CN112816711B - Molecular marker for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetus and application thereof - Google Patents
Molecular marker for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetus and application thereof Download PDFInfo
- Publication number
- CN112816711B CN112816711B CN202110285096.0A CN202110285096A CN112816711B CN 112816711 B CN112816711 B CN 112816711B CN 202110285096 A CN202110285096 A CN 202110285096A CN 112816711 B CN112816711 B CN 112816711B
- Authority
- CN
- China
- Prior art keywords
- palate
- heart disease
- congenital heart
- cleft lip
- neural tube
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 208000032170 Congenital Abnormalities Diseases 0.000 title claims abstract description 67
- 210000000276 neural tube Anatomy 0.000 title claims abstract description 64
- 208000002330 Congenital Heart Defects Diseases 0.000 title claims abstract description 48
- 208000028831 congenital heart disease Diseases 0.000 title claims abstract description 48
- 206010009260 Cleft lip and palate Diseases 0.000 title claims abstract description 46
- 208000016653 cleft lip/palate Diseases 0.000 title claims abstract description 46
- 210000003754 fetus Anatomy 0.000 title claims abstract description 43
- 206010061619 Deformity Diseases 0.000 title claims abstract description 40
- 238000003745 diagnosis Methods 0.000 title claims abstract description 32
- 239000003147 molecular marker Substances 0.000 title claims abstract description 10
- 102100028233 Coronin-1A Human genes 0.000 claims abstract description 40
- 101000860852 Homo sapiens Coronin-1A Proteins 0.000 claims abstract description 40
- 210000004369 blood Anatomy 0.000 claims abstract description 8
- 239000008280 blood Substances 0.000 claims abstract description 8
- 238000002360 preparation method Methods 0.000 claims abstract description 4
- 210000001808 exosome Anatomy 0.000 claims description 57
- 238000001514 detection method Methods 0.000 claims description 23
- 239000003153 chemical reaction reagent Substances 0.000 claims description 15
- 230000001605 fetal effect Effects 0.000 claims description 11
- 238000007901 in situ hybridization Methods 0.000 claims description 4
- 210000004381 amniotic fluid Anatomy 0.000 claims description 3
- 238000012165 high-throughput sequencing Methods 0.000 claims description 3
- 239000003550 marker Substances 0.000 claims description 3
- 238000004949 mass spectrometry Methods 0.000 claims description 3
- 238000003793 prenatal diagnosis Methods 0.000 claims description 3
- 238000003127 radioimmunoassay Methods 0.000 claims description 3
- 238000012360 testing method Methods 0.000 claims description 3
- 210000002700 urine Anatomy 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 abstract description 43
- 102000020630 Dynamin II Human genes 0.000 abstract description 42
- 108010044191 Dynamin II Proteins 0.000 abstract description 42
- 102000004169 proteins and genes Human genes 0.000 abstract description 41
- 102000004373 Actin-related protein 2 Human genes 0.000 abstract description 32
- 108090000963 Actin-related protein 2 Proteins 0.000 abstract description 32
- 210000005036 nerve Anatomy 0.000 abstract description 25
- 238000009609 prenatal screening Methods 0.000 abstract description 11
- 102100021236 Dynamin-1 Human genes 0.000 abstract description 9
- 230000002159 abnormal effect Effects 0.000 abstract description 4
- 101000817604 Homo sapiens Dynamin-1 Proteins 0.000 abstract description 3
- 108090001145 Nuclear Receptor Coactivator 3 Proteins 0.000 abstract description 3
- 102100022883 Nuclear receptor coactivator 3 Human genes 0.000 abstract description 3
- 238000003759 clinical diagnosis Methods 0.000 abstract description 3
- 238000007689 inspection Methods 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 238000002372 labelling Methods 0.000 description 26
- 238000000034 method Methods 0.000 description 26
- 210000005259 peripheral blood Anatomy 0.000 description 23
- 239000011886 peripheral blood Substances 0.000 description 23
- 210000002966 serum Anatomy 0.000 description 20
- 210000001519 tissue Anatomy 0.000 description 18
- 241000699670 Mus sp. Species 0.000 description 13
- 239000000523 sample Substances 0.000 description 12
- 238000001262 western blot Methods 0.000 description 12
- 230000036244 malformation Effects 0.000 description 11
- 230000035945 sensitivity Effects 0.000 description 11
- 210000000278 spinal cord Anatomy 0.000 description 11
- 210000001161 mammalian embryo Anatomy 0.000 description 10
- 230000035935 pregnancy Effects 0.000 description 10
- 239000000427 antigen Substances 0.000 description 8
- 108091007433 antigens Proteins 0.000 description 8
- 102000036639 antigens Human genes 0.000 description 8
- 238000013211 curve analysis Methods 0.000 description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- 238000002965 ELISA Methods 0.000 description 6
- 239000012634 fragment Substances 0.000 description 6
- 206010010356 Congenital anomaly Diseases 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 230000003828 downregulation Effects 0.000 description 5
- 210000002381 plasma Anatomy 0.000 description 5
- 108090000765 processed proteins & peptides Proteins 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 4
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 238000007622 bioinformatic analysis Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 3
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 102000003992 Peroxidases Human genes 0.000 description 3
- 230000005856 abnormality Effects 0.000 description 3
- 230000031018 biological processes and functions Effects 0.000 description 3
- 229960002685 biotin Drugs 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 239000011616 biotin Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000000703 high-speed centrifugation Methods 0.000 description 3
- 230000001272 neurogenic effect Effects 0.000 description 3
- 210000002569 neuron Anatomy 0.000 description 3
- 108040007629 peroxidase activity proteins Proteins 0.000 description 3
- 108060006184 phycobiliprotein Proteins 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000008925 shengjing Substances 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- 102000007469 Actins Human genes 0.000 description 2
- 108010085238 Actins Proteins 0.000 description 2
- 102100025222 CD63 antigen Human genes 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- 102100039611 Glutamine synthetase Human genes 0.000 description 2
- 101000934368 Homo sapiens CD63 antigen Proteins 0.000 description 2
- 101000888841 Homo sapiens Glutamine synthetase Proteins 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000027455 binding Effects 0.000 description 2
- 210000000981 epithelium Anatomy 0.000 description 2
- 210000004408 hybridoma Anatomy 0.000 description 2
- 238000012151 immunohistochemical method Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 210000004126 nerve fiber Anatomy 0.000 description 2
- 210000000933 neural crest Anatomy 0.000 description 2
- 230000001537 neural effect Effects 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000004393 prognosis Methods 0.000 description 2
- 230000006916 protein interaction Effects 0.000 description 2
- 239000012474 protein marker Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 239000012099 Alexa Fluor family Substances 0.000 description 1
- 108050007222 Coronin Proteins 0.000 description 1
- 201000010374 Down Syndrome Diseases 0.000 description 1
- 206010066054 Dysmorphism Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 101000817607 Homo sapiens Dynamin-2 Proteins 0.000 description 1
- 101001042351 Homo sapiens LIM and senescent cell antigen-like-containing domain protein 1 Proteins 0.000 description 1
- 101000734646 Homo sapiens Programmed cell death protein 6 Proteins 0.000 description 1
- 101001091538 Homo sapiens Pyruvate kinase PKM Proteins 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102100021754 LIM and senescent cell antigen-like-containing domain protein 1 Human genes 0.000 description 1
- 101001066400 Mesocricetus auratus Homeodomain-interacting protein kinase 2 Proteins 0.000 description 1
- 102000006833 Multifunctional Enzymes Human genes 0.000 description 1
- 108010047290 Multifunctional Enzymes Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 102100034785 Programmed cell death protein 6 Human genes 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 102100034911 Pyruvate kinase PKM Human genes 0.000 description 1
- 101000916532 Rattus norvegicus Zinc finger and BTB domain-containing protein 38 Proteins 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 102000013529 alpha-Fetoproteins Human genes 0.000 description 1
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 1
- 230000019552 anatomical structure morphogenesis Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000003766 bioinformatics method Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000007698 birth defect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 102000018123 coronin Human genes 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 125000002228 disulfide group Chemical group 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 238000010201 enrichment analysis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000008175 fetal development Effects 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- 230000002962 histologic effect Effects 0.000 description 1
- 102000047617 human DNM2 Human genes 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000003988 neural development Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 238000003921 particle size analysis Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000011870 unpaired t-test Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/38—Pediatrics
- G01N2800/385—Congenital anomalies
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/50—Determining the risk of developing a disease
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Microbiology (AREA)
- Pathology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Cell Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the technical field of biological medicines, and particularly relates to a molecular marker for prenatal noninvasive diagnosis of nerve tube deformity, congenital heart disease and cleft lip and palate fetuses and application thereof. One or more of the molecular markers, CORO1A, DNM2 and ACTR2 proteins for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetuses. The molecular marker for prenatal noninvasive diagnosis is applied to the preparation of products for prenatal screening, early warning, clinical diagnosis and biochemical inspection of neural tube deformity, congenital heart disease and cleft lip and palate fetuses. The invention discovers and proves that the abnormal expression of proteins (including CORO1A, DNM and ACTR 2) in the blood of pregnant women has close correlation with the occurrence of nerve tube deformity, congenital heart disease and cleft lip and palate fetus for the first time, the verified sample size is large, the result is accurate, and a novel approach is provided for prenatal screening, early warning and diagnosis of nerve tube deformity, congenital heart disease and cleft lip and palate fetus.
Description
Technical Field
The invention belongs to the technical field of medical biology, and particularly relates to a molecular marker (CORO 1A, DNM and ACTR2 proteins) for prenatal noninvasive diagnosis of nerve tube deformity, congenital heart disease and cleft lip and palate fetuses and application thereof.
Background
The neural tube deformity, congenital heart disease, cleft lip and palate and the like are common serious fetal development deformity in China, and seriously endanger the improvement of life quality of birth population. Therefore, the method for noninvasive diagnosis of the early embryo of the congenital deformity is researched, so that the diagnosis can be obtained before serious structural abnormality or irreversible damage, and a corresponding new strategy for early embryo treatment and prevention is formulated, so that the method has great significance for reducing the disability rate of the deformity and improving the population quality.
The establishment of a noninvasive early screening method for congenital malformations is always an ideal target for continuous pursuit. Although imaging techniques (ultrasound and MRI) are currently developed very rapidly, the time for diagnosing some congenital anomalies is advanced, and the requirements for early screening and diagnosis are still not met. Maternal serological examination is a noninvasive prenatal diagnosis method, is easy for pregnant women to accept, and is suitable for large-scale prenatal screening. Therefore, many scholars at home and abroad are devoted to the research of finding new diagnosis specific markers, but other birth defects are not clinically obtained to be used as diagnosis molecular markers except for neural tube deformity and Down syndrome which can be screened by using serum alpha fetoprotein.
With the rapid development of various histology technologies in recent years, a series of new technologies are integrated into high-throughput histology research, so that the screening work of disease diagnosis molecular markers is broken through, and an extremely important means is provided for the conversion medical research which is very important at present. The high-throughput histology technology is utilized to fully consider two factors of a mother and a fetus, a group of key molecules which are obviously changed are screened from a plurality of complex proteins, and the change rule is comprehensively analyzed, so that the molecular markers which are favorable for determining early diagnosis and prognosis judgment of diseases are also the trend of converting the complex molecular markers into clinical application in the future.
Disclosure of Invention
In view of the problems existing in the prior art, the invention aims to provide a molecular marker for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate and application thereof, wherein the molecular marker is CORO1A, DNM2 and ACTR2 proteins, can be singly or jointly used for prenatal screening of congenital deformity, and provides a new target point for treatment of neural tube deformity, congenital heart disease and cleft lip and palate.
In order to achieve the above purpose, the present invention adopts the following technical scheme.
The molecular marker for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetus is composed of one or more of CORO1A, DNM and ACTR2 proteins.
The molecular marker is applied to the preparation of products for prenatal screening, early warning, clinical diagnosis and biochemical inspection of neural tube deformity, congenital heart disease and cleft lip and palate fetuses.
Further, the product comprises a reagent, a kit, a chip, test paper and a high-throughput sequencing platform, and protein molecular markers related to prenatal noninvasive diagnosis of nerve tube deformity, congenital heart disease and cleft lip and palate fetuses are detected by mass spectrometry, PCR, in situ hybridization, fluorescence in situ hybridization, immunotransmission turbidimetry, radioimmunoassay and other related methods.
Further, specimens for prenatal screening, early warning, clinical diagnosis and biochemical examination of the nerve tube deformity, congenital heart disease and cleft lip and palate include pregnant woman blood (and exosomes thereof), urine (and exosomes thereof), amniotic fluid (and exosomes thereof), fetal specimens and the like.
Use of a reagent for detecting molecular markers for prenatal noninvasive diagnosis of a neural tube deformity, congenital heart disease, and cleft lip and palate fetus in the preparation of a prenatal noninvasive diagnostic tool for a fetus.
Further, the reagent for detecting the prenatal noninvasive diagnostic molecular markers of the neural tube deformity, congenital heart disease and cleft lip and palate fetus comprises a reagent capable of quantifying the above proteins.
Further, the reagent capable of quantifying the above protein may be a specific primer for a gene or transcript, or may be a specific recognition probe, or may include both a primer and a probe.
A tool for prenatal screening, early warning and diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetuses, said tool being capable of detecting the above molecular expression levels.
Further, the tools include molecular markers capable of quantifying prenatal noninvasive diagnosis proteins from neural tube deformity, congenital heart disease, and cleft lip and palate fetuses including CORO1A, DNM2 and ACTR2.
Further, the tools for prenatal screening, early warning and diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetuses include chips, kits, test paper and high throughput sequencing platforms.
A method of prenatal screening, early warning and diagnosis of nerve tube deformity, congenital heart disease and cleft lip and palate, the method comprising the steps of.
(1) A sample of the subject is obtained.
(2) Detecting the level of expression of said molecule in a sample from the subject.
(3) Correlating the measured trend of the molecular expression level with the disease correlation of the subject.
(4) The abnormal expression of the above molecules, compared to normal controls, indicates that the subject is at high risk of having a neural tube deformity, congenital heart disease, and cleft lip and palate.
The reagent for quantifying protein of the present invention can exert its function based on a known method other than antibody: such as mass spectrometry techniques including MRM and PRM, etc.
The reagent for quantifying protein of the present invention can exert its function based on a known method using an antibody: for example, chips (protein chips, microfluidic chips, etc.), digital single molecule immunoassays, ELISA, radioimmunoassays, immunonephelometry, immunohistochemistry, western blotting, etc. can be included.
The reagent for quantifying a protein of the present invention includes an antibody or a fragment thereof that specifically binds to a protein. Antibodies or fragments thereof of any structure, size, immunoglobulin class, origin, etc. may be used as long as it binds to the target protein. Antibodies or fragments thereof included in the detection products of the invention may be monoclonal or polyclonal. An antibody fragment refers to a portion of an antibody (a fragment of a portion) or a peptide containing a portion of an antibody that retains the binding activity of the antibody to an antigen. Antibody fragments may include F (ab ') 2, fab', fab, single chain Fv (scFv), disulfide bonded Fv (dsFv) or polymers thereof, dimerized V regions (diabodies), or CDR-containing peptides. The reagent for quantifying a protein of the present invention may include an isolated nucleic acid encoding an antibody or an amino acid sequence encoding an antibody fragment, a vector comprising the nucleic acid, and a cell carrying the vector.
Antibodies can be obtained by methods well known to those skilled in the art. For example, mammalian cell expression vectors retaining polypeptides of the whole or part of the target protein or polynucleotides encoding them are prepared as antigens. After immunization of an animal with an antigen, immune cells are obtained from the immunized animal and myeloma cells are fused to obtain hybridomas. Antibodies were then collected from the hybridoma cultures. Finally, monoclonal antibodies against the proteins of the invention can be obtained by performing antigen-specific purification of the obtained antibodies using the proteins or parts thereof used as antigens. Polyclonal antibodies can be prepared as follows: animals were immunized with the same antigens as above, blood samples were collected from the immunized animals, serum was separated from the blood, and antigen-specific purification of serum was performed using the above antigens. The antibody fragment may be obtained by treating the obtained antibody with an enzyme or by using sequence information of the obtained antibody.
Binding of the label to the antibody or fragment thereof may be carried out by methods commonly known in the art. For example, proteins or peptides may be fluorescently labeled as follows: the protein or peptide is washed with phosphate buffer, added with DMSO, buffer, etc., and the solution is then mixed and allowed to stand at room temperature for another 10 minutes. In addition, the labeling may use commercial labeling kits, such as biotin labeling kits, e.g., biotin labeling kit-NH 2, biotin labeling kit-SH (Dojindo Laboratories); alkaline phosphatase labeling kits such as alkaline phosphatase labeling kit-NH 2, alkaline phosphatase labeling kit-SH (Dojindo Laboratories); peroxidase labeling kits such as peroxidase labeling kit-NH 2, peroxidase labeling kit-NH 2 (Dojindo Laboratories); phycobiliprotein labeling kits such as phycobiliprotein labeling kit-NH 2, phycobiliprotein labeling kit-SH, B-phycoerythrin labeling kit-NH 2, B-phycoerythrin labeling kit-SH, R-phycoerythrin labeling kit-NH 2, R-phycoerythrin labeling kit SH (Dojindo Laboratories); fluorescent labeling kits such as fluorescein labeling kit-NH 2, hiLyte fluor (TM) 555 labeling kit-NH 2, hiLyte fluor (TM) 647 labeling kit-NH 2 (Dojindo Laboratories); and Dylight 547 and Dylight647 (Techno chemical Corp.), zenon (TM), alexa Fluor (TM) antibody labelling kit, qdot (TM) antibody labelling kit (Invitrogen Corporation) and EZ-marker protein labelling kit (Funakoshi Corporation). For proper labeling, a suitable instrument may be used to detect the labeled antibody or fragment thereof.
The sample of the present invention for detecting the expression level of the above-mentioned molecules is obtained by a routine technique in the art, preferably by a non-invasive or minimally invasive method.
The inventive samples may be (but are not limited to): pregnant woman blood, urine, amniotic fluid and malformed fetus or infant specimens. In a specific embodiment of the invention, the sample is from tissue of a subject.
The high-throughput proteomics detection platform is a special tool, and by comparing protein expression differences of patients with diseases and normal people, the abnormality of which protein expression is related to the diseases can be easily analyzed. Therefore, the knowledge that the abnormal expression of the molecules is related to nerve tube deformity, congenital heart disease and cleft lip and palate in high-throughput proteomics detection also belongs to the new application of the invention, and is also within the protection scope of the invention.
The kit of the present invention may comprise a plurality of different reagents suitable for practical use (e.g., for different detection methods), and is not limited to the reagents listed so far, as reagents for judging neural tube deformity, congenital heart disease, and cleft lip and palate based on the above molecular detection are included in the scope of the present invention.
In the context of the present invention, "prenatal screening, early warning and diagnosis of a neural tube deformity, congenital heart disease and cleft lip and palate fetus" includes determining whether the subject fetus has developed a neural tube deformity, congenital heart disease and cleft lip and palate, and determining whether the subject fetus is at risk of developing a neural tube deformity, congenital heart disease and cleft lip and palate.
Compared with the prior art, the invention has the following beneficial effects.
The invention discovers and proves that the abnormal expression of proteins (including CORO1A, DNM2 and ACTR 2) in the blood of pregnant women has close correlation with the occurrence of nerve tube deformity, congenital heart disease and cleft lip and palate fetus for the first time, and has the advantages of more verified sample size and accurate result.
The protein marker related to prenatal noninvasive diagnosis of the neural tube deformity, congenital heart disease and cleft lip and palate fetuses provided by the invention provides the service of prenatal diagnosis or risk monitoring of the neural tube deformity, congenital heart disease and cleft lip and palate fetuses, and the service of consulting diagnosis and prognosis is sold to hospitals and clinics cooperatively or independently.
The protein marker related to prenatal noninvasive diagnosis of the neural tube deformity, congenital heart disease and cleft lip and palate fetus provided by the invention provides a new way for prenatal screening, early warning and diagnosis of the neural tube deformity, congenital heart disease and cleft lip and palate fetus.
Drawings
FIGS. 1-3 are graphs showing that three differential proteins (CORO 1A, DNM and ACTR 2) with interactions were found by bioinformatic analysis using proteomic techniques to screen 33 differential expression proteins in the peripheral blood of neural tube deformities.
FIGS. 4-1 to 4-4 show that the quantitative detection and verification of three different expression proteins of CORO1A, DNM and ACTR2 are carried out in peripheral blood of the neural tube deformity pregnant mice on embryo day E12, day E14 and day E18 by using a Western-blot method, and the obvious low expression of CORO1A, DNM2 and ACTR2 in the peripheral blood of the neural tube deformity pregnant mice is proved.
FIGS. 5-1 to 5-4 show quantitative detection and verification of three differential expression proteins of CORO1A, DNM2 and ACTR2 in neural tube malformation spinal cord tissues of embryo E12 day and E18 day and in exosomes derived from peripheral blood embryo neural tissues of pregnant mice of embryo E18 day by using Western-blot method, and detection analysis of expression conditions of CORO1A, DNM2 and ACTR2 in neural tube malformation spinal cord tissues by using immunohistochemical method, and show that CORO1A, DNM2 and ACTR2 are obviously expressed in neural tube malformation.
FIGS. 6-1 to 6-3 show that the quantitative detection of CORO1A and DNM2 in the peripheral blood of the pregnant women with the malformation of the nerve ducts by ELISA method proves that the CORO1A and DNM2 are obviously low expressed in the peripheral blood of the pregnant women with the malformation of the nerve ducts and the change in early pregnancy is more obvious.
FIGS. 7-1 through 7-3 are results of ROC curve analysis showing that CORO1A and DNM2 are better for diagnosing neural tube deformity in early pregnancy, as shown by the diagnostic accuracy, sensitivity and specificity analysis of pregnant women with neural tube deformity fetuses by CORO1A and DNM 2.
Detailed Description
The invention will now be described in further detail with reference to the drawings and examples. The following examples are only illustrative of the present invention and are not intended to limit the scope of the invention. The experimental methods for which specific conditions are not specified in the examples are generally conducted under conventional conditions or under conditions recommended by the manufacturer. In the embodiment, an animal model is used to be sourced from the Shengjing hospital animal center, and the peripheral blood sample of the pregnant woman is sourced from the Shengjing birth queue sample library, so that the ethical committee approval (approval number: 2017PS 264K) of the Shengjing hospital affiliated to the university of medical science in China is obtained.
Example 1 screening of differentially expressed proteins and bioinformatic analysis in peripheral blood of neural tube-malformed pregnant mice using proteomic techniques.
Separating plasma exosomes of the neural tube deformed pregnant mice by adopting a super-high speed centrifugation method, and completing the related identification work of the exosomes. Proteomic screening was performed using non-labeled quantification techniques (label-free) in combination with bioinformatic analysis to initially screen for possible protein markers, as shown in FIGS. 1-3.
1. Plasma was isolated.
Collecting whole blood sample, mixing in EDTA anticoagulant tube, centrifuging at 4deg.C for 10min with 1600 Xg, collecting supernatant (blood plasma) into new EP tube, centrifuging at 16000 Xg for 10min to remove cell debris, and packaging into multiple centrifuge tubes.
2. Isolation and identification of exosomes.
Separating plasma exosomes by using a method of ultra-high speed centrifugation. The separation method comprises the following steps: 10000 Xg was centrifuged for 1h at 4℃and the supernatant was transferred to a new ultra-high speed centrifuge tube and centrifuged at 100000 Xg for 4h at 4 ℃. The supernatant was discarded and the exosomes were resuspended with 100 μl cold PBS. The isolated exosome particles were identified using three standard methods (transmission electron microscopy, particle size analysis and exosome marker proteins).
3. Proteomic detection and bioinformatic analysis.
Proteomics detection of 3 nerve tube malformed pregnant mice and 3 normal pregnant mice by a non-labeling quantitative technology (label-free) is carried out, the histology data is input into TBtools software to make a heat map, the bioinformatics analysis is carried out on the webgestalt website to carry out enrichment analysis of the gene GO biological process, and then protein interaction network is carried out on the differential proteins on the String website.
4. Exosome identification and proteome detection results.
(1) Exosome identification: the transmission electron microscope and particle diameter detection prove that the exosome has a peak at the diameter of 100nm, which indicates that the exosome extracted by the ultracentrifugation method has good purity. Then, western blot detection is carried out on Alix, CD63 and CD9 by using exosomes with different concentrations to identify the expression condition of the exosome markers, and the result shows that Alix, CD63 and CD9 bands show correct change trend along with the increase of the concentration gradient.
(2) Proteomic results: a total of 3 serum samples from E18 normal mice and 3 serum exosome samples from neural tube deformities were used for proteomic detection, a total of 63 enriched proteins were found, and 33 proteins were different between the normal and malformed groups. There were 7 proteins specifically expressed in NTDs, 12 proteins specifically expressed in the normal group, and 2 unknown proteins. In addition, 14 proteins were expressed in both NTDs and normal groups, with 1 protein being highly expressed in the nerve tube deformity and 13 proteins being under-expressed. GO analysis found an enrichment of 4 biological processes, respectively: modulating actin polymerization (depolymerization), modulating anatomical morphogenesis, reaction to oxygenates, and cellular structure assembly. Wherein ACTR2, cor 1A, DNM, AGT, GLUL, LIMS1, PDCD6, and PKM are involved in 2 and more biological processes, wherein ACTR2, cor 1A, GLUL, and DNM2 are associated with neural development and have not been reported in neural tube deformities. These 4 proteins were again analyzed for protein interactions on the String website, which showed that ACTR2, CORO1A, and DNM2 could be clustered into the same interacting network cluster.
Example 2 the expression of ACTR2, CORO1A and DNM2 was demonstrated in a neural tube malformed serum exosome.
1. The expression of ACTR2, cor 1A and DNM2 was verified in embryo E18 day serum exosomes.
And 19 samples which are not detected by histology and are used for 18 days of embryo E are selected for carrying out Western-blot expansion sample size verification. The assay uses Alix as an internal reference to detect ACTR2 at 45kDa, cor 1A at 57kDa and DNM2 at 98 kDa. A total of 19 samples from serum exosomes of pregnant mice (19 normal, 19 nerve tube deformities) were tested and found to be consistent with the trend of the histologic results, all under expressed in the malformed group. Statistical results also show that there are statistical differences in the low expression of ACTR2, cor o1A and DNM2 in the neural tube deformity group. The detection of CORO1A by ELISA also confirmed the consistency of the Western-blot results.
2. Expression of ACTR2, cor 1A and DNM2 in embryo E12, E14 and E16 day serum exosomes.
To verify if ACTR2, CORO1A and DNM2 were also altered in early gestation, the serum exosomes at E12, E14, E16 days were verified using Western-blot methods. The results found that the band expression and unpaired T test statistics were all shown to be significantly down-regulated in the neural tube dysmorphism group. This part of the results clearly shows that the trend of expression of these proteins in early gestation is consistent with day E18. In addition, the ELISA showed a significant downregulation trend for the CORO1A detection result of E12, and the consistency of the Western-blot results was also confirmed. This result in part defines that ACTR2, cor 1A and DNM2 also have a significant down-regulation trend during E12 to E16, suggesting that they may be diagnostic indicators of early-gestational neural tube deformity.
ACTR2, CORO1A and DNM2 found by the proteomics results above were validated in the neural tube malformation serum exosomes using the Western-blot method, as shown in FIGS. 4-1 through 4-4.
Example 3ACTR2, CORO1A and DNM2 were expressed in spinal cord tissue and embryonic neurogenic exosomes.
1. ACTR2, CORO1A and DNM2 expressed in spinal cord tissue.
In order to investigate the relationship between ACTR2, CORO1A and DNM2 and the occurrence of nerve tube deformity, the expression situation of the ACTR2, CORO1A and DNM2 in spinal cord tissues is clarified, the expression situation of the embryo E18 and E12 day spinal cord tissues is detected by using Western-blot, and beta-actin is used as an internal reference of tissue detection. The results show that ACTR2, CORO1A and DNM2 all appear to have statistically significant downregulation trends in spinal cord tissue of neural tube abnormalities consistent with serum exosome trends. Immunohistochemical staining showed ACTR2, cor 1A and DNM2 expression in neural tube malformation spinal cord tissue for E18 days, confirming whether or not these proteins were specifically expressed in neural tube tissue. ACTR2 is expressed in nerve cells, nerve epithelium, and on the neural crest side, and is significantly down-regulated in the neural tube malformation tissue; CORO1A is obviously expressed on nerve fibers and nerve cells, and has downregulation expression in nerve tube malformation tissues; DNM2 is expressed on nerve fibers, nerve epithelium, nerve cells, and neural crest, and shows down-regulation trend in nerve canal deformity tissues. Suggesting that changes in these 3 indices in serum exosomes are likely to be derived from changes in spinal cord tissue.
2. ACTR2, CORO1A and DNM2 were expressed in embryonic neurogenic exosomes.
In order to verify whether the low expression of ACTR2, CORO1A and DNM2 in the serum exosomes of the pregnant mice with the nerve tube deformity is caused by the low expression in the serum exosomes of the embryonic nerve origin, the serum exosomes of the pregnant mice are further separated, the separation kit combined by specific antibodies is used for separating the serum exosomes of the embryonic nerve origin, and the quantitative detection of ACTR2, CORO1A and DNM2 is carried out by using a Western-blot method, so that the results show that the expression of CORO1A and DNM2 is obviously reduced in the nerve tube deformity group, and the expression quantity of ACTR2 has no statistical difference between the nerve tube deformity group and the normal control group. The results suggest that reduced expression of CORO1A and DNM2 in serum exosomes of pregnant mice is due to reduced expression in exosomes produced by embryonic spinal cord tissue.
The above-described ACTR2, CORO1A and DNM2, which were verified in serum exosomes of pregnant mice, were verified in neural tube malformation spinal cord tissue and in embryonic neurogenic exosomes using Western-blot and immunohistochemical methods, as shown in FIGS. 5-1 through 5-4.
Example 4 diagnostic efficacy of CORO1A and DNM2 as molecular markers was validated in peripheral blood exosomes of neural tube deformities, congenital heart disease, and cleft lip and palate fetuses mothers.
1. A study sample was included.
Collecting peripheral blood of 19 nerve tube malformed fetus mothers, 100 congenital heart disease fetus mothers and 40 cleft lip and palate fetus mothers from Beijing birth queues, and taking 159 normal pregnant women with the age and gestational age similar to those of the pregnant women as normal control.
2. Isolation of exosomes.
Separating plasma exosomes by using a method of ultra-high speed centrifugation. The separation method comprises the following steps: 10000 Xg was centrifuged for 1h at 4℃and the supernatant was transferred to a new ultra-high speed centrifuge tube and centrifuged at 100000 Xg for 4h at 4 ℃. The supernatant was discarded and the exosomes were resuspended with 100 μl cold PBS.
3. ELISA detects the expression level of CORO1A and DNM 2.
50 Microliter of serum exosomes are diluted to 100 microliter in PBS liquid, then the human CORO1A ELISA kit and the human DNM2 ELISA kit are used, the operation is carried out according to the steps of the specification, after the reaction stopping solution is added, the detection is carried out at 450 nanometers by using a multifunctional enzyme-labeled instrument, and the expression quantity of CORO1A and DNM2 is calculated according to a standard curve.
4. CORO1A and DNM2 as molecular markers for diagnostic efficacy in peripheral blood exosomes of neural tube deformities, congenital heart disease and cleft lip and palate fetal mother.
DNM2 detection results show that the expression level of peripheral blood exosomes of the neural tube malformed fetus mother is obviously reduced, and compared with a normal pregnant woman group, the neural tube malformed fetus mother has statistical difference. ROC curve analysis DNM2 diagnosed neural tube deformity with a sensitivity of 73.68% and a specificity of 78.95%. Further analysis of the perigestational period of the neural tube malformed fetal mother shows that DNM2 has better diagnostic efficiency on early gestation period (12 weeks-18 weeks gestation), and the sensitivity and the specificity reach 100%. The sensitivity is 68.42 percent and the specificity is 89.47 percent for the diagnosis efficacy in the late pregnancy. The expression level of peripheral blood exosomes of the mother of the fetus with congenital heart disease is obviously reduced, and compared with the normal pregnant women, the expression level of peripheral blood exosomes of the mother of the fetus with congenital heart disease is statistically different. ROC curve analysis DNM2 diagnosed congenital heart disease with a sensitivity of 78.68% and a specificity of 80.65%. The expression level of peripheral blood exosomes of the cleft lip and palate fetal mother is also obviously reduced, and compared with a normal pregnant woman group, the expression level of peripheral blood exosomes of the cleft lip and palate fetal mother has statistical difference. ROC curve analysis DNM2 diagnosed neural tube deformity with a sensitivity of 70.38% and a specificity of 75.55%.
The detection result of CORO1A shows that the expression level of peripheral blood exosomes of the neural tube malformed fetal mother is also obviously reduced, and compared with a normal pregnant woman group, the expression level of peripheral blood exosomes of the neural tube malformed fetal mother is statistically different. ROC curve analysis cor o1A diagnosed neural tube deformity with a sensitivity of 73.68% and a specificity of 78.95%. Further analysis of the perigestational period of the neural tube malformed fetal mother revealed that CORO1A also had better diagnostic performance for early gestation (12-18 weeks gestation), sensitivity of 85.71% and specificity of 85.71%. The sensitivity is 75.00% and the specificity is 83.33% for the diagnosis efficacy in the late pregnancy. The expression level of peripheral blood exosomes of the mother of the fetus with congenital heart disease is obviously reduced, and compared with the normal pregnant women, the expression level of peripheral blood exosomes of the mother of the fetus with congenital heart disease is statistically different. ROC curve analysis cor o1A diagnosed congenital heart disease with a sensitivity of 80.11% and a specificity of 79.28%. The expression level of peripheral blood exosomes of the cleft lip and palate fetal mother is also obviously reduced, and compared with a normal pregnant woman group, the expression level of peripheral blood exosomes of the cleft lip and palate fetal mother has statistical difference. ROC curve analysis cor o1A diagnosed neural tube deformity with a sensitivity of 71.22% and a specificity of 77.78%.
For the molecular markers (CORO 1A and DNM 2) screened in the above animal model, ELISA was used to verify the peripheral blood exosomes of neural tube deformity, congenital heart disease and cleft lip and palate fetus mother, as shown in FIGS. 6-1 to 6-3 and 7-1 to 7-3.
Claims (5)
1. Use of a reagent for detecting prenatal noninvasive diagnostic molecular markers, said markers being one or more of the proteins CORO1A, DNM, in the manufacture of a product for prenatal diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetuses.
2. The use according to claim 1, wherein the product comprises reagents, kits, chips, test papers, high throughput sequencing platforms, detection of prenatal noninvasive diagnostic molecular markers for fetuses with neural tube deformity, congenital heart disease and cleft lip and palate by mass spectrometry, PCR, in situ hybridization, fluorescence in situ hybridization, immunonephelometry, radioimmunoassay.
3. The use of claim 1, wherein the diagnostic sample comprises pregnant woman blood and exosomes, urine and exosomes, amniotic fluid and exosomes, and fetal samples.
4. Use of a reagent for detecting a molecular marker for prenatal noninvasive diagnosis of a neural tube deformity, congenital heart disease or cleft lip and palate fetus in the preparation of a tool for prenatal noninvasive diagnosis of a neural tube deformity, congenital heart disease or cleft lip and palate fetus, said marker being composed of one or more of the proteins cor 1A, DNM.
5. The use of claim 4, wherein the means for detecting molecular markers for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease, and cleft lip and palate comprises means for quantitatively detecting molecular markers for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease, or cleft lip and palate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110285096.0A CN112816711B (en) | 2021-03-17 | 2021-03-17 | Molecular marker for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetus and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110285096.0A CN112816711B (en) | 2021-03-17 | 2021-03-17 | Molecular marker for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetus and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112816711A CN112816711A (en) | 2021-05-18 |
| CN112816711B true CN112816711B (en) | 2024-04-19 |
Family
ID=75863447
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110285096.0A Active CN112816711B (en) | 2021-03-17 | 2021-03-17 | Molecular marker for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetus and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112816711B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114019173B (en) * | 2021-11-17 | 2023-03-14 | 武汉大学 | Marker for predicting non-syndromic cleft lip and palate and application thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1852974A (en) * | 2003-06-09 | 2006-10-25 | 密歇根大学董事会 | Compositions and methods for treating and diagnosing cancer |
| EP2019318A1 (en) * | 2007-07-27 | 2009-01-28 | Erasmus University Medical Center Rotterdam | Protein markers for cardiovascular events |
| CN101541977A (en) * | 2006-09-19 | 2009-09-23 | 诺瓦提斯公司 | Biomarkers of target modulation, efficacy, diagnosis and/or prognosis for RAF inhibitors |
| KR20110135911A (en) * | 2011-12-05 | 2011-12-20 | 경북대학교 산학협력단 | Biomarker |
| CN103374627A (en) * | 2012-04-27 | 2013-10-30 | 中国科学院上海生命科学研究院 | Congenital heart defect (CHD)-related gene polycystic kidney disease 1 like l (PKD1L1) and application thereof |
| CN105699367A (en) * | 2016-01-19 | 2016-06-22 | 中国医科大学附属盛京医院 | Molecular marker and application of molecular marker in preparation of biological marker agent |
| CN110016504A (en) * | 2019-05-20 | 2019-07-16 | 广州市妇女儿童医疗中心 | Application, the product of neural tube malformation Prenatal Screening and method of the CDR1as in neural tube malformation Prenatal Screening |
| CN111944892A (en) * | 2020-08-24 | 2020-11-17 | 中国医科大学附属盛京医院 | Molecular marker for prenatal noninvasive diagnosis of cleft lip and palate and application thereof |
-
2021
- 2021-03-17 CN CN202110285096.0A patent/CN112816711B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1852974A (en) * | 2003-06-09 | 2006-10-25 | 密歇根大学董事会 | Compositions and methods for treating and diagnosing cancer |
| CN101541977A (en) * | 2006-09-19 | 2009-09-23 | 诺瓦提斯公司 | Biomarkers of target modulation, efficacy, diagnosis and/or prognosis for RAF inhibitors |
| EP2019318A1 (en) * | 2007-07-27 | 2009-01-28 | Erasmus University Medical Center Rotterdam | Protein markers for cardiovascular events |
| KR20110135911A (en) * | 2011-12-05 | 2011-12-20 | 경북대학교 산학협력단 | Biomarker |
| CN103374627A (en) * | 2012-04-27 | 2013-10-30 | 中国科学院上海生命科学研究院 | Congenital heart defect (CHD)-related gene polycystic kidney disease 1 like l (PKD1L1) and application thereof |
| CN105699367A (en) * | 2016-01-19 | 2016-06-22 | 中国医科大学附属盛京医院 | Molecular marker and application of molecular marker in preparation of biological marker agent |
| CN110016504A (en) * | 2019-05-20 | 2019-07-16 | 广州市妇女儿童医疗中心 | Application, the product of neural tube malformation Prenatal Screening and method of the CDR1as in neural tube malformation Prenatal Screening |
| CN111944892A (en) * | 2020-08-24 | 2020-11-17 | 中国医科大学附属盛京医院 | Molecular marker for prenatal noninvasive diagnosis of cleft lip and palate and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Neural Tube Defects: From a Proteomic Standpoint;Tania M. Puvirajesinghe;metabolites;20150317;第5卷(第1期);164-183 * |
| 神经管畸形发病机制研究进展与新策略;袁正伟;发育医学电子杂志;20200730;第8卷(第3期);205-206 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112816711A (en) | 2021-05-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6173387B2 (en) | Detection of intraamniotic infection | |
| US20100029006A1 (en) | Multiplexed diagnostic test for preterm labor | |
| JP6691617B2 (en) | Methods and compositions for providing an assessment of preeclampsia | |
| WO2011005893A9 (en) | Biomarkers and methods for detecting alzheimer's disease | |
| JP2018535433A (en) | Biomarkers for the detection of breast cancer in women with dense breast | |
| CN109891239B (en) | Methods and kits for providing preeclampsia assessment and prediction of preterm labor | |
| JP2019518225A (en) | Method and composition for predicting preterm birth | |
| JP2018205327A (en) | Method and composition for diagnosing preeclampsia | |
| WO2008092094A9 (en) | Method and materials for detection, diagnosis and management of ovarian cancer | |
| CN112816711B (en) | Molecular marker for prenatal noninvasive diagnosis of neural tube deformity, congenital heart disease and cleft lip and palate fetus and application thereof | |
| WO2016195051A1 (en) | Plasma biomarker panel for diagnosing pancreatic cancer | |
| JP6252949B2 (en) | Schizophrenia marker set and its use | |
| US20230400472A1 (en) | Combination of biomarkers of preterm delivery | |
| CN112946299B (en) | Application of product of quantitative FTL in preparation of preeclampsia diagnosis tool | |
| WO2013054984A1 (en) | Composition for diagnosis of lung cancer and diagnosis kit of lung cancer | |
| CN110656169B (en) | Diagnostic markers for atrial fibrillation | |
| CN112553332A (en) | Combined application of multiple genes in liver cancer diagnosis | |
| CN112695089A (en) | Combined diagnostic markers | |
| CN112680514A (en) | Marker for liver cancer diagnosis and application thereof | |
| CN117647645B (en) | Application of LBP, ATF6 and M-CSFR combination in preparation of product for diagnosing autoimmune liver disease and kit | |
| CN116643054A (en) | Non-syndrome type cleft lip and palate prenatal noninvasive diagnosis molecular marker C3 protein and application thereof | |
| WO2020091173A1 (en) | Biomarker for prediction of gestational diabetes and hypertension in pregnancy | |
| WO2009152601A1 (en) | Use of follistatin-related gene (flrg) as a marker for identifying a fetal abnormality | |
| CN114686582A (en) | Application of GDF15 and ITIH3 in prediction of spontaneous abortion in early pregnancy | |
| CN106048006B (en) | SMCHD1 is preparing the application in diagnosis of coronary heart disease product |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |