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Detecting and/or predicting miRNA markers or combinations thereof in human macrosomia and applications thereof

A marker, human technology, applied in the field of genetic engineering and clinical medicine, can solve problems that have not received corresponding attention, and achieve the effect of avoiding invasive diagnosis, improving sensitivity and specificity, and quantitatively accurate

Active Publication Date: 2021-08-31
NANJING HANWEI PUBLIC HEALTH RES INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of plasma miRNA in the early prediction and monitoring of macrosomia has not received corresponding attention. If stable specific plasma miRNA related to macrosomia can be found as a biomarker, and early prediction and monitoring kits for corresponding diseases can be developed , not only in the international leading position in this field, but also can create remarkable economic benefits, and will also be a strong impetus to the prevention and treatment of pregnancy-induced hypertension in my country

Method used

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  • Detecting and/or predicting miRNA markers or combinations thereof in human macrosomia and applications thereof
  • Detecting and/or predicting miRNA markers or combinations thereof in human macrosomia and applications thereof
  • Detecting and/or predicting miRNA markers or combinations thereof in human macrosomia and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Research Object Selection and Grouping Basis

[0043] From July 2010 to October 2012, the inventor collected blood samples from macrosomia patients and healthy control pregnant women who met the requirements from Huai’an First People’s Hospital Affiliated to Nanjing Medical University, and selected 80 cases of healthy controls and 80 cases of macrosomia who met the requirements were used as experimental subjects for Real-time PCR detection of miRNA expression. Specific sample classification criteria are as follows:

[0044] Group A: Healthy control group (n = 80, 20 for microarray screening, 30 for first-stage verification, and 30 for independent population verification):

[0045] 1. No other major systemic diseases.

[0046] Group B: macrosomia group (n = 80, 20 people for microarray screening, 30 people for phase one verification, 30 people for independent population verification):

[0047] 1. Clinically diagnosed as macrosomia;

[0048] 2. No other majo...

Embodiment 2

[0049] Example 2 TaqMan miRNA array screening

[0050] Prepare cDNA samples: a) Take 100 µl plasma; b) Add 900 µl TRIzol, shake and mix, centrifuge at 12,000 rpm at 4°C for 15 minutes, discard the waste liquid in the lower layer; c) Add 1.5 times the volume of supernatant absolute ethanol, shake and mix, Transfer to the spin column, centrifuge at 12000rpm for 15 seconds, and 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 500µl RPE buffer to the spin column, 10000rpm Centrifuge 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 50µl DEPC-treated water to the column and collect by centrifugation at 12,000rpm RNA; i) cDNA is then obtained by RNA reverse transcription reaction. The reverse transcription reaction system includes 4 μl 5×AMV buffer, 2 μl 10mM...

Embodiment 3

[0063] Example 3 Real-time PCR method for measuring plasma miRNA expression

[0064] The primers (Table 1) were designed for the quantitative Real-time PCR detection of each miRNAs in the plasma of 80 healthy controls and 80 macrosomia patients.

[0065] (1) Preparation of cDNA samples: a) Take 100 µl plasma; b) Add 900 µl TRIzol, shake and mix, centrifuge at 12,000 rpm at 4°C for 15 minutes, discard the lower layer of waste liquid; c) Add 1.5 times the volume of supernatant 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 500µl RPE buffer to the spin column , centrifuge at 10,000rpm for 15 seconds, 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 50µl DEPC-treated water ...

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Abstract

The invention discloses a miRNA marker related to human macrosomia or its combination and application, belonging to the fields of genetic engineering and clinical medicine. The marker is selected from one or more of hsa-miR-195-5p, hsa-miR-155-5p, and hsa-miR-221-3p. The marker has specificity and sensitivity for macrosomia, and can be used to prepare a kit for the early diagnosis or monitoring of macrosomia, which can be screened and diagnosed at an early stage, and can be repeatedly detected and easily monitored dynamically.

Description

technical field [0001] The invention belongs to the fields of genetic engineering and clinical medicine, and relates to the detection and / or prediction of miRNA markers or combinations thereof in human macrosomia and applications thereof. Background technique [0002] Among the indicators of children's growth and development, birth weight is an important indicator of the health status of newborns and an important factor affecting their quality of life. Abnormal birth weight of newborns will not only have adverse effects on the short-term health of infants, but also affect their health status in adulthood. Epidemiological surveys found that in the past ten years, the incidence of low birthweight infants in various parts of the world (Canada, Sweden, the United States, and China) has generally shown a downward trend, while the incidence of high birthweight infants has gradually increased. Health risks from birth weight. Among the high birth weight, newborns with a birth weig...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 吴炜汤秋勤江华陈丽平陈婷陈敏健陆春城夏彦恺
Owner NANJING HANWEI PUBLIC HEALTH RES INST CO LTD
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