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A serum microRNA marker related to the occurrence of human fetal growth restriction and its application

A technology of growth restriction and markers, applied in the fields of genetic engineering and clinical medicine, can solve problems that have not received corresponding attention

Active Publication Date: 2016-06-29
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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  • A serum microRNA marker related to the occurrence of human fetal growth restriction and its application
  • A serum microRNA marker related to the occurrence of human fetal growth restriction and its application
  • A serum microRNA marker related to the occurrence of human fetal growth restriction and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Embodiment 1 Research Object Selection and Grouping Basis

[0077] From July 2010 to January 2013, the inventor collected blood samples from fetal growth restriction cases and healthy control pregnant women who met the requirements from Huaian First People's Hospital Affiliated to Nanjing Medical University. A total of 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) were selected as subjects for Real-timePCR detection of miRNA expression. Specific sample classification criteria are as follows:

[0078] Group A: healthy control group (n=80, 10 people for microarray screening, 30 people for first-stage verification, 40 people for independent population verification):

[0079] 1. No other major systemic diseases.

[0080] Group B: fetal growth restriction group (n=80, 10 people for microarray screening, 30 people for phase one verification, 40 people for indepen...

Embodiment 2

[0083] Embodiment 2TaqManmiRNAarray screening

[0084] Preparation of cDNA samples: a) Take 100 μl of serum; b) Add 60 μl of lysate, vortex for 5 seconds, and let stand at room temperature for 3 minutes; c) Add 20 μl of protein solution, vortex for 5 seconds, stand at room temperature for 1 minute, and centrifuge at 11,000 g for 3 minutes; d) Take the above Clear to a new 2ml collection tube, add 270μl isopropanol, and vortex for 5s; e) Put the microRNA spin column in the collection tube, add the liquid in step d to the spin column, let stand at room temperature for 2min, centrifuge at 11000g for 30s, pour Discard the liquid in the collection tube and put the spin column back into the collection tube; f) Add 100 μl of eluent 1 to the microRNA spin column, centrifuge at 11,000 g for 30 seconds, discard the liquid in the collection tube, and put the spin column back into the collection tube; g) Add 700 μl Put the eluent 2 into the microRNA spin column, centrifuge at 11000g for 3...

Embodiment 3

[0098] Embodiment 3Real-timePCR method measures serum miRNA expression

[0099] Primers (Table 1) were designed for quantitative Real-time PCR detection of each miRNAs in the serum of 80 healthy controls and 80 patients with fetal growth restriction.

[0100] (1) Preparation of cDNA samples: a) Take 100 μl of serum; b) Add 60 μl of lysate, vortex for 5 s, and let stand at room temperature for 3 min; c) Add 20 μl of protein solution, vortex for 5 s, stand at room temperature for 1 min, centrifuge at 11,000 g for 3 min; d ) Take the supernatant to a new 2ml collection tube, add 270μl isopropanol, and vortex for 5s; e) Put the microRNA spin column in the collection tube, add the liquid in step d to the spin column, let stand at room temperature for 2min, and centrifuge at 11000g 30s, pour off the liquid in the collection tube, put the spin column back into the collection tube; f) Add 100μl eluent 1 to the microRNA spin column, centrifuge at 11000g for 30s, pour off the liquid in ...

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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

field of invention [0001] The invention belongs to the fields of genetic engineering and clinical medicine, and relates to serum 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 fetus whose birth weight is lower than two standard deviations of the mean weight for the same gestational age, or lower than the 10th percentile of the normal weight for the same age. FGR is an important perinatal complication, ranking second in the cause of perinatal death, and its perinatal mortality rate is 6-10 times that of normal infants, accounting for about 30% of perinatal deaths. 50% of perinatal infants with internal hypoxia are FGR, and according to statistics, the average incidence rate in my country is 6.39%. FGR can cause neonatal dyspnea, polycythemia, hypoglycemia, ventricular hemorrhage, hypothermia and other complications. FGR not only directly af...

Claims

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Application Information

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Patent Type & Authority Patents(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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