Tiny serum ribonucleic acid marker related to human fetal growth restriction and application thereof

A growth restriction, marker technology, applied in the fields of genetic engineering and clinical medicine, can solve problems such as not receiving corresponding attention

Active Publication Date: 2014-04-30
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of miRNA in the serum of pregnant women in the early diagnosis and monitoring of fetal growth restriction has not received corresponding attention. Diagnostic and monitoring kits will not only create remarkable economic benefits, but will also be a strong impetus to the prevention and treatment of fetal growth restriction in my country

Method used

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  • Tiny serum ribonucleic acid marker related to human fetal growth restriction and application thereof
  • Tiny serum ribonucleic acid marker related to human fetal growth restriction and application thereof
  • Tiny serum ribonucleic acid marker related to human fetal growth restriction and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Research object selection and grouping basis

[0077] The inventor collected blood samples of fetal growth restriction cases and healthy control pregnant women from hospitals such as Huai’an First People’s Hospital affiliated to Nanjing Medical University from July 2010 to January 2013. After sorting out the sample data, 80 healthy controls (average age of pregnant women: 26.39±3.42 days) and 80 cases of fetal growth restriction (average age of pregnant women: 27.47±4.15 days) that meet the requirements were selected as experimental subjects for Real-time PCR to detect miRNA expression. The specific sample classification standards are as follows:

[0078] Group A: Healthy control group (n=80, 10 people are screened by chip, 30 people are first-phase verification, 40 people are independent population verification):

[0079] 1. No other major systemic diseases.

[0080] Group B: Fetal growth restriction group (n=80, 10 people are screened by chip, 30 people are first-p...

Embodiment 2

[0083] Example 2 TaqMan miRNA array screening

[0084] Prepare cDNA samples: a) take 100μl serum; b) add 60μl lysate, vortex for 5s, and stand for 3min at room temperature; c) add 20μl protein solution, vortex for 5s, stand for 1min at room temperature, centrifuge at 11000g for 3min; d) take it up Clear to a new 2ml collection tube, add 270μl of isopropanol, vortex for 5s; e) Place the microRNA spin column in the collection tube, add the liquid from step d to the spin column, let stand at room temperature for 2min, centrifuge at 11000g for 30s, pour Remove the liquid in the collection tube and put the spin column back into the collection tube; f) add 100μl of eluent 1 to the microRNA spin column, centrifuge at 11000g for 30s, pour out the liquid in the collection tube, and put the spin column back into the collection tube; g) add 700μl Transfer the eluate 2 to the microRNA spin column, centrifuge at 11000g for 30s, pour out the liquid in the collection tube, and put the spin colu...

Embodiment 3

[0098] Example 3 Real-time PCR method to measure serum miRNA expression

[0099] Design primers (Table 1) to perform quantitative Real-time PCR detection of miRNAs on the sera of 80 healthy controls and 80 fetal growth restriction patients.

[0100] (1) Prepare cDNA samples: a) Take 100μl of serum; b) Add 60μl of lysate, vortex for 5s, and stand for 3min at room temperature; c) Add 20μl of protein solution, vortex for 5s, stand for 1min at room temperature, centrifuge at 11000g for 3min; d ) Take the supernatant to a new 2ml collection tube, add 270μl of isopropanol, and vortex for 5s; e) Place the microRNA spin column in the collection tube, add the liquid from step d to the spin column, let stand at room temperature for 2 minutes, and centrifuge at 11000g 30s, pour out the liquid in the collection tube and put the spin column back into the collection tube; f) Add 100μl of eluent 1 to the microRNA spin column, centrifuge at 11000g for 30s, pour out the liquid in the collection tub...

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Abstract

The invention belongs to the fields of genetic engineering and reproductive medicine, and discloses a tiny serum ribonucleic acid marker related to human fetal growth restriction and application thereof. The marker is a plurality of hsa-miR-10a, hsa-miR-320b, hsa-miR-409-3p, hsa-miR-483-3p and hsa-miR-675-5p. The marker has specificity and sensibility on the fetal growth restriction, can be applied to preparation of the kit for diagnosing and monitoring the fetal growth restriction, can repeatedly detect, and is easy to dynamically monitor the fetal growth restriction degree.

Description

Invention field [0001] The present invention belongs to the fields of genetic engineering and clinical medicine, and relates to serum microribonucleic acid (microRNA, miRNA) markers and applications thereof related to the occurrence of human fetal growth restriction. Background technique [0002] Fetal growth restriction (FGR) refers to a fetal birth weight lower than two standard deviations of the average weight of the same gestational age, or lower than the 10th percentile of the normal weight of the same age. FGR is an important perinatal complication and ranks the second cause of death in perinatal infants. Its perinatal mortality rate is 6-10 times that of normal infants. It accounts for about 30% of perinatal infant deaths. 50% of perinatal infants with internal hypoxia are FGR. According to statistics, the average incidence rate in my country is 6.39%. FGR can cause neonatal dyspnea, polycythemia, hypoglycemia, intraventricular hemorrhage, hypothermia and other complicatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 吴炜汤秋勤丁虹娟陈敏夏彦恺陆春城王心如
Owner NANJING MEDICAL UNIV
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