CN107058560A - Self-closing disease biomarker and its detection kit, application - Google Patents
Self-closing disease biomarker and its detection kit, application Download PDFInfo
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- CN107058560A CN107058560A CN201710341199.8A CN201710341199A CN107058560A CN 107058560 A CN107058560 A CN 107058560A CN 201710341199 A CN201710341199 A CN 201710341199A CN 107058560 A CN107058560 A CN 107058560A
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Classifications
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
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Abstract
The invention provides self-closing disease biomarker, including the grand genome signature of enteron aisle, it is embodied in the level of acidfast bacilli door etc., thus the diagnosis of self-closing disease will judge not against subjective experience, effective early warning not only can be provided to the morbidity of self-closing disease, and it can be made a definite diagnosis, greatly improve accuracy rate of diagnosis；Present invention also offers the composition of the kit for diagnosing self-closing disease, and prevention or treatment self-closing disease；Invention further provides self-closing disease biomarker, self-closing disease diagnostic kit and prevention or the application for the composition for treating self-closing disease；The system of the biological sample of self-closing disease is susceptible to suffer from invention still further provides screening, by this screening system, the biological sample for being susceptible to suffer from self-closing disease can be screened.
Description
Technical field
The invention belongs to biomedicine field, in particular it relates to self-closing disease biomarker and its detection reagent
Box, application.
Background technology
Self-closing disease (ASD, Autism Spectrum Disorders) or autism or autism-spectrum obstacle, be
A kind of hypotype of pervasive developmental disorders, common with male, onset is mainly shown as different degrees of speech in infantile period
Maldevelopment, Social disorder, interest are narrow and behavior is mechanical, there are about 3/4 patient with obvious Mental development
It is sluggish.According to U.S.'s diseases monitoring and prevention center (CDC) data, its incidence of disease 1/45 (i.e. 2.24%) in 2015.China still lacks
Weary national epidemiological survey data, if with 1/100 estimation, estimating China ASDs patient will be more than 10,000,000.
Self-closing disease (ASD) has a strong impact on the social function of infant as a kind of children's spirit disease, to infant family and society
It can bring heavy burden, but self-closing disease is estimated generally according to scale at present, no blood diagnosis method and effectively control
Treatment scheme, so diagnosis will judge by subjective experience, not only can not provide effective early warning to the morbidity of self-closing disease, and examine
Disconnected accuracy rate is very low.Therefore study self-closing disease pathogenesis and the biomarker of its order of severity, and and hour hands can be embodied
It is to treat disease, the key factor of mitigation burden on society that the cause of disease, which intervene and treat,.
The content of the invention
The technical problem to be solved in the present invention is to provide self-closing disease biomarker；Present invention also offers for examining
The kit of disconnected self-closing disease；The present invention further also provides a kind of system for the biological sample for screening and being susceptible to suffer from self-closing disease.
First aspect present invention provides self-closing disease biomarker, including the grand genome signature of enteron aisle, is embodied in
The level of acidfast bacilli door.
In a preference, the grand genome signature of enteron aisle be also embodied by actinomyces door, Chlamydia door, conveyor screw door,
β-deformation Gammaproteobacteria, Chlamydozoa, Dehalococcoidetes guiding principles, conveyor screw guiding principle, sour Thiobacillus mesh, Bifidobacterium mesh, clothing are former
Body mesh, coloring Zoopagales, Dehalococcoidales mesh, worm substance mesh, Erysipelothrix mesh, salt anaerobism Zoopagales, Nai Shi coccuses mesh,
Spirochaetale, Synergistales mesh, Thermoanaerobacterales mesh, amino acid Coccaceae, the sour shaft-like Cordycepps of sulphur,
Alkane eating bacteria section, bifidobacterium family, Brachyspira section, Cellulomonas section, Dehalococcoidaceae sections, the red spiral shell of outer sulphur
Revolve Cordycepps, worm Spiroplasmataceae, red Bacteriaceae, Gordonia bronchialis section, river Salmonella section, salt anaerobism Cordycepps, legion Cordycepps, methane grain Cordycepps,
Methanospirillum section, Methylobacterium section, Neisseriaceae, class Bacillaceae, clay Bacteriaceae, Peptococcaccae, core card body
Section, Spirochaetaceae, staphylococcaceae, syntrophism Cordycepps, syntrophism unit cell Cordycepps, acetobacter, Acidaminococcus, bifid bar
Pseudomonas, Brachyspira, Brevibacillus, pyrolysis cellulose Pseudomonas, Citrobacter, clo promise Pseudomonas, desulfiting bacterium
Category, desulfurization gemma Campylobacter, Desulfotomaculum, the new Pseudomonas of production hydrogen, big Faingold Pseudomonas, salt anaerobism Pseudomonas, cilium bacterium
Category, Salmonella, Sphaerochaeta category, syntrophism zygosaccharomyces, Tepidanaerobacter category, Tetracoccus,
Thermincola category, Treponema, Acaryochloris marina MBIC11017, acetobacter woodii DSM
1030th, Acidaminococcus fermentans, Acidaminococcus fermentans DSM 20731, Acidaminococcus
Intestini, Acinetobacter baumannii ATCC 17978, Advenella kashmirensis WT001,
Alcanivorax borkumensis SK2、Alicycliphilus denitrificans、Aliivibrio
unclassified、 Alkalilimnicola ehrlichii MLHE-1、Alkaliphilus oremlandii
OhILAs、Allochromatium vinosum DSM 180、Anaeromyxobacter sp.Fw 109-5、
Archaeoglobus veneficus SNP6, Arthrobacter subspecies FB24, Azoarcus sp.BH72, Azospirillum
Unclassified, Bacillaceae unclassified, Dipel, Bartonella tribocorum, bifid bar
Bacterium animalis, Bifidobacterium Bifidobacterium animalis subsp.Lactis, Bordetella bird tuberculosis
197N、 Borrelia burgdorferi group、Brachyspira murdochii DSM 12563、Brachyspira
pilosicoli 95/1000、Buchnera aphidicola、Burkholderia phymatum STM815、
Burkholderia pseudomallei 1710b、Burkholderia sp.CCGE1002、Burkholderia sp.
CCGE1003, Burkholderia xenovorans LB400, Caldicellulosiruptor, campylobacter hominis
ATCC BAA-381, campylobacter jejuni subsp.jejuni ICDCCJ07001, Candidatus
Accumulibacter phosphatis clade IIA str.UW-1、Candidatus Chloracidobacterium
thermophilum B、Candidatus Protochlamydia amoebophila UWE25、Cellulosilyticum
Lentocellum, Chlamydophila pneumoniae J138, Chlamydophila unclassified, fusobacterium
Genomosp.BVAB3, clostruidium botulinum Ba4str.657, clostruidium cellulolyticum,
Clostruidium cellulovorans 743B, clostruidium clariflavum DSM 19732, clostruidium
Kluyveri, clostruidium sp.BNL1100, clostruidium sticklandii DSM 519, clostruidium
It is tetani E88, clostruidium thermocellum, clostruidium thermocellum ATCC 27405, bar-shaped
Bacillus section unclassified, corynebacterium efficiens YS-314, Cupriavidus metallidurans
CH34、Dechlorosoma suillum PS、 Desulfitobacterium unclassified、
Desulfosporosinus acidiphilus SJ4、 Desulfosporosinus orientis DSM 765、
Desulfotomaculum acetoxidans DSM 771、 Desulfotomaculum reducens MI-1、
Eggerthella sp.YY7918、Eggerthella unclassified、 Erythrobacter litoralis
HTCC2594、Fibrobacter succinogenes subsp.succinogenes S85、Finegoldia magna
ATCC 29328, Gallionellaceae, Gardnerella vaginalis ATCC 14019, Gluconobacter oxydans
621H, Gordonia polyisoprenivorans, Hahella chejuensis KCTC 2396, helicobacter pylorus Pseudomonas
Cinaedi PAGU611, helicobacter pylorus Pseudomonas felis ATCC 49179, helicobacter pylorus Pseudomonas pylori F30, helicobacter pylorus
Pseudomonas pylori Puno120, Ignisphaera aggregans DSM 17230, Kangiella koreensis DSM
16069th, Kitasatospora setae KM-6054, genus lactubacillus johnsonii DPC 6026, Laribacter
Hongkongensis HLHK9, Legionnella longbeachae NSW150, Marinobacter adhaerens HP15,
Methanocorpusculum labreanum Z、Methanospirillum hungatei JF-1、
Methylobacterium nodulans ORS 2060、Methylotenera mobilis、Mobiluncus curtisii
ATCC 43063, Muricauda ruestringensis, Mycobacterium gilvum, Mycobacterium smegmatis
Str.MC2 155, mycoplasma bovis, mycoplasma bovis PG45, mycoplasma putrefaciens KS1, eisseria
Unclassified, Neisseriaceae unclassified, Neorickettsia sennetsu str.Miyayama, nitrous
Change Coccus halophilus Nc4, Nitrosomonadaceae unclassified, Nocardiaceae
unclassified、 Oceanobacillus iheyensis HTE831、Peptococcaceae unclassified、
Peptostreptococcaceae, Planctomyces limnophilus DSM 3776, Polaromonas sp. JS666,
Polynucleobacter necessarius, proteus mirabilis HI4320, Pseudogulbenkiania
Sp.NH8B, Pseudoxanthomonas spadix BD-a59, Ralstonia solanacearum PSI07, Rhod
opacus B4、Rhodospirillum photometricum DSM 122、Rhodothermus marinus
SG0.5JP17-172、Riemerella anatipestifer RA-GD、Saccharopolyspora erythraea NRRL
2338th, Selenomonas ruminantium subsp. Lactilytica, Shewanella denitrificans
OS217, Sphaerochaeta globus str.Buddy, Sphingomonas wittichii RW1, conveyor screw
Thermophila DSM 6192, staphylococcus aureus aureus subsp.aureus NCTC 8325,
Stenotrophomonas maltophilia, Streptococcaceae unclassified, streptococcus agalactiae A909, chain
Coccus suis ST3, streptomyces cattleya NRRL 8057=DSM 46488, streptomyces scabiei 87.22,
Syntrophobacter fumaroxidans MPOB、Tepidanaerobacter acetatoxydans Re1、
Thermoanaerobacterium、 Thermoanaerobacterium saccharolyticum、
It is Thermoanaerobacterium saccharolyticum JW/SL-YS485, Thermotoga lettingae TMO, close
Spirochaeta brennaborense DSM 12168, Treponema succinifaciens, Treponema
Succinifaciens DSM 2489, Treponema unclassified, vibrios unclassified, Vibrio vulnificus
One or more levels in CMCP6 and Xanthomonas oryzae pv.oryzae MAFF 311018.
Second aspect of the present invention, which is provided, detects that the agent combination of above-mentioned biomarker is self-closing for diagnosing in preparation
Purposes in the kit of disease.
Third aspect present invention provides a kind of self-closing disease diagnostic kit, including is marked suitable for the biology described in detection
The agent combination of will thing.
In a preference, the agent combination of the biomarker suitable for described in test right requirement 1 includes north
Jing Kang is the model CW2092S in century StoolGen faeces DNA extracts kits.
In a preference, the agent combination of the biomarker suitable for described in test right requirement 1 also includes
The kit and the Truseq DNA of Illumina companies of U.S. production that match Mo Feishier companies of the U.S. provide build storehouse kit.
In a preference, in addition to the analysis biomarker in vivo whether abnormal instrument.
Fourth aspect present invention provides described biomarker or described kit and is susceptible to suffer from self-closing disease in screening
Application in the system of biological sample.
Fifth aspect present invention provides Vibrio vulnificus CMCP6 and is preparing treatment or auxiliary treatment self-closing disease or its correlation
Purposes on the product of disease.
In a preference, described product is medicine or food.
Sixth aspect present invention provides a kind of composition, including Vibrio vulnificus CMCP6.
In a preference, the composition also includes Acaryochloris marina MBIC11017, Advenella
kashmirensis WT001、Alcanivorax borkumensis SK2、Alicycliphilus denitrificans、
Aliivibrio unclassified、Alkalilimnicola ehrlichii MLHE-1、 Allochromatium
Vinosum DSM 180, Anaeromyxobacter sp.Fw 109-5, Archaeoglobus veneficus SNP6, section
Bacterium belongs to subspecies FB24, Azoarcus sp.BH72, Azospirillum unclassified, Bartonella
Tribocorum, Bordetella bird tuberculosis 197N, Borrelia burgdorferi group, Buchnera
aphidicola、Burkholderia phymatum STM815、 Burkholderia pseudomallei 1710b、
Burkholderia sp.CCGE1002、Burkholderia sp. CCGE1003、Burkholderia xenovorans
LB400, campylobacter jejuni subsp.jejuni ICDCCJ07001, Candidatus Accumulibacter
phosphatis clade IIA str.UW-1、 Candidatus Chloracidobacterium thermophilum B、
Candidatus Protochlamydia amoebophila UWE25、Chlamydophila pneumoniae J138、
Chlamydophila unclassified, corynebacterium efficiens YS-314, Cupriavidus
metallidurans CH34、 Dechlorosoma suillum PS、Erythrobacter litoralis HTCC2594、
Gallionellaceae, Gluconobacter oxydans 621H, Gordonia polyisoprenivorans, Hahella
Chejuensis KCTC 2396, helicobacter pylorus Pseudomonas felis ATCC 49179, helicobacter pylorus Pseudomonas pylori F30, pylorus
Helicobacterium pylori Puno120, Ignisphaera aggregans DSM 17230, Kangiella koreensis
DSM 16069, Kitasatospora setae KM-6054, Laribacter hongkongensis HLHK9, Legionella
Belong to longbeachae NSW150, Marinobacter adhaerens HP15, ethanospirillum hungatei JF-
1st, Methylobacterium nodulans ORS 2060, Methylotenera mobilis, Mycobacterium gilvum,
Mycobacterium smegmatis str.MC2 155, eisseria unclassified, Neisseriaceae
Unclassified, Neorickettsia sennetsu str.Miyayama, Nitrosococcus halophilus Nc4,
Nitrosomonadaceae unclassified, Nocardiaceae unclassified, Planctomyces limnophilus
DSM 3776, Polaromonas sp.JS666, Polynucleobacter necessaries, proteus mirabilis
HI4320、Pseudogulbenkiania sp.NH8B、Pseudoxanthomonas spadix BD-a59、Ralstonia
Solanacearum PSI07, Rhod opacus B4, Rhodospirillum photometricum DSM 122,
Rhodothermus marinus SG0.5JP17-172、 Saccharopolyspora erythraea NRRL 2338、
Shewanella denitrificans OS217, Sphingomonas wittichii RW1, conveyor screw thermophila
DSM 6192, staphylococcus aureus aureus subsp.aureus NCTC 8325, Stenotrophomonas
Maltophilia, Streptococcaceae unclassified, streptomyces cattleya NRRL 8057=DSM 46488, streptomycete
Belong to scabiei 87.22, Syntrophobacter fumaroxidans MPOB, Thermotoga lettingae TMO, arc
One or more in bacterium unclassified and Xanthomonas oryzae pv.oryzae MAFF 311018；
In a preference, said composition also includes acceptable carrier or pharmaceutically acceptable carrier on food.
Seventh aspect present invention provides combinations of the above thing and is preparing treatment or auxiliary treatment self-closing disease or its related disease
Purposes on the product of disease.
In a preference, described product is medicine or food.
Eighth aspect present invention provides a kind of system for the biological sample for screening and being susceptible to suffer from self-closing disease, including：
Nucleic acid-extracting apparatus, the nucleic acid-extracting apparatus is used to extract nucleic acid from the biological sample and control sample
Sample；
Nucleotide sequence determining device, the nucleotide sequence determining device is connected with the nucleic acid-extracting apparatus, for institute
State sample of nucleic acid to be analyzed, to determine the sequence of the sample of nucleic acid；And
Judgment means, the judgment means are connected with the sequencing device, and the sequence pair based on the sample of nucleic acid is biological
The self-closing disease biomarker of claim 1 in sample and control sample contained by gut flora is analyzed, and is tied according to analysis
Fruit judges whether the biological sample is susceptible to suffer from self-closing disease.
In a preference, the nucleotide sequence determining device further comprises：
Library construction unit, the library construction unit is used to be based on the sample of nucleic acid, builds the text of the nucleic acid
Storehouse；And
Unit is sequenced, the sequencing unit is connected with the library construction unit, the library is sequenced, so as to
The sequence of the sample of nucleic acid is determined,
In a preference, the library construction unit further comprises：
PCR is expanded in module, the PCR amplifications module and is set specific primer, to utilize the specific primer,
The sample of nucleic acid is expanded.
Herein, term " biomarker " should be interpreted broadly, it include it is any can reflect abnormality appoint
What detectable Biological indicators, can include gene marker, species mark (planting mark/category mark) and function mark
Will thing (KO/OG marks), it has purposes widely, available for medical diagnosis on disease, judges staging or for commenting
The safety and efficacy of valency new drug or new treatment in target group.
The invention has the advantages that：
Present invention firstly discovers that causing a disease for self-closing disease is relevant with acidfast bacilli door etc., therefore by acidfast bacilli in enteron aisle etc.
As the detection mark of self-closing disease, the diagnosis of such self-closing disease will judge not against subjective experience, not only can be to self-closing disease
Morbidity effective early warning is provided, and it can be made a definite diagnosis, overcome the diagnosis of self-closing disease largely dependent on subjectivity
The defect of micro-judgment, greatly improves accuracy rate of diagnosis.
Embodiment
Unless specifically indicated, term used herein has the general sense in art of the present invention.
Below with reference to specific embodiment, the present invention will be described, it is necessary to illustrate, these embodiments are only
Bright property, and be not considered as limiting the invention.Unreceipted particular technique or condition in embodiment, according to routine
Experiment condition, such as Sambrook equimoleculars Cloning: A Laboratory Manual (Sambrook J＆Russell DW, Molecular
cloning：A laboratory manual, 2001), or according to the condition of manufacturer's specification suggestion.Agents useful for same or
The unreceipted production firm person of instrument, being can be by the conventional products of acquisition purchased in market.
Embodiment 1
The present embodiment tests and analyzes healthy population and the excrement sample of self-closing disease patient with the grand gene order-checking of shotgun
This, then carries out statistical check comparison, to determine the biomarker related to self-closing disease patient population.Comprise the following steps that：
1. sample source
Self-closing disease group：From Shenzhen Children's Hospital's Psychology Dept. and speech therapy section outpatient service self-closing disease infant, acquisition time
For 7~December in 2015.Inclusive criteria：1. according to international newest diagnostic criteria, i.e.,《Americanism obstacle diagnosis and statistics hand
Volume the 5th edition》(Diagnostic and Statistical Manual of Mental Disorders, 5th Edition,
DSM-5), be diagnosed as ASD " needs very support more " infant；2. age ＜ 14 years old；3. male or female.Exclusion standard：1. suffer from
There are other mental illnesses (such as schizophrenia)；2. other neurodevelopmental disorder diseases are suffered from；3. hereditary metabolic disorders disease is suffered from
Disease；4. the great physical disease history such as serious neurological disease and craniocerebral injury history is suffered from.
Control group：From children's health care section of Shenzhen Children's Hospital physical examination children.Inclusive criteria：1. be a cup too low disease, and body is good for
Health；2. organize the age with patient and sex is matched.Exclusion standard is with patient's group.It is all enter group Parents to this research know simultaneously
Sign informed consent form.
This research is ratified through Ethics Committee of Shenzhen Children's Hospital.
This research was divided to for two phases：Discovery phase and Qualify Phase.
The first phase is discovery phase, and 22 self-closing disease infants, man 18, female 4 are collected altogether；2~6 years old age, average (2
± 1) year.15 control children, man 8, female 7 are collected altogether；1~6 years old age, average (2 ± 1) year.Two groups of sexes
(Fisher is accurately examined, P ＞ 0.05), age (Student ' s T test, P ＞ 0.05) no significant difference.
The second phase is Qualify Phase, collects 22 self-closing disease infants, man 18, female 3；2~6 years old age, average (2 ±
1) year.16 control children, man 9, female 7 are collected altogether；1~6 years old age, average (2 ± 1) year.Two groups of sex (Fisher
Accurate to examine, P ＞ 0.05), age (Student ' s T test, P ＞ 0.05) no significant difference.
2. feces collection is handled with early stage
Certainly be about to excrement comes the feces collection basin specified to participant, is ready for disposable medical excrement sampling pipe, wears
Gloves, gather excrement, and collection is interrupted, it is to avoid the pollution caused by carelessness, once finding pollution, answer resampling, collection is completed
Quick freeze is stored in -80 DEG C of refrigerators afterwards.Relevant rudimentary information is collected, including：Name, sex, age, height, body weight, trouble
Medical history, home address, father and mother's contact method etc. are full and accurate《Health condition information table》；《Diet and habits and customs application form》Need
The data that emphasis is collected includes eating habit, if breast-feeding, father and mother's habits and customs, health status and smoking history, institute of family
Locate atmosphere, recent antibiotic, Yoghourt, probiotics uses.
After stool sample has been collected, Beijing health is used to be carried for the model CW2092S in century StoolGen faeces DNAs
Kit is taken to carry out the extraction of genomic DNA according to its specification.
3. grand gene sets up storehouse and sequencing
The DNA sample for taking 2ul to extract is quantified, and kit used matches what Mo Feishier companies provided for the U.S.
Kit (Qubit dsDNA BR assay kit).The Truseq DNA produced using Illumina companies of the U.S. build storehouse reagent
Box carries out building storehouse, and library inserts size is 200-500bp, and the kit is connected to the life of Beckman Coulter Inc. of the U.S.
On the automation SPRI work stations of production.After the completion of each library construction, produced using Agilent Technologies of the U.S.
The biological analysers of Agilent 2100 carry out Quality Control (QC, quality control).After QC, i.e. library concentration need to be
More than 3ng/ μ l, the library is sequenced for the later stage, and sequencing platform used is the microarray datasets of Illumina HiSeq 2500, is surveyed
Sequential mode is sequenced for 150bp both-ends.Each sample measures more than 1G data.
4. analysis of biological information and statistics
First, some sequences of poor quality or containing joint are filtered out.Secondly, the sequence of all measure compares the mankind
Genome sequence (referring to Hg19) is to reject artificially-contaminated sequence.Again, using MEGAN softwares (i.e. MEGAN5)
(http://ab.inf.uni-tuebingen.de/software/megan5/ is downloaded) v5 default parameters, it is grand to shotgun
Gene order-checking data carry out classification of flora (Taxonomic) and function (functional) profile analyses, obtain
Taxonomic profile results arrive boundary (super kingdom) door (phylum) guiding principle for annotation in each sample
(Class) mesh (order) section (family) category (genus) plants the bacteria abundance of (species).
The horizontal bacterium relative abundance of different classifications is calculated, then, it is contemplated that grand gene data meets negative binomial analysis, adopts
With based on negative binomial distribution statistical inspection model DESeq2 (DESeq2 use refers to document Love MI, Huber W,
Anders S.Moderated estimation of fold change and dispersion for RNA-seq data
With DESeq2 [J] .Genome Biology, 2014,15 (12)：550) it is, variant in disease group and control group to verify
Whether the bacterium of categorization levels is in disease and control group group significant difference.
This research is divided to two phases, i.e. discovery phase and Qualify Phase, and the standard of screening significant difference bacterium is to find and testing
Card stage, the P values of DESeq2 verifications are respectively less than 0.05,
As a result：
Research discovery has 4 doors, 4 guiding principles, 12 mesh, 26 sections, 22 category, and the bacterium of 143 kinds is in self-closing disease group
There were significant differences with control group, specific as shown in table 1.
The bacterium of what control group significant difference of the self-closing disease patient of table 1
Note：Discovery phase and the equal significant difference of Qualify Phase.P values ＜ 0.05, DESeq2 are examined.
As shown in Table 1, self-closing disease patient difference of gut flora compared with control group is more, and the bacterium of these differences can
With the mark as self-closing disease patient.
Claims (8)
1. self-closing disease biomarker, it is characterised in that including the grand genome signature of enteron aisle, be embodied in the water of acidfast bacilli door
It is flat；
It is optional, the grand genome signature of enteron aisle be also embodied by actinomyces door, Chlamydia door, conveyor screw door, β-deformation Gammaproteobacteria,
Chlamydozoa, Dehalococcoidetes guiding principles, conveyor screw guiding principle, sour Thiobacillus mesh, Bifidobacterium mesh, Chlamydiales, chomophoric bacterium
Mesh, Dehalococcoidales mesh, worm substance mesh, Erysipelothrix mesh, salt anaerobism Zoopagales, Nai Shi coccuses mesh, spirochaetale,
Synergistales mesh, Thermoanaerobacterales mesh, amino acid Coccaceae, the sour shaft-like Cordycepps of sulphur, alkane eating bacteria section,
Bifidobacterium family, Brachyspira section, Cellulomonas section, Dehalococcoidaceae sections, the red spiral Cordycepps of outer sulphur, worm are former
Body section, red Bacteriaceae, Gordonia bronchialis section, river Salmonella section, salt anaerobism Cordycepps, legion Cordycepps, methane grain Cordycepps, methanospirillum section,
Methylobacterium section, Neisseriaceae, class Bacillaceae, clay Bacteriaceae, Peptococcaccae, core card body section, Spirochaetaceae, Portugal
It is grape Coccaceae, syntrophism Cordycepps, syntrophism unit cell Cordycepps, acetobacter, Acidaminococcus, Bifidobacterium, Brachyspira, short
Bacillus, pyrolysis cellulose Pseudomonas, Citrobacter, clo promise Pseudomonas, Desulfitobacterium, desulfurization gemma Campylobacter spp
Category, Desulfotomaculum, production hydrogen new Pseudomonas, big Faingold Pseudomonas, salt anaerobism Pseudomonas, Leptothrix, Salmonella,
Sphaerochaeta category, syntrophism zygosaccharomyces, Tepidanaerobacter category, Tetracoccus, Thermincola category, it is close
Spirochaeta, Acaryochloris marina MBIC 11017, acetobacter woodii DSM 1030, amino acid coccus
Category fermentans, Acidaminococcus fermentans DSM 20731, Acidaminococcus intestini, Bao Man are motionless
Bacillus baumannii ATCC 17978, Advenella kashmirensis WT001, Alcanivorax
borkumensis SK2、Alicycliphilus denitrificans、Aliivibrio unclassified、
Alkalilimnicola ehrlichii MLHE-1、Alkaliphilus oremlandii OhILAs、
Allochromatium vinosum DSM 180、Anaeromyxobacter sp.Fw109-5、Archaeoglobus
Veneficus SNP6, Arthrobacter subspecies FB24, Azoarcus sp.BH72, Azospirillum unclassified, bud
Born of the same parents' Bacteriaceae unclassified, Dipel, Bartonella tribocorum, Bifidobacterium animalis, bifid
Bacillus Bifidobacterium animalis subsp.Lactis, Bordetella bird tuberculosis 197N, Borrelia
burgdorferi group、Brachyspira murdochii DSM 12563、Brachyspira pilosicoli 95/
1000、Buchnera aphidicola、Burkholderia phymatum STM815、Burkholderia
pseudomallei 1710b、Burkholderia sp.CCGE1002、Burkholderia sp.CCGE1003、
Burkholderia xenovorans LB400, Caldicellulosiruptor, campylobacter hominis ATCC BAA-
381st, campylobacter jejuni subsp.jejuni ICDCCJ07001, Candidatus Accumulibacter
phosphatis clade IIA str.UW-1、Candidatus Chloracidobacterium thermophilum B、
Candidatus Protochlamydia amoebophila UWE25、Cellulosilyticum lentocellum、
Chlamydophila pneumoniae J138, Chlamydophila unclassified, fusobacterium
Genomosp.BVAB3, clostruidium botulinum Ba4 str.657, clostruidium cellulolyticum,
Clostruidium cellulovorans 743B, clostruidium clariflavum DSM 19732, clostruidium
Kluyveri, clostruidium sp.BNL1100, clostruidium sticklandii DSM 519, clostruidium
It is tetani E88, clostruidium thermocellum, clostruidium thermocellum ATCC 27405, bar-shaped
Bacillus section unclassified, corynebacterium efficiens YS-314, Cupriavidus metallidurans
CH34、Dechlorosoma suillum PS、Desulfitobacterium unclassified、
Desulfosporosinus acidiphilus SJ4、Desulfosporosinus orientis DSM 765、
Desulfotomaculum acetoxidans DSM 771、Desulfotomaculum reducens MI-1、
Eggerthella sp.YY7918、Eggerthella unclassified、Erythrobacter litoralis
HTCC2594、Fibrobacter succinogenes subsp.succinogenes S85、Finegoldia magna
ATCC 29328, Gallionellaceae, Gardnerella vaginalis ATCC 14019, Gluconobacter oxydans
621H, Gordonia polyisoprenivorans, Hahella chejuensis KCTC 2396, helicobacter pylorus Pseudomonas
Cinaedi PAGU611, helicobacter pylorus Pseudomonas felis ATCC 49179, helicobacter pylorus Pseudomonas pylori F30, helicobacter pylorus
Pseudomonas pylori Puno120, Ignisphaera aggregans DSM 17230, Kangiella koreensis DSM
16069th, Kitasatospora setae KM-6054, genus lactubacillus johnsonii DPC 6026, Laribacter
Hongkongensis HLHK9, Legionnella longbeachae NSW 150, Marinobacter adhaerens HP 15,
Methanocorpusculum labreanum Z、Methanospirillum hungatei JF-1、
Methylobacterium nodulans ORS 2060、Methylotenera mobilis、Mobiluncus curtisii
ATCC 43063, Muricauda ruestringensis, Mycobacterium gilvum, Mycobacterium smegmatis
Str.MC2 155, mycoplasma bovis, mycoplasma bovis PG45, mycoplasma putrefaciens KS1, eisseria
Unclassified, Neisseriaceae unclassified, Neorickettsia sennetsu str.Miyayama, nitrous
Change Coccus halophilus Nc4, Nitrosomonadaceae unclassified, Nocardiaceae unclassified,
Oceanobacillus iheyensis HTE831、Peptococcaceae unclassified、
Peptostreptococcaceae, Planctomyces limnophilus DSM 3776, Polaromonas sp.JS666,
Polynucleobacter necessarius, proteus mirabilis HI4320, Pseudogulbenkiania
Sp.NH8B, Pseudoxanthomonas spadix BD-a59, Ralstonia solanacearum PSI07, Rhod
opacus B4、Rhodospirillum photometricum DSM 122、Rhodothermus marinus
SG0.5JP17-172、Riemerella anatipestifer RA-GD、Saccharopolyspora erythraea NRRL
2338th, Selenomonas ruminantium subsp.Lactilytica, Shewanella denitrificans
OS217, Sphaerochaeta globus str.Buddy, Sphingomonas wittichii RW1, conveyor screw
Thermophila DSM 6192, staphylococcus aureus aureus subsp.aureus NCTC 8325,
Stenotrophomonas maltophilia, Streptococcaceae unclassified, streptococcus agalactiae A909, chain
Coccus suis ST3, streptomyces cattleya NRRL 8057=DSM 46488, streptomyces scabiei 87.22,
Syntrophobacter fumaroxidans MPOB、Tepidanaerobacter acetatoxydans Re1、
Thermoanaerobacterium、Thermoanaerobacterium saccharolyticum、
It is Thermoanaerobacterium saccharolyticum JW/SL-YS485, Thermotoga lettingae TMO, close
Spirochaeta brennaborense DSM 12168, Treponema succinifaciens, Treponema
Succinifaciens DSM 2489, Treponema unclassified, vibrios unclassified, Vibrio vulnificus CMCP6
With one or more levels in Xanthomonas oryzae pv.oryzae MAFF 311018.
2. the agent combination of the biomarker in test right requirement 1 is preparing the use in being used to diagnose the kit of self-closing disease
On the way.
3. a kind of self-closing disease diagnostic kit, it is characterised in that including suitable for the biomarker described in test right requirement 1
Agent combination；
Optional, it is century that the agent combination of the biomarker suitable for described in test right requirement 1, which includes Beijing health,
Model CW2092S StoolGen faeces DNA extracts kits；
Optional, the agent combination of the biomarker suitable for described in test right requirement 1 also includes the U.S. and matches silent fly
The kit and the Truseq DNA of Illumina companies of U.S. production that your company of generation provides build storehouse kit；
It is optional, in addition to the analysis biomarker in vivo whether abnormal instrument.
4. the kit of the biomarker or claim 3 described in claim 1 is susceptible to suffer from the biological sample of self-closing disease in screening
System in application.
5. purposes of the Vibrio vulnificus CMCP6 on the product for preparing treatment or auxiliary treatment self-closing disease or its relevant disease；
Optional, described product is medicine or food.
6. a kind of composition, it is characterised in that including Vibrio vulnificus CMCP6；
Optional, the composition also includes Acaryochloris marina MBIC11017, Advenella
kashmirensis WT001、Alcanivorax borkumensis SK2、Alicycliphilus denitrificans、
Aliivibrio unclassified、Alkalilimnicola ehrlichii MLHE-1、Allochromatium
Vinosum DSM 180, Anaeromyxobacter sp.Fw109-5, Archaeoglobus veneficus SNP6, section are thin
Pseudomonas subspecies FB24, Azoarcus sp.BH72, Azospirillum unclassified, Bartonella
Tribocorum, Bordetella bird tuberculosis 197N, Borrelia burgdorferi group, Buchnera
aphidicola、Burkholderia phymatum STM815、Burkholderia pseudomallei 1710b、
Burkholderia sp.CCGE1002、Burkholderia sp.CCGE1003、Burkholderia xenovorans
LB400, campylobacter jejuni subsp.jejuni ICDCCJ07001, Candidatus Accumulibacter
phosphatis clade IIA str.UW-1、Candidatus Chloracidobacterium thermophilum B、
Candidatus Protochlamydia amoebophila UWE25、Chlamydophila pneumoniae J138、
Chlamydophila unclassified, corynebacterium efficiens YS-314, Cupriavidus
metallidurans CH34、Dechlorosoma suillum PS、Erythrobacter litoralis HTCC2594、
Gallionellaceae, Gluconobacter oxydans 621H, Gordonia polyisoprenivorans, Hahella
Chejuensis KCTC 2396, helicobacter pylorus Pseudomonas felis ATCC 49179, helicobacter pylorus Pseudomonas pylori F30, pylorus
Helicobacterium pylori Puno 120, Ignisphaera aggregans DSM 17230, Kangiella koreensis
DSM 16069, Kitasatospora setae KM-6054, Laribacter hongkongensis HLHK9, Legionnella
longbeachae NSW 150、Marinobacter adhaerens HP15、ethanospirillum hungatei JF-
1st, Methylobacterium nodulans ORS 2060, Methylotenera mobilis, Mycobacterium gilvum,
Mycobacterium smegmatis str.MC2 155, eisseria unclassified, Neisseriaceae
Unclassified, Neorickettsia sennetsu str.Miyayama, Nitrosococcus halophilus Nc4,
Nitrosomonadaceae unclassified, Nocardiaceae unclassified, Planctomyces limnophilus
DSM 3776, Polaromonas sp.JS666, Polynucleobacter necessaries, proteus mirabilis
HI4320、Pseudogulbenkiania sp.NH8B、Pseudoxanthomonas spadix BD-a59、Ralstonia
Solanacearum PSI07, Rhod opacus B4, Rhodospirillum photometricum DSM 122,
Rhodothermus marinus SG0.5JP17-172、Saccharopolyspora erythraea NRRL 2338、
Shewanella denitrificans OS217, Sphingomonas wittichii RW1, conveyor screw thermophila
DSM 6192, staphylococcus aureus aureus subsp.aureus NCTC 8325, Stenotrophomonas
Maltophilia, Streptococcaceae unclassified, streptomyces cattleya NRRL 8057=DSM 46488, streptomycete
Belong to scabiei 87.22, Syntrophobacter fumaroxidans MPOB, Thermotoga lettingae TMO, arc
One or more in bacterium unclassified and Xanthomonas oryzae pv.oryzae MAFF 311018；
Optional, said composition also includes acceptable carrier or pharmaceutically acceptable carrier on food.
7. a kind of composition as shown in claim 6 is preparing the product for the treatment of or auxiliary treatment self-closing disease or its relevant disease
On purposes；
Optional, described product is medicine or food.
8. a kind of system for screening the biological sample for being susceptible to suffer from self-closing disease, it is characterised in that including：
Nucleic acid-extracting apparatus, the nucleic acid-extracting apparatus is used to extract sample of nucleic acid from the biological sample and control sample；
Nucleotide sequence determining device, the nucleotide sequence determining device is connected with the nucleic acid-extracting apparatus, for the core
Acid sample is analyzed, to determine the sequence of the sample of nucleic acid；And
Judgment means, the judgment means are connected with the sequencing device, the sequence pair biological sample based on the sample of nucleic acid
Analyzed, sentenced according to analysis result with the self-closing disease biomarker of the claim 1 contained by gut flora in control sample
Whether the biological sample that breaks is susceptible to suffer from self-closing disease.
Optional, the nucleotide sequence determining device further comprises：
Library construction unit, the library construction unit is used to be based on the sample of nucleic acid, builds the library of the nucleic acid；And
Unit is sequenced, the sequencing unit is connected with the library construction unit, the library is sequenced, to determine
The sequence of sample of nucleic acid is stated,
Optionally, the library construction unit further comprises：
PCR is expanded in module, the PCR amplifications module and is set specific primer, to utilize the specific primer, to described
Sample of nucleic acid is expanded.
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| CN109182577A (en) * | 2018-09-25 | 2019-01-11 | 深圳市英马诺生物科技有限公司 | Self-closing disease biomarker and its application |
| CN109266733A (en) * | 2018-09-25 | 2019-01-25 | 深圳市英马诺生物科技有限公司 | Self-closing disease intestinal flora virulence factor gene and its application |
| WO2020140848A1 (en) * | 2018-12-30 | 2020-07-09 | 深圳碳云智能数字生命健康管理有限公司 | Intestinal microbe sequencing data processing method and device, storage medium, and processor |
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| CN109182577A (en) * | 2018-09-25 | 2019-01-11 | 深圳市英马诺生物科技有限公司 | Self-closing disease biomarker and its application |
| CN109266733A (en) * | 2018-09-25 | 2019-01-25 | 深圳市英马诺生物科技有限公司 | Self-closing disease intestinal flora virulence factor gene and its application |
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| CN113166811A (en) * | 2018-11-06 | 2021-07-23 | 斯大利卡拉公司 | Metabolic profiling for diagnosing subtype ASD phenotype 1 in patients with idiopathic autism spectrum disorders |
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| CN111455076A (en) * | 2020-04-17 | 2020-07-28 | 济南市儿童医院 | Oral flora microorganisms for childhood autism assessment |
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