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Preterm birth risk prediction using change in blood microbial community

A microorganism and risk technology, applied in the field of compositions for diagnosing the risk of preterm birth, can solve problems such as non-specificity and insensitive methods

Pending Publication Date: 2021-02-12
EWHA UNIV IND COLLABORATION FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The general approach to predict preterm birth is to identify groups of women who need special attention obstetrically, demographically, and by various syndromes, but the problem is that this method is not sensitive and specific
[0007] With recent developments in gene sequencing technology, it has become possible to study the human microbiome in various regions of the body, but until now, there has been little aim to identify the association between the microbiome in the blood (microbiome) and the risk of preterm birth and to diagnose the risk of preterm birth Research
Moreover, no such blood microbial characterization has been conducted in Korean pregnant women who experienced preterm labor and preterm premature rupture of membranes

Method used

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  • Preterm birth risk prediction using change in blood microbial community
  • Preterm birth risk prediction using change in blood microbial community
  • Preterm birth risk prediction using change in blood microbial community

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077]Example 1: Selection of experimental materials

[0078]Analysis of microbiological groups in the blood of un pregnant women, pregnant pregnant women and pregnant pregnant women, selected 41 unsophiliated women in the Pear Flower Women's University Wooden Hospital, 20 pregnant women and 21 pregnant women in pregnant women. The normal foot-filled childbirth group was selected for pregnant women who experienced the feet childbirth after 37 weeks of pregnancy. When the pregnant woman occurs or the symptoms of fetal membrane, the parent blood is collected in a test tube containing EDTA, and plasma is separated and stored at -70 ° C.

[0079]Only the single-weef production data of 25-42 weeks, excluded multi-child, dead production, malformed, chronic hypertension, placenta preamp, and placental peeling.

[0080]Regardless of pregnant women from healthy women without pregnancy-related diseases.

[0081]All processes and processes of the experiment were approved by the institutional review commit...

Embodiment 2

[0082]Example 2: DNA extraction and 16S rRNA sequencing

[0083]Bacterial DNA was extracted from the blood samples of Example 1 using the PowerMax Soil DNA separation kit (Mobio, Carlsbad, CA, US) using the PowerMax Soil DNA separation kit (Mobio, Carlsbad, CA, USA).

[0084]The 16S RRNA gene V3-V4 high variable region (519F-816R) of the bacterial genomic DNA (519F-816R) was amplified by the ILLUMINA 16S Acer. The fusion primer sequence used for amplified strip codes is as follows

[0085]16S_V3_F primers (SEQ ID NO: 1)

[0086]5'-tcgcggcagcgtcagatgtgtataagagagacagcctacgggnggcwgcAg-3 '

[0087]16S_V4_F primers (SEQ ID NO: 2)

[0088]5'-gtctcgtgggctcgggagatgtgtataagagagaggactachvggtatctaatcc -3.

[0089]The library is prepared using the PCR product in accordance with the MISEQ System Guide (Illumina) and quantitatively quantitatively using Qiaxpert (Qiagen, Hilden, Germany). After extracting and quantifying the PCR product, the equimolar ratio of each sample was analyzed using Miseq (Illumina) to analyze...

Embodiment 3

[0090]Example 3: Analysis of bacteria composition

[0091]The sequence information obtained in Example 2 was treated using Miseq (Illumina) according to barcode algorithm and primer sequence. Use the analysis program MDX-Pro Ver.1 (MD Healthcare, Seoul, South Korea) to classify (TAXOMICASSIGNMENT).

[0092]For the result of the serial number analysis, the high quality reading is selected after the reading length (= 300 bp) and the quality score (the average Phred Score = 20) are selected. Use the sequence clustering algorithm CD-HIT to cluster the operation classification unit. Subsequently, UCLST and QIIME are classified using Uclst and QIIME according to the 16SRNA sequence database of Greengenes 8.15.13. All 16SRNA sequences are allocated to the classification level based on the sequence similarity.Figure 4a to Figure 4fBy stacking the strip of the problem of 6 bacterial communities in the blood of women, women in PTB women and PTB women. If a sequence or redundant sequence is lacking ...

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Abstract

The present invention relates to prediction of preterm birth by using a change in blood microbiome and, more particularly, provides a composition for diagnosing a preterm birth risk by detecting strains of Weissella spp., Thermacetogenium spp., Faecalibacterium spp., Clostridium spp., or Lactobacillus spp. in the blood of pregnant women.

Description

Technical field[0001]The present invention relates to a composition, a diagnostic kit and a diagnostic method for diagnosing the risk of preterm birth by detecting the microbiome in the blood of pregnant women. More particularly, the present invention relates to detection of Weissella sp., Thermacetogenium sp., Lactobacillus sp. or Clostridium sp. in the blood of pregnant women. ) To diagnose the risk of preterm birth, a composition, a diagnostic kit and a diagnostic method.Background technique[0002]Preterm birth (PTB) usually refers to the delivery between 20 and 37 weeks of gestation. Premature newborns have a higher risk of death. Preterm birth accounts for about 60% of all infant deaths. Even surviving infants are also due to neurological development disorders and breathing. Systemic complications, postnatal growth retardation, etc. require intensive care. In addition, serious long-term or short-term diseases have a high incidence. About 70% of preterm births are caused by prete...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/689
CPCC12Q1/6883C12Q1/689C12Q2531/113
Inventor 金英珠俞泳我李庆珠杜智恩权恩震朴美惠
Owner EWHA UNIV IND COLLABORATION FOUND
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