CN112410443B - Biological population marker for identifying elderly individuals and acquisition method and application thereof - Google Patents
Biological population marker for identifying elderly individuals and acquisition method and application thereof Download PDFInfo
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Abstract
The invention relates to a biological population marker for identifying elderly individuals, the biological population marker comprising: escherichia coli, mycobacterium ondarium Alistipes onderdonkii, mycobacterium sandi Alistipes shahii, mycobacterium vaccae Alistipes finegoldii, mycobacterium Alistipes sp AP11, bacteroides faecalis Bacteroides caccae, enterobacter visceral odor bacillus Odoribacter splanchnicus, bacillus natto Pyramidobacter piscolens, paralopecuroides gordonii Parabacteroides goldsteinii and Pediococcus pseudopolis Subdellovirens sp 4_3_54A2FAA. The biological population markers are determined by comparing, analyzing and verifying the differences of the abundance of various intestinal microorganisms in faecal samples of elderly individuals and non-elderly individuals (healthy children or healthy individuals with equivalent ages), and the probability that the individuals are in an ultra-high age state can be determined by using the biological population markers.
Description
Technical Field
The invention relates to the technical field of biomarkers, in particular to a biological population marker for identifying elderly individuals, and an acquisition method and application thereof.
Background
With the development of social economy, the life expectancy of Chinese is steadily increased, and the proportion of the elderly is continuously increased. Many sub-health states such as debilitation (frailty) and inflammatory aging (scaling) often occur in the elderly, so that the health level of the elderly is improved, and thus the promotion of the social and economic participation of the elderly has become an important subject in the current generation. Among the various options for improving the sub-health status of the elderly, the microecological method is a relatively mild, low invasive and low risk intervention, but this requires a thorough and intensive study of the flora characteristics and other relevant factors of the relevant population, in particular the elderly. The related factors of the advanced age comprise genetics, socioeconomic, environment and the like, and more evidence shows that the intestinal flora is closely related to the advanced age. Intestinal microorganisms play an important role in intestinal epithelial cells, including forming a microbial barrier to prevent colonization by pathogenic bacteria, participation in immunomodulation and metabolic function. There are studies showing that an imbalance in intestinal flora can lead to injury to the body in different forms, and pathogenic microorganisms can cause inflammatory reactions in the intestinal tract by activating recognition receptors, adsorbing, secreting enterotoxins or invading the body. Changes in the number, structure and stability of intestinal microorganisms, especially imbalance in the flora, alter normal physiological functions and thus cause intestinal diseases, affecting health and even life expectancy.
Our macro genome study of elderly people shows that intestinal flora characteristics of elderly individuals show an overall worsening trend compared to control groups, and obvious characteristics include an increase in escherichia coli, a decrease in two main probiotics, bifidobacterium bifidum Faecalibacterium prausnitzii and bifidobacterium longum, and an increase in lipopolysaccharide (lipopolysaccharide) synthesis pathway and an anionic antibacterial peptide (Cationic antimicrobial peptide) resistance pathway in functional elements. Thus, we speculate that among highly enriched microorganisms in elderly individuals, multiple species are closely related to the sub-health status of the elderly. Thus, the senior flora marker not only provides a means for identifying the senior individual, but also can produce a microecological diagnostic intervention preparation for inflammatory aging.
Disclosure of Invention
The invention aims at overcoming the defects in the prior art and provides a biological population marker for identifying an elderly individual, an acquisition method and an identification method thereof.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
a first aspect of the present invention provides a biomarker for identifying elderly individuals, the biomarker comprising:
escherichia coli, oon's further fungus Alistipes onderdonkii, further fungus of the genus Mycobacterium griseus Alistipes shahii, further fungus of the genus Verticillium Alistipes finegoldii, further fungus of the genus Mycobacterium griseus Alistipes sp AP11, bacteroides faecalis Bacteroides caccae, enterobacter visceri Odoribacter splanchnicus, bacillus natto Pyramidobacter piscolens, paralobacter gossypii Parabacteroides goldsteinii and Pediococcus pseudopolis Subdivisii sp 4_3_54A2FAA;
wherein the relative abundance interval of the Escherichia coli is 6.51X10 according to the 95% confidence interval -2 ～1.14×10 -1 The relative abundance interval of the arbuscular mycorrhizal fungi Alistipes onderdonkii is 2.54 multiplied by 10 -2 ～4.81×10 -2 The relative abundance interval of the amygdalina griseus shahii is 1.31 multiplied by 10 -2 ～2.00×10 -2 The relative abundance interval of the Verticillium Var Alistipes finegoldii is 3.66 multiplied by 10 -3 ～6.51×10 -3 The relative abundance interval of the amycolatopsis Alistipes_sp_AP11 is 3.14X10 -3 ～5.71×10 -3 The relative abundance interval of the bacteroides faecalis Bacteroides caccae is 1.53 multiplied by 10 -2 ～3.37×10 -2 The relative abundance interval of the visceral odor bacillus Odoribacter splanchnicus is 8.75x10 -3 ～1.46×10 -2 The relative abundance interval of the fishy smell cone bacillus Pyramidobacter piscolens is 1.06 multiplied by 10 -3 ～1.04×10 -2 The relative abundance interval of the parabacteroides goiter Parabacteroides goldsteinii is 2.79 multiplied by 10 -3 ～7.18×10 -3 The relative abundance interval of the pseudo-micrococcus Subdingranulosum sp_4_3_54A2FAA is 2.61 multiplied by 10 -3 ～7.18×10 -3 。
A second aspect of the present invention provides a method for obtaining a biomarker as described above, comprising the steps of:
s1, determining various microorganism abundance of the biological population markers in a fecal sample of an elderly individual;
s2, respectively comparing the abundance of various microorganisms of the biological population marker determined in the S1 with the abundance of the microorganisms in a control group, and obtaining the biological population marker according to a comparison result;
wherein the control group comprises a group of stool samples from children of the subject and a group of stool samples from healthy subjects consistent with the age of the children of the subject.
Preferably, the step S1 includes:
s1-1, obtaining sequencing data of a nucleic acid sequence in a fecal sample of an individual, wherein the sequencing data comprises a plurality of reads;
s1-2, determining assembly fragments contained by various microorganisms of the biological population markers;
s1-3, respectively determining the abundance of each assembly fragment in the gene set according to the sequencing data, wherein the abundance of each assembly fragment contained in each microorganism of the biological population marker is respectively determined;
s1-4, determining the abundance of various microorganisms of the biological population marker according to the abundance of the determined assembly fragments.
Preferably, the step S1-2 includes:
and respectively comparing the assembled fragments in the gene set with the reference sequences of various microorganisms, and if the similarity between the assembled fragments and the reference sequences of one microorganism is greater than or equal to 90%, the assembled fragments are derived from the microorganism.
Preferably, the abundance of the microorganism is the median or average of the abundance of all assembled fragments comprised by such microorganism.
A third aspect of the present invention provides the use of a biomarker as described above in identifying an elderly individual, the step of identifying an elderly individual comprising:
b1, determining various microbial abundances of the biological population markers in a fecal sample of the individual;
b2, respectively comparing the various microorganism abundances of the biological population markers determined in the B1 with the relative abundance intervals thereof, and determining the state of the individual according to the obtained comparison result;
wherein the states include an advanced state and a non-advanced state.
Preferably, the step B1 includes:
b1-1, obtaining sequencing data of a nucleic acid sequence in a fecal sample of the individual, the sequencing data comprising a plurality of reads;
b1-2, determining assembled fragments comprised by the various microorganisms of the biomarker;
b1-3, respectively determining the abundance of each assembly fragment in the gene set according to the sequencing data, including respectively determining the abundance of each assembly fragment contained by each microorganism of the biological population marker;
b1-4, determining the abundance of various microorganisms of the biological population marker according to the abundance of the determined assembly fragments.
Preferably, the step B1-2 comprises:
and respectively comparing the assembled fragments in the gene set with the reference sequences of various microorganisms, and if the similarity between the assembled fragments and the reference sequences of one microorganism is greater than or equal to 90%, the assembled fragments are derived from the microorganism.
Preferably, the abundance of the microorganism is the median or average of the abundance of all assembled fragments comprised by such microorganism.
Preferably, the step B2 includes:
and B1, if the abundance of each microorganism in the biological population marker determined in the step B1 falls into a relative abundance interval, determining that the state of the individual is an advanced state.
Compared with the prior art, the invention has the following technical effects:
the biological population markers of the invention are determined by the inventor through comparative analysis and verification of the difference of the abundance of various intestinal microorganisms in fecal samples of elderly individuals and non-elderly individuals (healthy children or healthy individuals of comparable ages). The biological population markers are significantly enriched in stool samples in the elderly group compared to stool samples in the control group, wherein the significant enrichment means that the abundance of each microorganism contained in the biological population markers is statistically significantly higher or significantly, substantially higher in the elderly group than in the control group. Thus, the biomarker can determine the probability that an individual is in an ultra-advanced state.
Drawings
FIG. 1 is a schematic illustration of a test assay procedure for screening for identification of biomarkers in an embodiment of the invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other.
The invention is further described below with reference to the drawings and specific examples, which are not intended to be limiting.
Example 1
Biological markers are cellular, biochemical or molecular changes that can be detected from biological media. Biological agents include various body fluids, tissues, cells, feces, hair, breath, and the like.
The abundance of a microorganism refers to the abundance of that microorganism in a population of microorganisms, e.g., the abundance of that microorganism in a population of intestinal microorganisms, and can be expressed as the content of that microorganism in the population.
A first aspect of the present embodiments provides a biomarker for identifying elderly individuals, the biomarker comprising:
escherichia coli, oon's further fungus Alistipes onderdonkii, further fungus of the genus Mycobacterium griseus Alistipes shahii, further fungus of the genus Verticillium Alistipes finegoldii, further fungus of the genus Mycobacterium griseus Alistipes sp AP11, bacteroides faecalis Bacteroides caccae, enterobacter visceri Odoribacter splanchnicus, bacillus natto Pyramidobacter piscolens, paralobacter gossypii Parabacteroides goldsteinii and Pediococcus pseudopolis Subdivisii sp 4_3_54A2FAA;
wherein the relative abundance interval of the Escherichia coli is 6.51X10 according to the 95% confidence interval -2 ～1.14×10 -1 The relative abundance interval of the arbuscular mycorrhizal fungi Alistipes onderdonkii is 2.54 multiplied by 10 -2 ～4.81×10 -2 The relative abundance interval of the amygdalina griseus shahii is 1.31 multiplied by 10 -2 ～2.00×10 -2 The relative abundance interval of the Verticillium Var Alistipes finegoldii is 3.66 multiplied by 10 -3 ～6.51×10 -3 The relative abundance interval of the amycolatopsis Alistipes_sp_AP11 is 3.14X10 -3 ～5.71×10 -3 The relative abundance interval of the bacteroides faecalis Bacteroides caccae is 1.53 multiplied by 10 -2 ～3.37×10 -2 The relative abundance interval of the visceral odor bacillus Odoribacter splanchnicus is 8.75x10 -3 ～1.46×10 -2 The relative abundance interval of the fishy smell cone bacillus Pyramidobacter piscolens is 1.06 multiplied by 10 -3 ～1.04×10 -2 The relative abundance interval of the parabacteroides goiter Parabacteroides goldsteinii is 2.79 multiplied by 10 -3 ～7.18×10 -3 The relative abundance interval of the pseudo-micrococcus Subdingranulosum sp_4_3_54A2FAA is 2.61 multiplied by 10 -3 ～7.18×10 -3 。
The biological population markers of the invention are determined by the inventor through comparative analysis and verification of the difference of the abundance of various intestinal microorganisms in fecal samples of elderly individuals and non-elderly individuals (healthy children or healthy individuals of comparable ages). The biological population markers are significantly enriched in stool samples in the elderly group compared to stool samples in the control group, wherein the significant enrichment means that the abundance of each microorganism contained in the biological population markers is statistically significantly higher or significantly, substantially higher in the elderly group than in the control group. Thus, the biomarker can determine the probability that an individual is in an ultra-advanced state.
The embodiment provides an application of the biological population marker in identifying elderly individuals, wherein the step of identifying the elderly individuals comprises the following steps:
b1, determining various microbial abundances of the biological population markers in a fecal sample of the individual:
obtaining sequencing data of a nucleic acid sequence in a fecal sample of the individual, the sequencing data comprising a plurality of reads; determining assembly fragments comprised by each microorganism in the biomarker; determining the abundance of each assembly segment in the gene set according to the sequencing data, wherein the method comprises the steps of determining the abundance of each assembly segment contained in each microorganism in the biological population marker; and determining the abundance of each microorganism in the biological population marker according to the determined abundance of the assembled fragments.
The sequencing data is obtained by sequencing the nucleic acid sequence in the sample, and the sequencing can be selected from, but is not limited to, semiconductor sequencing technology platforms such as PGM, ion Proton, BGISEQ-100 platform, sequencing-while-synthesis technology platforms such as Hiseq, miseq sequence platform of Illumina company and single molecule real-time sequencing platform such as PacBio sequence platform. The sequencing mode can be single-ended sequencing or double-ended sequencing, and the obtained off-machine data is the fragment which is read out and called reads.
The assembly fragments contained in various microorganisms in the determined biological population markers are judged whether the assembly fragments are from a microorganism according to the similarity degree of the assembly fragments and a microorganism reference sequence by performing Blat alignment on the assembly fragments in the gene set and the microorganism reference sequence. The reference sequence refers to a predetermined sequence, and may be any reference template of a biological class to which a sample to be tested belongs or is included in the sample to be tested, for example, if the target is a microorganism in the sample to be tested, the reference sequence may select a reference genome of various microorganisms in an NCBI database and/or a DACC intestinal genome disclosed in HMP and MetaHIT projects, further, a resource library including more reference sequences may be preconfigured, for example, a sequence which is closer to each other is selected or determined to be assembled according to factors such as a state, a region, and the like of an individual from which the sample to be tested is derived as the reference sequence. Determining the assembled fragments comprised by each microorganism in the biomarker comprises: and comparing the assembled fragments in the gene set with the reference sequences of various microorganisms respectively, and determining that the assembled fragments with the similarity of more than or equal to 90% with the reference sequences of one microorganism are from the microorganism. More strictly, it is determined that assembled fragments with greater than or equal to 95% similarity to the reference sequence of a microorganism are derived from that microorganism.
The alignment can be performed by using known alignment software, such as SOAP, BWA, teraMap, etc., and in the process of the alignment, the alignment parameters are generally set, one or a pair of reads (reads) is set to allow s base mismatches (mismatch) at most, for example, s is set to be less than or equal to 2, and if there are more than s bases in the reads that are mismatched, the reads cannot be aligned to the assembled fragment. The obtained comparison result contains comparison conditions of each reading segment and each assembly segment, including information such as whether the reading segment can compare with one or some assembly segments, whether only one assembly segment is compared with a plurality of assembly segments, whether the reading segment is compared with the assembly segments, and whether the reading segment is compared with the assembly segments. The comparison was performed using a SOAPalign 2.21, setting the parameters to-r 2-m 100-x 1000.reads can be aligned with gene sets in two parts: a) Uniquely comparing the read sections of the previous assembly fragment, and calling the read sections as unique reads (U); b) The reads were termed multiplex reads (M) by aligning the multiple assembled fragments. For a given assembled fragment G, i.e., gene G in the gene set, its abundance is Ab (G), which is associated with the unique reads and multiple reads, ab (U) and Ab (M) in the above formula are the abundance of unique reads and multiple reads, respectively, of the assembled fragment G. Each multiplex reads, having a unique gene abundance coefficient Co, can be calculated using the following formula assuming that one multiplex read is aligned with the upper N assembled fragments:that is, for such multiplexing, the abundance of unique reads of the N genes (i.e., assembled fragments of the gene set) to which they are alignedAnd as denominator.
According to one embodiment of the invention, in the step of determining the abundance of each microorganism in the biomarker, said abundance being the median or average of the abundance of all the assembled fragments comprised by that microorganism, respectively, depending on the determined abundance of the assembled fragments.
B2, respectively comparing the various microorganism abundances of the biological population markers determined in the B1 with the relative abundance intervals thereof, and determining the states of the individuals according to the obtained comparison result, wherein the states comprise an advanced state and a non-advanced state:
if a predetermined confidence interval for the abundance of a microorganism in the biomarker population has an intersection with a predetermined confidence interval for the abundance in the elderly group and a predetermined confidence interval for the abundance in the control group, then the predetermined interval for that microorganism is determined to be part of the difference set of the two. The confidence interval refers to an estimated interval of the overall parameter constructed from the sample statistics. In statistics, the confidence interval (confidence interval) of a probability sample is an interval estimate of some overall parameter of the sample. The confidence interval reveals the extent to which the true value of this parameter falls around the measurement with a certain probability. The confidence interval gives the degree of confidence in the measured value of the parameter being measured, i.e. the "certain probability" as required before. This probability is referred to as the confidence level. According to an embodiment of the invention, the predetermined confidence intervals are all 95% confidence intervals, and the predetermined intervals of the Escherichia coli in the biological population markers are 6.51X10% -2 ～1.14×10 -1 The predetermined interval of the arbuscular kansui Alistipes onderdonkii in the biological population marker is 2.54 multiplied by 10 -2 ～4.81×10 -2 The method comprises the steps of carrying out a first treatment on the surface of the The predetermined interval of Bacteroides faecalis Bacteroides caccae in the biological population marker is 1.53X10 -2 ～3.37×10 -2 The method comprises the steps of carrying out a first treatment on the surface of the The preset interval of the microorganism Alistipes shahii in the biological population marker is 1.31X10% -2 ～2.00×10 -2 The method comprises the steps of carrying out a first treatment on the surface of the The predetermined interval of the visceral odor bacillus Odoribacter splanchnicus in the biological population marker is 8.75x10 -3 ～1.46×10 -2 . It should be noted that, depending on the purpose or requirement, there may be different requirements for determining the confidence level of the individual status result, and those skilled in the art may select different significance levels (α), i.e. different probabilities of possible mistakes, so that the determined individual status has a confidence level of 1- α. For example, using the present embodiment, it is determined that an individual is 95% reliable in the determined state.
The method is based on detecting the abundance of various microorganisms in biological population markers in a stool sample of an individual, comparing the abundance of various microorganisms determined by detection with the abundance of the various microorganisms in a control group, and determining the probability that the individual is an elderly individual or a non-elderly individual according to the obtained comparison result.
All or part of the steps of any of the above methods for determining the status of an individual using a biomarker may be performed using an apparatus/system comprising detachable corresponding unit function modules, or the method may be programmed, stored on a machine-readable medium, which is run using a machine.
An apparatus for determining the status of an individual using the biomarker in any of the above embodiments of the present invention is provided according to an embodiment of the present invention, the apparatus being configured to implement all or part of the steps of the method for determining the status of an individual using the biomarker in any of the above embodiments of the present invention, the apparatus comprising: a data input unit for inputting data; a data output unit for outputting data; a processor for executing an executable program, the executing the executable program including performing the method for determining the status of an individual in the above-described embodiment of the present inventor; and the storage unit is connected with the data input unit, the data output unit and the processor and is used for storing data, wherein the executable program is included. The above description of the technical features and advantages of the method for determining the status of an individual using a biomarker in any embodiment of the present invention also applies to the apparatus of this aspect of the present invention, and will not be described in detail here.
According to another embodiment of the present invention, there is provided a method for classifying a plurality of individuals using the biomarker for a biological population according to any of the embodiments of the present invention described above, the method comprising: determining the status of each individual by using the method for determining the status of an individual in any of the embodiments of the present invention described above, respectively; and classifying each individual according to the obtained state of each individual. The method can distinguish a plurality of individuals or a plurality of unknown stool samples according to different states of the individuals, and is convenient for classification and marking management. In addition, the above description of the technical features and advantages of the method for determining the status of an individual using the biomarker in any embodiment of the present invention also applies to the apparatus of this aspect of the present invention, and will not be described in detail herein.
The medicine or the functional food of the embodiment is reasonably and effectively applied to the determined biological population markers, supports the growth of beneficial intestinal bacteria and/or inhibits potential pathogenic bacteria of the intestinal tract, can prevent the defect of the intestinal barrier, improves and restores the intestinal microecological structure, and has important significance for assisting in reducing the blood endotoxin level and/or improving the sub-health state of the old.
The method for producing or screening the microecology capable of improving the health level and the life quality of the old can be used for screening by reasonably and effectively applying the determined biological population markers, can obtain the medicine capable of supporting the growth of beneficial intestinal bacteria and/or inhibiting potential pathogenic bacteria of the intestinal tract, can prevent the defect of the intestinal barrier, improves and restores the microecological structure of the intestinal tract, and has important significance for assisting in reducing the blood endotoxin level and/or relieving the sub-health state.
The method and/or apparatus of the present invention will be described in detail below with reference to specific examples. Unless otherwise indicated, the reagents, sequences (adaptors, tags and primers), software and instrumentation involved in the examples below are conventional commercial products or open source, such as the transcriptome library construction kit purchased for Illumina.
Example 2
In this example, the inventors developed a correlation analysis of intestinal microbiota composition from fecal samples of 116 elderly people and 232 orthodontics and 77 healthy individuals of comparable ages to elderly people. In general, the sequencing data for each sample in the control group described above was downloaded for a total of about 1084.87Gb. Quantitative metagenomic analysis showed 14,993 genes exhibited significant differences (FDR < 0.05) in a large number of patients and healthy controls.
1. Subject recruitment
116 elderly people and 232 children thereof (1-2 children for each elderly person) were recruited from Jiangsu Qidong people hospitals, and 77 healthy controls corresponding to the ages of the elderly people were recruited by Shanghai tenth people hospitals. Fecal samples were collected from subjects, each sample yielding on average 5Gb high quality sequencing data.
2. Determination and identification of biological population markers
Referring to the experimental procedure of fig. 1, identification of relevant biomarkers of advanced age was determined.
2.1 basic processing of sequencing data
Sequencing raw data has been subjected to quality control and de-hosting, but there are many short reads in the data, pairs of reads less than 60nt in length in the raw data are filtered.
3. Screening for biological population markers
All 116 elderly samples and 232 child samples were divided into experimental and control group 1. There were 61 species of significant difference between the control and the elderly group found using a threshold (fdr < 0.05). Of these, 40 were enriched in the elderly and 21 were enriched in the control group.
As can be seen from Table 1, escherichia coli and further Escherichia coli Alistipes onderdonkii have intersections between 95% confidence intervals of abundance in the offspring group and in the elderly individual group, and when individual status determination is performed using a marker containing all or a part of these three, the intersection part of 95% confidence intervals of abundance in the two groups of these three microorganisms is removed to obtain a predetermined interval judged by comparison based on their abundance, the predetermined interval of Escherichia coli being 6.51X10 -2 ～1.14×10 -1 The predetermined interval of the arbuscular mycorrhizal fungi Alistipes onderdonkii is 2.54 multiplied by 10 -2 ～4.81×10 -2 。
Table 1 comparison of the senior group with the offspring group enriched species in all senior individuals
Example 3
To verify that the species markers determined in example 1 were able to act as biological population markers, 116 elderly people and 77 healthy individuals of the same age as the children of the elderly people were used for verification. Wherein the determination of the abundance of a species is performed with reference to the steps of the above examples. The resulting differential species were cross-validated against the results obtained in example 1, and the resulting intersection species were the species group marker from which the most abundant 10 species were selected in this patent.
In addition, the inventors further carried out a state verification test on a large number of stool samples of elderly people with any one, two, three or all four, three combinations and four combinations of the other 4 microorganisms determined in example 1 as markers, wherein the state can be judged to be identical to or more than 90% of the recorded state by the method of this example.
The foregoing description is only illustrative of the preferred embodiments of the present invention and is not to be construed as limiting the scope of the invention, and it will be appreciated by those skilled in the art that equivalent substitutions and obvious variations may be made using the description and illustrations of the present invention, and are intended to be included within the scope of the present invention.
Claims (10)
1. A biomarker for identifying elderly individuals, the biomarker comprising:
escherichia coli, oon's further fungus Alistipes onderdonkii, further fungus of the genus Mycobacterium griseus Alistipes shahii, further fungus of the genus Verticillium Alistipes finegoldii, further fungus of the genus Mycobacterium griseus Alistipes sp AP11, bacteroides faecalis Bacteroides caccae, enterobacter visceri Odoribacter splanchnicus, bacillus natto Pyramidobacter piscolens, paralobacter gossypii Parabacteroides goldsteinii and Pediococcus pseudopolis Subdivisii sp 4_3_54A2FAA;
wherein the relative abundance interval of the Escherichia coli is 6.51X10 according to the 95% confidence interval -2 ～1.14×10 -1 The relative abundance interval of the arbuscular mycorrhizal fungi Alistipes onderdonkii is 2.54 multiplied by 10 -2 ～4.81×10 -2 The relative abundance interval of the amygdalina griseus shahii is 1.31 multiplied by 10 -2 ～2.00×10 -2 The relative abundance interval of the Verticillium Var Alistipes finegoldii is 3.66 multiplied by 10 -3 ～6.51×10 -3 The relative abundance interval of the amycolatopsis Alistipes_sp_AP11 is 3.14X10 -3 ～5.71×10 -3 The relative abundance interval of the bacteroides faecalis Bacteroides caccae is 1.53 multiplied by 10 -2 ～3.37×10 -2 The relative abundance interval of the visceral odor bacillus Odoribacter splanchnicus is 8.75x10 -3 ～1.46×10 -2 The relative abundance interval of the fishy smell cone bacillus Pyramidobacter piscolens is 1.06 multiplied by 10 -3 ～1.04×10 -2 The relative abundance interval of the parabacteroides goiter Parabacteroides goldsteinii is 2.79 multiplied by 10 -3 ～7.18×10 -3 The relative abundance interval of the pseudo-micrococcus Subdingranulosum sp_4_3_54A2FAA is 2.61 multiplied by 10 -3 ～7.18×10 -3 。
2. A method of obtaining a biomarker for a biological population according to claim 1, comprising the steps of:
s1, determining various microorganism abundance of the biological population markers in a fecal sample of an elderly individual;
s2, respectively comparing the abundance of various microorganisms of the biological population marker determined in the S1 with the abundance of the microorganisms in a control group, and obtaining the biological population marker according to a comparison result;
wherein the control group comprises a group of stool samples from children of the subject and a group of stool samples from healthy subjects consistent with the age of the children of the subject.
3. The method according to claim 2, wherein the step S1 comprises:
s1-1, obtaining sequencing data of a nucleic acid sequence in a fecal sample of an individual, wherein the sequencing data comprises a plurality of reads;
s1-2, determining assembly fragments contained by various microorganisms of the biological population markers;
s1-3, respectively determining the abundance of each assembly fragment in the gene set according to the sequencing data, wherein the abundance of each assembly fragment contained in each microorganism of the biological population marker is respectively determined;
s1-4, determining the abundance of various microorganisms of the biological population marker according to the abundance of the determined assembly fragments.
4. The method according to claim 3, wherein the step S1-2 comprises:
and respectively comparing the assembled fragments in the gene set with the reference sequences of various microorganisms, and if the similarity between the assembled fragments and the reference sequences of one microorganism is greater than or equal to 90%, the assembled fragments are derived from the microorganism.
5. The method according to claim 2, wherein the abundance of the microorganism is the median or average of the abundance of all assembled fragments comprised by the microorganism.
6. Use of a biomarker according to claim 1, in the identification of elderly individuals, wherein the step of identifying elderly individuals comprises:
b1, determining various microbial abundances of the biological population markers in a fecal sample of the individual;
b2, respectively comparing the various microorganism abundances of the biological population markers determined in the B1 with the relative abundance intervals thereof, and determining the state of the individual according to the obtained comparison result;
wherein the states include an advanced state and a non-advanced state.
7. The use according to claim 6, wherein said step B1 comprises:
b1-1, obtaining sequencing data of a nucleic acid sequence in a fecal sample of the individual, the sequencing data comprising a plurality of reads;
b1-2, determining assembled fragments comprised by the various microorganisms of the biomarker;
b1-3, respectively determining the abundance of each assembly fragment in the gene set according to the sequencing data, including respectively determining the abundance of each assembly fragment contained by each microorganism of the biological population marker;
b1-4, determining the abundance of various microorganisms of the biological population marker according to the abundance of the determined assembly fragments.
8. The use according to claim 7, wherein said step B1-2 comprises:
and respectively comparing the assembled fragments in the gene set with the reference sequences of various microorganisms, and if the similarity between the assembled fragments and the reference sequences of one microorganism is greater than or equal to 90%, the assembled fragments are derived from the microorganism.
9. The use according to claim 6, wherein the abundance of a microorganism is the median or average of the abundance of all assembled fragments comprised by that microorganism.
10. The use according to claim 6, wherein step B2 comprises:
and B1, if the abundance of each microorganism in the biological population marker determined in the step B1 falls into a relative abundance interval, determining that the state of the individual is an advanced state.
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