KR102363092B1 - Predicting or Diagnosing Composition for Risk of Obesity Using Human Intestinal Microbiome, Diagnosing Kit, Method For Providing Information, And Screening Method For Drugs For Preventing Or Treating Obesity Using The Same - Google Patents

Abstract

The present invention relates to a composition for predicting or diagnosing the risk of obesity using an intestinal microorganism, a kit for predicting or diagnosing the risk of obesity comprising the same, a method for providing information for predicting or diagnosing the risk of obesity, and a screening method for preventing or treating obesity, Blautia Obe Um ( Blautia obeum ) Species strain, Veillonella dispar ( Veillonella dispar ) Species strain and Lachnospiraceae ) Any one or more strains selected from the group consisting of any one or more strains selected from the group consisting of strains An agent capable of detecting, or an agent capable of detecting any one or more strains selected from the group consisting of a bacteroides uniformis species strain and a Clostridium sp. strain, together with the agent, or lichenel Predicting or diagnosing the risk of obesity through an agent capable of detecting any one or more strains selected from the group consisting of Rasae ( Rikenellaceae ) and strains and strains of Anaerostipes species, and screening for prevention or treatment of obesity to make it possible

Description

Predicting or Diagnosing Composition for Risk of Obesity Using Human Intestinal Microbiome, Diagnosing Kit, Method For Providing Information, And Screening Method For Drugs For Preventing Or Treating Obesity Using The Same}

The present invention relates to a composition for predicting or diagnosing the risk of obesity using an intestinal microorganism, a diagnostic kit for predicting or diagnosing the risk of obesity using the same, a method for predicting or diagnosing obesity risk using the same, and a screening method for preventing or treating obesity using the same.

With the development of next-generation sequencing techniques, it has become possible to quickly and accurately measure the types and relative amounts of microorganisms living in an individual's gut. In particular, this technology makes it possible to find out the type and relative amount of microorganisms inhabiting an individual's intestine using the 16S ribosomal RNA gene variable region sequence, which is a conserved region common to intestinal microbes, and the information can be used as personal disease information or information. Research is being conducted to determine the association with biomarkers related to disease information.

Korea Patent No. 1445243 relates to a composition for predicting or diagnosing metabolic and inflammatory diseases using the intestinal bacterial community, and includes the prediction or diagnosis of obesity, but Akkermansia muciniphila as a microorganism that is associated with obesity. ), Bacteroides spp. , Eubacterium spp. , Roseburia spp. , Methanobrevibacter smithii ( Methanobrevibacter smithii ), Methanosphaera stadtmanae ( Methanosphaera stadtmanae ) ), Ruminococcus obeum and Phascolarctobacterium are used, but the results obtained from fecal samples obtained from 15 pairs of twins are not generalized. There were limits.

Korean Patent Publication No. 2019-0039502 discloses, in the treatment of obesity-related diseases, Prevotellaceae , Flavonifractor , Clostridium IV and Butyricicoccus from the genus consisting of It is described to increase the selected bacterial population, or to increase the selected bacterial population from the genera consisting of Coriobacteriaceae , Lactobacillaceae and Rikenellaceae .

Korean Patent No. 1940445 proposes a method for diagnosing diabetes through metagenome analysis using extracellular vesicles secreted from bacteria from blood or urine samples as a sample.

However, the results presented in the above patents are biomarker microorganisms that can distinguish between the high-risk group of obesity and the normal group, a combination of the microorganisms, a characteristic nucleotide sequence to identify the microorganisms, and the increase/decrease pattern of microorganisms in the high-risk group In, there was a difference from the study results conducted by the present inventors who identified the association with obesity based on the microbiome obtained from 890 Koreans, and therefore the present inventors completed the present invention based on the results of this study.

Korean Patent No. 1445243 Korean Patent Publication No. 2019-0039502 Korean Patent No. 1940445

The present invention can detect any one or more strains selected from the group consisting of Blautia obeum species strains, Veillonella dispar species strains and Lachnospiraceae family strains. It is to provide a composition for predicting or diagnosing obesity risk using intestinal microorganisms, including the preparation.

Another object of the present invention is to provide a kit for predicting or diagnosing obesity risk, including a composition for predicting or diagnosing the risk of obesity using the intestinal microorganism.

In addition, the present invention from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject Blautia obeum species strain, Veillonella dispar species strain and Lachnospiraceae ( Lachnospiraceae ) family strain detecting any one or more strains selected from the group consisting of; And from the genomic DNA of microorganisms obtained from the intestinal-derived sample of the test subject, Blautia obeum spp. strain, Veillonella dispar spp. strain and Lachnospiraceae family consisting of strains By comparing the increase or decrease of any one or more strains selected from the normal group, Blautia obeum in the genomic DNA of the microorganism is reduced when the strain is reduced, or Veillonella dispar ( Veillonella dispar ) species When the strain is increased, or when the ratio of Lachnospiraceae and the strain is reduced, diagnosing the test subject as an obesity or obesity risk group; predicting or diagnosing obesity risk using intestinal microorganisms, including This is to provide a method of providing information for

In addition, the present invention from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject Blautia obeum species strain, Veillonella dispar species strain, Lachnospiraceae ( Lachnospiraceae ) and strains , Bacteroides uniformis ( bacteroides uniformis ) species strains, Clostridium genus strains, Rikenellaceae family strains and Anaerostipes ( Anaerostipes ) Phase of two or more strains selected from strains detecting the mass; And the result obtained in the detecting step, calculating the obesity risk of the test subject by a multivariate linear model predicting an obesity group using the relative amount of the two or more strains as a variable; This is to provide an information provision method for risk prediction or diagnosis.

In addition, the present invention from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject Blautia obeum species strain, Veillonella dispar species strain, Lachnospiraceae ( Lachnospiraceae ) and strains , Bacteroides uniformis ( bacteroides uniformis ) species strains, Clostridium genus strains, Rikenellaceae family strains and Anaerostipes ( Anaerostipes ) Detecting the species strains; And Veillonella dispar in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject If the ratio of the species strain is increased, or Blautia obeum species strain, Lachnospira ( Lachnospiraceae ) and strains, Bacteroides uniformis ( bacteroides uniformis ) species strains, Clostridium genus strains, Rikenellaceae family strains and Anaerostipes ( Anaerostipes ) The ratio of strains In order to provide an information providing method for predicting or diagnosing diabetes risk using intestinal microorganisms, including the step of diagnosing the test subject as a renal disease or renal disease risk group when reduced.

In addition, the present invention comprises the steps of administering a candidate substance for the prevention or treatment of obesity to a non-human animal; From the genomic DNA of microorganisms obtained from the intestinal-derived sample before and after the candidate material treatment, Blautia obeum species strain, Veillonella dispar species strain and Lachnospiraceae family detecting any one or more strains selected from the group consisting of strains; And in the genomic DNA of microorganisms before and after treatment with the candidate material, Blautia obeum spp. strain, Veillonella dispar spp. strain and Lachnospiraceae family strain. By comparing the increase or decrease of any one or more strains selected from, if the Blautia obeum species strain is increased, or Veillonella dispar ) If the strain is reduced, or lark It is to provide a screening method for preventing or treating obesity using intestinal microorganisms, including; determining the candidate substance as a preventive or therapeutic agent for obesity in the case of Lachnospiraceae and strain is increased.

The present invention can detect any one or more strains selected from the group consisting of Blautia obeum species strains, Veillonella dispar species strains and Lachnospiraceae family strains. It relates to a composition for predicting or diagnosing the risk of obesity using intestinal microorganisms, including the preparation.

The present invention also relates to a kit for predicting or diagnosing the risk of obesity, comprising the composition for predicting or diagnosing the risk of obesity using the intestinal microorganism.

In addition, the present invention from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject Blautia obeum species strain, Veillonella dispar species strain and Lachnospiraceae ( Lachnospiraceae ) family strain detecting any one or more strains selected from the group consisting of; And from the genomic DNA of microorganisms obtained from the intestinal-derived sample of the test subject, Blautia obeum spp. strain, Veillonella dispar spp. strain and Lachnospiraceae family consisting of strains By comparing the increase or decrease of any one or more strains selected from the normal group, Blautia obeum in the genomic DNA of the microorganism is reduced when the strain is reduced, or Veillonella dispar ( Veillonella dispar ) species If the strain is increased, or if the Lachnospiraceae and strain is reduced, diagnosing the test subject as an obesity or obesity risk group; for predicting or diagnosing the risk of obesity using intestinal microorganisms, including It is about the method of providing information.

In addition, the present invention from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject Blautia obeum species strain, Veillonella dispar species strain, Lachnospiraceae ( Lachnospiraceae ) and strains , Bacteroides uniformis ( bacteroides uniformis ) species strains, Clostridium genus strains, Rikenellaceae family strains and Anaerostipes ( Anaerostipes ) Phase of two or more strains selected from strains detecting the mass; And the result obtained in the detecting step, calculating the obesity risk of the test subject by a multivariate linear model predicting an obesity group using the relative amount of the two or more strains as a variable; It relates to a method of providing information for risk prediction or diagnosis.

In addition, the present invention from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject Blautia obeum species strain, Veillonella dispar species strain, Lachnospiraceae ( Lachnospiraceae ) and strains , Bacteroides uniformis ( bacteroides uniformis ) species strains, Clostridium genus strains, Rikenellaceae family strains and Anaerostipes ( Anaerostipes ) Detecting the species strains; And Veillonella dispar in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject If the ratio of the species strain is increased, or Blautia obeum species strain, Lachnospira ( Lachnospiraceae ) and strains, Bacteroides uniformis ( bacteroides uniformis ) species strains, Clostridium genus strains, Rikenellaceae family strains and Anaerostipes ( Anaerostipes ) The ratio of strains It relates to a method for providing information for predicting or diagnosing diabetes risk using intestinal microbes, including; diagnosing the test subject as a renal disease or renal disease risk group when reduced.

In addition, the present invention comprises the steps of administering a candidate substance for the prevention or treatment of obesity to a non-human animal; From the genomic DNA of microorganisms obtained from the intestinal-derived sample before and after the candidate material treatment, Blautia obeum species strain, Veillonella dispar species strain and Lachnospiraceae family detecting any one or more strains selected from the group consisting of strains; And in the genomic DNA of microorganisms before and after treatment with the candidate material, Blautia obeum spp. strain, Veillonella dispar spp. strain and Lachnospiraceae family strain. By comparing the increase or decrease of any one or more strains selected from, if the Blautia obeum species strain is increased, or Veillonella dispar ) If the strain is reduced, or lark It relates to a screening method for preventing or treating obesity using intestinal microorganisms, comprising the step of determining the candidate substance as a preventive or therapeutic agent for obesity when the number of Lachnospiraceae and strains is increased.

In the present invention, based on the body mass index (BMI) of 890 Korean subjects, those of less than 18.5 were classified into the underweight group, those of 18.5 or more and less than 25 were classified as the normal group, and those of more than 25 were classified into the obese group, and the average A biomarker for predicting or diagnosing the risk of obesity by searching for microorganisms whose ratio decreases or increases in the order of the obese group, the normal group, and the underweight group, and among the discovered microorganisms, the microorganisms that show a statistically significant difference in ratio between the obese group and the normal group strain was identified.

Biomarker strains for predicting or diagnosing the risk of obesity showing a statistically significant difference between the obese group and the normal group include Blautia obeum strains, Veillonella dispar strains, Lachnospiraceae family strains, Bacteroides uniformis species strains, Clostridium genus strains, Rikenellaceae family strains and Anaerostipes species It may be any one selected from the group consisting of strains or a combination of two or more strains.

By comparing the increase or decrease of the strain, the increase or decrease of the strain means an increase or decrease in the relative ratio of the biomarker strains in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract, or the obesity group, the normal group, or It may mean an increase or decrease in the number of bacteria of the biomarker strains in the low-weight group or an absolute value indicating the same, for this purpose, the relative ratio of the biomarker microorganisms in the obese group, the normal group or the low-weight group, the number of bacteria or the range of the absolute value indicating the same can be stored as a database in advance.

Among the biomarker strains, Blautia obeum spp. strains, preferably Blautia obeum spp. strains identified by the 16S rRNA sequence of SEQ ID NO: 1, are normal and underweight groups. It was significantly lower in the obese group compared to the obese group. Therefore, when the genomic DNA of the microorganism obtained from the test subject's intestinal-derived sample is lower than the normal group, the Blautia obeum species strain can be predicted or diagnosed as having a high risk of obesity. In addition, if the Blautia obeum species strain is higher than that of the obese group, it is predicted or diagnosed as a normal group or a low-weight group, and thus the risk of obesity may be predicted or diagnosed as low.

Among the biomarker strains, Veillonella dispar strains, preferably Veillonella dispar identified by the 16S rRNA nucleotide sequence of SEQ ID NO: 2, strains in the normal group and underweight group It was significantly higher in the obese group than in the obese group. Therefore, when the Veillonella dispar strain is higher than that of the normal group in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract, it can be predicted or diagnosed as having a high risk of obesity. In addition, Veillonella dispar ( Veillonella dispar ) When the strain is low compared to the obese group, it is predicted or diagnosed as a normal group or a low-weight group, and thus the risk of obesity can be predicted or diagnosed as low.

Among the biomarker strains, Lachnospiraceae and strains, preferably Lachnospiraceae identified by the 16S rRNA nucleotide sequence of SEQ ID NO: 3, are significantly lower in the obese group than in the normal group. appear. Therefore, when the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is Lachnospiraceae and the strain is lower than that of the normal group, it can be predicted or diagnosed as having a high risk of obesity. In addition, if the Lachnospiraceae and strain is high compared to the obese group, it can be predicted or diagnosed as having a low risk of obesity.

Among the biomarker strains, Bacteroides uniformis species strains, preferably Bacteroides uniformis identified by the 16S rRNA sequence of SEQ ID NO: 4, strains are normal and underweight. It was significantly lower in the obese group than in the obese group. Therefore, when the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is lower than that of the normal group, the Bacteroides uniformis species strain can be predicted or diagnosed as having a high risk of obesity. In addition, when the Bacteroides uniformis species strain is higher than the obese group, it is predicted or diagnosed as a normal group or a low-weight group, so that the obesity risk can be predicted or diagnosed as low.

Among the biomarker strains, Clostridium sp. strain, preferably Clostridium sp. strain identified by the 16S rRNA sequence of SEQ ID NO: 5, is significantly lower in the obese group than in the normal group. Therefore, when the genomic DNA of the microorganism obtained from the test subject's intestinal-derived sample is lower than that of the normal group, the Clostridium spp. strain can be predicted or diagnosed as having a high risk of obesity. In addition, if the Clostridium sp. strain is higher than the obese group, it can be predicted or diagnosed as having a low risk of obesity.

Among the biomarker strains, Rikenellaceae strains, preferably SEQ ID NO: 6 to SEQ ID NO: 16, identified by any one sequence selected from the rRNA nucleotide sequence of SEQ ID NO: 16 Rikenellaceae family strains are normal It was significantly lower in the obese group than in the group and the underweight group. Therefore, when the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is lower than that of the normal group, Rikenellaceae can be predicted or diagnosed as having a high risk of obesity. In addition, if the Rikenellaceae family strain is higher than the obese group, it is predicted or diagnosed as a normal group or a low-weight group, and thus the risk of obesity can be predicted or diagnosed as low.

Among the biomarker strains, Anaerostipes species strain, preferably SEQ ID NO: 17 to SEQ ID NO: 19 Anaerostipes identified by any one sequence selected from the rRNA nucleotide sequence ( Anaerostipes ) The species strain was significantly lower in the obese group than in the normal group. Therefore, when the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is Anaerostipes , when the strain is lower than that of the normal group, it can be predicted or diagnosed as having a high risk of obesity. In addition, when the Anaerostipes strain is high compared to the obese group, the risk of obesity can be predicted or diagnosed as low.

The 16S rRNA nucleotide sequence of SEQ ID NOs: 1 to 19 is an ASV nucleotide sequence capable of identifying each of the biomarker strains, that is, an amplicon sequence variant nucleotide sequence. In particular, the 16S rRNA nucleotide sequence of SEQ ID NOs: 1 to 5 was first identified as a nucleotide sequence capable of identifying each biomarker strain. Therefore, the biomarker strain identified by any one sequence selected from the ASV nucleotide sequences of SEQ ID NO: 1 to SEQ ID NO: 5 is a strain belonging to each species or genus, or is molecularly biologically related to strains of each species or genus known in the art. These are the first strains to be identified in the clearly distinct intestine of Koreans.

In the present invention, 'risk prediction' refers to determining whether a patient is likely to develop a disease, delaying the onset or preventing the onset of the disease through special and appropriate management of a patient with a high risk of disease, or the most appropriate treatment It can be used clinically to make treatment decisions by choosing a modality. In addition, 'diagnosis' means confirming the presence or characteristics of a pathological condition, and for the purpose of the present invention, diagnosis may mean confirming whether or not the onset of .

As an agent capable of detecting the strain provided as a biomarker in the present invention, a protein, nucleic acid, lipid, glycolipid, glycoprotein or sugar (monosaccharide, disaccharide, oligosaccharide, etc.) specifically present in the corresponding strain in the sample, etc. Primers, probes, antisense oligonucleotides, aptamers, antibodies, and the like capable of specifically detecting the same organic molecule may be used.

For example, when the agent for detecting the strain is a primer, it is preferable that the primer specifically detects the genomic sequence (eg, 16S rRNA) of the microorganism and does not specifically bind to the genomic sequence of another strain.

In the present invention, the term 'primer' refers to 7 to 50 nucleic acid sequences capable of forming a base pair complementary to the template strand and functioning as a starting point for copying the template strand. Primers are usually synthesized but can also be used on naturally occurring nucleic acids. The sequence of the primer does not necessarily have to be exactly the same as the sequence of the template, but is sufficiently complementary so that it can hybridize with the template. Additional features that do not change the basic properties of the primer may be incorporated. Examples of additional features that may be incorporated include, but are not limited to, methylation, encapsulation, substitution of one or more nucleic acids with homologs, and modifications between nucleic acids.

In the present invention, '16s rRNA' is an rRNA constituting the 30S subunit of the prokaryotic ribosome, and the nucleotide sequence is largely conserved, while high nucleotide sequence diversity appears in some sections. In particular, since there is little diversity among homogeneous species while diversity appears among other species, prokaryotes can be usefully identified by comparing the sequences of 16S rRNA.

In the present invention, the primer can be used to amplify the conserved 16S rRNA sequence of the microorganism, and the presence of the microorganism can be detected by whether a desired product is generated as a result of the sequence amplification. A sequence amplification method using a primer may use various methods known in the art. For example, polymerase chain reaction (PCR), reverse transcription-polymerase chain reaction (RT-PCR), multiplex PCR, touchdown PCR, hot start PCR, nested PCR, Booster PCR, real-time PCR, differential display PCR (DD-PCR), rapid amplification of cDNA ends (RACE), inverse polymerase chain reaction , vectorette PCR, TAIL-PCR (thermal asymmetric interlaced PCR), ligase chain reaction, repair chain reaction, transcription-mediated amplification, self-maintaining sequence cloning, selective amplification of the target sequence can be used, The scope of the present invention is not limited thereto.

Also, for example, when the agent for detecting the strain is an antibody, the microorganism can be detected using an immunological method based on an antigen-antibody reaction. Analysis methods for this include western blot, ELISA (enzyme linked immunosorbent asay), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, and rocket immunoelectrolysis. Electrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, fluorescence activated cell sorter (FACS), protein chip, etc., but are not limited thereto.

In addition, molecular and immunological methods widely used in the art can be used to detect the microorganisms of the present invention.

The composition comprising the agent capable of detecting the strain of the present invention may be implemented in the form of a diagnostic kit and provided as a kit for predicting or diagnosing obesity risk.

The diagnostic kit includes a primer, a probe, an antisense oligonucleotide, an aptamer, and a detection agent such as an antibody for detecting the microorganisms, as well as one or more other component compositions, solutions, or devices suitable for the analysis method. can

For example, in the present invention, a diagnostic kit including a primer specific for a corresponding microorganism may be a diagnostic kit including essential elements for performing an amplification reaction such as PCR and the like. The diagnostic kit for PCR includes a test tube or other suitable container, reaction buffer, deoxynucleotides (dNTPs), enzymes such as Taq-polymerase reverse transcriptase, DNase, RNAse inhibitors, DEPC-water, sterile water. and the like.

In the present invention, the test subject's intestinal-derived sample may be preferably a fecal sample.

In order to detect microorganisms from a sample from the intestinal tract of a subject, general amplification techniques known in the art, for example, polymerase chain reaction, reverse transcription-polymerase chain reaction, multiplex PCR, touchdown PCR, hot start PCR, four Steed PCR, Booster PCR, Real-Time PCR, Fractional Display PCR, Rapid Amplification of cDNA Ends, Inverse PCR, Vectoret PCR, TAIL-PCR, Ligase Chain Reaction, Repair Chain Reaction, transcription-mediated amplification, self-maintaining sequence cloning, A selective amplification reaction of a target sequence may be used, but the scope of the present invention is not limited thereto.

In addition, immunological methods based on general antigen-antibody reactions known in the art, for example, Western blot, ELISA, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, locate immunoelectrophoresis, Tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, protein chip, etc. may be used, but the scope of the present invention is not limited thereto.

The present invention Blautia obeum ( Blautia obeum ) species strain, Veillonella dispar ( Veillonella dispar ) species strain and Lachnospiraceae ) In the group consisting of any one or more strains selected from the group consisting of strains An agent capable of detecting any one or more strains selected, or together with it, any one or more strains selected from the group consisting of a bacteroides uniformis species strain and a Clostridium sp. strain can be detected Through an agent that can detect any one or more strains selected from the group consisting of an agent that can It can be used as a diagnostic composition, a kit for predicting or diagnosing obesity risk including the same, a method for providing information for predicting or diagnosing obesity risk, and a screening method for preventing or treating obesity.

1 is a graph showing the intestinal microbial distribution of 890 Koreans.
2 is a boxplot showing the relative proportion distribution of the Blautia obeum species strain identified by the ASV of SEQ ID NO: 1 in the obese group, the normal group and the low-weight group.
3 is an obese group, a normal group and a low weight group Veillonella dispar ( Veillonella dispar ) All strains belonging to the species and Veillonella dispar identified by the ASV of SEQ ID NO: 2 Relative ratio distribution of the species strain is a boxplot showing
4 is an obese group, a normal group and a low-weight group Lachnospiraceae All strains belonging to the family and Lachnospiraceae identified by the ASV of SEQ ID NO: 3 Boxplot showing the relative ratio distribution of the family strain am.
5 is an obese group, a normal group and a low-weight group in the Bacteroides uniformis ( bacteroides uniformis ) species identified by the ASV of the total strain and SEQ ID NO: 4 Bacteroides uniformis ( bacteroides uniformis ) relative of the species strain This is a boxplot showing the ratio distribution.
6 is a boxplot showing the relative ratio distribution of all strains belonging to the genus Clostridium and the ASV of SEQ ID NO: 5 in the obese group, the normal group and the low-weight group.
7 is a boxplot showing the relative proportion distribution of all strains belonging to the family Rikenellaceae in the obese group, the normal group and the low-weight group.
8 is a boxplot showing the relative proportion distribution of all strains belonging to the Anaerostipes species in the obese group, the normal group and the low-weight group.
9 is Blautia obeum ( Blautia obeum ) species strain of Table 1, Veillonella dispar ( Veillonella dispar ) Species strain and Lachnospiraceae ( Lachnospiraceae ) By constructing a multivariate linear model using the relative amounts of the strain, This is a boxplot predicting the difference between the normal group and the obese group.
10 is a boxplot predicting the difference between the normal group and the obese group by constructing a multivariate linear model using the relative amounts of all biomarker microorganisms in Table 1.

Hereinafter, the present invention will be described in detail by way of experimental examples and examples. However, the following examples are merely illustrative of the present invention, and the present invention is not limited by the following examples.

Experimental Example 1: Research subject and sample collection

Fecal samples were collected from 890 Koreans participating in the health checkup. To minimize changes in fecal microorganisms, fecal samples were collected using the OMNIgene-GUT kit (DNA Genotek, Ontario, Canada) and stored at room temperature until DNA extraction.

In addition, during the health check-up, questionnaires about lifestyle and physical measurements were performed, and the body mass coefficient was calculated by dividing the weight (kg) by the square of the height (m) from the weight and height results.

Experimental Example 2: DNA Extraction

DNA was extracted from the fecal sample of Experimental Example 1 using the bead-beating extraction method, and DNA was extracted using the QIAamp DNA Stool Mini Kit (Qiagen, Hilden, Germany).

Experimental Example 3: 16S rRNA gene sequencing

A library targeting the V3-V4 hypervariable region of the 16S rRNA gene was prepared using the DNA extracted in Experimental Example 2, and the sequence of the portion was sequenced using Illumina MiSeq 2x300 (Illumina, CA, USA).

Experimental Example 4: Sequence analysis and annotation

The sequencing result in Experimental Example 3 was converted into an amplicon sequence variant (ASV) table using the QIIME2 DADA2 module. Each ASV corresponds to a partial sequence of 16S rRNA, and each can be used as an indicator for detecting a specific microorganism. To identify the microbial phylogeny, the microbial phylogeny was analyzed using the Naive Bayesian classifier of QIIME2 and the GreenGene 13.8 database, and the microbial species unit annotation was performed.

Experimental Example 5: Correlation analysis between intestinal microbes and body mass index

Based on the body mass index (BMI) of 890 health check-up subjects, those below 18.5 were classified as underweight, those above 18.5 and below 25 were classified as normal, and those above 25 were classified as obese.

For each ASV nucleotide sequence representing a specific microbial species, the average ratio of the total intestinal microflora in the obese group, the normal group, and the low-weight group is obtained, and the average ratio is sequentially increased or decreased in the obese group, the normal group and the low-weight group. was selected. When the ratio of the selected microorganisms showed a statistically significant difference between the obese group and the normal group, it was defined as a biomarker microorganism for predicting obesity risk or for diagnosis. One-way ANOVA was used for statistical analysis, and a significant difference was shown between the obese group and the normal group by comparing the box plots as in FIGS. 2 to 8 .

Experiment result

The distribution of intestinal microbes in 890 Koreans is shown in FIG. 1 . 890 Korean intestinal microbes Bacteroides, Prevotella, Picalibacterium, Unclassified Lachnospiraceae, Unclassified Luminococci, Oscillospira, Luminococcus, Parabacteroides, Suterella, Coprococcus and Classified as other, the relative ratio of Bacteroides was shown by arranging from left to right from high to low subjects.

As biomarker strains for predicting or diagnosing the risk of obesity that show a statistically significant difference in ratio between the obese group and the normal group, the Blautia obeum species strain in Table 1 below, Veillonella dispar ) species strains, Lachnospiraceae family strains, Bacteroides uniformis species strains, Clostridium genus strains, Rikenellaceae family strains and Anaerostifes ( Anaerostipes ) species strains were selected.

division system between the obese group and the normal group
P-value Blautia obeum ASV Bell 0.003872 Veillonella dispar ASV Bell 0.010611 Lachnospiraceae ASV class 0.038421 Bacteroides uniformis
( bacteroides uniformis ) ASV Bell 0.004944 Clostridium ASV inside 0.034308 Rikenellaceae class 0.002120 Anaerostipes Bell 0.044725

Looking at the relative proportion distribution of the Blautia obeum species strain identified by the ASV of SEQ ID NO: 1 in the obese group, the normal group and the low weight group of FIG. 2, the Blautia identified by the ASV of SEQ ID NO: 1 Obeum ( Blautia obeum ) In the case of the strain, the obese group showed a significantly lower ratio compared to the normal group or the underweight group, and furthermore, the obese group showed a significantly lower ratio compared to the underweight group. Therefore, when the ratio of the Blautia obeum species strain identified by the ASV of SEQ ID NO: 1 in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is lower than that of the normal group, the risk of obesity is predicted or diagnosed can do. In addition, when the ratio of the Lachnospira spp . strain identified by the ASV of SEQ ID NO: 1 is higher than that of the obese group, it can be predicted or diagnosed as having a low risk of obesity corresponding to the underweight group or the normal group.

The relative proportion distribution of all strains belonging to Veillonella dispar species and ASV of SEQ ID NO: 2 in the obese group, normal group and low weight group of FIG. 3 and Veillonella dispar species strain Looking at the, the relative ratio of the total strains belonging to the Veillonella dispar species has no significant difference for each group, but in the case of the Veillonella dispar species strain identified by the ASV of SEQ ID NO: 2 Compared to the normal group, the obese group showed a significantly higher ratio, and further, the obese group showed a significantly higher ratio than the underweight group. Therefore, when the Veillonella dispar species strain identified by the ASV of SEQ ID NO: 2 in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is higher than that of the normal group, it can be predicted or diagnosed as having a high risk of obesity. there is. In addition, when the Veillonella dispar species strain identified by the ASV of SEQ ID NO: 2 is lower than that of the obese group, it can be predicted or diagnosed as having a low risk of obesity by corresponding to the normal group or the underweight group.

In the obese group, normal group and low weight group of FIG. 4, Lachnospiraceae identified by the entire strain belonging to the family and the ASV of SEQ ID NO: 3 Lachnospiraceae Looking at the relative ratio distribution of the strain, Lach The relative ratio of the total strains belonging to the family Nospira ( Lachnospiraceae ) There is no significant difference for each group, but Lachnospiraceae identified by the ASV of SEQ ID NO: 3 In the case of the family strain, it is significant in the obese group compared to the normal group represents a low ratio. Therefore, if the ratio of Lachnospiraceae and strain identified by ASV of SEQ ID NO: 3 in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject is lower than that of the normal group, the risk of obesity is high It can be predicted or diagnosed there is. In addition, when the ratio of Lachnospiraceae and strains identified by the ASV of SEQ ID NO: 3 is high compared to the obese group, it can be predicted or diagnosed as having a low risk of obesity.

In the obese group, normal group and low weight group of FIG. 5 , the entire strain belonging to the Bacteroides uniformis species and the Bacteroides uniformis species strain identified by the ASV of SEQ ID NO: 4 Relative of the strain Looking at the ratio distribution, the relative ratio of the total strain belonging to the Bacteroides uniformis species is not significantly different for each group, but identified by the ASV of SEQ ID NO: 3 Bacteroides uniformis ( bacteroides uniformis ) In the case of the species strain, the obese group showed a significantly lower ratio than the normal group, and further, the obese group showed a significantly lower ratio compared to the underweight group. Therefore, if the Bacteroides uniformis species strain identified by the ASV of SEQ ID NO: 4 in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is lower than that of the normal group, the risk of obesity can be predicted or diagnosed. can In addition, when the Bacteroides uniformis species strain identified by the ASV of SEQ ID NO: 4 is higher than that of the obese group, it can be predicted or diagnosed as having a low risk of obesity by corresponding to a normal group or a low-weight group.

In the obese group, normal group and low weight group of Figure 6, Clostridium ( Clostridium ) The entire strain belonging to the genus and Clostridium identified by the ASV of SEQ ID NO: 5 Looking at the relative ratio distribution of the genus strain, Clostridium ( Clostridium ) The relative ratio of all strains belonging to the genus does not have a significant difference for each group, but the Clostridium spp. strain identified by ASV of SEQ ID NO: 5 shows a significantly lower ratio in the obese group than in the normal group. Therefore, when the ratio of the Clostridium genus strain identified by the ASV of SEQ ID NO: 5 in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject is lower than that of the normal group, it can be predicted or diagnosed as having a high risk of obesity. . In addition, when the ratio of the Clostridium sp. strain identified by the ASV of SEQ ID NO: 5 is high compared to the obese group, it can be predicted or diagnosed as having a low risk of obesity.

7 is an obese group, a normal group and a low-weight group. Looking at the relative ratio distribution of all strains belonging to the family Rikenellaceae identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 6 to SEQ ID NO: 16 , For the entire strain belonging to the family Rikenellaceae identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 6 to SEQ ID NO: 16, a significantly lower ratio in the obese group than in the normal group or the underweight group , and furthermore, the ratio was significantly lower in the obese group than in the underweight group. Therefore, the ratio of all strains belonging to the family Rikenellaceae identified by any one ASV sequence selected from the nucleotide sequences of SEQ ID NO: 6 to SEQ ID NO: 16 in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is normal If it is low compared to the group, it can be predicted or diagnosed as having a high risk of obesity. In addition, when the ratio of Rikenellaceae and the entire strain identified by the ASV of SEQ ID NOs: 6 to 16 is higher than that of the obese group, it can be predicted or diagnosed as having a low risk of obesity by corresponding to the underweight group or the normal group.

8 is an obese group, a normal group and a low weight group, Anaerostipes identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 17 to SEQ ID NO: 19 Relative ratio of total strains belonging to the strain Looking at the distribution, Anaerostipes identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 17 to SEQ ID NO: 19 In the case of the entire strain belonging to the strain, it is significant in the obese group compared to the normal group. represents a low ratio. Therefore, in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract, Anaerostipes identified by any one ASV sequence selected from the nucleotide sequences of SEQ ID NO: 17 to SEQ ID NO: 19 of the entire strain belonging to the strain When the ratio is lower than that of the normal group, it can be predicted or diagnosed as having a high risk of obesity. In addition, if the ratio of the entire strain belonging to the Anaerostipes species strain identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 17 to SEQ ID NO: 19 is higher than that of the obese group, the risk of obesity is low. predictable or diagnosable.

Among the biomarker microorganisms in Table 1, Blautia obeum spp. strain, Veillonella dispar spp. strain and Lachnospiraceae multivariate linear model to predict the obesity group by combining the strains and strains. 9 shows the results of predicting the difference between the normal group and the obese group when configuring . 9 is a configuration of a Bayesian Ridge model for predicting an obesity group from the relative amounts of the three types of biomarker microorganisms, and the Bayesian Ridge model is a type of linear model. Among the variables constituting the linear model, meaningless variables are automatically excluded. way. In the above method, the BayesianRidge function of the Python package scikit-learn was used. According to FIG. 9 , it can be confirmed that the risk of diabetes can be predicted or diagnosed more clearly when the three types of biomarker microorganisms are combined (P value = 4.3 X 10 ^-5 ).

10 is a result of predicting the risk of obesity using the Bayesian Ridge model using the entire biomarker microorganisms of Table 1 as in FIG. It can be confirmed that it can be clearly predicted (P value = 3.3 X 10 ^-7 ).

<110> KOREA FOOD RESEARCH INSTITUTE <120> Predicting or Diagnosing Composition for Risk of Obesity Using Human Intestinal Microbiome, Diagnosing Kit, Method For Providing Information, And Screening Method For Drugs For Preventing Or Treating Obesity Using The Same <130> HPC9313 <160> 19 <170> KoPatentIn 3.0 <210> 1 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Blautia obeum ASV <400> 1 tggggaatat tgcacaatgg gggaaaccct gatgcagcga cgccgcgtga aggaagaagt 60 atctcggtat gtaaacttct atcagcaggg aagatagtga cggtacctga ctaagaagcc 120 ccggctaact acgtgccagc agccgcggta atacgtaggg ggcaagcgtt atccggattt 180 actgggtgta aagggagcgt agacggactg gcaagtctga tgtgaaaggc gggggctcaa 240 cccctggact gcattggaaa ctgttagtct tgagtgccgg agaggtaagc ggaattccta 300 gtgtagcggt gaaatgcgta gatattagga ggaacaccag tggcgaaggc ggcttactgg 360 acggtaactg acgttgaggc tcgaaagcgt ggggagcaaa ca 402 <210> 2 <211> 427 <212> DNA <213> Artificial Sequence <220> <223> Veillonella dispar ASV <400> 2 tggggaatct tccgcaatgg acgaaagtct gacggagcaa cgccgcgtga gtgatgacgg 60 ccttcgggtt gtaaagctct gttaatcggg acgaaaggcc ttcttgcgaa tagttagaag 120 gattgacggt accggaatag aaagccacgg ctaactacgt gccagcagcc gcggtaatac 180 gtaggtggca agcgttgtcc ggaattattg ggcgtaaagc gcgcgcaggc ggattggtca 240 gtctgtctta aaagttcggg gcttaacccc gtgatgggat ggaaactgcc aatctagagt 300 atcggagagg aaagtggaat tcctagtgta gcggtgaaat gcgtagatat taggaagaac 360 accagtggcg aaggcgactt tctggacgaa aactgacgct gaggcgcgaa agccagggga 420 gcgaacg 427 <210> 3 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Lachnospiraceae ASV <400> 3 tggggaatat tgcacaatgg gggaaaccct gatgcagcga cgccgcgtga gcgaagaagt 60 atttcggtat gtaaagctct atcagcaggg aagataatga cggtacctga ctaagaagcc 120 ccggctaact acgtgccagc agccgcggta atacgtaggg ggcaagcgtt atccggattt 180 actgggtgta aagggagcgt agacggcaag gcaagtctga tgtgaaaacc cagggcttaa 240 ccctgggact gcattggaaa ctgtctggct cgagtgccgg agaggtaagc ggaattccta 300 gtgtagcggt gaaatgcgta gatattagga agaacaccag tggcgaaggc ggcttactgg 360 acggtaactg acgttgaggc tcgaaagcgt ggggagcaaa ca 402 <210> 4 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Bacteroides uniformis ASV <400> 4 tgaggaatat tggtcaatgg acgagagtct gaaccagcca agtagcgtga aggatgactg 60 ccctatgggt tgtaaacttc ttttatacgg gaataaagtg aggcacgtgt gcctttttgt 120 atgtaccgta tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggagcgt aggcggacgc ttaagtcagt 240 tgtgaaagtt tgcggctcaa ccgtaaaatt gcagttgata ctgggtgtct tgagtacagt 300 agaggcaggc ggaattcgtg gtgtagcggt gaaatgctta gatatcacga agaactccga 360 ttgcgaaggc agcttgctgg actgtaactg acgctgatgc tcgaaagtgt gggtatcaaa 420 ca 422 <210> 5 <211> 405 <212> DNA <213> Artificial Sequence <220> <223> Clostridium ASV <400> 5 tggggaatat tgcgcaatgg gggcaaccct gacgcagcaa cgccgcgtgc aggaagaagg 60 tcttcggatt gtaaactgtt gtcgcaaggg aagaagacag tgacggtacc ttgtgagaaa 120 gtcacggcta actacgtgcc agcagccgcg gtaatacgta ggtgacaagc gttgtccgga 180 tttactgggt gtaaagggcg cgtaggcgga ctgtcaagtc agtcgtgaaa taccggggct 240 taaccccggg gctgcgattg aaactgacag ccttgagtat cggagaggaa agcggaattc 300 ctagtgtagc ggtgaaatgc gtagatatta ggaggaacac cagtggcgaa ggcggctttc 360 tggacgacaa ctgacgctga ggcgcgaaag tgtggggagc aaaca 405 <210> 6 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 6 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggatgacgg 60 ctctatgagt tgtaaactgc ttttgtacga gggtaaaccc ggatacgtgt atccggctga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 ttcaagcgtt atccggattt attgggttta aagggtgcgt aggcggtttg ataagttaga 240 ggtgaaatac cggtgcttaa caccggaact gcctctaata ctgttgaact agagagtagt 300 tgcggtaggc ggaatgtatg gtgtagcggt gaaatgctta gagatcatac agaacaccga 360 ttgcgaaggc agcttaccaa actatatctg acgttgaggc acgaaagcgt ggggagcaaa 420 ca 422 <210> 7 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 7 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggatgacgg 60 ctctatgagt tgtaaactgc ttttgtacga gggtaaaccc ggatacgtgt atccggctga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 ttcaagcgtt atccggattt attgggttta aagggtgcgt aggcggtttg ataagttaga 240 ggtgaaatac cggtgcttaa caccggaact gcctctaata ctgttgagct agagagtagt 300 tgcggtaggc ggaatgtatg gtgtagcggt gaaatgctta gagatcatac agaacaccga 360 ttgcgaaggc agcttaccaa actatatctg acgttgaggc acgaaagcgt ggggagcaaa 420 ca 422 <210> 8 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 8 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggaagacgg 60 ctctatgagt tgtaaactgc ttttgtacga gggtaaacgc tcttacgtgt aagagcctga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccaagcgtt atccggattt attgggttta aagggtgcgt aggcggtttg ataagttaga 240 ggtgaaatac cggtgcttaa caccggaact gcctctaata ctgttgaact agagagtagt 300 tgcggtaggc ggaatgtatg gtgtagcggt gaaatgctta gagatcatac agaacaccga 360 ttgcgaaggc agcttaccaa actatatctg acgttgaggc acgaaagcgt ggggagcaaa 420 ca 422 <210> 9 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 9 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggaagacgg 60 ctctatgagt tgtaaactgc ttttgtacga gggtaaactc acctacgtgt aggtgactga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 ttcaagcgtt atccggattt attgggttta aagggtgcgt aggcggtttg ataagttaga 240 ggtgaaatcc cggggcttaa ctccggaact gcctctaata ctgttagact agagagtagt 300 tgcggtaggc ggaatgtatg gtgtagcggt gaaatgctta gagatcatac agaacaccga 360 ttgcgaaggc agcttaccaa actatatctg acgttgaggc acgaaagcgt ggggagcaaa 420 ca 422 <210> 10 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 10 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggatgacgg 60 ctctatgagt tgtaaactgc ttttgtacga gggtaaacgc agatacgtgt atctgtctga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 ttcaagcgtt atccggattt attgggttta aagggtgcgt aggcggtttg ataagttaga 240 ggtgaaattt cggggctcaa ccctgaacgt gcctctaata ctgttgagct agagagtagt 300 tgcggtaggc ggaatgtatg gtgtagcggt gaaatgctta gagatcatac agaacaccga 360 ttgcgaaggc agcttaccaa actatatctg acgttgaggc acgaaagcgt ggggagcaaa 420 ca 422 <210> 11 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 11 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggaagacgg 60 ctctatgagt tgtaaactgc ttttgtacga gagtaaacgc tcttacgtgt aagagcctga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccaagcgtt atccggattt attgggttta aagggtgcgt aggcggtttg ataagttaga 240 ggtgaaatac cggtgcttaa caccggaact gcctctaata ctgttgaact agagagtagt 300 tgcggtaggc ggaatgtatg gtgtagcggt gaaatgctta gagatcatac agaacaccga 360 ttgcgaaggc agcttaccaa actatatctg acgttgaggc acgaaagcgt ggggagcaaa 420 ca 422 <210> 12 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 12 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggaagacgg 60 ctctatgagt tgtaaactgc ttttgtacta gggtaaacgc ttctacgtgt aggagcctga 120 aagtatagta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccaagcgtt atccggattt attgggttta aagggtgcgt aggcggtttg ataagttaga 240 ggtgaaatac cggggctcaa ctccggaact gcctctaata ctgttgaact agagagtagt 300 tgcggtaggc ggaatgtatg gtgtagcggt gaaatgctta gagatcatac agaacaccga 360 ttgcgaaggc agcttaccaa actatatctg acgttgaggc acgaaagcgt ggggagcaaa 420 ca 422 <210> 13 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 13 tgaggaatat tggtcaatgg acggaagtct gaaccagcca tgccgcgtgc aggaagacgg 60 ctctatgagt tgtaaactgc ttttgtacga gggtaaacgc agatacgtgt atctgcctga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccaagcgtt atccggattt attgggttta aagggtgcgt aggcggttta gtaagtcagc 240 ggtgaaattt tggtgcttaa caccaaacgt gccgttgata ctgctgggct agagagtagt 300 tgcggtaggc ggaatgtatg gtgtagcggt gaaatgctta gagatcatac agaacaccga 360 ttgcgaaggc agcttaccaa actatatctg acgttgaggc acgaaagcgt ggggagcaaa 420 ca 422 <210> 14 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 14 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggatgaagg 60 tgctatgcat tgtaaactgc ttttgtacga gggtaaatgc aggtacgtgt acctgtttga 120 aagtatcgta cgaataaggg tcggctaact ccgtgccagc agccgcggta atacggagga 180 cccgagcgtt atccggattt attgggttta aagggtgcgt aggcggatta gtaagttaga 240 ggtgaaagct cgatgctcaa catcgaaatt gcctctgata ctgttagtct agagtatagt 300 tgcggaaggc ggaatgtgtg gtgtagcggt gaaatgctta gatatcacac agaacaccga 360 ttgcgaaggc agctttccaa gctattactg acgctgatgc acgaaagcgt ggggagcgaa 420 ca 422 <210> 15 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 15 tgaggaatat tggtcaatgg acggaagtct gaaccagcca tgccgcgtgc aggatgaatg 60 tgctatgcat tgtaaactgc ttttgtacga gggtaaacac agatacgcgt atctgcttga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggtgcgt aggctgtttt ttaagttaga 240 ggtgaaagct cgacgctcaa cgtcgaaatt gcctctgata ctgagagact agagtgtagt 300 tgcggaaggc ggaatgtgtg gtgtagcggt gaaatgctta gatatcacac agaacaccga 360 ttgcgaaggc agctttccaa gctattactg acgctgaggc acgaaagcgt ggggagcgaa 420 ca 422 <210> 16 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Rikenellaceae <400> 16 tgaggaatat tggtcaatgg acgcaagtct gaaccagcca tgccgcgtgc aggatgaagg 60 tgctatgcat tgtaaactgc ttttgtacga gggtaaattc agatacgtgt atctgtttga 120 aagtatcgta cgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggtgcgt aggcggtttg ataagttaga 240 ggtgaaagct cgatgctcaa catcgaaatt gcctctgata ctgtcagact agagtgtagt 300 tgcggaaggc ggaatgtgtg gtgtagcggt gaaatgctta gatatcacac agaacaccga 360 ttgcgaaggc agctttccaa gctattactg acgctgaggc acgaaagcgt gggtagcgaa 420 ca 422 <210> 17 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Anaerostipes <400> 17 tggggaatat tgcacaatgg gggaaaccct gatgcagcga cgccgcgtga aggaagaagt 60 atttcggtat gtaaacttct atcagcaagg aagaaaatga cggtacttga ctaagaagcc 120 ccggctaaat acgtgccagc agccgcggta atacgtatgg ggcaagcgtt atccggattt 180 actgggtgta aagggagcgt aggcggtaag acaagtcaga agtgaaaggc tggggctcaa 240 ccctgggact gcttttgaaa ctgtctaact agagtgcagg agaggtaagt ggaattccta 300 gtgtagcggt gaaatgcgta gatattagga ggaacaccag tggcgaaggc ggcttactgg 360 actgtaactg acgctgaggc tcgaaagcgt ggggagcaaa ca 402 <210> 18 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Anaerostipes <400> 18 tagggaatat tgcacaatgg gggaaaccct gatgcagcga cgccgcgtga aggaagaagt 60 atttcggtat gtaaacttct atcagcaagg aagaaaatga cggtacttga ctaagaagcc 120 ccggctaaat acgtgccagc agccgcggta atacgtatgg ggcaagcgtt atccggattt 180 actgggtgta aagggagcgt aggcggcatg gcaagtcaga agtgaaagcc tggggctcaa 240 ccccggaatt gcttttgaaa ctgtcaggct agagtgtcgg aggggtaagc ggaattccta 300 gtgtagcggt gaaatgcgta gatattagga ggaacaccgg tggcgaaggc ggcttactgg 360 acgattactg acgctgaggc tcgaaagcgt ggggagcaaa ca 402 <210> 19 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Anaerostipes <400> 19 tagggaatat tgcacaatgg gggaaaccct gatgcagcga cgccgcgtga aggatgaagt 60 atttcggtat gtaaacttct atcagcaagg aagaaaatga cggtacttga ctaagaagcc 120 ccggctaaat acgtgccagc agccgcggta atacgtatgg ggcaagcgtt atccggattt 180 actgggtgta aagggagcgt aggcggtatg gcaagtcaga agtgaaagcc tggggctcaa 240 ccccggaatt gcttttgaaa ctgtcaaact agagtgtcgg aggggtaagc ggaattccta 300 gtgtagcggt gaaatgcgta gatattagga ggaacaccgg tggcgaaggc ggcttactgg 360 acgaccactg acgctgaggc tcgaaagcgt ggggagcaaa ca 402

Claims (17)

Veillonella dispar containing the 16S rRNA nucleotide sequence of SEQ ID NO: 2 A composition for predicting or diagnosing obesity risk using an intestinal microorganism, comprising an agent capable of detecting a species strain. According to claim 1, Blautia obeum containing the 16S rRNA nucleotide sequence of SEQ ID NO: 1 Species strain, and Lachnospiraceae containing the 16S rRNA nucleotide sequence of SEQ ID NO: 3 ( Lachnospiraceae ) family strain A composition for predicting or diagnosing obesity risk using intestinal microorganisms, characterized in that it further comprises an agent capable of detecting The method of claim 2, wherein in the preparation, Bacteroides uniformis ( bacteroides uniformis ) species strain comprising the 16S rRNA sequence of SEQ ID NO: 4, Clostridium containing the 16S rRNA sequence of SEQ ID NO: 5 ) genus strain, SEQ ID NO: 6 to SEQ ID NO: 16S rRNA nucleotide sequence of SEQ ID NO: 16 Rikenellaceae comprising any one sequence selected from the family strain, and SEQ ID NO: 17 to SEQ ID NO: 19 16S rRNA nucleotide sequence selected from Anaerostipes ( Anaerostipes ) A composition for predicting or diagnosing obesity risk using intestinal microorganisms, further comprising an agent capable of detecting a strain comprising any one of the sequences. delete delete delete According to any one of claims 1 to 3, wherein the agent capable of detecting the strain is a primer, probe, antisense oligonucleotide, aptamer or antibody specific to the strain. A composition for risk prediction or diagnosis. The composition for predicting or diagnosing obesity risk using intestinal microorganisms according to claim 7, wherein the primer is a primer capable of amplifying the 16S rRNA of the strain. A kit for predicting or diagnosing obesity risk, comprising the composition of any one of claims 1 to 3. Detecting the Veillonella dispar strain comprising the 16S rRNA base sequence of SEQ ID NO: 2 from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject; and
Comparing the increase or decrease of the Veillonella dispar species strain comprising the 16S rRNA nucleotide sequence of SEQ ID NO: 2 with a normal group; Provide information necessary for predicting or diagnosing obesity risk using intestinal microorganisms, including As a method for
The obesity or obesity risk group is a method, characterized in that satisfying the following selection criteria:
Compared to the normal group, the Veillonella dispar species strain containing the 16S rRNA nucleotide sequence of SEQ ID NO: 2 in the genomic DNA of the test subject's microorganism is increased. 11. The method of claim 10, wherein from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject, the 16S rRNA base sequence of SEQ ID NO: 1 Blautia obeum species strain comprising the nucleotide sequence of SEQ ID NO: 1, and 16S rRNA of SEQ ID NO: 3 Detecting the Lachnospiraceae and strain containing the nucleotide sequence; And Blautia obeum species strain comprising the 16S rRNA base sequence of SEQ ID NO: 1, and Lachnospiraceae containing the 16S rRNA base sequence of SEQ ID NO: 3 Normal increase or decrease in strains Comparing with the group; further comprising,
The obesity or obesity risk group is a method, characterized in that satisfying the following selection criteria:
Compared to the normal group, the Veillonella dispar species strain comprising the 16S rRNA base sequence of SEQ ID NO: 2 in the genomic DNA of the test subject's microorganism is increased,
Compared to the normal group, the strain of Blautia obeum containing the 16S rRNA nucleotide sequence of SEQ ID NO: 1 in the genomic DNA of the test subject's microorganism is reduced, and
Compared to the normal group, Lachnospiraceae containing the 16S rRNA nucleotide sequence of SEQ ID NO: 3 in the genomic DNA of the test subject's microorganism is reduced and the strain. According to claim 11, Bacteroides uniformis comprising the 16S rRNA base sequence of SEQ ID NO: 4 from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject ( bacteroides uniformis ) species strain, 16S rRNA of SEQ ID NO: 5 Clostridium genus strain comprising a nucleotide sequence, SEQ ID NO: 6 to SEQ ID NO: 16S rRNA comprising any one sequence selected from rRNA nucleotide sequence Rikenellaceae ( Rikenellaceae ) and strains, and SEQ ID NOs: 17 to Detecting a strain of Anaerostipes comprising any one sequence selected from the 16S rRNA nucleotide sequence of SEQ ID NO: 19; And Bacteroides uniformis ( bacteroides uniformis ) species strain comprising the 16S rRNA sequence of SEQ ID NO: 4, Clostridium genus strain comprising the 16S rRNA sequence of SEQ ID NO: 5, SEQ ID NO: 6 to sequence No. 16 Rikenellaceae comprising any one sequence selected from the 16S rRNA nucleotide sequence, and strain, and SEQ ID NO: 17 to SEQ ID NO: 19 Ana comprising any one sequence selected from the rRNA nucleotide sequence Erostipes ( Anaerostipes ) Comparing the increase and decrease of the species strain with the normal group; further comprising,
The obesity or obesity risk group is a method, characterized in that satisfying the following selection criteria:
Compared to the normal group, the Veillonella dispar species strain containing the 16S rRNA nucleotide sequence of SEQ ID NO: 2 in the genomic DNA of the test subject's microorganism is increased,
Compared to the normal group, the Blautia obeum species strain comprising the 16S rRNA base sequence of SEQ ID NO: 1 in the genomic DNA of the test subject's microorganism is reduced,
Compared to the normal group, the Lachnospiraceae and strains containing the 16S rRNA nucleotide sequence of SEQ ID NO: 3 in the genomic DNA of the test subject's microorganism are reduced,
Compared to the normal group, Bacteroides uniformis containing the 16S rRNA nucleotide sequence of SEQ ID NO: 4 in the genomic DNA of the test subject's microorganism is reduced in the strain,
Compared to the normal group, the Clostridium spp. strain containing the 16S rRNA nucleotide sequence of SEQ ID NO: 5 in the genomic DNA of the test subject's microorganism is reduced,
Compared to the normal group, Rikenellaceae containing any one sequence selected from the 16S rRNA nucleotide sequence of SEQ ID NO: 6 to SEQ ID NO: 16 in the genomic DNA of the test subject's microorganism is reduced, and
Compared to the normal group, Anaerostipes containing any one sequence selected from the 16S rRNA nucleotide sequence of SEQ ID NO: 17 to SEQ ID NO: 19 in the genomic DNA of the test subject's microorganism is reduced. From the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject, the Blautia obeum species strain containing the 16S rRNA sequence of SEQ ID NO: 1, Baylonella containing the 16S rRNA sequence of SEQ ID NO: 2 Dispar ( Veillonella dispar ) species strain, and Lachnospiraceae comprising the 16S rRNA nucleotide sequence of SEQ ID NO: 3 , detecting the relative amount of the family strain; and
The result obtained in the detecting step, calculating the obesity risk of the test subject by a multivariate linear model for predicting the obesity group using the relative amount of the strains as a variable; How to provide information for diagnosis. From the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject, the Blautia obeum species strain containing the 16S rRNA sequence of SEQ ID NO: 1, Baylonella containing the 16S rRNA sequence of SEQ ID NO: 2 Dispar ( Veillonella dispar ) species strain, Lachnospiraceae containing the 16S rRNA sequence of SEQ ID NO: 3 and strains, Bacteroides uniformis containing the 16S rRNA sequence of SEQ ID NO: 4 ) Species strain, Clostridium sp. strain comprising the 16S rRNA nucleotide sequence of SEQ ID NO: 5, SEQ ID NO: 6 to SEQ ID NO: 16S rRNA nucleotide sequence comprising any one sequence selected from the ricenella bird ( Rikenellaceae ) and strains, and SEQ ID NO: 17 to SEQ ID NO: 19 Anaerostipes containing any one sequence selected from the rRNA nucleotide sequence of SEQ ID NO: Detecting the relative amount of the species strain; and
The result obtained in the detecting step, calculating the obesity risk of the test subject by a multivariate linear model for predicting the obesity group using the relative amount of the strains as a variable; How to provide information for diagnosis. delete delete delete

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