Recurrent spontaneous abortion relevant microRNA and applications thereof

A technology for repeated spontaneous abortions and uses, applied in the fields of biotechnology and medicine, can solve the problems of RM identification and prediction methods that are poorly understood

Inactive Publication Date: 2017-01-11
SHANGHAI INST OF PLANNED PARENTHOOD RES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In summary, it can be seen from the existing research that little is known about the identification and prediction methods of RM

Method used

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  • Recurrent spontaneous abortion relevant microRNA and applications thereof
  • Recurrent spontaneous abortion relevant microRNA and applications thereof
  • Recurrent spontaneous abortion relevant microRNA and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0088] The preparation of the miRNA chip can adopt conventional manufacturing methods of biochips known in the art. For example, if the solid phase carrier is a modified glass slide or silicon wafer, and the 5' end of the probe contains amino-modified poly dT strings, the oligonucleotide probe can be formulated into a solution, and then spotted with a spotting instrument. The miRNA chip of the present invention can be obtained by arranging in a predetermined sequence or array on a modified glass slide or a silicon chip, and then fixing it by standing overnight. If the nucleic acid does not contain amino modification, its preparation method can also refer to: "Gene Diagnosis Technology-Non-Radioactive Operation Manual" edited by Wang Shenwu; J.L.erisi, V.R.Iyer, P.O.BROWN. Exploring the metabolic and genetic control of gene expression on a genomic scale. Science, 1997; 278:680 and Ma Liren, edited by Jiang Zhonghua. Biochip. Beijing: Chemical Industry Press, 2000, 1-130.

[00...

Embodiment 1

[0107] Example 1 Screening of Serum Markers

[0108] method:

[0109] Patients with recurrent miscarriage who visited the family planning clinic of the hospital were selected, and research subjects were recruited according to the following inclusion criteria. After informed consent of the research subjects, 15 cases of recurrent miscarriage were collected.

[0110]

[0111] 1. Real-time fluorescent quantitative PCR detection of expression level of hsa-miR-3074-5p

[0112] 1) Experimental reagents and their preparation

[0113] (1) RT primers: provided by Ribobio (RN: R10031.2), operated according to the instructions, the lyophilized powder was mixed with RNase-free water to prepare a 5 μM primer storage solution, and then stored in aliquots; before use , diluted 10 times (45 μl water + 5 μl storage solution), and prepared as 500 nM RT primer working solution.

[0114] (2) RT enzyme: MulV Reverse Transcriptase, a product of Roche Company.

[0115] (3) 10×PCR Buffer: a pr...

Embodiment 2

[0154] Embodiment 2 prepares miRNA chip

[0155] The miRNA sequence (SEQ ID NO: 1) provided by the present invention is converted into a complementary sequence, and a 10-20nt connection sequence is added to both ends of the sequence according to the GC ratio and other characteristics of the generated sequence; the core sequence is different, and the connection sequence is also different. The junction sequence can be randomly generated by the program, and the probes formed by the junction sequence and the core sequence meet the following conditions:

[0156] 1) In the probe sequence, the number of the same nucleotide (A, C, G, T) cannot exceed 50% of the total number of sequences;

[0157] 2) The number of any continuous A, T or C, G cannot exceed 25% of the total number of sequences;

[0158] 3) G and C content account for 40%-60% of the total number of sequences;

[0159] 4) The probe sequence cannot self-hybridize, that is, the length of the complementary segment in the pr...

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Abstract

The invention relates to a microRNA marker for diagnosing the natural recurrent abortion, and in particular relates to a microRNA marker hsa-miR-3074 used for distinguishing the tissue suffering from natural recurrent abortion and the normal tissue. The inspection proves that the specific microRNA marker can be used for effectively diagnosing and / or predicating the natural recurrent abortion. The invention further relates to applications of the marker microRNA hsa-miR-3074 and a kit used for detecting the natural recurrent abortion.

Description

technical field [0001] The invention relates to the fields of biotechnology and medicine. Specifically, the present invention relates to the method and application of hsa-miR-3074-5p gene in identifying RM pathological pregnancy. Background technique [0002] The establishment and maintenance of normal human pregnancy needs to go through a series of important steps such as embryo implantation, placenta formation, and embryonic development, which are coordinated and regulated by different systems at multiple levels. Any obstacle in any link will lead to adverse pregnancy outcomes such as spontaneous abortion. In 2005, European Society of Human Reproduction and Embryology (ESHRE, European Society of Human Reproduction and Embryology) defined "miscarriage" as: (1) a pregnancy that was positive in a urine pregnancy test or elevated in blood HCG levels, but ultrasound or histology failed to confirm pregnancy The situation is called "biochemical loss", which generally occurs befo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113
CPCC12N15/113C12N2310/141C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 张璇顾艳孙兆贵王建梅杨倩王健
Owner SHANGHAI INST OF PLANNED PARENTHOOD RES
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