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

A technology for congenital heart disease and neural tube defects, applied in the field of medicine and biology, can solve problems such as the lack of clinical application of diagnostic molecular markers

Pending Publication Date: 2021-05-18
SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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  • 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, other birth defects have not been clinically recognized. Applied diagnostic molecular markers

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Screening of differentially expressed proteins in the peripheral blood of pregnant mice with neural tube defects using proteomics technology and bioinformatics analysis.

[0044] Ultrahigh-speed centrifugation was used to separate plasma exosomes from pregnant mice with neural tube defects, and the identification of exosomes was completed. Use label-free quantitative technology (label-free) for proteomics screening, combined with bioinformatics analysis, to initially screen possible protein markers, such as Figure 1-3 shown.

[0045] 1. Separation of plasma.

[0046] 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.

[0047] 2. Isolation and identification of exosome...

Embodiment 2

[0054] Example 2 Verify the expression of ACTR2, CORO1A and DNM2 in serum exosomes of neural tube defects.

[0055] 1. The expression of ACTR2, CORO1A and DNM2 was verified in the serum exosomes of embryonic day E18.

[0056] 19 pairs of embryonic E18 days samples other than omics detection were selected for Western-blot expansion sample size verification. The validation used Alix as an internal control to detect ACTR2 at 45 kDa, CORO1A at 57 kDa and DNM2 at 98 kDa. A total of 19 pairs of samples (19 normal and 19 neural tube defects) derived from serum exosomes of pregnant mice were tested, and they were found to be consistent with the trend of omics results, all of which were low-expressed in the abnormal group. The statistical results also showed that there were statistical differences in the low expression of ACTR2, CORO1A and DNM2 in the neural tube defect group. The detection of CORO1A by ELISA also confirmed the consistency of Western-blot results.

[0057] 2. The ex...

Embodiment 3

[0060] Example 3 Expression of ACTR2, CORO1A and DNM2 in spinal cord tissue and embryonic neurogenic exosomes.

[0061] 1. The expression of ACTR2, CORO1A and DNM2 in spinal cord tissue.

[0062] In order to explore the relationship between ACTR2, CORO1A and DNM2 and the occurrence of neural tube defects, and to clarify their expression in the spinal cord tissue, Western-blot was used to detect the expression in the spinal cord tissue of embryonic E18 and E12 days, and β-actin was used as the tissue Internal reference for detection. The results showed that consistent with the trend of serum exosomes, ACTR2, CORO1A, and DNM2 all showed a statistically significant down-regulation trend in spinal cord tissues with neural tube defects. Immunohistochemical staining showed the expression of ACTR2, CORO1A and DNM2 in the spinal cord tissues of neural tube defects on day E18, to clarify whether these proteins were specifically expressed in neural tube tissues. ACTR2 is expressed in ...

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Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to a molecular marker for antenatal noninvasive diagnosis of a fetal with neural tube malformation, congenital heart disease or cleft lip and palate and application of the molecular marker. The molecular marker is composed of one or more selected from CORO1A, DNM2 and ACTR2 proteins for antenatal noninvasive diagnosis of fetuses with neural tube malformation, congenital heart disease and cleft lip and palate. The molecular marker for antenatal noninvasive diagnosis is applied to preparation of products for antenatal screening, early warning, clinical diagnosis and biochemical inspection of fetuses with neural tube malformation, congenital heart disease and cleft lip and palate. According to the invention, the close correlation between the abnormal expression of the proteins (including CORO1A, DNM2 and ACTR2) in the blood of the pregnant woman and the occurrence of the fetal with the neural tube deformity, the congenital heart disease and the cleft lip and palate is found and confirmed for the first time, the quantity of verified samples is large, the result is accurate, and a new way is provided for the prenatal screening, early warning and diagnosis of the fetal with the neural tube deformity, the congenital heart disease and the cleft lip and palate.

Description

technical field [0001] The invention belongs to the technical field of medicine and biology, and specifically relates to molecular markers (CORO1A, DNM2 and ACTR2 proteins) used for prenatal non-invasive diagnosis of neural tube defects, congenital heart disease and cleft lip and palate fetuses and applications thereof. Background technique [0002] Neural tube defects, congenital heart disease, and cleft lip and palate are common major fetal developmental malformations in my country, which seriously endanger the improvement of the quality of life of the birth population. Therefore, research on methods for early non-invasive diagnosis of these congenital malformations enables them to be diagnosed before severe structural abnormalities or irreversible damage, and formulate corresponding new strategies for early treatment and prevention of embryos. is of great significance. [0003] Establishing a non-invasive early screening method for congenital malformations has always bee...

Claims

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Application Information

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IPC IPC(8): G01N33/68C12Q1/6883C12N15/11
CPCG01N33/6893C12Q1/6883C12Q2600/158G01N2800/385G01N2800/50G01N2800/52
Inventor 袁正伟王衍夫马玲王勤博
Owner SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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