CN104195146A - Microbial marker of liver cirrhosis, and application - Google Patents
Microbial marker of liver cirrhosis, and application Download PDFInfo
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Abstract
The invention discloses a microbial marker of liver cirrhosis, and an application. The microbial marker is one or more selected from one microorganism of Veillonella atypica ACS-134-V-Col7a enriched in a patient group suffering the liver cirrhosis or two microorganisms of Bacteroides uniformis ATCC8492 and Clostridiales enriched in a healthy group. The invention also relates to a drug for treating the liver cirrhosis. A drug for treating the liver cirrhosis can promote or increase one or two selected from the two microorganisms of Bacteroides uniformis ATCC8492 and Clostridiales enriched in the healthy group, and/or suppress or clear the microorganism of Veillonella atypica ACS-134-V-Col7a enriched in the patient group suffering the liver cirrhosis. The method detects relative abundance of the microbial marker in intestinal flora and compares the obtained relative abundance value and a predetermined cutoff value. The invention also relates to a beneficial microorganism combination or functional food. The beneficial microorganism combination or functional food comprises one or two of the two microorganisms of Bacteroides uniformis ATCC8492 and Clostridiales enriched in the healthy group. The invention also relates to a kit for detecting the liver cirrhosis, monitoring a treatment process, or producing and screening the drug. The kit is used for detecting the microbial marker.
Description
Technical field
The present invention relates to genetically engineered and biomedicine field, be specifically related to microbial markers and the application thereof of liver cirrhosis.
Background technology
Liver cirrhosis (Liver cirrhosis) is clinical common chronic progressive external hepatopathy, or diffusivity hepatic injury that repeated action form long-term by one or more causes of disease.Liver cirrhosis is the pathology in late period of many hepatic diseases, is by due to a variety of causes such as viral hepatitis, alcoholism, malnutrition, cholestasis, schistosomicide, cycle penalty, is characterized in hepatocellular degeneration and necrosis.Early stage liver cirrhosis by timely control, can reverse or no longer progress, and late period by irreversible, have a strong impact on patient's quality of life, even threat to life.
In China, the modal cause of disease is viral hepatitis, is mainly hepatitis B, is secondly hepatitis C.And the harm of these cirrhotic complications such as spontaneous peritonitis, liver ascites, hepatorenal syndrome, hepatogenic encephalopathy, esophageal varix, hemorrhage, primary hepatocarcinoma is also obvious to all.In the face of modern so many treatment means, but there is a big chunk liver cirrhosis patient still agonizing, still suffering sensual torment, psychological fear and spiritual wrecking every day.
Liver cirrhosis patient is due to portal hypertension, and liver function obstacle causes abnormal bile secretion, and detoxification ability declines, and the factors such as host resistance reduction cause intestinal barrier function to be destroyed, and microbial environment changes, alteration of intestinal flora.Yet the phylogeny of enteric microorganism and the variation of functional component relevant to liver cirrhosis process are not clear.Although some studies (Garcia et al2004, Wiest et al2012, Bass et al2010) change that shows enteric microorganism play an important role in cirrhotic complications in latter stage (as spontaneous bacterial peritonitis and hepatogenic encephalopathy), and induce early stage hepatic diseases and promote liver injury effect (as alcoholic liver disease and non-alcoholic fatty liver disease), the clear and definite associated still the unknown between intestinal microflora and people's hepatic pathology.There is research (Yi JH et al1999, Paik YH et al2003) show to be due to Intestinal Mucosal Injury in Patients Undergoing bacterium excessive multiplication and to produce intracellular toxin, suppress intestinal epithelial cells protein synthesis, there is morphologic change in intestinal microvillus, intestinal cells plasma membrane is abnormal, cytoskeleton weave construction changes, on IBB carrier positions or cytoskeleton, carrier anchor point changes, cause amino acid and carbohydrate passing through the transportation defect of jejunum brush border film, yet its concrete mechanism await further research.
Liver cirrhosis and patients with alcoholic liver disease have disclosed the similar substantial change (Chen Y et al2011, Yan A W.et al2011) in an enteric microorganism by 16S rRNA order-checking research.In enteric microorganism, how phylogeny mechanism changes still unintelligible.
Summary of the invention
The present invention aims to provide the knowledge that more Intestinal Flora in Patients with Cirrhosis are modified, and provides microbial markers targetedly, for early discovery disease provides a kind of noninvasive method.Using microbe mark rationally and effectively, help the growth of intestinal beneficial bacterium (as bifidus bacillus), suppress enteron aisle potentially pathogenic organism (as enterobacteriaceae lactobacteriaceae) etc., can stop the damaged of gut barrier, improve and recover intestinal microecology structure, for reducing blood level of endotoxin, the clinical symptom that alleviates liver cirrhosis is significant.
The object of this invention is to provide liver cirrhosis microbial markers and application.
A microbial markers, is selected from one or more in the microorganism Veillonella atypica ACS-134-V-Col7a of enrichment in liver cirrhosis patient group or two kinds of microorganism Bacteroides uniformis ATCC8492, Clostridiales of the enrichment of healthy group.Enrichment refers to by missing gene finds significant difference gene, and homology is greater than the cluster that 95% gene carries out.
Described microbial markers has the microorganism that is selected from enrichment in liver cirrhosis patient group simultaneously, and is selected from one or both in two kinds of microorganisms of healthy group enrichment.
The medicine for the treatment of liver cirrhosis, described medicine promotes or increases described one or both in healthy group enrichment two kinds of microorganism Bacteroides uniformis ATCC8492, Clostridiales of being selected from, and/or suppresses or remove the described microorganism Veillonella atypica ACS-134-V-Col7a that is selected from enrichment in liver cirrhosis patient group.
The method of production or screening of medicaments, described medicine promotes or increases described one or both in healthy group enrichment two kinds of microorganism Bacteroides uniformis ATCC8492, Clostridiales of being selected from, and/or suppresses or remove the described microorganism Veillonella atypica ACS-134-V-Col7a that is selected from enrichment in liver cirrhosis patient group.
Before described pharmacological agent or intervention, in detected object intestinal microflora, whether be enriched with described microbial markers.
Described detecting step comprises:
A, from object ight soil, extract DNA sample;
B, for DNA sample, build sequencing library;
C, sequencing library is carried out to high-flux sequence, obtain sequencing result;
D, according to sequencing result, determine in the intestinal microflora of described object, whether there is described microbial markers.
Described steps d further comprises: the relative abundance for the microbial markers in intestinal microflora detects, and by resulting relative abundance value and predetermined cutoff value, compares.
In step c, according to sequencing result, obtain gene clustering method, propose the method for a kind of grand gene species (MGS).
Beneficial microorganism combination or functional foodstuff, one or both in two kinds of microorganism Bacteroides uniformis ATCC8492, Clostridiales that contain the health group enrichment described in being selected from.
Detect liver cirrhosis, monitor therapy process, or the test kit of production, screening of medicaments, for detection of described microbial markers.
Beneficial effect of the present invention: diagnosis or treatment liver cirrhosis patient are provided, or susceptible liver cirrhosis crowd's microbial markers, can effectively monitor Susceptible population's (comprising that familial inheritance and foeign element cause) of liver cirrhosis or find early stage patient, and the result for the treatment of of monitoring liver cirrhosis; The medicine that this microbial markers is used for the treatment of to liver cirrhosis, or produce or screen the pharmaceutical methods for the treatment of liver cirrhosis; The beneficial microorganism combination or the functional foodstuff that utilize microbial markers are provided; Test kit utilizing microbial markers etc. is provided.
Accompanying drawing explanation
Fig. 1 is experimental analysis schematic flow sheet;
Fig. 2 a, Fig. 2 b are that grand genome species taxonomy is learned schematic diagram;
The MGS of Chinese liver cirrhosis patient and healthy population enrichment in 2a.Gene in a MGS, if after blastN and the contrast of known genome database, if the coverage >90% of two sequences in contrast, the similarity >95% of cover part, so just just classifies this to the genome of being compared this MGS.Do not reach the MGS of threshold values, be referred to minimum categorization levels, at least 80% gene is consistent with best BlastP classification; This standard is applicable to all higher levels of classification.
In 2b, the classification classification of 58 samples is associated with Dane's enteron aisle gene enrichment phase.
Embodiment
Below in conjunction with drawings and Examples, embodiment of the present invention are described in detail, but it will be understood by those skilled in the art that following drawings and Examples are only for the present invention is described, rather than the restriction to scope of the present invention.With the following detailed description of preferred embodiment, it is obvious that various objects of the present invention and favourable aspect will become to those skilled in the art with reference to the accompanying drawings.In the present invention, except as otherwise noted, otherwise Science and Technology noun used herein has the implication that those skilled in the art understand conventionally.And various laboratory operation steps used herein are widely used conventional steps in corresponding field.
In embodiment, fecal microorganism group and functional component feature are described in the association analysis research that we carry out whole intestinal microflora microorganism from the faecal samples of 98 liver cirrhosis patients, 83 normal healthy controls.Generally speaking, we obtain the high-quality sequencing data of about 860Gb, have built liver cirrhosis reference gene collection.This gene set comprises 2,690,000 genes, and the 36.1%th, before unpub.Quantitative grand genome analysis is presented in a large amount of patients and normal healthy controls group, and 75,245 genes present significant difference (fdr<0.0001).The gene of most can classify as 66 gene clusters that represent bacterial species, wherein in liver cirrhosis patient group main enrichment be 1 MGS (metagenomic species, table 3), that main enrichment is 2 MGS (table 2) in healthy group.The first stage of embodiment is the discovery stage: the enteric microorganism composition of 98 liver cirrhosis patients and 83 normal healthy controls groups and changing function stage; Subordinate phase is Qualify Phase: the accuracy of 25 liver cirrhosis patients and 31 normal healthy controls group checking first stage results.
Embodiment 1: sample collection and DNA extraction
Liver cirrhosis patient is from the first affiliated hospital of Hangzhou Zhejiang University, coupling normal healthy controls group is volunteer, experiment has gathered 181 routine faecal samples altogether, the faecal samples of 98 Chinese liver cirrhosis patients and 83 normal Chineses' faecal samples, wherein each individual fresh excreta sample is divided into 200mg/ part, totally 5 parts, immediately-80 ℃ of refrigerator freezings are preserved.
In the faecal samples of 98 Chinese liver cirrhosis patients and 83 normal Chineses' faecal samples, extract total DNA.Phenol trichloromethane is processed and is extracted DNA method extraction DNA.
Embodiment 2: build library and order-checking
DNA builds storehouse and is undertaken by the operational guidance of apparatus manufacturer (Illumina).PE2*100bp order-checking is carried out in library.Illumina HiSeq2000 (Illumina, San Diego, CA) platform checks order to the library of 181 samples.Each sample mean produces 4.74Gb (sd. ± 2.04Gb) high quality sequencing result, amounts to 858Gb sequencing data amount.
With reference to the experiment flow of Fig. 1, identify the related microorganisms mark of liver cirrhosis, wherein elliptical step or details are well known to those skilled in the art, and several important steps are described below described in the several embodiment of face.
Embodiment 3: the evaluation of biomarker
The base conditioning of 3.1 sequencing datas
After obtaining the sequencing data of first-phase 181 samples, it is filtered, Quality Control is undertaken by following standard: a) remove the reads that is greater than 3 N bases; B) remove the N50reads of inferior quality (Q20); C) remove and surpass the base of 10 inferior quality (Q2) or specify afterbody N base number.The sequence of losing paired reads is considered to wall scroll reads for assembling.
3.2 obtain a set of liver cirrhosis microorganism group gene set
Grand genome biomarker main body is gene and corresponding function, therefore need to assemble and predictive genes sequencing sequence, and de-redundancy, builds nonredundancy with reference to gene set.With SOAPdenovo software, all samples reads is assembled into contigs (assembling fragment).The unassembled reads of sample is merged and carries out the assembling from new (de novo).By 61.68% of total reads number, produce 4,400,000 contigs (minimal segment length is 500bp) eventually.These contigs overall lengths 11.1Gb, N50 length range is 1,673～48,822bp, mean length is 8,644bp.
In order to predict each sample microbial gene of 181 samples, we adopt the method in MetaHIT human intestine genome research.MetaGene program is greater than the open reading frame (ORFs) of 100bp from predicting 13,371,697 length.The ORFs overall length of prediction is 9,495,923,532bp, accounts for 90.28% of contigs total length.In ORFs, 1,047,855 (54.6%) is complete gene, and 869,808 (45.4%) is incomplete.By removing unnecessary ORFs, set up nonredundancy " LC gene set ", be defined as pairing rear consistent with the short ORFs of 90% pairing over 95%.Final nonredundancy liver cirrhosis enteron aisle gene set comprises 2,668,468 ORFs, and mean length 750bp, 42% reads can compare gene pool.
By our LC gene set and 3 other enteric microorganism gene sets contrasts, MetaHIT, HMP and T2D.During comparison, all predictive genes are used identical standard.MetaHIT storehouse comprises 3,452,726 genes, and HMP is containing 4,768,112 genes, and T2D is containing 2,148,029 gene.Four gene sets have 674,131 total genes.LC, MetaHIT, HMP, T2D gene set comprise respectively 794,647, Isosorbide-5-Nitrae 29,517,2,620,096,623,570 unique gene.
Gene and nonredundancy gene set from LC, T2D, MetaHIT gene pool merge, and analyze subsequently.Do not comprise HMP enteron aisle gene set, because it comprises sanger, 454 or Illumina16S sequence, except whole grand genomic data, it is produced by control group healthy population, but not patient.Storehouse is containing 5,382,817 genes, and wherein 797,690 is that other three gene pools are total.Gene in LC gene set has 63.9% to be present in other one or two gene sets, and the 37.1%th, unique (unique) gene.In LC gene set and T2D gene set, there is great difference in Chinese's gene.
3.3 biological abundance analyses
SOAPalign2.21 for coupling for the genomic paired-end clean of redundancy reads, parameter Wei – r2 – m200 – x1000.Reads and redundancy genome alignment, may be divided into two portions: a) Unique reads (U): reads only with a genome alignment; These genes are defined as unique reads.B) Multiple reads (M): if these genomes same species, reads and more than one genome alignment; We are defined as unique reads by these reads.If different plant species, we are defined as multiple reads.
For species S, if abundance is Ab (S), may be relevant with the shared reads of M to the peculiar reads of U, assessment mode is as follows:
Ab(S)＝Ab(U)+Ab(M)
Ab(U)＝U/l
Ab (U) and Ab (M) are respectively the abundance of unique and multiple, and l represents genome length.Each multiple reads, has special species coefficient Co; Make we suppose M} has relevant N in different plant species, then calculates by the following method CO:
For these reads, we add the unique abundance of N as standard.
3.4 gene enrichment analysis
When terminal gene can calculate to it abundance of gene on same gene.If the end that matches on a gene only has a read to align, by inspection by previous unjustified translational domain or do not read read with assembling sequence mate.Matched if the gene number reading has been verified, if do not had, so just ignored.
When calculating the abundance of gene, we use the strategy that biological enrichment analysis is identical.For given gene G, its abundance is Ab (G), may be relevant with the shared reads of M to the exclusive reads of U, and assessment mode is as follows:
Ab(S)＝Ab(U)+Ab(M)
Ab(U)＝U/l
Ab (U) and Ab (M) are respectively the abundance of unique and multiple, and l represents genome length.Each multiple reads, has endemic species coefficient Co; Make we suppose M} has relevant N in different plant species, then calculates by the following method CO:
For these reads, we add the unique abundance of N as standard.
Embodiment 4 association analysis/screening species markers (microbial markers)
In order to study the dependency of the enteron aisle metagenomics of normal people's (83 example) and liver cirrhosis patient (98 example), in the gene set after merging, done the research of a dependency.On gene set based on 181 samples, identify the gene of different abundance, by the Wilcoxon rank test of the multiple check in conjunction with Benjamini Hochberg, test.Use a very strict threshold value (fdr<0.0001) to find that the significant difference gene between healthy group and liver cirrhosis group has 75245.Adopt median test to find wherein 49830 gene more enrichments in patient with liver cirrhosis, 25415 genes are organized enrichment in health.On these 75245 gene basises, be PCA and analyze, be presented at that between healthy group and liver cirrhosis group, there were significant differences.
For the species marker of exploring and understanding is relevant to liver cirrhosis, according to abundance table, gene is divided into groups.With having given the gene abundance of species similar in individuality, and the same species gene significant difference of Different Individual, so the gene of same species can be by abundance effective cluster of being correlated with.Consequent bunch represents grand genome species (MGS).In order to analyze on the whole a large amount of grand genomic datas structure, reduce quantity of information and carry out classified description, the research about grand genome species (MGS) cluster that the Le Chatelier etc. of take delivered on Nature in 2013 is foundation.Briefly, first by the gene abundance of all individualities, calculate different genes Spearman's correlation coefficient between two, by the genes involved cluster of given threshold values (single cluster).The gene of same species is classified as to a class, and having set threshold values is rho>0.7.In order to correct for the first time the partial loss of classification, carried out classification for the second time, this subseries is used is the mean value of best front 50 the gene abundance of dependency in every group.If the Spearman's correlation coefficient between mean value is greater than 0.85, just by these two combinations also.
This part use be respectively 49,830 patient with liver cirrhosis genes and 25,415 healthy population genes.21,423 genes in healthy population 25,415 genes for the first time 51 gene to 2702 gene Clusterings become 43 bunches, and 51 genes that classification forms are for the second time to 38 bunches of 2970 genomic constitutions.31386 the first step 51-300 gene Clusterings in liver cirrhosis patient 49,830 genes become 60 bunches, and 51 genes to 5755 of second step gene Clustering forms 28 bunches.
In order to prove that the gene in gene cluster belongs to genome and consistent with MGS classification annotation, 6006 genomes (with reference to the effective DACC enteron aisle genome with reference to genome and HMP, MetaHIT in the NCBI of the third edition of ending in August, 2012) have been carried out to blastN and blastP analysis.After blastN, be greater than 80% spike gene and compare on genome, the part in comparison accounts for 90% of shorter ORF, and similarity reaches 95%, and MGS is assigned to genome.6 " health " and 24 " liver cirrhosis " MGS are referred to kind level (Fig. 2 a and table 1).BlastP analyzes annotation for remaining MGS, as its 50 missing gene is greater than 80% unanimously with classification, is referred to corresponding genus, order categorization levels.All 36 residue species, can be referred to given genus and species or order except 1.Marker gene evenly annotates has verified cluster quality, is applicable to whole MGS gene, has completed the calculating of 66 MGS by 50 spike genes.
Table 1 species marker
The MGS essence of exploring species level annotation, we have set up reference library, collect 114 disclosed active chain coccuses, fusobacterium, milk-acid bacteria, Wei Er Shi Coccus and Megasphaera gene set, be mainly the microorganism that oral cavity or enteron aisle are separated.With Blast and T value comparison MGS, 16 similar liver cirrhosis MGS are classified as a class, note some:
(1) reach 95% homology & 90% fraction of coverage (Q);
(2) average homology (R);
(3) theaverage coverage (S);
(4)T＝Q*R*S。
Most of liver cirrhosis patient MGS (15/28) that this standard obtains are from oral cavity, but have 6 from enteron aisle or ight soil, comprise ileum list species.For further explaining the source of the MGS of liver cirrhosis enrichment, we with BlastN by them and 3 grand genome alignments of effective ileum.
75,245 heterogenic great majority of significance difference (70%) are clustered into 66MGS.38 21,423 genes that MGS comprises healthy individual wherein, 31,386 genes (table 1) that 28MGS comprises patient.Result shows that spike gene is only present in homologous gene group before, is present in all homologous gene groups after more order-checkings, can be used for thus a large amount of MGS.There is significant difference in the abundance of Healthy People and liver cirrhosis patient 66MGS.
Embodiment 5 checkings
In order to confirm the discovery in embodiment 4,31 Healthy Peoples in further comparatively validate group and the MGS abundance of 21 liver cirrhosis patients, the analysis of the extraction of DNA and order-checking and gene abundance is carried out according to embodiment 1, embodiment 2 and embodiment 3.Although reduced reliability of statistics because study population's number reduces, the present invention still finds to exist significant difference (q<0.05).
Be enriched in healthy population 38MGS, only have small portion (15.8%) to process by BlastN (95%), overlap (90%) the species development system information that is assigned to.Result reflection lacks enteron aisle bacterial strain separated and order-checking.In addition the 38MGS that, surprisingly Chinese Healthy is rich in is present in (Fig. 2 b) in Denmark crowd simultaneously.Due in whole gene abundance function, the composition of bacterial flora is widely different, and we check China and Denmark crowd's 38MGS.Report wherein 33MGS (86.8%) and q<10 before
-3chinese population corresponding one by one, 26MGS (78.8%) is similar to Denmark crowd.In addition, there are significant difference (q≤10 in 15MGS Chinese population gene abundance
-11), 10MGS (66.7%) is corresponding one by one with the Denmark crowd of identical threshold values.These discoveries show may with Continent of Europe Healthy People enteron aisle Community Facies seemingly.This points out the gene abundance of our healthier people and liver cirrhosis patient; There is before in addition report to point out fat and inflammatory bowel patient gene abundance is lower.In 38 MGS of healthy population enrichment, have at least 2 MGS to trace back to the categorization levels of bacterial classification, wherein as shown in following table (table 2).
The species marker of enrichment in table 2 healthy population
| MGS | Taxonomic?assignment |
| H_4 | Bacteroides?uniformis?ATCC8492 |
| H_7 | Clostridiales |
Form sharp contrast with the MGS of Healthy People, most patient MGS (accounting for 24 in 28) can compare species.Owing to changing and making this species diversity be difficult to discover separately, represent that enteric microorganism composition is highly revised.
In liver cirrhosis patient, one of them species of enrichment are depicted as following table (table 3): atypia Wei Rong bacterium Veillonella atypica, bacterium can cause infecting as spontaneous bacterial peritonitis, liver abscess, intractable respiratory tract infection, the acquired character infection of joint, endocarditis and microbemia.Potential pathogen replaces and contributes to lack the butyric acid production bacterium that bile makes little position enteron aisle become more loose health, causes conversely the life-threatening infection relevant to liver cirrhosis.
The species marker of table 3 liver cirrhosis patient enrichment
| MGS | Taxonomic?assignment |
| L_4 | Veillonella?atypica?ACS-134-V-Col7a |
Utilize above-mentioned species marker to diagnose, treat liver cirrhosis patient; monitor therapy process; or production screening of medicaments, functional foodstuff, probiotic bacterium; known to those skilled in the art the knowing such as the test kit of the above-mentioned species marker of production testing and device, all within protection scope of the present invention.
Species mark can be selected from one or more in the species marker of enrichment in the species marker of liver cirrhosis patient enrichment or healthy population.Preferably, for liver cirrhosis patient or Susceptible population, species mark in table 3 should be detected by enrichment, simultaneously in table 2 species marker not by enrichment.
In treatment plan, preferred, make the species mark growth in table 3 be inhibited or remove, make in table 2 species marker by enrichment.
Claims (10)
1. a microbial markers, is characterized in that, is selected from one or more in microorganism Veillonella atypica ACS-134-V-Col7a, Bacteroides uniformis ATCC 8492, Clostridiales.
2. microbial markers according to claim 1, it is characterized in that thering is Veillonella atypica ACS-134-V-Col7a simultaneously and be selected from one or both of microorganism Bacteroides uniformis ATCC 8492, Clostridiales.
3. a medicine for the treatment of liver cirrhosis, it is characterized in that, one or both in described medicine promotion or increase Bacteroides uniformis ATCC 8492, Clostridiales, and/or suppress or removing microorganism Veillonella atypica ACS-134-V-Col7a.
4. produce or the method for screening of medicaments for one kind, it is characterized in that, one or both in described medicine promotion or increase Bacteroides uniformis ATCC 8492, Clostridiales, and/or suppress or removing microorganism Veillonella atypica ACS-134-V-Col7a.
5. method according to claim 4, is characterized in that, before described pharmacological agent or intervention, whether is enriched with the microbial markers described in claim 1 or 2 in detected object intestinal microflora.
6. method according to claim 5, is characterized in that, described detecting step comprises:
A, from object ight soil, extract DNA sample;
B, for DNA sample, build sequencing library;
C, sequencing library is carried out to high-flux sequence, obtain sequencing result;
D, according to sequencing result, determine in the intestinal microflora of described object, whether there is described microbial markers.
7. method according to claim 6, is characterized in that, described steps d further comprises: the relative abundance for the microbial markers in intestinal microflora detects, and by resulting relative abundance value and predetermined cutoff value, compares.
8. method according to claim 6, is characterized in that, in described step c, according to sequencing result, obtains gene clustering method, proposes the method for a kind of grand gene species (MGS).
9. beneficial microorganism combination or a functional foodstuff, is characterized in that, contains and be selected from one or both in microorganism Bacteroides uniformis ATCC 8492, Clostridiales.
10. detect liver cirrhosis, a monitor therapy process, or the test kit of production, screening of medicaments, it is characterized in that, for detection of microbial markers as described in claim 1.
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| CN107217088A (en) * | 2016-03-22 | 2017-09-29 | 浙江中医药大学 | Ankylosing spondylitis microbial markers |
| CN107217088B (en) * | 2016-03-22 | 2020-08-11 | 浙江中医药大学 | Ankylosing spondylitis microbial markers |
| CN107832584A (en) * | 2017-09-18 | 2018-03-23 | 中国科学院深圳先进技术研究院 | Genetic analysis method, apparatus, equipment and the storage medium of grand genome |
| CN107832584B (en) * | 2017-09-18 | 2021-06-22 | 中国科学院深圳先进技术研究院 | Gene analysis method, device, equipment and storage medium of metagenome |
| WO2019071516A1 (en) * | 2017-10-12 | 2019-04-18 | Perfect (China) Co., Ltd. | Biomarkers for chronic hepatitis b and use thereof |
| CN111051532A (en) * | 2017-10-12 | 2020-04-21 | 完美（中国）有限公司 | Biomarker for chronic hepatitis B and application thereof |
| CN112119167A (en) * | 2018-04-24 | 2020-12-22 | 深圳华大生命科学研究院 | Biomarkers for depression and uses thereof |
| CN112119167B (en) * | 2018-04-24 | 2024-01-12 | 深圳华大生命科学研究院 | Biomarker for depression and application thereof |
| CN109371147A (en) * | 2018-11-01 | 2019-02-22 | 任志刚 | A kind of enteric microorganism gene marker and its application for distinguishing liver cancer and non-liver cancer |
| CN111549155A (en) * | 2020-05-13 | 2020-08-18 | 朱彧 | Analysis technology for predicting liver diseases through relative abundance of intestinal flora |
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