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Microorganism associated with recurrent abortion and application thereof

A technology for recurrent miscarriage and microorganisms, applied in the directions of microorganisms, microorganism-based methods, and microorganism determination/examination, etc., which can solve the problems of ineffective treatment of patients, unknown causes of recurrent miscarriage patients, etc., and achieve the effect of improving the quality of life.

Active Publication Date: 2019-04-02
THE FIRST AFFILIATED HOSPITAL OF BENGBU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In recent years, with the development of society, food, radiation, air quality and other factors have affected the human body, and the incidence of recurrent miscarriage has increased year by year. The problem of early pregnancy tissue loss caused by maternal and embryonic reasons has gradually been effectively solved, but there are still nearly 1 The cause of 2 / 2 patients with recurrent miscarriage is unknown, so that such patients cannot be effectively treated from the etiology. How to achieve early detection and early prediction of embryonic development stop, and apply certain drugs to intervene in the occurrence or evaluation of recurrent miscarriage The therapeutic effect of a certain drug has become a current research hotspot

Method used

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  • Microorganism associated with recurrent abortion and application thereof

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Embodiment approach

[0026] As a kind of preferred embodiment, comprise the following steps:

[0027] (1) Collection and processing of samples: collect relevant vaginal secretion samples, use kits for DNA extraction, and obtain nucleic acid samples;

[0028] (2) Library construction and sequencing: use high-throughput sequencing for DNA library construction and sequencing, so as to obtain the nucleic acid sequences of microorganisms contained in the sample;

[0029] (3) Determine specific microbial nucleic acid sequences related to recurrent miscarriage through bioinformatics analysis methods.

[0030] First, obtain the sequencing data of the nucleic acid sequence in the individual's vaginal secretion sample, the sequencing data includes a plurality of reads; assemble the reads to obtain a gene set, the gene set includes a plurality of assembled fragments, the The assembly fragments in the gene set are non-redundant sequences; the assembly fragments contained in the various microorganisms in the ...

Embodiment 1

[0050] Example 1 Screening of intestinal flora associated with unexplained recurrent miscarriage

[0051] 1. Collection of samples

[0052] Sixteen patients with unexplained recurrent abortion in early pregnancy were selected as the experimental group (URSA), and 20 cases of normal pregnant women were selected as the control group, and the latter were strictly matched. The use of all specimens was signed informed consent by the patients or their clients, and was reviewed and approved by the Ethics Committee.

[0053] 2. 16S rRNA sequencing

[0054] 2.1 Extraction of DNA

[0055] Bacterial DNA was extracted from vaginal secretions using a DNA extraction kit, and the operating steps were performed according to the instructions.

[0056] 2.2 DNA sample purity and concentration determination

[0057] Genomic DNA was detected by 1% agarose gel electrophoresis.

[0058] 2.3 PCR amplification and product purification

[0059] TransGen AP221-02 (TransStart Fastpfu DNA Polymerase...

Embodiment 2

[0073] The preparation of the diagnostic kit of embodiment 2 unexplained recurrent miscarriage

[0074] According to the correlation between Anaerococcus hydrogenalis and unexplained recurrent miscarriage, unexplained recurrent miscarriage can be diagnosed by detecting the abundance of Anaerococcus hydrogenalis in the sample. Accordingly, the present invention provides a method for diagnosing unexplained recurrent miscarriage based on detecting the abundance of Anaerococcus hydrogenalis Kit for sexual abortion. The kit components are as follows: DNA extraction reagent, primer pair for specific detection of Anaerococcus hydrogenalis 16SrRNA, reaction buffer, deoxynucleotide triphosphates (dNTPs), Taq-polymerase reverse transcriptase, DNase, RNAse inhibitor, DEPC-water, sterile water, SYBR Green fluorescent dye.

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Abstract

The present invention discloses a microorganism associated with recurrent abortion and an application thereof. The specific microorganism is Anaerococcus hydrogenalis and abundance of the Anaerococcushydrogenalis in patients with unexplained recurrent abortion is significantly increased, suggesting that the Anaerococcus hydrogenalis can be used as a microorganism marker applied in an early diagnosis of the unexplained recurrent abortion.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a microorganism related to recurrent abortion and its application. Background technique [0002] Recurrent miscarriage (habitual abortion) refers to two or more consecutive spontaneous abortions. Most recurrent miscarriages occurred in the first trimester, and most of the recurrent miscarriage patients terminated their pregnancy at the same gestational days. The causes of miscarriage in early pregnancy are complex, often due to abnormal uterine development, endocrine reasons, physical and chemical factors in the surrounding environment, etc., but recurrent miscarriage without a specific cause accounts for about 50%, and is collectively classified as recurrent miscarriage of unknown cause clinically. abortion. In recent years, with the progress of immunology research, recurrent miscarriage caused by immune factors has gradually been accepted by colleagues at home and abroad, and it is...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/04G01N33/569C12R1/01
CPCC12Q1/689G01N33/56916G01N2800/368
Inventor 焦雪娟王立群宋倩倩
Owner THE FIRST AFFILIATED HOSPITAL OF BENGBU MEDICAL COLLEGE
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