CN108841974A - Extract the method that fecal sample 16s rRNA is monitored in infant's intestinal microecology maturity - Google Patents
Extract the method that fecal sample 16s rRNA is monitored in infant's intestinal microecology maturity Download PDFInfo
- Publication number
- CN108841974A CN108841974A CN201810687215.3A CN201810687215A CN108841974A CN 108841974 A CN108841974 A CN 108841974A CN 201810687215 A CN201810687215 A CN 201810687215A CN 108841974 A CN108841974 A CN 108841974A
- Authority
- CN
- China
- Prior art keywords
- maturity
- sample
- infant
- enteric microorganism
- rrna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16H—HEALTHCARE INFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR THE HANDLING OR PROCESSING OF MEDICAL OR HEALTHCARE DATA
- G16H20/00—ICT specially adapted for therapies or health-improving plans, e.g. for handling prescriptions, for steering therapy or for monitoring patient compliance
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16H—HEALTHCARE INFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR THE HANDLING OR PROCESSING OF MEDICAL OR HEALTHCARE DATA
- G16H20/00—ICT specially adapted for therapies or health-improving plans, e.g. for handling prescriptions, for steering therapy or for monitoring patient compliance
- G16H20/60—ICT specially adapted for therapies or health-improving plans, e.g. for handling prescriptions, for steering therapy or for monitoring patient compliance relating to nutrition control, e.g. diets
-
- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16H—HEALTHCARE INFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR THE HANDLING OR PROCESSING OF MEDICAL OR HEALTHCARE DATA
- G16H50/00—ICT specially adapted for medical diagnosis, medical simulation or medical data mining; ICT specially adapted for detecting, monitoring or modelling epidemics or pandemics
- G16H50/20—ICT specially adapted for medical diagnosis, medical simulation or medical data mining; ICT specially adapted for detecting, monitoring or modelling epidemics or pandemics for computer-aided diagnosis, e.g. based on medical expert systems
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medical Informatics (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Primary Health Care (AREA)
- Epidemiology (AREA)
- Analytical Chemistry (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Databases & Information Systems (AREA)
- Nutrition Science (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Data Mining & Analysis (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses the methods that extraction fecal sample 16s rRNA is monitored in infant's intestinal microecology maturity, including the processes such as the analysis of sample collection, 16s-rRNA gene sequencing, sequencing data, enteric microorganism maturity and enteron aisle cell age calculate, user report push.It is put forward for the first time this monitoring index of enteric microorganism maturity, enteric microorganism maturity refers to infant enteric microorganism and its similitude of mother enteric microorganism in composition, maturity is higher, think that flora development is better, the index can react the immunity level of infant, the present invention is analyzed by experimental study and data, under room temperature environment, method by detecting fecal sample 16s rRNA V3-V4 hypervariable region, realize the assessment of infant enteric microorganism, its sampling is convenient, it is fast to analyze speed, it is at low cost, and assessment result and existing research are consistent, make it possible that consumer level enteric microorganism detects.
Description
Technical field
The present invention relates to biotechnology technical fields, and specially extraction fecal sample 16s rRNA is in the micro- life of infant's enteron aisle
The method of state maturity monitoring.
Background technique
Human intestinal microorganisms are colonized in enteron aisle, and quantity is more than 10 times or more of human body own cells, to nutrients
Matter metabolism, human body itself development, the immune and generation of disease etc. are all played the role of extremely important.It is micro- to infant's enteron aisle
Biology the study found that early stage enteric microorganism constitute, baby most important to the development of later period enteric microorganism and personal health
The destruction of child period enteric microorganism will lead to a variety of diseases such as asthma, diabetes, self-closing disease, asthma and generate.Furthermore more
The transformation of nonrandomness is presented in research report, infant enteric microorganism with advancing age.By being surveyed using 16s rRNA
Sequence versatility analyzes enteric microorganism sample in human feces, and discovery infant enteric microorganism is not broken between 0-3 years old
Long development, its final enteric microorganism, which is constituted, is intended to adult.Fredrik et al. utilizes macro genome shotgun method sequencing technologies point
Fecal sample is analysed, finds the mother of the enteric microorganism of the length of 12 month infants and they compared with newborn and 4 month infants
It is more alike.The research also found, different production methods（Such as dig woman's production and natural labor）And feeding patterns（Breast-feeding, formula milk
Powder nursing etc.）The development of infant enteric microorganism is had a major impact.Derrick et al. is from pregnant woman and their 4-6 week old babies
The different parts of body（Enteron aisle, oral cavity, nostril, skin and vagina）Micro-biological samples are acquired, the analysis found that, newborn is not
Enteric microorganism with physical feeling constitutes the very strong homology of presentation.However after 6 weeks, microorganism is constituted and function is presented greatly
The amplification and diversity of amplitude.This phenomenon shows that baby microorganism constitutes and will send out from baby due to 4 to 6 weeks
Amplitude variation, is the material time section of Microorganism colonization.
Currently, there are mainly two types of enteric microorganism detection methods：16s rRNA and macro genome.16s rRNA coding
Gene order shares 9 conserved regions and 9 hypervariable regions, by that can do Phylogenetic diversity of bacteria point to 9 hypervariable region sequencing analysis
Analysis and metabolic pathway analysis.Then the gene of tiny organisms all in sample is all sequenced for macro genome, and sequencing data passes through
It after splicing, compares and annotates with reference flora database, and then carry out Phylogenetic diversity of bacteria analysis and metabolic pathway analysis.
Present invention firstly provides this monitoring indexes of enteric microorganism maturity.Enteric microorganism maturity refers to infant
Enteric microorganism and its similitude of mother enteric microorganism in composition.Maturity is higher, it is believed that flora development is better, this refers to
Mark can react the immunity level of infant.Although the generation of influence and complex disease of the enteric microorganism to infantile health is
There are many researchs and document report, but most researchs are still in laboratory level, are hardly formed the conversion of commercial product.With
Laboratory research is different, commercial product need to solve user be in convenient sampling, sample stablize save, shorten haulage time and
Sequencing and data-analysis time and cost.Wherein, selecting suitable enteric microorganism detection mode is exactly one of to challenge.Although macro
Genome detection method can capture all microorganisms that may be present in sample, but for the 16s rRNA that compares, it is macro
Genome species annotation information is only more than 16s rRNA on belonging to level, and 16s rRNA can be in door, guiding principle, mesh, section, category level
Obtain more information.In addition to this, macro genome detection method sequencing cost is high, the time is sequenced, and long, calculating complexity etc. causes
This method is difficult to apply in business.
Summary of the invention
The purpose of the present invention is to provide extract fecal sample 16s rRNA to monitor in infant's intestinal microecology maturity
Method, to take into account cost and detection accuracy, under room temperature environment by detection 16s rRNA V3-V4 hypervariable region domain analysis
Infant's intestinal flora is constituted, and calculates enteric microorganism maturity, to solve to propose under room temperature environment in above-mentioned background technique,
By 16s rRNA in high-throughput two generations sequencing detection fecal sample, analysis infant's intestinal flora is constituted, and to the micro- life of enteron aisle
The problem of state maturity is continued to monitor, and infant enteric microorganism development condition and body immunity situation are assessed.
To achieve the above object, the present invention provides the following technical solutions：Fecal sample 16s rRNA is extracted in infant's intestines
The method of road Tiny ecosystem maturity monitoring, includes the following steps：
S1：User applies for item detection to company；
S2：Company audits user information, and sends sample detector to user；
S3：User receives sample detector, carries out fecal sample acquisition, sample binding and questionnaire and fills in；
S4：The pattern detection center of company is returned in fecal sample express delivery by user；
S5：Company's pattern detection center carries out DNA extraction, establishes database；
S6：The sequencing of the hypervariable region V3-V4 is carried out using microarray dataset Illumima MiSeq sequenator；
S7：It is analyzed using QIIME v1.7.0 software lower machine data original to 16s rRNA；
S8：Enteric microorganism maturity calculates, and generates examining report；
S9：It will test report and be in electronic form sent to user hand generator terminal, convenient for checking report.
Preferably, sample collection includes 2000 couples of mothers and sons, totally 4000 fecal samples and corresponding questionnaire, Mei Geyong
The fecal sample collector that the company of receiving is sent at family, one prepares for baby, and one prepares for mother, furthermore user is needed to sweep
The two dimensional code progress sample binding and questionnaire retouched on collector are filled in, and the acquisition and transport of entire sample are at normal temperature
It carries out, without using ice chest and cold chain transportation.
Preferably, DNA is extracted and is used paramagnetic particle method DNA purification technique in the step S5.
Preferably, specific method includes in the step S7：
S701：Initial data pretreatment：Sample joint sequence is removed, and both-end sequencing sequence is spliced into single sequence, according to
Barcode sequence distinguishes different sample sequences, filters low quality sequence；
S702：OTU（Activity classification unit）Classification and statistics：Using clustering method, according to the similarity threshold of sequence 97% by sequence
Column are divided into different OTU, and each OTU is considered as a microbial species, and carries out species annotation using SILVA database, this
Outside, quantity is sequenced to each sample and OTU number counts；
S703：Diversity analysis：Analyze the alpha diversity of each sample（Chao1, observed species, Shannon are more
Sample sex index, Simpson diversity indices）；
S704：Function and metabolic pathway prediction：Using macro genome prediction technique（Tax4Fun-r software package）To 16s rRNA into
The functional prediction of row.
Preferably, maturation is represented with infant enteric microorganism and its mother's enteric microorganism similitude in the step S8
Spend index（Jaccard likeness coefficient）, specific formula for calculation is as follows：
Jaccard(A,B) indicate enteric microorganism maturity,A Indicate infant OTU number,B Indicate mother OTU number；Root
According to 2000 big-sample datas, linear fit model is constructed, the true enterobacteriaceae of infant is predicted by enteric microorganism maturity
Age：
AgeIndicating infant's enteron aisle cell age, x indicates coefficient, Jaccard(A,B) Indicate enteric microorganism maturity.
Preferably, the report content includes baby's enteron aisle age, maturation index, the Pseudomonas detected and every kind of bacterium
Abundance, beneficial bacterium nocuousness bacterial content and the crowd's lateral comparison of category, prebiotic bacterial content and crowd's lateral comparison, how is important Pseudomonas
Influence baby's health, protein, lipid, carbohydrate metabolism ability, and the baby's health raising side provided according to report
Case and dietary recommendation.
Compared with prior art, the beneficial effects of the invention are as follows：
Present invention firstly provides this monitoring index of enteric microorganism maturity, enteric microorganism maturity refers to infant's enteron aisle
Microorganism and its similitude of mother enteric microorganism in composition, maturity are higher, it is believed that flora development is better, which can
The immunity level of infant is reacted, the present invention is analyzed by experimental study and data, under room temperature environment, by detecting excrement
The method of sample 16s rRNA V3-V4 hypervariable region realizes the assessment of infant enteric microorganism, samples convenient, analysis speed
Spend fast, at low cost, and assessment result and existing research are consistent, make it possible that consumer level enteric microorganism detects.
Detailed description of the invention
Fig. 1 is sampling process and data analysis flowcharts of the invention；
Fig. 2 is mobile phone terminal electronic report figure of the invention；
Fig. 3 is the relational graph at infant's age and enteric microorganism maturity of the invention；
Relational graph of the Fig. 4 between baby Different Month group crowd of the invention and mother's group and 4 kinds of alpha diversities；
Relationship of the Fig. 5 between different production methods and enteric microorganism maturity and 4 kinds of alpha diversities of the invention；
Fig. 6 is that two pairs of mother and sons' enteric microorganism data of the different production methods of the present invention are compared relational graph.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
In the specific embodiment of the invention：Refering to fig. 1,
Firstly, sample collection；2000 couples of mothers and sons are collected, totally 4000 fecal samples and corresponding questionnaire；Each user will
The fecal sample collector that the company of receiving sends, one prepares for baby, and one prepares for mother, and user wipes what household provided
Son picks mung bean size excrement and is fitted into containing in the collection tube for saving liquid, and express delivery mode returns center of a sample, furthermore needs user
Two dimensional code on scanning collection device carries out sample binding and questionnaire is filled in, and the acquisition and transport of entire sample are in room temperature
Lower progress, without using ice chest and cold chain transportation, questionnaire problem is shown in Table 1：
Table 1：Questionnaire data
| Questionnaire problem | Problem number | Detailed description |
| Baby's personal information | 5 | Gender, monthly age, height, weight, wean age etc. |
| Mother's personal information | 3 | Age, city of residence, fertility cycle etc. |
| Production method | 1 | Caesarean birth, natural labor |
| Feeding patterns | 6 | Breast-feeding, formula milk nursing, mixed feeding（Milk powder+breast milk）, table meal, table canteen species, whether drink Yoghourt etc. |
| Health status | 13 | Asthma, rhinitis, allergy, inflammatory enteritis, influenza, diarrhea, indigestion, anaemia etc. |
| Medicining condition | 7 | Antibiotic, cold drug, Infantile diarrhea, Medilac-Vita, dioctahedral smectite, cold drug etc. |
| Living environment | 3 | Whether inhabitation crowd has siblings, hygienic habit |
| Stool characteristics | 4 | Defecation frequency, stool color, stool etc.. |
Step 2：16s-rRNA gene sequencing；DNA of bacteria, which extracts, in excrement uses paramagnetic particle method DNA purification technique；Sequencing fragment：
The hypervariable region V3-V4, microarray dataset Illumima MiSeq sequenator；
Step 3：Sequencing data analysis：The original lower machine data of 16s rRNA are analyzed using QIIME v1.7.0 software：Packet
It includes：
（1）Initial data pretreatment：Sample joint sequence is removed, and both-end sequencing sequence is spliced into single sequence, according to
Barcode sequence distinguishes different sample sequences, filters low quality sequence；
（2）OTU（Activity classification unit）Classification and statistics：Using clustering method, according to the similarity threshold of sequence 97% by sequence
Different OTU are divided into, each OTU is considered as a microbial species, and carries out species annotation using SILVA database, in addition,
Quantity is sequenced to each sample and OTU number counts；
（3）Diversity analysis：Analyze the alpha diversity of each sample（Chao1, observed species, Shannon multiplicity
Sex index, Simpson diversity indices）；
（4）Function and metabolic pathway prediction：Using macro genome prediction technique（Tax4Fun-r software package）16s rRNA is carried out
Functionality prediction；
Step 4：Enteric microorganism maturity and enteron aisle cell age calculate：With infant enteric microorganism and its micro- life of mother's enteron aisle
Object similitude represents maturation index（Jaccard likeness coefficient）, specific formula for calculation is as follows：
Jaccard(A,B) indicate enteric microorganism maturity,A Indicate infant OTU number,B Indicate mother OTU number；Root
According to 2000 big-sample datas, linear fit model is constructed, the true enterobacteriaceae of infant is predicted by enteric microorganism maturity
Age：
AgeIndicating infant's enteron aisle cell age, x indicates coefficient, Jaccard(A,B) Indicate enteric microorganism maturity；
Step 5：User report push：After 16s rRNA sequencing analysis is completed, confluence analysis questionnaire data, mother's enteron aisle are micro-
Biological data and child's enteric microorganism data produce report；Examining report will push to mother's cell phone application, user
Report can be checked in mobile phone terminal；Report content includes baby's enteron aisle age, maturation index, the Pseudomonas detected and every kind of bacterium
The abundance of category, beneficial bacterium nocuousness bacterial content and crowd's lateral comparison, prebiotic bacterial content and crowd's lateral comparison, how is important Pseudomonas
Influence baby's health, protein, lipid, carbohydrate metabolism ability, and the baby's health raising side provided according to report
Case and dietary recommendation；Fig. 2 is shown in partial report displaying；
To sum up：Inventive result is summarized：
（1）Enteric microorganism maturity and monthly age relationship：By to 2000 Different Month infant's enteric microorganism maturity
Analysis, the results showed that, enteric microorganism maturity constantly increases with advancing age, and increases quickly 0-12 monthly age,
Increase after 12 monthly ages and gradually slow down, final smooth-out, concrete outcome is shown in Fig. 3（Wherein abscissa indicates infant's monthly age,
Ordinate indicates enteric microorganism maturity）；
It analyzes to 2000 pairs of mother and sons' enteric microorganism alpha diversities, finds 4 kinds of alpha diversity indexes as infant's age increases
And increase, and move closer to the alpha diversity of mother enteric microorganism；Concrete outcome is shown in Fig. 4（Abscissa indicates five groups of crowds, point
Not Wei infant's 0-3 monthly age, 4-8 monthly age, 9-12 monthly age, 13-37 monthly age and mother, ordinate indicates 4 different alpha diversities
Index）；
（2）In order to determine whether production method significantly affects enteric microorganism, constituency 0-3 monthly age sample carries out analysis covariance point
Analysis；26.32% is caesarean birth in sample, and 73.68% is natural labor；The result shows that different production methods significantly affect enteric microorganism
Mature indicator（p=0.04）, the enteric microorganism maturity mean value of surgical neonate is 0.27, lower than natural labor baby 0.04, but
This phenomenon does not embody in other four classes alpha diversity indexs；Concrete outcome is shown in Fig. 5（Abscissa indicates different production methods
（C-section:Caesarean birth, Vaginally born：Natural labor）, A figure ordinate expression enteric microorganism maturity, BCDE figure point
4 class alpha diversity indexes are not indicated）；
（3）Two pairs of mother and sons' enteric microorganism data compare：Choose two pairs of mothers and sons enteric microorganism data of different production methods into
Row compares（See Fig. 6）；Baby was 2 monthly ages at the practical monthly age, one of them is caesarean birth production（Fig. 6-B）, enteric microorganism
Maturity is 0.25；Another is natural labor production（Fig. 6-A）, enteric microorganism maturity is 0.38, and two babies are at this
Significant difference is presented in index；Fig. 6 middle circle indicates the classification of enteric microorganism category level, 4 sample Pseudomonas union numbers
It is 130；Abundance of the red histogram graph representation baby on the Pseudomonas, blue histogram graph representation mother are rich on the Pseudomonas
Degree；Two groups of sample lateral comparisons, it is clear that natural labor baby is more some more than the Pseudomonas number of caesarean birth baby；Outermost layer circle
Different colours indicate the information in bacterium difference gate layer face, wherein it is Firmicutes that all sample accountings are maximum（Firmicutes）
And Proteobacteria（Proteobacteria）, this is consistent with other document reports.
In addition, we show 4 sample abundance rankings near 5 preceding Pseudomonas, the results showed that, either caesarean birth is also
It is natural labor, maximum baby's Pseudomonas abundance is Bifidobacterium, but natural labor baby's Bifidobacterium abundance is 75.26%, is cut open the belly
Producing baby is then 44.10%, is much smaller than natural labor.The Bifidobacterium Pseudomonas important as infant's enteron aisle, plays antiallergy, anti-sense
Dye, improves nutrition, promotes the important function such as immunity adjustment function of intestinal canal.And caesarean birth baby Bifidobacterium is too low, shows it
Body immunity and intestinal health are more much lower than natural labor baby, this is consistent with literature research at this stage, illustrate that the present invention can
To monitor baby's immunity level well.
In summary：Present invention firstly provides this monitoring index of enteric microorganism maturity, enteric microorganism maturity
Refer to infant enteric microorganism and its similitude of mother enteric microorganism in composition, maturity is higher, it is believed that flora hair
It is better to educate, which can react the immunity level of infant, to the assessment of infant enteric microorganism at present to be confined to Pseudomonas more
The various dimensions indexs such as abundance, beneficial bacterial content, harmful bacterial content, from different directions assess intestinal flora, there is no one
Can represent with it is recapitulative, and the index of sustainable detection and measurement reacts infant's intestinal flora development and body immune
Power is horizontal, analyzes by experimental study and data, present invention firstly provides enteric microorganism maturity Testing index, in room temperature ring
Under border, by the method for detection fecal sample 16s rRNA V3-V4 hypervariable region, the assessment of infant enteric microorganism is realized,
Its sampling is convenient, analysis speed is fast, at low cost, and assessment result and existing research are consistent, detect consumer level enteric microorganism
It is possibly realized.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (6)
1. extracting the method that fecal sample 16s rRNA is monitored in infant's intestinal microecology maturity, which is characterized in that including
Following steps：
S1：User applies for item detection to company；
S2：Company audits user information, and sends sample detector to user；
S3：User receives sample detector, carries out fecal sample acquisition, sample binding and questionnaire and fills in；
S4：The pattern detection center of company is returned in fecal sample express delivery by user；
S5：Company's pattern detection center carries out DNA extraction, establishes database；
S6：The sequencing of the hypervariable region V3-V4 is carried out using microarray dataset Illumima MiSeq sequenator；
S7：It is analyzed using QIIME v1.7.0 software lower machine data original to 16s rRNA；
S8：Enteric microorganism maturity calculates, and generates examining report；
S9：It will test report and be in electronic form sent to user hand generator terminal, convenient for checking report.
2. extracting fecal sample 16s rRNA as described in claim 1 in the side that infant's intestinal microecology maturity monitors
Method, which is characterized in that sample collection includes 2000 couples of mothers and sons, totally 4000 fecal samples and corresponding questionnaire, Mei Geyong
The fecal sample collector that the company of receiving is sent at family, one prepares for baby, and one prepares for mother, furthermore user is needed to sweep
The two dimensional code progress sample binding and questionnaire retouched on collector are filled in, and the acquisition and transport of entire sample are at normal temperature
It carries out, without using ice chest and cold chain transportation.
3. extracting fecal sample 16s rRNA as described in claim 1 in the side that infant's intestinal microecology maturity monitors
Method, which is characterized in that DNA, which is extracted, in the step S5 uses paramagnetic particle method DNA purification technique.
4. extracting fecal sample 16s rRNA as described in claim 1 in the side that infant's intestinal microecology maturity monitors
Method, which is characterized in that specific method includes in the step S7：
S701：Initial data pretreatment：Sample joint sequence is removed, and both-end sequencing sequence is spliced into single sequence, according to
Barcode sequence distinguishes different sample sequences, filters low quality sequence；
S702：OTU（Activity classification unit）Classification and statistics：Using clustering method, according to the similarity threshold of sequence 97% by sequence
Column are divided into different OTU, and each OTU is considered as a microbial species, and carries out species annotation using SILVA database, this
Outside, quantity is sequenced to each sample and OTU number counts；
S703：Diversity analysis：Analyze the alpha diversity of each sample（Chao1, observed species, Shannon are more
Sample sex index, Simpson diversity indices）；
S704：Function and metabolic pathway prediction：Using macro genome prediction technique（Tax4Fun-r software package）To 16s rRNA into
The functional prediction of row.
5. extracting fecal sample 16s rRNA as described in claim 1 in the side that infant's intestinal microecology maturity monitors
Method, which is characterized in that represent maturation with infant enteric microorganism and its mother's enteric microorganism similitude in the step S8
Spend index（Jaccard likeness coefficient）, specific formula for calculation is as follows：
Jaccard(A,B) indicate enteric microorganism maturity,A Indicate infant OTU number,B Indicate mother OTU number；Root
According to 2000 big-sample datas, linear fit model is constructed, the true enterobacteriaceae of infant is predicted by enteric microorganism maturity
Age：
AgeIndicating infant's enteron aisle cell age, x indicates coefficient, Jaccard(A,B) Indicate enteric microorganism maturity.
6. extracting fecal sample 16s rRNA as described in claim 1 in the side that infant's intestinal microecology maturity monitors
Method, which is characterized in that the report content includes baby's enteron aisle age, maturation index, the Pseudomonas detected and every kind of bacterium
Abundance, beneficial bacterium nocuousness bacterial content and the crowd's lateral comparison of category, prebiotic bacterial content and crowd's lateral comparison, how is important Pseudomonas
Influence baby's health, protein, lipid, carbohydrate metabolism ability, and the baby's health raising side provided according to report
Case and dietary recommendation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810687215.3A CN108841974A (en) | 2018-06-28 | 2018-06-28 | Extract the method that fecal sample 16s rRNA is monitored in infant's intestinal microecology maturity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810687215.3A CN108841974A (en) | 2018-06-28 | 2018-06-28 | Extract the method that fecal sample 16s rRNA is monitored in infant's intestinal microecology maturity |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108841974A true CN108841974A (en) | 2018-11-20 |
Family
ID=64200602
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810687215.3A Pending CN108841974A (en) | 2018-06-28 | 2018-06-28 | Extract the method that fecal sample 16s rRNA is monitored in infant's intestinal microecology maturity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108841974A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109706235A (en) * | 2019-01-29 | 2019-05-03 | 广州康昕瑞基因健康科技有限公司 | A kind of the detection and analysis method and its system of intestinal microflora |
| CN110097928A (en) * | 2019-04-17 | 2019-08-06 | 广东省微生物研究所（广东省微生物分析检测中心） | A kind of prediction technique and prediction model based on intestinal flora prediction tissue micronutrient levels |
| CN110111897A (en) * | 2019-05-24 | 2019-08-09 | 天益健康科学研究院（镇江）有限公司 | A method of obtaining genital tract flora sequencer address |
| CN110144415A (en) * | 2019-04-23 | 2019-08-20 | 大连大学 | One kind introducing milk cow health and immunity level method based on intestinal flora prediction |
| CN111161794A (en) * | 2018-12-30 | 2020-05-15 | 深圳碳云智能数字生命健康管理有限公司 | Intestinal microorganism sequencing data processing method and device, storage medium and processor |
| CN111383730A (en) * | 2020-03-09 | 2020-07-07 | 圣湘生物科技股份有限公司 | Method, device, equipment and storage medium for generating detection report |
| CN111445957A (en) * | 2020-02-25 | 2020-07-24 | 北京德润良品健康科技有限公司 | Mother-infant flora transfer monitoring system and method |
| CN111986732A (en) * | 2020-07-24 | 2020-11-24 | 贵州师范大学 | Method for analyzing mandarin duck feeding habits by using mandarin duck excrement |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160326574A1 (en) * | 2013-11-01 | 2016-11-10 | Washington University | Methods to establish and restore normal gut microbiota function of subject in need thereof |
| CN107506582A (en) * | 2017-08-16 | 2017-12-22 | 周礼红 | A kind of health risk forecasting system based on enteric microorganism |
-
2018
- 2018-06-28 CN CN201810687215.3A patent/CN108841974A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160326574A1 (en) * | 2013-11-01 | 2016-11-10 | Washington University | Methods to establish and restore normal gut microbiota function of subject in need thereof |
| CN107506582A (en) * | 2017-08-16 | 2017-12-22 | 周礼红 | A kind of health risk forecasting system based on enteric microorganism |
Non-Patent Citations (1)
| Title |
|---|
| 魏向倩: "婴儿生后粪便微生物区系的演变", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111161794A (en) * | 2018-12-30 | 2020-05-15 | 深圳碳云智能数字生命健康管理有限公司 | Intestinal microorganism sequencing data processing method and device, storage medium and processor |
| CN111161794B (en) * | 2018-12-30 | 2024-03-22 | 深圳碳云智能数字生命健康管理有限公司 | Intestinal microorganism sequencing data processing method, device, storage medium and processor |
| CN109706235A (en) * | 2019-01-29 | 2019-05-03 | 广州康昕瑞基因健康科技有限公司 | A kind of the detection and analysis method and its system of intestinal microflora |
| CN110097928A (en) * | 2019-04-17 | 2019-08-06 | 广东省微生物研究所（广东省微生物分析检测中心） | A kind of prediction technique and prediction model based on intestinal flora prediction tissue micronutrient levels |
| CN110097928B (en) * | 2019-04-17 | 2022-03-11 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Prediction method and prediction model for predicting tissue trace element content based on intestinal flora |
| CN110144415A (en) * | 2019-04-23 | 2019-08-20 | 大连大学 | One kind introducing milk cow health and immunity level method based on intestinal flora prediction |
| CN110111897A (en) * | 2019-05-24 | 2019-08-09 | 天益健康科学研究院（镇江）有限公司 | A method of obtaining genital tract flora sequencer address |
| CN111445957A (en) * | 2020-02-25 | 2020-07-24 | 北京德润良品健康科技有限公司 | Mother-infant flora transfer monitoring system and method |
| CN111445957B (en) * | 2020-02-25 | 2024-05-24 | 北京德润良品健康科技有限公司 | Mother and infant flora transfer monitoring system and method |
| CN111383730A (en) * | 2020-03-09 | 2020-07-07 | 圣湘生物科技股份有限公司 | Method, device, equipment and storage medium for generating detection report |
| CN111986732A (en) * | 2020-07-24 | 2020-11-24 | 贵州师范大学 | Method for analyzing mandarin duck feeding habits by using mandarin duck excrement |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108841974A (en) | Extract the method that fecal sample 16s rRNA is monitored in infant's intestinal microecology maturity | |
| CN109706235A (en) | A kind of the detection and analysis method and its system of intestinal microflora | |
| Vallès et al. | Microbial succession in the gut: directional trends of taxonomic and functional change in a birth cohort of Spanish infants | |
| Kadir et al. | Prevalence of Candida species in Turkish children: relationship between dietary ıntake and carriage | |
| CN110144415A (en) | One kind introducing milk cow health and immunity level method based on intestinal flora prediction | |
| Levin et al. | Urban-rural differences in dental caries of 5-year old children in Scotland | |
| CN1840206A (en) | Model construction of human flora-associated piggy and molecular method for detecting flora in intestine tract of baby pig | |
| CN106202989A (en) | A kind of method obtaining child's individuality biological age based on oral microbial community | |
| CN105326852B (en) | Beta glucan is used for upper respiratory tract infection symptoms and mental health | |
| Guo et al. | Distinct stage changes in early-life colonization and acquisition of the gut microbiota and its correlations with volatile fatty acids in goat kids | |
| CN107058490A (en) | The analysis method of enteron aisle and oral cavity flora diversity and otherness | |
| CN102770154B (en) | The probiotic bifidobacterium compositions conformed to secretor blood group state | |
| Foxman et al. | Use of the microbiome in the practice of epidemiology: a primer on-omic technologies | |
| CN111370069A (en) | Human intestinal flora detection method and device and storage medium | |
| CN106446599A (en) | Method for screening oral pathogenic biomarkers of infant caries | |
| CN111334590A (en) | Kit for identifying colorectal cancer and application thereof | |
| Theodosiou et al. | The infant pharyngeal microbiomes: origin, impact and manipulation | |
| Manus et al. | Interactions with alloparents are associated with the diversity of infant skin and fecal bacterial communities in Chicago, United States | |
| CN109008958A (en) | Intestinal flora research method based on fecal filtration and transplantation | |
| CN104997813A (en) | Preparation method of probiotic microecological preparation | |
| Quijada et al. | Dietary supplementation with sugar beet fructooligosaccharides and garlic residues promotes growth of beneficial bacteria and increases weight gain in neonatal lambs | |
| CN107541496A (en) | A kind of human esophageal carcinoma cell line and its application | |
| CN102781466B (en) | The probiotic bifidobacterium compositions conformed to secretor blood group state | |
| Li et al. | Community Structure and Ecological Network's Changes of Vaginal Microbiome in Women Right After Delivery | |
| Smith et al. | Exploring the effects of weaning age on adult infectious disease mortality among 18th–19th century Italians |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181120 |