CN109182577A - Self-closing disease biomarker and its application - Google Patents
Self-closing disease biomarker and its application Download PDFInfo
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Abstract
The invention discloses self-closing disease biomarker and its applications.Wherein, self-closing disease biomarker of the invention is intestinal flora virulence factor gene.Based on test individual and the intestinal flora virulence factor gene polymorphic detection of normal healthy controls as a result, effectively can determine whether test individual is susceptible to suffer from self-closing disease.
Description
Technical field
The invention belongs to biomedicine fields, in particular it relates to self-closing disease biomarker and its application.
Background technique
Self-closing disease (ASDs, Autism Spectrum Disorders) or autism or autism-spectrum obstacle are
A kind of hypotype of pervasive developmental disorders, common with male, onset is mainly shown as different degrees of speech hair in infantile period
Educate that obstacle, Social disorder, interest are narrow and behavior is mechanical, it is slow with apparent Mental development that there are about 3/4 patients
It is stagnant.According to U.S.'s diseases monitoring and prevention center (CDC) data, its disease incidence 1/45 (i.e. 2.24%) in 2015.China still lacks
National epidemiological survey data, if estimating China self-closing disease patient will be more than 10,000,000 with 1/100 estimation.Self-closing disease conduct
A kind of children's spirit disease, seriously affects the social function of infant, also can bring heavy burden to infant family and society.
However, self-closing disease is assessed generally according to scale at present, without blood diagnosis method and effective therapeutic scheme,
Diagnosis will judge by subjective experience in this way, cannot not only provide the morbidity of self-closing disease effective early warning, but also accuracy rate of diagnosis
It is very low.Therefore, self-closing disease pathogenesis and its relevant biomarker of disease are studied, and then is intervened in time for the cause of disease
It is the key factor treated disease, mitigate burden on society with treatment.
But for the research of self-closing disease pathogenesis and its relevant biomarker of disease, there are also to be added at this stage
By force.
Summary of the invention
The present invention is directed at least solve one of the technical problems existing in the prior art.For this purpose, one object of the present invention
It is to propose a kind of relevant biomarker of self-closing disease.
It should be noted that the present invention is following research, discovery and work based on inventor and completes:
Inventor has found in the course of the research, the generally existing function of intestinal canal disorder of self-closing disease patient, and function of intestinal canal disorder
The symptom of self-closing disease may be aggravated, in turn, a series of enteric microorganism has been carried out to self-closing disease patient and Healthy People and has deeply been ground
Study carefully.Wherein, inventor has detected the fecal sample enterobacteriaceae of self-closing disease patient and control group by the macro gene order-checking of shotgun
Group's polymorphism, all there were significant differences for the total number of bacteria and bacteria abundance for finding the two, and further studies, and surprisingly finds,
The polymorphism of the intestinal flora virulence factor gene of self-closing disease patient is significantly higher than Healthy People.
Inventor further verifies above-mentioned discovery, determines the intestinal flora based on test individual and normal healthy controls
Virulence factor gene polymorphic detection is as a result, effectively can determine whether test individual is susceptible to suffer from self-closing disease.Inventor recognizes as a result,
For that can examine using intestinal flora virulence factor gene as self-closing disease biomarker for clinical and scientific research etc. to be various
Self-closing disease detection and screening under disconnected and non-diagnostic situation.
As a result, in the first aspect of the present invention, the present invention provides a kind of self-closing disease biomarkers.It is according to the present invention
Embodiment, the self-closing disease biomarker are intestinal flora virulence factor gene.As described above, right based on test individual and health
According to intestinal flora virulence factor gene polymorphic detection as a result, effectively can determine whether test individual is susceptible to suffer from self-closing disease.
According to an embodiment of the invention, the self-closing disease biomarker shows themselves in that the intestinal flora poison of self-closing disease patient
The polymorphism of power factor gene is significantly higher than Healthy People.
Herein, term " biomarker " shall be understood in a broad sense comprising any to be able to reflect appointing for abnormality
What detectable Biological indicators may include gene marker, species marker (kind marker/category marker) and function mark
Object (KO/OG marker) can be used for medical diagnosis on disease, judge staging or for evaluating with very extensive purposes
The safety and efficacy of new drug or new treatment in target group.
Term " virulence factor " is to instigate microorganism with the host of particular species or establish itself inside it and enhance
It causes the characteristic (i.e. gene product) of the potentiality of disease.Virulence factor includes bacteriotoxin, the cell table of mediating bacterial attachment
Face protein, the protein of Cell surface carbohydrate and protection bacterium, and potentially contribute to the hydrolysis of Pathogenicity of Bacteria
Enzyme.
Expression way " polymorphism of intestinal flora virulence factor gene " refer to intestinal flora virulence contained in sample because
Subbase because specific diversity, wherein the type of intestinal flora virulence factor gene contained in polymorphism height, that is, sample is more.
In the second aspect of the present invention, the present invention also provides be adapted to detect for mentioned-above self-closing disease biomarker
Purposes of the reagent in preparation self-closing disease detection kit.It is self-closing using being adapted to detect in the kit prepared as a result,
The reagent of disease biomarker can carry out intestinal flora virulence factor gene to biological sample to be measured and normal healthy controls sample
Polymorphic detection, in turn, it is self-closing that the polymorphic differences result based on the two can effectively judge whether biological sample to be measured is susceptible to suffer from
Disease.
In the third aspect of the present invention, the present invention provides a kind of self-closing disease detection kits.Implementation according to the present invention
Example, which includes the reagent for being adapted to detect for mentioned-above self-closing disease biomarker.Institute is utilized as a result,
State the reagent for being adapted to detect for self-closing disease biomarker in kit, can to biological sample to be measured and normal healthy controls sample into
The polymorphic detection of row intestinal flora virulence factor gene, in turn, the polymorphic differences result based on the two can effectively judge
Whether biological sample to be measured is susceptible to suffer from self-closing disease.
According to an embodiment of the invention, the reagent for being adapted to detect for mentioned-above self-closing disease biomarker includes:
Extracting genome DNA is carried out suitable for the fecal specimens to test individual and macro gene sets up the reagent in library and sequencing.It is as a result,
The polymorphic detection that can carry out intestinal flora virulence factor gene to biological sample to be measured and normal healthy controls sample is based in turn
The polymorphic differences result of the two can effectively judge whether biological sample to be measured is susceptible to suffer from self-closing disease.
Some specific examples according to the present invention, the fecal specimens to test individual carry out extracting genome DNA
Reagent is faeces DNA extracts kit, and preferably Beijing health is that the StoolGen faeces DNA that the article No. in century is CW2092S extracts
Kit.Fecal specimens extracting genome DNA effect is good as a result, so that the testing result of self-closing disease biomarker is more quasi-
It is really reliable.
According to some embodiments of the present invention, it is Illumina that the macro gene, which sets up library and the reagent of sequencing,
HiSeq2500 microarray dataset builds library and sequencing reagent.The testing result of self-closing disease biomarker more accurately may be used as a result,
It leans on.
According to some embodiments of the present invention, the kit use the following steps are included:
Extracting genome DNA is carried out to biological sample to be measured and normal healthy controls sample respectively, to obtain genomic DNA；
Macro gene is carried out for the genomic DNA and sets up library and sequencing, to obtain macro gene order-checking data；
Determine the intestinal flora of the biological sample to be measured and normal healthy controls sample respectively based on macro gene order-checking data
The polymorphism of virulence factor gene；
Determine whether the biological sample to be measured is susceptible to suffer from self-closing disease based on polymorphism definitive result, wherein the life to be measured
The polymorphism of the intestinal flora virulence factor gene of object sample is significantly higher than normal healthy controls sample, is that the biological sample to be measured is easy
Suffer from the instruction of self-closing disease.
The detection to the self-closing disease biomarker of biological sample to be measured can be effectively realized using the kit as a result,
And result is accurate and reliable.
According to an embodiment of the invention, the biological sample to be measured and normal healthy controls sample are fecal specimens, come respectively
From test individual and Healthy People.
Some specific examples according to the present invention carry out the macro gene using the macro genome sequencing method of shotgun and set up library
And sequencing, the macro gene order-checking of shotgun (Shotgun metagenome sequencing) being capable of high-precision reacting bacteria
Type and abundance.
According to some embodiments of the present invention, the macro gene is carried out using 2500 microarray dataset of Illumina HiSeq
Set up library and sequencing.Sequencing result is accurate as a result, so that the testing result of self-closing disease biomarker is more accurate and reliable.
Some specific examples according to the present invention determine the biological sample to be measured based on macro gene order-checking data respectively
With the polymorphism of the intestinal flora virulence factor gene of normal healthy controls sample, further comprise: by the macro gene order-checking number
Virulence factor gene database (virulence factor database, http://www.mgc.ac.cn/ are arrived according to comparing
VFs/main.htm), so as to obtain each sample intestinal flora virulence factor gene polymorphism.Self-closing disease biology mark as a result,
The testing result of will object is accurate and reliable.
According to some embodiments of the present invention, determine whether the biological sample to be measured is susceptible to suffer from based on polymorphism definitive result
Self-closing disease further comprises: by multidimensional variance analysis to the intestinal flora in the biological sample to be measured and normal healthy controls sample
The polymorphism of virulence factor gene carries out significant difference analysis, and carries out sparse offset minimum binary to significant difference analysis result and sentence
It does not analyze, to determine whether the polymorphism of intestinal flora virulence factor gene of the biological sample to be measured is significantly higher than health
Control sample.The testing result of self-closing disease biomarker is more accurate and reliable as a result,.
In conclusion self-closing disease biomarker and self-closing disease detection kit of the invention can be effective for screenings
It is susceptible to suffer from the biological sample of self-closing disease.To which in the fourth aspect of the present invention, it is raw that the present invention also provides mentioned-above self-closing diseases
The application of object marker or self-closing disease detection kit in the biological sample that screening is susceptible to suffer from self-closing disease.
Further, in the fifth aspect of the invention, the biological sample of self-closing disease is susceptible to suffer from the present invention provides a kind of screening
System.According to an embodiment of the invention, the system includes:
Self-closing disease biomarker detection device, the self-closing disease biomarker detection device are to be measured for detecting respectively
Mentioned-above self-closing disease biomarker in biological sample and normal healthy controls sample, and determine the intestinal flora virulence of the two
The polymorphism of factor gene；And
Analytical equipment, the analytical equipment are connected with the self-closing disease biomarker detection device, for from it is described from
It closes disease biomarker detection device and receives intestinal flora virulence factor in the biological sample to be measured and normal healthy controls sample
Gene polynorphisms definitive result, and it is self-closing based on the polymorphism definitive result to determine whether the biological sample to be measured is susceptible to suffer from
Disease, wherein the polymorphism of the intestinal flora virulence factor gene of the biological sample to be measured is significantly higher than normal healthy controls sample, is
The biological sample to be measured is susceptible to suffer from the instruction of self-closing disease.
According to an embodiment of the invention, the system that the biological sample of self-closing disease is susceptible to suffer from using above-mentioned screening, it can be effective
Screening is susceptible to suffer from the biological sample of self-closing disease, and the result for detecting screening is accurate, reliably, favorable repeatability.
Some specific examples according to the present invention, the biological sample to be measured and normal healthy controls sample are fecal specimens,
Respectively from test individual and Healthy People.
According to some embodiments of the present invention, the self-closing disease biomarker detection device includes: that faeces DNA extracts list
Member；Macro gene sets up library and sequencing unit；And polymorphism determination unit, wherein the polymorphism determination unit is suitable for being based on
Macro gene order-checking data determine the intestinal flora virulence factor gene of the biological sample to be measured and normal healthy controls sample respectively
Polymorphism.
According to an embodiment of the invention, the macro gene sets up library and sequencing unit is adapted for carrying out the macro genome of shotgun and surveys
Sequence.
Some specific examples according to the present invention, using 2500 microarray dataset of Illumina HiSeq as the macro base
Because setting up library and sequencing unit.Sequencing data is accurate as a result, and the testing result of self-closing disease biomarker is more accurate and reliable.
According to some embodiments of the present invention, in the polymorphism determination unit, based on macro gene order-checking data point
The polymorphism of the intestinal flora virulence factor gene of the biological sample to be measured and normal healthy controls sample is not determined, is further wrapped
Include: by the macro gene order-checking comparing to virulence factor gene database (virulence factor database,
Http:// www.mgc.ac.cn/VFs/main.htm), so as to obtain each sample intestinal flora virulence factor gene it is polymorphic
Property.The testing result of self-closing disease biomarker is accurate and reliable as a result,.
Other embodiments according to the present invention are determined in the analytical equipment based on the polymorphism definitive result
Whether the biological sample to be measured is susceptible to suffer from self-closing disease: by multidimensional variance analysis to the biological sample to be measured and
The polymorphism of intestinal flora virulence factor gene in normal healthy controls sample carries out significant difference analysis, and analyzes significant difference
As a result sparse partial least squares discriminant analysis is carried out, to determine the intestinal flora virulence factor gene of the biological sample to be measured
Polymorphism whether be significantly higher than normal healthy controls sample.The testing result of self-closing disease biomarker is more accurate and reliable as a result,.
Present invention firstly discovers that causing a disease for self-closing disease is related with the polymorphism of intestinal flora virulence factor gene, therefore by intestines
Detection marker of the road flora virulence factor gene as self-closing disease.Self-closing disease biomarker energy according to an embodiment of the present invention
It is enough to be detected effective for the self-closing disease of any diagnosis and non-diagnostic (such as scientific research) purposes, specifically, based on to biology to be measured
Sample and normal healthy controls sample carry out the polymorphic detection of intestinal flora virulence factor gene, and the polymorphic differences based on the two
As a result it can effectively judge whether biological sample to be measured is susceptible to suffer from self-closing disease.And result is accurate, reliably, favorable repeatability.As a result,
The diagnosis of self-closing disease will judge not against subjective experience, not only can provide effective early warning to the morbidity of self-closing disease, but also can be right
It is made a definite diagnosis, and overcomes the defect that the diagnosis of self-closing disease largely judges dependent on subjective experience, greatly improves and examine
Disconnected accuracy rate.
Additional aspect and advantage of the invention will be set forth in part in the description, and will partially become from the following description
Obviously, or practice through the invention is recognized.
Detailed description of the invention
Above-mentioned and/or additional aspect of the invention and advantage will become from the description of the embodiment in conjunction with the following figures
Obviously and it is readily appreciated that, in which:
Fig. 1 is discovery phase self-closing disease and control group intestinal flora virulence factor gene comparison diagram in embodiment 1, and A figure is aobvious
Show that virulence factor gene can significantly distinguish disease group and control group (multidimensional variance analysis, p < 0.006)；B figure shows standardization
Polymorphism comparing result afterwards, self-closing disease group intestinal flora virulence factor polymorphism are significantly higher than control group (* *, rank sum test P
Value < 0.01).
Fig. 2 is Qualify Phase self-closing disease and control group intestinal flora virulence factor gene comparison diagram in embodiment 1, and A figure is aobvious
Show that virulence factor gene can significantly distinguish disease group and control group (multidimensional variance analysis, p < 0.0002)；B figure shows standardization
Polymorphism comparing result afterwards, self-closing disease group intestinal flora virulence factor polymorphism are significantly higher than control group (*, rank sum test P value
<0.05)。
Specific embodiment
The embodiment of the present invention is described below in detail.The embodiments described below with reference to the accompanying drawings are exemplary, only
It is used to explain the present invention, and is not considered as limiting the invention.Particular technique or condition are not specified in embodiment, according to
Technology or conditions described in document in the art (such as write with reference to J. Pehanorm Brooker etc., " the molecule gram that Huang Peitang etc. is translated
Grand experiment guide ", the third edition, Science Press) or carry out according to product description.Production is not specified in agents useful for same or instrument
Manufacturer person, being can be with conventional products that are commercially available, such as can purchase from Illumina company.
Embodiment 1
1 object and method
1.1 research object
Patient group comes from Shenzhen Children's Hospital's Psychology Dept. and speech therapy section outpatient service self-closing disease infant, acquisition time are
7~December in 2015.It is included in standard: 1. according to international newest diagnostic criteria, i.e. " Americanism obstacle diagnosis and statistic handbook the
5 editions " (Diagnostic and Statistical Manual of Mental Disorders, 5th Edition, DSM-
5) it, is diagnosed as " self-closing disease " and needs the infants supported very more；2. age < 14 year old；3. male or female.Exclusion criteria: it 1. suffers from
Other mental diseases (such as schizophrenia)；2. suffering from other neurodevelopmental disorder diseases；3. suffering from hereditary metabolic disorders；
4. suffering from the great physical disease history such as serious neurological disease and craniocerebral injury history.Control group comes from Shenzhen Children's Hospital's children's health care
Section physical examination children.It is included in standard: 1. listless disease, health；2. being matched with patient group age and gender.Exclusion criteria is same
Patient group.It is all enter group Parents to this research know and sign informed consent form.This research is through Shenzhen Children's Hospital's human relations
The approval of the reason committee.
This research was divided to for two phases, and the first phase is discovery phase, collected 22 infants, male 18, female 4 altogether；Age 2~6 years old,
Average (2 ± 1) year.15 control children, male 8, female 7 are collected altogether；Age 1~6 years old, average (2 ± 1) year.Two groups of genders
(Fisher is accurately examined, P > 0.05), age (Student ' s T test, P > 0.05) no significant difference.The second phase
For Qualify Phase, 22 infants, male 18, female 3 are collected；Age 2~6 years old, average (2 ± 1) year.16 control youngsters are collected altogether
Child, male 9, female 7；Age 1~6 years old, average (2 ± 1) year.Two groups of genders (Fisher is accurately examined, P > 0.05), ages
(Student ' s T test, P > 0.05) no significant difference
1.2 acquisition excrement and pre-processing
Certainly be about to excrement comes specified feces collection basin to participant, is ready for disposable medical excrement sampling pipe, wears
Gloves acquire excrement, and acquisition is interrupted, pollution caused by avoiding because of carelessness, once discovery pollution, answers resampling, acquisition is completed
Quick freeze is stored in -80 DEG C of refrigerators afterwards.Collect relevant rudimentary information, comprising: name, gender, age, height, weight, illness
History, home address, parent's contact method etc. full and accurate " health condition information table "；" diet and living habit application form " needs emphasis
The data of collection includes eating habit, if breast-feeding, parent's living habit, health status and smoking history, atmosphere locating for family
It encloses, recent antibiotic, Yoghourt, probiotics uses.
After stool sample has been collected, Beijing health is used to extract for the StoolGen faeces DNA of the model CW2092S in century
Kit carries out the extraction of genomic DNA according to its specification.
1.3 macro genes set up library and sequencing
Quantify DNA using NanoDrop 2000, total amount needs to reach 5ug or more, while taking 200ng base
Because group DNA carries out electroresis appraisal；Genomic DNA 5ug is taken (to be subject to qubit quantitative result, the body of 130ul is supplied with TE
Product), macro genome DNA is interrupted into 180bp using Covaris；It is eluted after interrupting with EB 85ul；End-filling adds A,
Adjunction head, 2% Ago-Gel choose target fragment (QIAGEN Gel Kit, 20ul EB elution), and Qubit is quantitative, then
High-flux sequence is carried out with llumina HiSeq2500 microarray dataset.
1.4 analysis of biological information and statistics
Firstly, filtering out sequence some of poor quality or containing connector.Secondly, the sequence of all measurements compares mankind's base
Because a group sequence (referring to Hg19) is to reject artificially-contaminated sequence.Again, using the default parameters of newest MEGAN software v5,
Blast is carried out to the macro gene order-checking data of shotgun to compare to virulence factor related gene database (virulence
Factor-related gut microbiota genes:virulence factor database, http: //
www.mgc.ac.cn/VFs/main.htm).Obtain the abundance of each sample intestinal flora virulence factor related gene.
Then, by multidimensional variance analysis judge disease group and control group virulence factor gene whether significant difference, with
And with being shown after sparse partial least squares discriminant analysis (PLSDA), the virulence factor gene of disease group and control group it is more
Whether significant difference is analyzed with rank sum test to state property.
2 results
2.1 discovery phase
The first phase is discovery phase, collects 22 infants, male 18, female 4 altogether；Age 2~6 years old, average (2 ± 1) year.
15 control children are collected altogether.Inventor has found that intestinal flora virulence factor related gene can significantly distinguish self-closing disease patient and just
Often control (Figure 1A, multidimensional variance analysis, p < 0.006), and the polymorphism of self-closing disease group intestinal flora virulence factor is significantly higher than
Control group (value < 0.01 Figure 1B, * *, rank sum test P).
2.2 Qualify Phase
The second phase is Qualify Phase, collects 22 infants, male 18, female 3 altogether；Age 2~6 years old, average (2 ± 1) year.
16 control children, male 9, female 7 are collected altogether；Age 1~6 years old, average (2 ± 1) year.As a result see Fig. 2.Inventor has found intestines
Road flora virulence factor related gene can significantly distinguish self-closing disease patient and normal control (Fig. 2A, multidimensional variance analysis, p <
0.0002), and the polymorphism of self-closing disease group intestinal flora virulence factor be significantly higher than control group (Fig. 2 B, *, rank sum test P value <
0.05)。
3 conclusions
Inventors have found that self-closing disease Intestinal Mucosal Injury in Patients Undergoing flora virulence factor gene can significantly distinguish disease group and control group, and
The polymorphism of self-closing disease group intestinal flora virulence factor is significantly higher than control group, thus self-closing disease Intestinal Mucosal Injury in Patients Undergoing flora virulence factor
Gene can be used as the marker of self-closing disease patient.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
Centainly refer to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be any
One or more embodiment or examples in can be combined in any suitable manner.
Although an embodiment of the present invention has been shown and described, it will be understood by those skilled in the art that: not
A variety of change, modification, replacement and modification can be carried out to these embodiments in the case where being detached from the principle of the present invention and objective, this
The range of invention is defined by the claims and their equivalents.
Claims (10)
1. a kind of self-closing disease biomarker is intestinal flora virulence factor gene.
2. self-closing disease biomarker according to claim 1, which is characterized in that the self-closing disease biomarker performance
Are as follows: the polymorphism of the intestinal flora virulence factor gene of self-closing disease patient is significantly higher than Healthy People.
3. being adapted to detect for the reagent of self-closing disease biomarker of any of claims 1 or 2 in preparation self-closing disease detection kit
In purposes.
4. a kind of self-closing disease detection kit, which is characterized in that raw including being adapted to detect for self-closing disease of any of claims 1 or 2
The reagent of object marker.
5. self-closing disease detection kit according to claim 4, which is characterized in that described to be adapted to detect for claims 1 or 2
The reagent of the self-closing disease biomarker include: suitable for the fecal specimens of test individual carry out extracting genome DNA with
And macro gene sets up the reagent in library and sequencing,
Optionally, it is that faeces DNA extracts reagent that the fecal specimens to test individual, which carry out the reagent of extracting genome DNA,
Box, preferably Beijing health are the StoolGen faeces DNA extracts kit that the article No. in century is CW2092S,
Optionally, the macro gene set up the reagent of library and sequencing be 2500 microarray dataset of Illumina HiSeq build library and
Sequencing reagent.
6. self-closing disease detection kit according to claim 4, which is characterized in that the use of the kit includes following
Step:
Extracting genome DNA is carried out to biological sample to be measured and normal healthy controls sample respectively, to obtain genomic DNA；
Macro gene is carried out for the genomic DNA and sets up library and sequencing, to obtain macro gene order-checking data；
Determine the intestinal flora virulence of the biological sample to be measured and normal healthy controls sample respectively based on macro gene order-checking data
The polymorphism of factor gene；
Determine whether the biological sample to be measured is susceptible to suffer from self-closing disease based on polymorphism definitive result, wherein the biological sample to be measured
The polymorphism of the intestinal flora virulence factor gene of product is significantly higher than normal healthy controls sample, is that the biological sample to be measured is susceptible to suffer from certainly
The instruction of disease is closed,
Optionally, the biological sample to be measured and normal healthy controls sample are fecal specimens, respectively from test individual and health
People,
Optionally, the macro gene is carried out using the macro genome sequencing method of shotgun and sets up library and sequencing,
Optionally, the macro gene is carried out using 2500 microarray dataset of Illumina HiSeq and sets up library and sequencing,
Optionally, the enterobacteriaceae of the biological sample to be measured and normal healthy controls sample is determined respectively based on macro gene order-checking data
Group virulence factor gene polymorphism, further comprise: by the macro gene order-checking comparing to virulence factor gene number
It is each to obtain according to library (virulence factor database, http://www.mgc.ac.cn/VFs/main.htm)
The polymorphism of the intestinal flora virulence factor gene of sample,
Optionally, determine whether the biological sample to be measured is susceptible to suffer from self-closing disease and further comprises based on polymorphism definitive result: logical
Multidimensional variance analysis is crossed to the polymorphic of the intestinal flora virulence factor gene in the biological sample to be measured and normal healthy controls sample
Property carry out significant difference analysis, and sparse partial least squares discriminant analysis is carried out to significant difference analysis result, to determine
Whether the polymorphism for stating the intestinal flora virulence factor gene of biological sample to be measured is significantly higher than normal healthy controls sample.
7. self-closing disease biomarker of any of claims 1 or 2 or the described in any item self-closing disease detections of claim 4-6
Application of the kit in the biological sample that screening is susceptible to suffer from self-closing disease.
8. a kind of screen the system for being susceptible to suffer from the biological sample of self-closing disease characterized by comprising
Self-closing disease biomarker detection device, the self-closing disease biomarker detection device for detecting biology to be measured respectively
Self-closing disease biomarker of any of claims 1 or 2 in sample and normal healthy controls sample, and determine the intestinal flora of the two
The polymorphism of virulence factor gene；And
Analytical equipment, the analytical equipment are connected with the self-closing disease biomarker detection device, are used for from the self-closing disease
Biomarker detection device receives the intestinal flora virulence factor gene in the biological sample to be measured and normal healthy controls sample
Polymorphism definitive result, and determine whether the biological sample to be measured is susceptible to suffer from self-closing disease based on the polymorphism definitive result,
Wherein, the polymorphism of the intestinal flora virulence factor gene of the biological sample to be measured is significantly higher than normal healthy controls sample, is institute
The instruction that biological sample to be measured is susceptible to suffer from self-closing disease is stated,
Optionally, the biological sample to be measured and normal healthy controls sample are fecal specimens, respectively from test individual and health
People.
9. system according to claim 8, which is characterized in that the self-closing disease biomarker detection device includes:
Faeces DNA extraction unit；
Macro gene sets up library and sequencing unit；And
Polymorphism determination unit,
Wherein, the polymorphism determination unit be suitable for based on macro gene order-checking data determine respectively the biological sample to be measured with
The polymorphism of the intestinal flora virulence factor gene of normal healthy controls sample,
Optionally, the macro gene sets up library and sequencing unit is adapted for carrying out the macro gene order-checking of shotgun,
Optionally, library and sequencing unit are set up as the macro gene using 2500 microarray dataset of Illumina HiSeq,
Optionally, in the polymorphism determination unit, the biological sample to be measured is determined respectively based on macro gene order-checking data
The polymorphism of product and the intestinal flora virulence factor gene of normal healthy controls sample further comprises: by the macro gene order-checking
Comparing is to virulence factor gene database (virulence factor database, http://www.mgc.ac.cn/
VFs/main.htm), so as to obtain each sample intestinal flora virulence factor gene polymorphism.
10. system according to claim 8, which is characterized in that in the analytical equipment, determined based on the polymorphism
As a result determine whether the biological sample to be measured is susceptible to suffer from self-closing disease and further comprises: by multidimensional variance analysis to the life to be measured
The polymorphism of intestinal flora virulence factor gene in object sample and normal healthy controls sample carries out significant difference analysis, and to significant
Variance analysis result carries out sparse partial least squares discriminant analysis, to determine the intestinal flora virulence of the biological sample to be measured
Whether the polymorphism of factor gene is significantly higher than normal healthy controls sample.
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