Prevalence of epsilon toxin-producing Clostridium perfringens isolates among patients with multiple sclerosis and neuromyelitis optica spectrum disorder in Iran

Background Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobuli...

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Published inBMC neurology Vol. 25; no. 1; pp. 234 - 12
Main Authors Azimirad, Masoumeh, Hamtaeigashti, Sara, Raeisi, Hamideh, Looha, Mehdi Azizmohammad, Gholami, Fatemeh, Naser Moghadasi, Abdorreza, Zali, Mohammad Reza, Yadegar, Abbas
Format Journal Article
LanguageEnglish
Published London BioMed Central 31.05.2025
BioMed Central Ltd
BMC
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ISSN1471-2377
1471-2377
DOI10.1186/s12883-025-04251-z

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Abstract Background Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls. Methods Between September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS ( n  = 33) and NMOSD ( n  = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx -encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis. Results Totally, 11 C. perfringens were isolated from MS ( n  = 4), NMOSD ( n  = 2), and healthy ( n  = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients’ risk of developing MS and NMOSD can be predicted using these predictors. Conclusions Our findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD.
AbstractList Abstract Background Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls. Methods Between September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS (n = 33) and NMOSD (n = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx-encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis. Results Totally, 11 C. perfringens were isolated from MS (n = 4), NMOSD (n = 2), and healthy (n = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients’ risk of developing MS and NMOSD can be predicted using these predictors. Conclusions Our findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD.
BackgroundClostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls.MethodsBetween September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS (n = 33) and NMOSD (n = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx-encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis.ResultsTotally, 11 C. perfringens were isolated from MS (n = 4), NMOSD (n = 2), and healthy (n = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients’ risk of developing MS and NMOSD can be predicted using these predictors.ConclusionsOur findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD.
Background Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls. Methods Between September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS ( n  = 33) and NMOSD ( n  = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx -encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis. Results Totally, 11 C. perfringens were isolated from MS ( n  = 4), NMOSD ( n  = 2), and healthy ( n  = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients’ risk of developing MS and NMOSD can be predicted using these predictors. Conclusions Our findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD.
Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls.BACKGROUNDClostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls.Between September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS (n = 33) and NMOSD (n = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx-encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis.METHODSBetween September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS (n = 33) and NMOSD (n = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx-encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis.Totally, 11 C. perfringens were isolated from MS (n = 4), NMOSD (n = 2), and healthy (n = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients' risk of developing MS and NMOSD can be predicted using these predictors.RESULTSTotally, 11 C. perfringens were isolated from MS (n = 4), NMOSD (n = 2), and healthy (n = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients' risk of developing MS and NMOSD can be predicted using these predictors.Our findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD.CONCLUSIONSOur findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD.
Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls. Between September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS (n = 33) and NMOSD (n = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx-encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis. Totally, 11 C. perfringens were isolated from MS (n = 4), NMOSD (n = 2), and healthy (n = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients' risk of developing MS and NMOSD can be predicted using these predictors. Our findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD.
Background Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls. Methods Between September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS (n = 33) and NMOSD (n = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx-encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis. Results Totally, 11 C. perfringens were isolated from MS (n = 4), NMOSD (n = 2), and healthy (n = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients' risk of developing MS and NMOSD can be predicted using these predictors. Conclusions Our findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD. Keywords: Clostridium perfringens, Epsilon toxin, Multiple sclerosis, Neuromyelitis optica, NMOSD, ETX, Prediction models
Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in demyelinating CNS disorders. Here, we investigated the prevalence of C. perfringens and anti-ETX immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody levels in stool and sera of patients with multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD) compared to healthy controls. Between September 2019 and June 2020, 43 stool and 43 sera specimens were collected from MS (n = 33) and NMOSD (n = 10) patients, and 49 samples from healthy subjects. Stool samples were cultured for the presence of C. perfringens and PCR was used to detect etx-encoding gene. The level of serum IgG and IgM against ETX was examined using competitive ELISA. The predictive value of sex, age, ETX IgG and IgM levels, and C. perfringens was assessed using multivariable prediction models, with all models trained on the full set of variables. Independent contributions were evaluated separately using logistic regression (LR) analysis. Totally, 11 C. perfringens were isolated from MS (n = 4), NMOSD (n = 2), and healthy (n = 5) subjects. The etx gene was detected in 1/4 (25%) isolates from MS and 2/2 (100%) from NMOSD patients, while none of the isolates from healthy subjects carried the gene. The analysis of serum IgG and IgM level against ETX showed no significant difference between patients and healthy subjects. Among the prediction models used, LR and Naïve Bayes (NB), trained on all predictor variables, exhibited the highest statistical accuracy for MS diagnosis, with AUC = 0.71 and AUC = 0.93, respectively. Among individual parameters, sex (AUC = 0.63) and IgG level (AUC = 0.57) were the most reliable predictors. The nomogram prediction demonstrated that the probability of patients' risk of developing MS and NMOSD can be predicted using these predictors. Our findings indicate a potential association between ETX-producing C. perfringens strains and MS/NMOSD diseases, as its presence was detected in a subset of patient samples. However, no significant differences in anti-ETX antibody levels were observed between control and patients. Our prediction models highlighted key demographic and serological factors that may contribute to MS diagnosis. These results highlight the need for further research to meticulously decipher the biological interplay of C. perfringens and its ETX toxin with the development of MS and NMOSD.
ArticleNumber 234
Audience Academic
Author Zali, Mohammad Reza
Azimirad, Masoumeh
Naser Moghadasi, Abdorreza
Looha, Mehdi Azizmohammad
Raeisi, Hamideh
Hamtaeigashti, Sara
Gholami, Fatemeh
Yadegar, Abbas
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  givenname: Mehdi Azizmohammad
  surname: Looha
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  givenname: Abbas
  orcidid: 0000-0002-2135-7581
  surname: Yadegar
  fullname: Yadegar, Abbas
  email: a.yadegar@sbmu.ac.ir, babak_y1983@yahoo.com
  organization: Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences
BackLink https://www.ncbi.nlm.nih.gov/pubmed/40450217$$D View this record in MEDLINE/PubMed
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Issue 1
Keywords ETX
Epsilon toxin
Multiple sclerosis
Neuromyelitis optica
Prediction models
NMOSD
Clostridium perfringens
Language English
License 2025. The Author(s).
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Snippet Background Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved...
Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved in...
Background Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved...
BackgroundClostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be involved...
Abstract Background Clostridium perfringens epsilon toxin (ETX) is a potent neurotoxin that crosses the blood-brain barrier (BBB), and has been suggested to be...
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StartPage 234
SubjectTerms Adult
Antibodies, Bacterial - blood
Bacterial toxins
Bacterial Toxins - immunology
Bayesian analysis
Blood-brain barrier
Chemical properties
Chronic illnesses
Clostridium
Clostridium Infections - epidemiology
Clostridium perfringens
Clostridium perfringens - isolation & purification
Clostridium perfringens - metabolism
Demyelination
Development and progression
Diagnosis
Disease
Enzyme-linked immunosorbent assay
Enzymes
Epsilon toxin
ETX
Feces - microbiology
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Immunoglobulin G
Immunoglobulin G - blood
Immunoglobulin M
Immunoglobulin M - blood
Immunoglobulins
Iran - epidemiology
Male
Measurement
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Medicine & Public Health
Medicine, Experimental
Middle Aged
Multiple sclerosis
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Neomycin
Neurochemistry
Neurology
Neuromyelitis
Neuromyelitis optica
Neuromyelitis Optica - blood
Neuromyelitis Optica - epidemiology
Neuromyelitis Optica - microbiology
Neurosurgery
Neurotoxic agents
NMOSD
Physiological aspects
Prediction models
Prevalence
Regression analysis
Risk factors
Statistics
Toxins
Young Adult
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Title Prevalence of epsilon toxin-producing Clostridium perfringens isolates among patients with multiple sclerosis and neuromyelitis optica spectrum disorder in Iran
URI https://link.springer.com/article/10.1186/s12883-025-04251-z
https://www.ncbi.nlm.nih.gov/pubmed/40450217
https://www.proquest.com/docview/3216559774
https://www.proquest.com/docview/3214326475
https://pubmed.ncbi.nlm.nih.gov/PMC12125731
https://doaj.org/article/16457b744faa44b6addb9ae6cca8f919
Volume 25
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