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Corin gene variant and application thereof

A gene variant and mutation technology, applied in the field of clinical medicine and molecular biology, can solve the problems of unclear pathogenesis and unclear pathogenic mechanism of patients, and achieve highly specific effects.

Inactive Publication Date: 2016-07-06
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, only 5% of hypertensive patients can be clearly diagnosed as being caused by a specific genetic defect, and the pathogenic mechanism of most patients is still unclear
Hypertensive disorders in pregnancy (also known as pregnancy-induced hypertension syndrome, Pregnancy-induced hypertension syndrome) are related factors including vascular growth factor receptors, angiotensin and estrogen, etc., but the specific pathogenesis is still unclear

Method used

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  • Corin gene variant and application thereof
  • Corin gene variant and application thereof
  • Corin gene variant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Collection of blood samples.

[0027] Research objects: Patients with hypertension and pregnancy-induced hypertension were taken as the research objects. The diagnostic criteria of hypertension were based on the 1999 WHO / ISH blood pressure classification and standards. The clinical diagnostic criteria of pregnancy-induced hypertension are as follows: hypertension occurs after 20 weeks of pregnancy, and the random blood pressure is ≥140 / 90mmHg twice at intervals of 4 hours. The normal control group had no history of essential hypertension, chronic liver disease, kidney disease, diabetes, heart disease, and hyperthyroidism, and no underlying diseases such as smoking, drinking, blood transfusion, and immunotherapy.

[0028] Sample collection: venous blood collection using EDTA anticoagulant tubes, centrifugation at 3000 rpm for 10 minutes at room temperature, separation of plasma and blood cells, aliquoting and storage at -80°C for later use.

Embodiment 2

[0029] Example 2: Extraction and PCR amplification of genomic DNA.

[0030] Extraction of genomic DNA from blood cells: Use the whole blood DNA extraction kit (QIAampDNAMiniandBloodMiniKit) produced by Qiagen, USA, and follow the instructions in the kit to extract genomic DNA from the sample, and store it at -20°C.

[0031] Design of PCR primers: According to the sequence of human Corin gene in Genbank, the Primer5.0 software was used to design Corin exon amplification primers. The specific primer sequences are shown in Table 1.

[0032]

[0033]

[0034]

[0035]

[0036] PCR amplification and sequence analysis: PCR amplification conditions: pre-denaturation at 94°C for 6 min; denaturation at 94°C for 50 s, extension at 58°C for 32 s, annealing at 72°C for 32 s, and extension at 72°C for 8 min after 35 cycles. PCR products were sequenced and analyzed. The sequencing results were compared with the human Corin mRNA sequence (NM_006587.3) on the NCBI website using C...

Embodiment 3

[0037] Example 3: Identification of Corin gene mutations or polymorphisms and research on their relationship with diseases.

[0038] In order to judge whether the found Corin gene variation is mutation or polymorphism, the distribution of Corin gene variation in normal population and hypertensive patients was analyzed by PCR product sequencing or quantitative PCR. Taking normal people and hypertensive patients as research objects (n>100 cases / group), peripheral blood was collected, genomic DNA was extracted, the exon where the gene was changed was amplified, and the PCR product was sequenced and analyzed. The sequencing results were compared with the human Corin mRNA sequence (NM_006587.3) in the NCBI website using Chromas2.31 software.

[0039] If the corresponding variation is not found in the normal population, it indicates that the variation is a gene mutation; if the variation frequency in the normal population is greater than or equal to 1%, it indicates that the variati...

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Abstract

The invention discloses a Corin gene variant and application thereof.Specifically, compared with a Corin gene shown in SEQ ID NO:1, the Corin gene variant has any of variations including c.G1203A, c.C2753T, c.C2928T and c.G1747T, and a polypeptide encoded through the variant has any of variations including p.R349H, p.P866S, p.T924M and p.R530S.The variant and the polypeptide can be used for preparing a hypertension diagnosis reagent and a gestational hypertension diagnosis reagent, detecting and screening people on a large scale and finding whether a testee suffers from hypertension sensibilization; by detecting peripheral blood, the pathogenesis of patients suffering from hypertension and gestational hypertension can be clearly obtained, and the variant is easy and convenient to use, high in specificity and capable of providing guidance for clinical treatment.

Description

technical field [0001] The invention belongs to the fields of clinical medicine and molecular biology, and relates to a Corin gene variant and its application in the preparation of a diagnostic reagent for hypertension and a diagnostic reagent for hypertensive disorders in pregnancy. Background technique [0002] Hypertension (Hypertension) is a common cardiovascular disease. According to the World Health Organization (WHO), about 1 / 3 of adults worldwide suffer from high blood pressure. Hypertension also increases the incidence of high-mortality diseases such as thrombosis, acute myocardial infarction, stroke, renal failure, and heart failure. The social and economic burdens of these diseases are enormous. With the development of society and economy in our country, the risk factors of hypertension and cardiovascular disease such as hyperlipidemia and obesity are also increasing day by day. Hypertension and cardiovascular disease have become the main health problems in modern...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/57C12N9/64C12Q1/68G01N33/573
CPCC12N9/6421C12Q1/6883C12Q2600/156G01N33/573G01N2333/96425G01N2800/321G01N2800/368
Inventor 吴庆宇董宁征张玥
Owner SUZHOU UNIV
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