Method for diagnosing head and neck cancer by bacterial metagenomic analysis

文档序号：1315896 发布日期：2020-07-10 浏览：7次中文

阅读说明：本技术 通过细菌宏基因组分析来诊断头颈癌的方法 (Method for diagnosing head and neck cancer by bacterial metagenomic analysis ) 是由金润根于 2018-04-26 设计创作，主要内容包括：本发明涉及通过细菌宏基因组分析诊断头颈癌的方法,并且更具体地涉及通过使用来源于正常个人和受试者的样品执行细菌宏基因组分析经由分析来源于特定细菌的细胞外囊泡的含量的增加或减少来诊断头颈癌的方法。从环境中存在的细菌分泌的细胞外囊泡引起局部慢性炎症或吸收到人体中,因此可直接影响癌症的发生,且难以在其症状发作前及早诊断出头颈癌,因此难以有效治疗。因此,根据本发明,可通过使用来源于人体的样品对来源于细菌的细胞外囊泡进行宏基因组分析诊断头颈癌的风险,且因此使得能够及早诊断并预测头颈癌的风险群,从而通过适当治疗延迟头颈癌发作或预防头颈癌发作,并且甚至在头颈癌发作后能够实施对头颈癌的早期诊断,从而降低头颈癌的发生率并提高治疗效果。(The present invention relates to a method for diagnosing head and neck cancer through bacterial metagenomic analysis, and more particularly, to a method for diagnosing head and neck cancer through analysis of increase or decrease in the content of extracellular vesicles derived from specific bacteria by performing bacterial metagenomic analysis using samples derived from normal individuals and subjects. Extracellular vesicles secreted from bacteria present in the environment cause local chronic inflammation or are absorbed into the human body, and thus may directly affect the occurrence of cancer, and it is difficult to diagnose head and neck cancer early before the onset of its symptoms, and thus it is difficult to effectively treat it. Therefore, according to the present invention, it is possible to diagnose the risk of head and neck cancer by metagenomic analysis of extracellular vesicles derived from bacteria using a human-derived sample, and thus to enable early diagnosis and prediction of a risk group of head and neck cancer, thereby delaying the onset of head and neck cancer or preventing the onset of head and neck cancer through appropriate treatment, and enabling early diagnosis of head and neck cancer to be performed even after the onset of head and neck cancer, thereby reducing the incidence of head and neck cancer and improving the therapeutic effect.)

1. A method of providing information for diagnosing head and neck cancer, the method comprising:

(a) extracting DNA from extracellular vesicles isolated from a sample from a normal individual and a sample from a subject;

(b) using a nucleic acid comprising SEQ ID NO: 1 and SEQ ID NO: 2, performing Polymerase Chain Reaction (PCR) on the extracted DNA; and

(c) by sequencing the products of the PCR, the content of the bacterial-derived extracellular vesicles from the subject sample is increased or decreased compared to the content of the bacterial-derived extracellular vesicles from a sample from a normal individual and determined.

2. The method of claim 1, wherein process (c) comprises comparing and determining:

the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of Cyanobacteria (Cyanobacteria) and Fusobacteria (Fusobacteria) is increased or decreased.

3. The method of claim 1, wherein process (c) comprises comparing and determining:

the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of halobacteriaceae (halobacteriaceae), chloroplasts (chloroplasts), fusobacteriaceae (fusobacteria) and proteobacteria (epsilon proteobacteria) is increased or decreased.

4. The method of claim 1, wherein process (c) comprises comparing and determining:

increased or decreased content of extracellular vesicles derived from one or more bacteria selected from the group consisting of the order halophiles (Halobacteriales), the order bifidobacteria (bifidobacteria), the order trichoderma (Streptophyta), the order pseudomonales (pseudomonas), the order Oceanospirillales (Oceanospirillales), the order clostridiales (fusobacteriacees) and the order campylobacter (campylobacter).

5. The method of claim 1, wherein process (c) comprises comparing and determining:

the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of pseudomonaceae (pseudomonas adaceae), Halobacteriaceae (Halobacteriaceae), oxalobacteriaceae (oxaloabacteriaceae), Halomonadaceae (Halomonadaceae), comamoniaceae (comamoniaceae), lactobacillaceae (L acetobacteriaceae), pratensitaceae (paranovotellaceae), clostridiaceae (Fusobacteriaceae), and campylobacter (campylobacter asiaticae) is increased or decreased.

6. The method of claim 1, wherein process (c) comprises comparing and determining:

the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of cupriasis (Cupriavidus), Chromohalobacter (Chromohalobacter), Pseudomonas (Pseudomonas), Acinetobacter (Acinetobacter), waterborne bacillus (endobacter), lactobacillus (L acetobacter), Veillonella (Veillonella), clostridium (Fusobacterium), Actinomyces (Actinomyces), Prevotella (Prevotella), Megasphaera (Megasphaera), Campylobacter (Campylobacter) and Oribacterium (Oribacterium) is increased or decreased.

7. The method of claim 1, wherein the sample of normal individuals and the sample of subjects is saliva.

8. The method of claim 1, wherein process (c) comprises comparing and determining:

an increased or decreased content of extracellular vesicles derived from one or more bacteria selected from the group consisting of the phylum Cyanobacteria (Cyanobacteria) and the phylum fusobacterium (Fusobacteria);

an increased or decreased content of extracellular vesicles derived from one or more bacteria selected from the group consisting of halobacteriaceae (halobacteriaceae), chloroplasts (chloroplasts), fusobacteriaceae (fusobacteria) and proteobacteria (epsilon proteobacteria);

(ii) an increased or decreased content of extracellular vesicles derived from one or more bacteria selected from the group consisting of the order of halophiles (halobacterales), the order of bifidobacteria (bifidobacteria), the order of trichoderma (Streptophyta), the order of pseudomonas (pseudomonas), the order of marine spirochetes (Oceanospirillales), the order of clostridia (Fusobacteriales) and the order of campylobacter (campylobacter);

the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of Pseudomonas (Pseudomonas), Halobacteriaceae (Halobacteriaceae), Oxalobacter (Oxalobacter), Halomonas (Halomonadaceae), Comamonas (Comamoneaceae), Lactobacillus (L Acobacteriaceae), Pasteurellaceae (Paraprevotellaceae), Clostridium (Fusobacteriaceae) and Campylobacter (Campylobacter) is increased or decreased, or

The content of extracellular vesicles derived from one or more bacteria selected from the group consisting of cupriasis (Cupriavidus), Chromohalobacter (Chromohalobacter), Pseudomonas (Pseudomonas), Acinetobacter (Acinetobacter), waterborne bacillus (endobacter), lactobacillus (L acetobacter), Veillonella (Veillonella), clostridium (Fusobacterium), Actinomyces (Actinomyces), Prevotella (Prevotella), Megasphaera (Megasphaera), Campylobacter (Campylobacter) and Oribacterium (Oribacterium) is increased or decreased.

9. The method of claim 8, wherein in process (c), an increase in the content of extracellular vesicles as compared to a sample derived from a normal individual is diagnosed as head and neck cancer:

extracellular vesicles from bacteria of the Fusobacteria (fusobacterobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of clostridia (fusobacteriaceia) and proteobacteria (epsilon proteobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of Fusobacteriales (Fusobacteriales) and campylobacter (campylobacter);

extracellular vesicles derived from one or more bacteria selected from the group consisting of the Pseudomonadaceae family (L Acobacteriaceae), the Papaveraceae family (Paraprevotateceae), the Clostridiaceae family (Fusobateriaceae), and the Campylobacter family (Campylobacter eraceae), or

Extracellular vesicles derived from one or more bacteria selected from the group consisting of lactobacillus (L actinobacillus), Veillonella (Veillonella), clostridium (Fusobacterium), Actinomyces (Actinomyces), Prevotella (Prevotella), Megasphaera (Megasphaera), Campylobacter (Campylobacter) and Oribacterium (oribacter).

10. The method of claim 8, wherein in process (c), a reduction in the content of extracellular vesicles as follows is diagnosed as head and neck cancer, as compared to a sample derived from a normal individual:

extracellular vesicles derived from bacteria of the phylum Cyanobacteria (Cyanobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of halobacteriaceae (halobacteriia) and chloroplasts (Chloroplast);

extracellular vesicles derived from one or more bacteria selected from the group consisting of the order halophiles (Halobacteriales), the order bifidobacteria (bifidobacteria), the order trichoderma (Streptophyta), the order pseudomonas (pseudomonas) and the order marine spirochaeta (oceanosporilales);

extracellular vesicles derived from one or more bacteria selected from the group consisting of Pseudomonadaceae (pseudomonas adaceae), Halobacteriaceae (Halobacteriaceae), oxalobacteriaceae (Oxalobacteraceae), halomonas (Halomonadaceae), and comamonoconadaceae (comamoonadaceae); or

Extracellular vesicles derived from one or more bacteria selected from the group consisting of cuprias (Cupriavidus), Chromohalobacter (chromahalobacter), Pseudomonas (Pseudomonas), Acinetobacter (Acinetobacter), and aquaticum (enterobacter).

11. A method of diagnosing head and neck cancer, the method comprising:

(a) extracting DNA from extracellular vesicles isolated from a sample from a normal individual and a sample from a subject;

(b) using a nucleic acid comprising SEQ ID NO: 1 and SEQ ID NO: 2, performing Polymerase Chain Reaction (PCR) on the extracted DNA; and

12. The method of claim 11, wherein process (c) comprises comparing and determining:

the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of Cyanobacteria (Cyanobacteria) and Fusobacteria (Fusobacteria) is increased or decreased.

13. The method of claim 11, wherein process (c) comprises comparing and determining:

14. The method of claim 11, wherein process (c) comprises comparing and determining:

15. The method of claim 11, wherein process (c) comprises comparing and determining:

16. The method of claim 11, wherein process (c) comprises comparing and determining:

17. The method of claim 11, wherein the sample of normal individuals and the sample of subjects is saliva.

18. The method of claim 11, wherein process (c) comprises comparing and determining:

19. The method of claim 18, wherein in process (c), an increase in the content of extracellular vesicles as compared to a sample derived from a normal individual is diagnosed as head and neck cancer:

extracellular vesicles from bacteria of the Fusobacteria (fusobacterobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of clostridia (fusobacteriaceia) and proteobacteria (epsilon proteobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of Fusobacteriales (Fusobacteriales) and campylobacter (campylobacter);

20. The method of claim 18, wherein in process (c), a reduction in the content of extracellular vesicles as follows is diagnosed as head and neck cancer, as compared to a sample derived from a normal individual:

extracellular vesicles derived from bacteria of the phylum Cyanobacteria (Cyanobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of halobacteriaceae (halobacteriia) and chloroplasts (Chloroplast);

Technical Field

The present invention relates to a method for diagnosing head and neck cancer by bacterial metagenomic analysis, and more particularly, to a method for diagnosing head and neck cancer by performing bacterial metagenomic analysis using a sample derived from a normal individual and a sample derived from a subject to analyze an increase or decrease in the content of extracellular vesicles derived from a specific bacterium, and the like.

Background

The head and neck means a portion from the brain to the upper chest and includes the oral cavity, the larynx, the pharynx, the nasal cavity, and the like, and cancer occurring in these organs is called head and neck cancer. Among them, oral cancer is cancer occurring in the oral cavity, which mainly originates from squamous cells constituting the mucosa around the oral cavity. As risk factors for oral cancer, smoking, oral hygiene, persistent chronic irritation, and the like are known. Pharyngeal cancer is a malignant tumor occurring in the pharyngeal mucosa, and smoking, drinking, viral infection, and the like are known risk factors. According to data published by the korean Central Cancer Registry (Korea Central Cancer Registry), the number of head and neck Cancer patients per year is 3,000, and the occurrence frequency thereof is ranked eighth among cancers, in addition to thyroid Cancer. Diagnosis of head and neck cancer is determined by excising tissue from the tumor for biopsy when cancer is suspected. In addition, in order to determine the extent of cancer, tests such as computed tomography, magnetic resonance imaging examination, and positron emission tomography are performed.

Meanwhile, it is known that the number of symbiotic microorganisms in the human body is 100 trillion, which is 10 times of the number of human cells, and the number of genes of microorganisms exceeds 100 times of the human gene. Microbiota (microbiota) is a microbial community that includes bacteria, archaea and eukaryotes present in a given habitat. It is known that the intestinal microbiota plays a crucial role in human physiological phenomena and has a major impact on human health and disease through interaction with human cells. Bacteria coexisting in the human body secrete nano-scale vesicles to exchange information about genes, proteins, low molecular weight compounds, and the like with other cells. The mucosa forms a physical barrier membrane that does not allow particles having a size of 200nm or more to pass through, and thus bacteria symbiotic in the mucosa cannot pass through, but extracellular vesicles derived from bacteria have a size of about 100nm or less, and thus relatively freely pass through the mucosa and are absorbed into the human body.

Metagenomics (also referred to as environmental genomics) may be an analytics for metagenomic data obtained from samples collected from the environment (korean patent publication No. 2011-073049). Recently, bacterial compositions of human microbiota were listed using a method based on 16s ribosomal RNA (16 srna) base sequences, and 16s rDNA base sequences, which are genes of 16s ribosomal RNA, were analyzed using a Next Generation Sequencing (NGS) platform. However, with respect to the occurrence of head and neck cancer, there is no report on a method of identifying the cause of head and neck cancer from human-derived materials (e.g., saliva) and diagnosing head and neck cancer by analyzing the genome present in vesicles derived from bacteria.

Disclosure of Invention

[ problem ] to

The present inventors extracted genes from bacteria-derived extracellular vesicles present in saliva as a sample derived from a normal individual and a sample derived from a subject, and conducted metagenomic analysis in this regard in order to diagnose the cause and risk of head and neck cancer in advance, and as a result, identified bacteria-derived extracellular vesicles that can be used as a causative agent of head and neck cancer, thereby completing the present invention based on this.

Accordingly, it is an object of the present invention to provide a method for providing information for diagnosing head and neck cancer, a method for predicting the risk of head and neck cancer onset, and the like, by metagenomic analysis of extracellular vesicles derived from bacteria.

However, the technical objects of the present invention are not limited to the above objects, and other technical objects not mentioned will be clearly understood by those of ordinary skill in the art from the following description.

[ solution ]

In order to achieve the above object of the present invention, there is provided a method of providing information for head and neck cancer diagnosis, the method including the processes of:

(a) extracting DNA from extracellular vesicles isolated from a sample from a normal individual and a sample from a subject;

(b) using a nucleic acid comprising SEQ ID NO: 1 and SEQ ID NO: 2, performing Polymerase Chain Reaction (PCR) on the extracted DNA; and

The present invention also provides a method for diagnosing head and neck cancer, comprising the steps of:

(a) extracting DNA from extracellular vesicles isolated from a sample from a normal individual and a sample from a subject;

(b) using a nucleic acid comprising SEQ ID NO: 1 and SEQ ID NO: 2, performing Polymerase Chain Reaction (PCR) on the extracted DNA; and

The present invention also provides a method of predicting the risk of head and neck cancer, the method comprising the steps of:

(a) extracting DNA from extracellular vesicles isolated from a sample from a normal individual and a sample from a subject;

(b) using a nucleic acid comprising SEQ ID NO: 1 and SEQ ID NO: 2, performing Polymerase Chain Reaction (PCR) on the extracted DNA; and

(c) determining an increase or decrease in the content of the bacterial-derived extracellular vesicles in the subject-derived sample, as compared to the content of the bacterial-derived extracellular vesicles in a normal individual-derived sample, by sequencing the products of the PCR.

In one embodiment of the invention, the sample may be saliva.

In another embodiment of the present invention, in the process (c), the head and neck cancer may be diagnosed by comparing and determining an increase or decrease in the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of the order halophiles (halobacilli), the order bifidobacteria (bifidobacteria), the order trichoderma (Streptophyta), the order pseudomonas (pseudomonas), the order marine spirillum (Oceanospirillales), the order clostridia (Fusobacteriales), and the order campylobacter (campylobacter).

In another embodiment of the present invention, in the process (c), the head and neck cancer may be diagnosed by comparing and determining an increase or decrease in the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of pseudomonas (pseudomonas adaceae), Halobacteriaceae (Halobacteriaceae), oxalobacteriaceae (oxalobobacteriaceae), Halomonadaceae (haloonadaceae), comamonoconadaceae (comamoonadaceae), lactobacillaceae (L actacilaceae), paranovotellaceae (paranovolaceae), clostridiaceae (Fusobacteriaceae), and campylobacter (campylobacter sphaeraceae).

In another embodiment of the present invention, in the process (c), the head and neck cancer may be diagnosed by comparing and determining an increase or decrease in the content of extracellular vesicles derived from one or more bacteria selected from the group consisting of cupriasis (Cupriavidus), Chromohalobacter (Chromohalobacter), Pseudomonas (Pseudomonas), Acinetobacter (Acinetobacter), aquaticus (aquabacter), lactobacillus (L acetobacter), Veillonella (Veillonella), clostridium (Fusobacterium), Actinomyces (Actinomyces), Prevotella (Prevotella), Megasphaera (Megasphaera), Campylobacter (Campylobacter), and Oribacterium (oribacter).

In one embodiment of the present invention, process (c) may comprise comparing and determining:

In another embodiment of the present invention, in the process (c), the increase in the content of extracellular vesicles as follows may be diagnosed as head and neck cancer, as compared to a sample derived from a normal individual:

extracellular vesicles from bacteria of the Fusobacteria (fusobacterobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of clostridia (fusobacteriaceia) and proteobacteria (epsilon proteobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of Fusobacteriales (Fusobacteriales) and campylobacter (campylobacter);

In another embodiment of the present invention, in the process (c), the following decrease in the content of extracellular vesicles can be diagnosed as head and neck cancer, as compared with a sample derived from a normal individual:

extracellular vesicles derived from bacteria of the phylum Cyanobacteria (Cyanobacteria);

extracellular vesicles derived from one or more bacteria selected from the group consisting of halobacteriaceae (halobacteriia) and chloroplasts (Chloroplast);

[ advantageous effects ]

Extracellular vesicles secreted by bacteria present in the environment are absorbed by the human body, and thus may directly affect the occurrence of cancer, and it is difficult to diagnose head and neck cancer early before the onset of symptoms, and thus it is difficult to effectively treat it. Therefore, according to the present invention, the etiology and risk of head and neck cancer can be diagnosed by metagenomic analysis of extracellular vesicles derived from bacteria using a human-derived sample, and thus a population at risk of head and neck cancer can be diagnosed early, thereby delaying the onset of head and neck cancer or preventing the onset of head and neck cancer through appropriate management. In addition, head and neck cancer can be diagnosed early even after the occurrence of head and neck cancer, thereby reducing the incidence rate of head and neck cancer and improving the therapeutic effect, and metagenomic analysis enables patients diagnosed with head and neck cancer to avoid exposure to the etiology thus predicted, so that the progression of cancer can be improved, or the recurrence of head and neck cancer can be prevented.

Drawings

Fig. 1A illustrates an image showing a distribution pattern of bacteria and Extracellular Vesicles (EV) over time after oral administration of intestinal bacteria and Extracellular Vesicles (EV) derived from bacteria to mice, and fig. 1B illustrates an image showing a distribution pattern of bacteria and EV after oral administration of mice, and at 12 hours, saliva and various organs were extracted.

Fig. 2 is a result showing the distribution of Extracellular Vesicles (EV) derived from bacteria, which is significant in terms of diagnostic performance on the gate level, by isolating vesicles derived from bacteria from saliva of head neck cancer patients and normal individuals and then performing metagenomic analysis.

Fig. 3 is a result showing the distribution of Extracellular Vesicles (EV) derived from bacteria, which is significant in diagnostic performance on a class level, by isolating vesicles derived from bacteria from saliva of cervical cancer patients and normal individuals and then performing metagenomic analysis.

Fig. 4 is a result showing the distribution of Extracellular Vesicles (EV) derived from bacteria, which is significant in terms of the diagnostic performance on the eye level, by isolating vesicles derived from bacteria from saliva of head neck cancer patients and normal individuals and then performing metagenomic analysis.

Fig. 5 is a result showing the distribution of Extracellular Vesicles (EV) derived from bacteria, which is significant in terms of diagnostic performance at a scientific level, by isolating vesicles derived from bacteria from saliva of cervical cancer patients and normal individuals and then performing metagenomic analysis.

Fig. 6 is a result showing the distribution of Extracellular Vesicles (EV) derived from bacteria, which is significant in diagnostic performance on a genus level, by isolating vesicles derived from bacteria from saliva of head neck cancer patients and normal individuals and then performing metagenomic analysis.

Detailed Description

The present invention relates to a method for diagnosing head and neck cancer by metagenomic analysis of bacteria. The inventors of the present invention extracted genes from the extracellular vesicles derived from bacteria using samples of normal individuals and samples derived from subjects, performed metagenomic analysis thereof, and identified the extracellular vesicles derived from bacteria that can serve as causative agents of head and neck cancer.

Accordingly, the present invention provides a method of providing information for diagnosing head and neck cancer, the method comprising:

(a) extracting DNA from extracellular vesicles isolated from a sample of a normal individual and a sample of a subject;

(b) using a nucleic acid comprising SEQ ID NO: 1 and SEQ ID NO: 2, performing Polymerase Chain Reaction (PCR) on the extracted DNA; and

As used herein, the term "head and neck cancer" refers to cancer occurring in organs such as the oral cavity, larynx, pharynx, nasal cavity, and the like, and includes oral cancer, salivary gland cancer, pharynx cancer, and nasal cavity cancer.

As used herein, the term "head and neck cancer diagnosis" refers to determining whether a patient is at risk for head and neck cancer, whether the risk for head and neck cancer is relatively high, or whether head and neck cancer has already occurred. The method of the present invention can be used to delay head and neck cancer onset (which is a patient with a high risk of head and neck cancer) or to prevent head and neck cancer onset by specific and appropriate care for the particular patient. In addition, the method can also be used clinically to determine treatment by selecting the most appropriate treatment method through early diagnosis of head and neck cancer.

As used herein, the term "metagenome" refers to the entire genome of all viruses, bacteria, fungi, etc., included in an isolated region such as soil, animal intestines, etc., and is mainly used as a concept of genome, which explains the identification of many microorganisms at a time using a sequencer to analyze non-cultured microorganisms. In particular, metagenome does not refer to the genome of one species, but to a mixture of genomes, including the genomes of all species of an environmental unit. This term stems from the idea that: when a species is defined in the course of biological evolution into omics (omics), various species and an existing one functionally interact to form a complete species. Technically, it is the subject of a technique that analyzes all DNA and RNA, regardless of species, using rapid sequencing to identify all species in one environment and to verify interactions and metabolism. In the present invention, bacterial metagenomic analysis is performed using extracellular vesicles derived from bacteria isolated from, for example, serum.

In the present invention, the sample of the normal individual and the sample of the subject may be saliva, but the present invention is not limited thereto.

In one embodiment of the present invention, metagenomic analysis is performed on extracellular vesicles derived from bacteria, and in fact, by analysis at the phylum, class, mesh, family and genus levels, extracellular vesicles derived from bacteria are identified that can serve as causes of head and neck cancer episodes.

More specifically, in one embodiment of the present invention, the content of extracellular vesicles derived from bacteria belonging to the phyla Cyanobacteria (cyanobacter) and Fusobacteria (fusobacterobacteria) significantly differs between head and neck cancer patients and normal individuals as a result of bacterial metagenomic analysis at the phylum level of extracellular vesicles present in saliva samples derived from subjects (see example 4).

More specifically, in one embodiment of the present invention, the content of extracellular vesicles derived from bacteria belonging to the classes halobacteriaceae (halobacteriaceae), chloroplasts (chloroplasts), fusobacteriaceae (fusobacteria) and proteobacteria (epislonobacteria) differs significantly between patients with head and neck cancer and normal individuals as a result of bacterial metagenomic analysis of extracellular vesicles present in saliva samples derived from subjects at class level (see example 4).

More specifically, in one embodiment of the present invention, the content of extracellular vesicles derived from bacteria belonging to the order of halophiles (halobacilli), bifidobacteria (bifidobacteria), trichoderma (Streptophyta), pseudomonas (pseudomonales), oceanic spirobacteriales (Oceanospirillales), clostridia (Fusobacteriales) and campylobacter (campylobacter) significantly differs between head and neck cancer patients and normal individuals as a result of bacterial metagenomic analysis of extracellular vesicles present in saliva samples derived from subjects at the mesh level (see example 4).

More specifically, in one embodiment of the present invention, the content of extracellular vesicles derived from bacteria belonging to the Pseudomonadaceae (pseudomonas adaceae), Halobacteriaceae (Halobacteriaceae), oxalobacteriaceae (oxalobobacteriaceae), Halomonadaceae (haloonadaceae), comamodiaceae (comamonoaceae), lactobacillaceae (L actobacteriaceae), prasudaceae (paranovotaleae), clostridiaceae (Fusobacteriaceae), and campylobacter (campylobacter) significantly differs between head and neck cancer patients and normal individuals as a result of bacterial metagenomic analysis of extracellular vesicles present in saliva samples derived from subjects at the scientific level (see example 4).

More specifically, in one embodiment of the present invention, the content of extracellular vesicles derived from bacteria belonging to the genera cuprianidus (cupriavirdus), Chromohalobacter (Chromohalobacter), Pseudomonas (Pseudomonas), Acinetobacter (Acinetobacter), aquaticum (enterobacter), lactobacillus (L acetobacter), Veillonella (Veillonella), clostridium (Fusobacterium), Actinomyces (Actinomyces), Prevotella (Prevotella), Megasphaera (Megasphaera), Campylobacter (Campylobacter), and Oribacterium (Oribacterium) significantly differs between cancer patients and normal individuals as a result of bacterial metagenomic analysis of extracellular vesicles present in saliva samples derived from subjects at the genus level (see example 4).

By way of example, it is demonstrated that the distribution variables of the identified bacterial-derived extracellular vesicles can be used to predict the onset of head and neck cancer.

[ modes for carrying out the invention ]

Hereinafter, the present invention will be described with reference to exemplary embodiments to assist understanding of the present invention. However, these examples are provided for illustrative purposes only, and are not intended to limit the scope of the present invention.

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Method for diagnosing head and neck cancer by bacterial metagenomic analysis

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