Serum micro ribonucleic acid marker related to human fetal growth restriction and application thereof

A growth restriction and marker technology, applied in the fields of genetic engineering and clinical medicine, can solve the problem that the application in the early diagnosis and monitoring of growth restriction in children has not received corresponding attention.

Active Publication Date: 2013-07-10
夏彦恺
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the application of miRNA in serum in the early diagnosis and monitoring of fetal growth restriction has not received corresponding attention. If stable specific serum miRNA related to the onset of fetal growth restriction can be found as a biomarker, and the diagnosis o...

Method used

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  • Serum micro ribonucleic acid marker related to human fetal growth restriction and application thereof
  • Serum micro ribonucleic acid marker related to human fetal growth restriction and application thereof
  • Serum micro ribonucleic acid marker related to human fetal growth restriction and application thereof

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Experimental program
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Embodiment 1

[0075] Embodiment 1 Research Object Selection and Grouping Basis

[0076] From July 2009 to October 2011, 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: 26.76±3.25 years old) and 80 cases of fetal growth restriction (average age of pregnant women: 27.56±4.35 years old) who met the requirements were selected as experimental subjects for real-time PCR detection of miRNA expression. Specific sample classification criteria are as follows:

[0077] Group A: healthy control group (n=80, 20 people for microarray screening, 20 people for first-stage verification, 40 people for independent population verification):

[0078] 1. No other major systemic diseases.

[0079] Group B: fetal growth restriction group (n=80, 20 people for chip screening, 20 people for phas...

Embodiment 2

[0082] Example 2 TaqMan miRNA array screening

[0083] Preparation of cDNA samples: a) Take 100 μl of serum; b) Add 900 μl of TRIzol, shake and mix, centrifuge at 12,000 rpm at 4°C for 15 minutes, and discard the waste liquid in the lower layer; c) Add 1.5 times the volume of supernatant, shake and mix, transfer To the spin column, centrifuge at 12000rpm for 15 seconds, discard the lower waste liquid; d) add 700μl RWT buffer to the spin column, centrifuge at 10000rpm for 15 seconds, discard the lower waste liquid. e) Add 500 μl of RPE buffer to the spin column, centrifuge at 10,000 rpm for 15 seconds, and discard the lower layer. f) Repeat e. g) Add the spin column to a new 2ml tube and centrifuge at 10,000rpm for 1 minute to remove the RPE buffer. h) Add 50 μl of DEPC-treated water to the column and collect RNA by centrifugation at 12000 rpm. i) Then cDNA is obtained by RNA reverse transcription reaction. The reverse transcription reaction system included 4 μl 5×AMV buffe...

Embodiment 3

[0094] Embodiment 3Real-time PCR method measures serum miRNA expression

[0095] 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.

[0096] (1) Preparation of cDNA samples: a) Take 100 μl of serum; b) Add 900 μl of TRIzol, shake and mix, centrifuge at 12,000 rpm at 4°C for 15 minutes, and discard the waste liquid in the lower layer; c) Add 1.5 times the volume of supernatant in absolute ethanol, shake and mix Mix well, transfer to the spin column, centrifuge at 12000rpm for 15 seconds, discard the lower waste liquid; d) add 700μl RWT buffer to the spin column, centrifuge at 10000rpm for 15 seconds, discard the lower waste liquid. e) Add 500 μl of RPE buffer to the spin column, centrifuge at 10,000 rpm for 15 seconds, and discard the lower layer. f) Repeat e. g) Add the spin column to a new 2ml tube and centrifuge at 10,000rpm for 1 minute to remove the RPE...

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Abstract

The invention belongs to the field of gene engineering and clinical medicine, and discloses a serum micro ribonucleic acid marker related to human fetal growth restriction and application thereof. The marker is more of hsa-miR-212, hsa-miR-597 and hsa-let-7d. The marker is specific and sensitive to fetal growth restriction, and can be used for preparing diagnostic or monitor reagents for fetal growth restriction.

Description

field of invention [0001] The invention belongs to the fields of genetic engineering and clinical medicine, and relates to serum microRNA (miRNA) markers related to human fetal growth restriction and applications thereof. Background technique [0002] Fetal growth restriction (FGR) refers to fetal birth weight below two standard deviations of the mean weight for the same gestational age, or below the 10th percentile of normal weight for the same age. FGR not only directly affects the fetus and neonate, but also alters the susceptibility of adults to disease. FGR can cause neonatal dyspnea, polycythemia, hypoglycemia, ventricular hemorrhage, hypothermia and other complications. Barker et al. conducted a longitudinal study of 13,517 males and females born in the University Hospital of Helsinki from 1924 to 1944 and found that these populations were born with low birth weight and accelerated weight gain in childhood, which made adult coronary heart disease, type II diabetes, ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/113C12N15/11
Inventor 夏彦恺吴炜王心如许倩江华黄璐汤秋勤吕述彦傅广波
Owner 夏彦恺
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