CN110283754B - Intestinal microbial flora for evaluating rheumatoid arthritis symptom state and prognosis evaluation - Google Patents
Intestinal microbial flora for evaluating rheumatoid arthritis symptom state and prognosis evaluation Download PDFInfo
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Abstract
The invention provides an intestinal microbial flora for evaluating the symptom state of rheumatoid arthritis and prognosis evaluation, wherein the intestinal microbial flora comprises two intestinal bacteria of intestinal butyric acid producing bacteria and intestinal non-zymocyte; wherein the enterobacteria comprise 12 microorganisms of the genus butyric acid bacteria and the enterobacteria comprise 33 microorganisms of the genus non-zymobacteria. The invention firstly proposes that butyric acid producing bacteria and non-zymogenic bacteria in the intestinal flora influence the net yield of intestinal butyric acid by producing/consuming intestinal butyric acid, thereby influencing the inflammatory state and prognosis of RA. The results of the verification of the large sample show that the total abundance ratio of the butyric acid producing bacteria to the non-zymogenic bacteria is obtained through the fecal metagenome detection, so that the inflammatory state and the prognosis condition of RA can be evaluated, and the method has higher sensitivity and specificity.
Description
Technical Field
The invention relates to the field of microbiology, in particular to an intestinal microbial flora for evaluating rheumatoid arthritis symptom state and prognosis evaluation.
Background
Rheumatoid Arthritis (RA) is a chronic, inflammatory synovitis-predominant autoimmune disease of unknown etiology. It is characterized by that it possesses multiple joints of hand and foot small joint, symmetrical and invasive arthritis, and often has the condition that the external organ of joint is affected by serum rheumatoid factor and is positive, so that it can result in joint deformity and function loss. The prevalence rate is 0.33%, about 500 thousands of patients exist in China, and most of the patients are young and middle-aged women, and the disability rate is 75% without formal treatment. It is usually treated with steroidal anti-inflammatory drugs and joint glucocorticoid injection. The current treatment method can only control the disease condition but not cure RA, and the treatment is a continuous and repeated process, so that patients need to follow up regularly, evaluate the disease activity degree, the progress condition and the drug side effect in time, and consider whether to change the treatment scheme according to the actual condition.
Unlike initial diagnostic assessment of RA, current assessment of the prognosis and disease activity status of rheumatoid arthritis patients is mainly judged by ACR criteria (ACR20, ACR50, ACR70), which are comprehensively judged based on observations and questionnaire scores. The method changes relatively to physiology and lags behind, and for patients with poor prognosis, the judgment method is relatively lagged behind, so that the inflammation state of the organism of the RA patient cannot be reflected in time, and the method is not beneficial to timely adjustment of treatment and rehabilitation schemes.
At present, the research finds that the imbalance of intestinal flora plays an important role in the occurrence and development of autoimmune diseases such as rheumatoid arthritis. On one hand, the abnormal proliferation of certain intestinal bacteria is closely related to the occurrence of RA, for example, Prevotella copri can increase the occurrence probability of RA, and Porphyromonas gingivalis can mediate the generation of citrullinated protein, thereby promoting the generation of anti-citrullinated protein antibody (ACPA) and initiating the cross reaction to self-antigen; on the other hand, butyric acid-producing bacteria in the intestinal tract, such as Clostridium (Clostridium), Eubacterium (Eubacterium), Vibrio butyricum (Butyrivibrio), Roseburia (Roseburia) and faecalis (Faecalibacterium), promote the immune balance in the intestinal tract by producing butyric acid. The main manifestations are as follows: 1) butyric acid provides an energy source for epithelial cells and participates in the regeneration and repair of intestinal epithelial tissue cells; 2) as a signal molecule of an organism, the polypeptide can recognize membrane receptors of GPR41, GPR43 and GPR109A, further activates a downstream signal path, and influences the expression of inflammatory factors and chemokines; 3) activating nuclear receptor PPAR gamma, inhibiting NF-kB signal channel, and inhibiting inflammatory factor expression; 4) inducing Treg cells to differentiate, promoting acetylation of Foxp3promoter regions, and up-regulating Foxp3 expression; 5) butyric acid inhibits dendritic cells and macrophages, and inhibits differentiation of Th17 cells and Th1 cells; 6) promoting the expression of tight junction protein, promoting the expression of mucin in intestinal epithelial goblet cells, and keeping the intestinal epithelial barrier complete; 7) the pH value of the intestinal tract is low, thereby being beneficial to the colonization of intestinal probiotics and maintaining the steady state of the intestinal flora. Meanwhile, a considerable amount of non-zymogens such as Achromobacter, Burkholderia, Pseudomonas and the like in the intestinal tract do not generate short-chain fatty acids by fermentation, and the short-chain fatty acids (such as butyric acid and the like) can be used as a carbon source, so that the fluctuation of the content of the butyric acid in the intestinal tract is also influenced to a certain extent. Therefore, the intestinal flora can indirectly reflect the immune state of the organism of the RA patient.
Meanwhile, gram-negative bacilli which can not utilize glucose in a fermentation mode exist in intestinal tracts, are represented by Pseudomonas, Acinetobacter and atotrophomonas, are called non-fermentation bacilli and are usually conditional pathogenic bacteria. The intestinal non-zymocyte does not take carbohydrate as an energy source, and does not degrade the carbohydrate through metabolic pathways other than fermentation (short-chain fatty acid is not generated), but obtains energy from various small molecular compounds. In the gut, short chain fatty acids such as butyric acid may provide an energy source for these bacteria.
At present, no relevant report that butyric acid bacteria and non-zymocyte in intestinal tracts are jointly used for evaluating the conditions and the prognosis of rheumatoid arthritis exists.
Disclosure of Invention
The invention aims to provide an intestinal microbial flora for evaluating the symptom state and prognosis of rheumatoid arthritis.
Another objective of the invention is to provide a marker for evaluating the symptom state and prognosis of rheumatoid arthritis.
In order to achieve the object, the invention provides an intestinal microbial flora for evaluating the symptom state and prognosis of rheumatoid arthritis, wherein the intestinal microbial flora comprises intestinal butyric acid producing bacteria and intestinal non-zymocyte;
wherein the enterotoxigenic butyric acid bacteria comprise 12 microorganisms of the genus butyric acid bacteria and the enterotoxigenic non-fermentative bacteria comprise 33 microorganisms of the genus non-fermentative bacteria;
the 12 butyric acid producing bacteria are Anaerostipes, Acetobacter, Clostridium, Fusobacterium, Megasphaera, Treponema, Brachyspira, Filifactor, Butyrivibrio, Peptoniphilius, Anaerotigum, and Roseburia; the 33 non-fermentable bacteria genera are Achromobacter, Burkholderia, Pseudomonas, Ralstonia, Robiginitalea, Stenotrophoromas, Tenacibaculum, Ornithobacter, Cupriavidus, Delftia, Sphingomonas, Variovorax, Alcaligenes, Gramela, Zobellia, Curvibacter, Gilvibacter, Limnobitetans, Alicyphilius, rdella, Roseomonas, Polaribacter, Paraburkderia, Rhodoferax, Hydrogenophaghaga, Flavobacterium, Camocytoga, Azotobacter, Nolabebens, Ramlibacter, Brundimonas, Castanomonas and Commornella.
The Anaerostipes comprises Anaerostipes hadrus and Anaerostipes caccae;
said Acetobacter comprises Acetobacter woodii;
the Clostridium includes Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium botulinum, Clostridium butyricum, Clostridium carboxidigorans, Clostridium cellulovorans, Clostridium difficile, Clostridium kluyveri, Clostridium perfringens, Clostridium saccharolyticum, Clostridium saccharoperbutyricum, Clostridium sporogenes, Clostridium sycambium, Clostridium sytemum, Clostridium tetani;
the Fusobacterium includes Fusobacterium goniodiaformans, Fusobacterium mortiferum, Fusobacterium nuclear, Fusobacterium periodonticum, Fusobacterium ulcerans, and Fusobacterium variatum;
said Megasphaera includes Megasphaera elsdenii, Megasphaera genomics, Megasphaera micucifloris;
said Treponema comprises Treponema phagedenis, Treponema vincentii;
said Brachyspira comprises Brachyspira hypodysenteriae, Brachyspira murdochii, Brachyspira pilosicolii;
the Filifactor comprises Filifactor alocci;
the Butyrivibrio includes Butyrivibrio crossutus, Butyrivibrio fibrilolvens, Butyrivibrio proteoclasius;
said Peptoniphilus includes Peptoniphilus duerdenii, Peptoniphilus harei, Peptoniphilus laciimalis;
the antiaerotigum comprises antiaerotigum propionicum;
said Roseburia comprises Roseburia hominis, Roseburia intestinalis, Roseburia inulinvorans;
said Achromobacter includes Achromobacter densificans, Achromobacter xylosoxidans;
the Burkholderia comprises Burkholderia cenocecia, Burkholderia gladioli, Burkholderia lata, Burkholderia metallica, Burkholderia Multivorans, Burkholderia oklahomensis, Burkholderia pseudolei, Burkholderia bacterium;
said Pseudomonas comprises Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas chlororaphis, Pseudomonas cichororii, Pseudomonas citronello, Pseudomonas aeruginosa, Pseudomonas kongensis, Pseudomonas aeruginosa, Pseudomonas mendonula, Pseudomonas aeruginosa, Pseudomonas strain, Pseudomonas;
the Ralstonia includes Ralstonia inertiosa, Ralstonia mannitolytica, Ralstonia pickettii, Ralstonia solanacearum;
the Robiginitalea comprises Robiginitalea _ biformata;
said Stenotrophoromonas includes Stenotrophoromonas acidaminiphila, Stenotrophoromonas maltophia, Stenotrophoromonas rhizophila;
the Tenacibaculum comprises Tenacibaculum dicentrarrchi, Tenacibaculum jejuense;
the Ornithobacterium includes Ornithobacterium rhizothrahale;
said Cupriavidus includes Cupriavidus gillardii, Cupriavidus malllidans, Cupriavidus necator, Cupriavidus pinatubenis, Cupriavidus taiwanensis;
the Delftia comprises Delftia acidiovarans and Delftia tsuuruhatensis;
said Sphingomonas includes Sphingomonas panacis, Sphingomonas taxi;
the variovax comprises variovax paradoxus, variovax boronicumulans;
said Alcaligenes includes Alcaligenes faecalis;
said Zobellia comprises Zobellia galactantivorans;
said Alicyclophilus comprises Alicyclophilus dentifrices;
said Bordetella includes Bordetella bronchialis, Bordetella genomosp.13, Bordetella genomosp.9, Bordetella hinzii, Bordetella holmesii, Bordetella petrii, Bordetella pseudohinzii;
the Roseomonas comprises Roseomonas gilardii;
the Parabrukholderia includes Parabrukholderia caribensis, Parabrukholderia phymatum, Parabrukholderia sprentiae, Parabrukholderia xenovorans;
the Rhodoferax includes Rhodoferax antarcticas, Rhodoferax ferrireducens, Rhodoferax saidenbachensis;
said Hydrogenophaga includes Hydrogenophaga crassostreae;
the Flavobacterium includes Flavobacterium columnare, Flavobacterium johnsoniae;
said Capnocytophaga comprises Capnocytophaga haemolytica, Capnocytophaga leadbeteri, Capnocytophaga sputigena, Capnocytophaga stomatis;
the Azotobacter comprises Azotobacter chromaccum, Azotobacter vinelandii;
the Nonlabens comprises Nonlabens spongiae;
the Ramlibacter comprises Ramlibacter tatataouinensis;
the Brevundimonas comprises Brevundimonas diminuta, Brevundimonas naejangsanensis, Brevundimonas subvibrioides and Brevundimonas veneris;
said Castellaniella comprises Castellaniella defrarans;
the Commonas includes Commonas aquatica, Commonas kerstersii, Commonas serivorans, Commonas testosteroni.
In a second aspect, the invention provides a marker for evaluating the symptom state and prognosis of rheumatoid arthritis, wherein the marker is the Ratio (Ratio) of total abundance of butyric acid producing bacteria and non-fermenting bacteria in intestinal tracts of a human body,
the ratio is in the range of 12.584 + -0.071, which indicates the risk of exacerbation of rheumatoid arthritis, and the ratio is lower than 12.394, and is evaluated as exacerbation of rheumatoid arthritis inflammation or poor prognosis of rheumatoid arthritis.
For example, the inflammatory state and prognosis of RA can be assessed by performing metagenomic detection on the feces of a patient to obtain the total abundance ratio of butyric acid producing bacteria to non-zymogenic bacteria. Specifically, the detection method is fecal 16S rRNA gene amplicon sequencing or Shotgun whole genome sequencing.
The evaluation method is based on the following mechanisms: the butyric acid producing bacteria and the non-zymogenic bacteria influence the immune state of patients with rheumatoid arthritis by influencing the net yield of butyric acid. In a third aspect, the invention provides an application of the marker Ratio in the aspects of rheumatoid arthritis diagnosis and prognosis evaluation.
In one embodiment of the invention, 40 rheumatoid arthritis patients and 29 healthy human specimens were included. The anti-cyclic citrullinated peptide antibody is an autoantibody taking synthesized Cyclic Citrullinated Polypeptide (CCP) as an antigen. The anti-CCP antibody is not only an early diagnosis and diagnosis index of RA, but also is related to the severity of diseases, and is a sensitive index for distinguishing invasive RA from non-invasive RA. CCP-positive patients are more susceptible to developing more severe joint bone destruction than antibody-negative patients. 40 cases of rheumatoid arthritis patients who were included in the present invention were classified into anti-CCP negative group and anti-CCP positive group according to the serum anti-cyclic citrulline antibody.
Through Illumina high-throughput sequencing of the intestinal flora of the sample, 12 butyric acid producing bacteria Acetobacter, Anaerostipes, Clostridium, Fusobacterium, Megasphaera, Treponema, Brachyspira, Filifactor, Butyrivibrio, Peptophilus, Anaerostignam, Roseburia and 33 non-fermenting bacteria Achromobacter, Burkholderia, Pseudomonas, Rattonia, Robinitialia, Stenotromomonas, Tenacibaculum, Ornithobacter, Cupriavidus, Delftia, Sphingomonas, Variovorona, Alcaligenes, Grameholla, Zymobacteria, Curviabacter, Gimbiellaceae, Bombochaeta, Bombopogora, Achillea, Achillobacter, Achilles, Achill.
Non-targeted metabolite analysis of the samples revealed that fecal butyrate content was significantly higher in healthy population than in RA patients, and that fecal butyrate content was significantly lower in patients with DAS28 ≥ 2.6 than in patients with DAS28<2.6 in the RA population (fig. 3). DAS28 is the activity score for rheumatoid arthritis, with less than 2.6 indicating remission.
By the technical scheme, the invention at least has the following advantages and beneficial effects:
the invention firstly proposes that butyric acid producing bacteria and non-zymogenic bacteria in the intestinal flora influence the net yield of intestinal butyric acid by producing/consuming intestinal butyric acid, thereby influencing the inflammatory state and prognosis of RA. The results of the verification of the large sample show that the total abundance ratio of the butyric acid producing bacteria to the non-zymogenic bacteria is obtained through the fecal metagenome detection, so that the inflammatory state and the prognosis condition of RA can be evaluated, and the method has higher sensitivity and specificity. In addition, the intestinal flora is related to the change of the immune condition of the organism and can reflect the inflammatory state of the organism in time. The evaluation of the activity condition of the traditional rheumatoid disease is mainly judged by ACR standards (ACR20, ACR50 and ACR70), and the method is comprehensively judged based on observation and questionnaire scoring. This approach is relatively delayed with respect to physiological changes, and for patients with poor prognosis, this approach does not reflect the inflammatory status of the body of RA patients in a timely manner. Based on the detection method, the sample collection is convenient, the body does not need to be damaged, the sensitivity is high, the change of the immune state of the body can be found as early as possible by regular detection, the treatment and the timely adjustment of the rehabilitation scheme are carried out, and the aggravation and the deterioration of diseases are avoided.
Drawings
FIG. 1 shows the difference between the abundance of butyric acid-producing bacteria and non-fermentative bacteria and the abundance Ratio (Ratio) of two groups of bacteria in the intestinal tracts of healthy people, CCP antibody negative patients and CCP antibody positive patients in example 2 of the present invention. Among them, HC, healthy control group; anti-CCP (-), CCP antibody negative group; anti-CCP (+), CCP antibody positive group.
FIG. 2A shows the correlation between the genera and total abundance Ratio (Ratio) of butyric acid producing bacteria and nonfermentation bacteria in the intestinal tract and the clinical index of rheumatoid arthritis in example 3 of the present invention. ESR, erythrocyte sedimentation rate, among others; CRP, C-reactive protein; das28.esr and das28.crp, rheumatoid arthritis score; RF, rheumatoid factor; Anti-CCP, Anti-cyclic citrullinated polypeptide antibody; rho >0, positively correlated; rho <0, negative correlation; note p-value < 0.05; FIG. 2B is a linear regression curve of the Ratio of total abundance (Ratio) of butyric acid-producing bacteria to non-zymophyte bacteria and each index of RA.
FIG. 3 is a graph showing the difference in fecal butyric acid content between healthy and RA groups in example 4 of the present invention. Wherein, A: the content of short-chain fatty acid in the excrement of healthy people and RA patients is different. Butyric acid content significantly differs in the intestinal tracts of healthy and RA patients (p ═ 0.012); b: the difference in fecal butyrate content among the intestinal tracts of patients with different degrees of disease. DAS28, activity score for rheumatoid arthritis, less than 2.6 indicates remission; DJC, joint deformity, DJC (-), no joint deformity; DJC (+), deformity of the joint. C: the content of fecal butyric acid of patients without joint deformity is higher than that of patients with joint deformity.
FIG. 4 is a graph showing the accuracy of evaluation of inflammatory conditions (anti-CCP antibodies) by butyric acid producing bacteria and non-fermentative bacteria in the intestine in example 5 of the present invention.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. Unless otherwise indicated, the examples follow conventional experimental conditions, such as the Molecular Cloning handbook, Sambrook et al (Sambrook J & Russell DW, Molecular Cloning: a Laboratory Manual,2001), or the conditions as recommended by the manufacturer's instructions.
Example 1 enrichment of butyric acid-producing bacteria in feces of patients with rheumatoid arthritis and CCP antibody positivity
The anti-cyclic citrullinated peptide antibody is an autoantibody taking Cyclic Citrullinated Polypeptide (CCP) as an antigen, has higher sensitivity and specificity on Rheumatoid Arthritis (RA), and is a highly specific index for early diagnosis of RA. In this example, stool samples from 29 healthy persons and 40 initially untreated RA patients (15 CCP-negative and 25 CCP-positive) were subjected to metagenomic sequencing using Illumina hiseq 2000 high throughput sequencing technology, low quality data filtration was performed using FastQC (Version 0.11.5), Trimmomatic (Version 0.33, option: SLIDNGWINDOW: 4:20MINLEN:40), FASTX-Toolkit (Version 0.0.13, option: q 20-p 80), human DNA sequence filtration was performed using BWA, and repetitive sequences caused by amplification were removed using PREQ (constraints-verbern 1-deep _ min 2). The filtered sequences were species annotated with Kraken to remove species present in less than 10% of the samples. Differential species (wilcoxon rank-sum test p-value. ltoreq.0.05, FDR. ltoreq.0.3) were screened between groups of samples based on rank-sum test, in which differential colonies of anti-CCP antibody positive group (ACCP +) and anti-CCP antibody negative group (ACCP-) were concentrated in two broad groups of butyric acid-producing bacteria and non-fermenting bacteria (Table 1). A large number of butyric acid bacteria such as 11 species of Clostridium and fusarium were included in the enriched differential flora of the antibody negative patient group (table 1, black underlined). The enriched gut flora of the antibody positive patient group comprises a large number of non-fermentable bacteria, e.g. Achromobacter, Bordetella, Pseudomonas, Stenotrophoromonas, Sphingomonas and 19 species of the genus Variovorax (Table 1, black bold italics). The intestinal non-zymocyte does not take carbohydrate as an energy source, and does not degrade the carbohydrate through metabolic pathways other than fermentation (short-chain fatty acid is not generated), but obtains energy from various small molecular compounds. In the gut, short chain fatty acids such as butyric acid may provide an energy source for these bacteria. Thus, the non-fermenters may compete with the intestinal epithelial cells for the consumption of butyric acid. It was therefore concluded that the intestinal flora has a major influence on the inflammatory state of the body of rheumatoid patients by influencing the net production of butyric acid in the intestine.
Example 2 the abundance ratio of butyric acid bacteria/non-zymocyte in the excrement of patients with mild rheumatoid arthritis is obviously increased
According to literature reports and genomic information, the present invention detects 12 genera Anaerosperms, Acetobacter, Clostridium, Fusobacterium, Megasphaera, Treponema, Brachyspira, Filifoctor, Butyrivibrio, Peptophilus, Anerolignum, Rosebula, and non-fermenting bacteria 33 genera Achromobacter, Burkholderia, Pseudomonas, Ralstonia, Robiginia, Stenotropophomonas, Tenacibaculum, Ornithobacter, Cupriavidus, Delftia, Sphingononas, Variogenes, Alcaligenes, Merella, Zobacter, Klebsiella, or Klebsiella, or Klebsiella, or Kl. The abundance statistics of the two broad groups of flora show that the distribution of the median total abundance of the butyric acid producing bacteria among three groups, namely a control group HC (healthy people), a CCP antibody negative group anti-CCP (-), and a CCP antibody positive group anti-CCP (+) is as follows: CCP negative group > control group > CCP positive group (fig. 1, a); the distribution of the median of the total abundance of the non-zymophytes among the three groups is as follows: CCP negative group < control group < CCP positive group. The distribution has no significant difference (figure 1, A), but compared with a CCP antibody positive patient, a CCP antibody negative patient has a significantly increased ratio of total abundance of butyric acid producing bacteria to total abundance of non-zymogenic bacteria in intestinal tract (figure 1, C), and the result proves that the intestinal flora can reflect the disease degree of the organism of the RA patient and influence the prognosis degree of RA through the net yield of butyric acid.
Example 3 correlation of butyric acid producing bacteria and non-zymocyte in intestinal tract and clinical index of rheumatoid arthritis
In this example, the correlation analysis of the abundance of each of the mentioned butyric acid producing bacteria and non-zymogenic bacteria and clinical indexes of blood sedimentation (ESR), C-reactive protein (CRP), das28.ESR, das28.CRP, Rheumatoid Factor (RF) and Anti-CCP (Anti-CCP) of RA patients shows that there is a strong correlation between the butyric acid producing bacteria and the CCP indexes of RA patients (fig. 2A, table 2), and that clinical indexes indicating the severity of diseases such as 11 butyric acid producing bacteria, Rheumatoid Factor (RF) and Anti-cyclic citrullinated peptide antibody (Anti-CCP) are negatively correlated, but the non-zymogenic bacteria are contrary to them. According to the invention, the ratio (ratio) of the butyric acid bacteria to the total abundance of the non-zymophyte is used as an index, and the correlation analysis of the ratio and the clinical index of the RA patient shows that the characteristic is in negative correlation with all clinical indexes (figure 2B), which suggests that the butyric acid bacteria has a potential effect of relieving the inflammation of the RA patient by generating butyric acid. The correlation between the abundance of the genus and the clinical index is shown in Table 2.
TABLE 2 correlation of the abundance of the genus and clinical indices
Example 4 fecal butyric acid levels are elevated in healthy and mild RA patients
The content of butyric acid in the RA group was significantly reduced and the content of other short-chain fatty acids (acetic acid, propionic acid, isobutyric acid, valeric acid, isovaleric acid) was not significantly changed as detected by the content of short-chain fatty acids in feces samples of 31 healthy persons and 36 RA patients (22 with deformed joints and 14 without deformed joints) treated for more than two years (fig. 3, a). Fecal butyrate content was also found to correlate with disease activity (DAS28) and joint deformity status. In patients with low disease activity levels (DAS28<2.6), fecal butyrate levels were significantly higher than in patients with intermediate-high disease activity levels (DAS28 ≧ 2.6) (FIG. 3, B). The content of fecal butyric acid in patients without joint deformity is higher than that in patients with joint deformity (fig. 1, C). This result confirms that the reduction of the butyric acid producing bacteria in the intestinal tract and its butyric acid product is closely related to the occurrence and severity status of rheumatoid arthritis.
Example 5 accuracy of butyric acid producing bacteria and nonfermented bacteria for prognosis evaluation
Two groups of anti-CCP antibody negative/positive patients in example 2 are taken as study objects, the anti-CCP antibody negative patients are considered to be patients with good prognosis and low inflammation state level, the total abundance ratio of differential butyric acid bacteria and non-zymogenic bacteria screened from intestinal tracts is taken as an observation index, the classification and prediction of the patient prognosis and the organism inflammation state are carried out, the AUC is 0.757, CI:0.5946-0.9201 is analyzed by ROC, the optimal cut-off value of the ratio is 12.584, the prediction accuracy is 75.7%, and the method can be used for evaluating disease prognosis and inflammation state (figure 4).
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (2)
1. The marker is used for evaluating the symptom state of rheumatoid arthritis and evaluating the prognosis, and is characterized in that the marker is butyric acid bacteria and non-zymocyte in intestinal tracts of organisms;
the ratio of the total abundance of the butyric acid bacteria in the intestinal tract and the non-zymocyte in the intestinal tract is less than or equal to 12.584, which indicates that the rheumatoid arthritis has exacerbation risk;
wherein the enterotoxigenic butyric acid bacteria consist of 12 microorganisms of the genus butyric acid bacteria and the enterotoxigenic non-fermentative bacteria consist of 33 microorganisms of the genus non-fermentative bacteria;
the 12 butyric acid producing bacteria are Anaerostipes, Acetobacter, Clostridium, Fusobacterium, Megasphaera, Treponema, Brachyspira, Filifactor, Butyrivibrio, Peptoniphilius, Anaerotigum, and Roseburia; the 33 non-fermentable bacteria genera are Achromobacter, Burkholderia, Pseudomonas, Ralstonia, Robiginitalea, Stenotrophoromas, Tenacibaculum, Ornithobacter, Cupriavidus, Delftia, Sphingomonas, Variovorax, Alcaligenes, Gramela, Zobellia, Curvibacter, Gilvibacter, Limnobitetans, Alicyphilius, rdella, Roseomonas, Polaribacter, Paraburkderia, Rhodoferax, Hydrogenophaghaga, Flavobacterium, Camocytoga, Azotobacter, Nolabebens, Ramlibacter, Brundimonas, Castanomonas and Commornella.
2. The marker of claim 1, wherein said Anaeromotipes comprises Anaeromotipes hadrus, Anaeromotipes caccae;
said Acetobacter comprises Acetobacter woodii;
the Clostridium includes Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium botulinum, Clostridium butyricum, Clostridium carboxidigorans, Clostridium cellulovorans, Clostridium difficile, Clostridium kluyveri, Clostridium perfringens, Clostridium saccharolyticum, Clostridium saccharoperbutyricum, Clostridium sporogenes, Clostridium sycambium, Clostridium sytemum, Clostridium tetani;
the Fusobacterium includes Fusobacterium goniodiaformans, Fusobacterium mortiferum, Fusobacterium nuclearum, Fusobacterium periodicium, Fusobacterium ulcerans, and Fusobacterium variatum;
said Megasphaera includes Megasphaera elsdenii, Megasphaera genomics, Megasphaera micuciformis;
said Treponema comprises Treponema phagedenis, Treponema vincentii;
said Brachyspira comprises Brachyspira hypodysenteriae, Brachyspira murdochii, Brachyspira pilosicolii;
the Filifactor comprises Filifactor alocci;
the Butyrivibrio includes Butyrivibrio crossutus, Butyrivibrio fibrilolvens, Butyrivibrio proteoclasius;
said Peptoniphilus includes Peptoniphilus duerdenii, Peptoniphilus harei, Peptoniphilus laciimalis;
the antiaerotigum comprises antiaerotigum propionicum;
said Roseburia comprises Roseburia hominis, Roseburia intestinalis, Roseburia inulinvorans;
said Achromobacter includes Achromobacter densificans, Achromobacter xylosoxidans;
the Burkholderia comprises Burkholderia cenocecia, Burkholderia gladioli, Burkholderia lata, Burkholderia metallica, Burkholderia Multivorans, Burkholderia oklahomensis, Burkholderia pseudolei, Burkholderia bacterium;
said Pseudomonas comprises Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas chlororaphis, Pseudomonas cichororii, Pseudomonas citronello, Pseudomonas aeruginosa, Pseudomonas kongensis, Pseudomonas aeruginosa, Pseudomonas mendonula, Pseudomonas aeruginosa, Pseudomonas strain, Pseudomonas;
the Ralstonia includes Ralstonia inertiosa, Ralstonia mannitolytica, Ralstonia pickettii, Ralstonia solanacearum;
the Robiginitalea comprises Robiginitalea biformata;
said Stenotrophoromonas includes Stenotrophoromonas acidaminiphila, Stenotrophoromonas maltophia, Stenotrophoromonas rhizophila;
the Tenacibaculum comprises Tenacibaculum dicentrarrchi, Tenacibaculum jejuense;
the Ornithobacterium includes Ornithobacterium rhizothrahale;
said Cupriavidus includes Cupriavidus gillardii, Cupriavidus malllidans, Cupriavidus necator, Cupriavidus pinatubenis, Cupriavidus taiwanensis;
the Delftia comprises Delftia acidiovarans and Delftia tsuuruhatensis;
said Sphingomonas includes Sphingomonas panacis, Sphingomonas taxi;
the variovax comprises variovax paradoxus, variovax boronicumulans;
said Alcaligenes includes Alcaligenes faecalis;
said Zobellia comprises Zobellia galactantivorans;
said Alicyclophilus comprises Alicyclophilus dentifrices;
said Bordetella includes Bordetella bronchialis, Bordetella genomosp.13, Bordetella genomosp.9, Bordetella hinzii, Bordetella holmesii, Bordetella petrii, Bordetella pseudohinzii;
the Roseomonas comprises Roseomonas gilardii;
the Parabrukholderia includes Parabrukholderia caribensis, Parabrukholderia phymatum, Parabrukholderia sprentiae, Parabrukholderia xenovorans;
the Rhodoferax includes Rhodoferax antarcticas, Rhodoferax ferrireducens, Rhodoferax saidenbachensis;
said Hydrogenophaga includes Hydrogenophaga crassostreae;
the Flavobacterium includes Flavobacterium columnare, Flavobacterium johnsoniae;
said Capnocytophaga comprises Capnocytophaga haemolytica, Capnocytophaga leadbeteri, Capnocytophaga sputigena, Capnocytophaga stomatis;
the Azotobacter comprises Azotobacter chromaccum, Azotobacter vinelandii;
the Nonlabens comprises Nonlabens spongiae;
the Ramlibacter comprises Ramlibacter tatataouinensis;
the Brevundimonas comprises Brevundimonas diminuta, Brevundimonas naejangsanensis, Brevundimonas subvibrioides and Brevundimonas veneris;
said Castellaniella comprises Castellaniella defrarans;
the Commonas includes Commonas aquatica, Commonas kerstersii, Commonas serivorans, Commonas testosteroni.
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