantages that gene expression abnormality of piRNAs in blood of pregnant women is discovered and confirmed for thefirst time to be closely related to occurrence of the fetuses with cleft lip and palate, neural tube malformation and congenital heart disease, verified sample sizes are large, results are accurate,and a novel way is provided for prenatal screening, early warning and diagnosis of the fetuses with cleft lip and palate, neural tube malformation and congenital heart disease.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to molecular markers and applications thereof for prenatal non-invasive diagnosis of cleft lip and palate, neural tube defects and congenital heart disease. Background technique [0002] Cleft lip and palate, neural tube defects, and congenital heart disease are common major fetal developmental malformations in my country, including among the 12 malformations routinely monitored by the World Health Organization, which seriously endanger the improvement of the quality of my country's birth population. Therefore, it is the key direction of birth defect research at home and abroad to study the methods of early non-invasive diagnosis of these birth defects, so that they can be diagnosed before serious structural abnormalities or irreversible damage, and to formulate corresponding new strategies for early treatment and prevention of embryos. It is of great significance to...

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 Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 袁正伟贾杉杉顾卉魏晓伟马巍刘丹罗文婷
Owner SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com