Method of diagnosis and treating gastrointestinal and neurological diseases associated with species of genus clostridium

Abstract

The invention includes a method of diagnosis and treating autism associated with an overgrowth of beta2-toxin-gene-positive Clostridium perfringens in the gut of an autistic subject. In one embodiment, the method comprises administering to the subject (e.g. a subject having an overgrowth of beta2-toxin-gene-positive Clostridium perfringens in the gut) one or more agents to reduce or eliminate beta2-toxin-gene positive Clostridium perfringens in the subject so as to relieve one or more symptoms of autism.

1 . A method of treating autism associated with an overgrowth of beta2-toxin-gene-positive  Clostridium perfringens  in the gut of an autistic subject comprising administering to the subject having an overgrowth of  Clostridium perfringens  positive for beta2-toxin gene one or more agents to reduce or eliminate beta2-toxin-gene positive  Clostridium perfringens  in the subject so as to relieve one or more symptoms of autism; thereby, treating autism associated with an overgrowth of beta2-toxin-gene-positive  Clostridium perfringens  in the autistic subject. 
     
     
         2 . The method of  claim 1 , wherein the overgrowth of beta2-toxin-gene-positive  Clostridium perfringens  is an overrepresentation or excess of beta2-toxin-gene-positive  Clostridium perfringens  vegetative cells, spores or a combination thereof, in the gut of the autistic subject. 
     
     
         3 . The method of  claim 2 , wherein the overgrowth of beta2-toxin-gene-positive  Clostridium perfringens  is detected as an overrepresentation or excess of  Clostridium perfringens  vegetative cells,  Clostridium perfringens  spores or combination thereof, in the feces of an autistic subject compared to a control subject or population of control subjects. 
     
     
         4 . The method of  claim 1 , wherein the control subject or population of control subjects has about 1.5×10 3  colony forming units of beta2-toxin-gene-positive  Clostridium perfringens  per gram dry weight of fecal specimen. 
     
     
         5 . (canceled) 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 1 , wherein the overgrowth of beta2-toxin-gene-positive  Clostridium perfringens  in the gut of an autistic subject is positively correlated with an overgrowth of  Clostridium perfringens.    
     
     
         8 . The method of  claim 7 , wherein the overgrowth of  Clostridium perfringens  results in at least a 3-fold increase in  Clostridium perfringens  colony forming units in the autistic subject over a control subject or a population of control subjects. 
     
     
         9 .- 17 . (canceled) 
     
     
         18 . The method of  claim 1 , wherein the beta2-toxin-gene-positive  Clostridium perfringens  in the gut of an autistic subject additionally comprises a  Clostridium perfringens  alpha-toxin gene. 
     
     
         19 . The method of  claim 1 , wherein the beta2-toxin-gene-positive  Clostridium perfringens  in the gut of an autistic subject is free of a  Clostridium perfringens  alpha-toxin gene. 
     
     
         20 .- 32 . (canceled) 
     
     
         33 . The method of  claim 1 , wherein the beta2-toxin-gene-positive  Clostridium perfringens  in the gut of an autistic subject produces a  Clostridium perfringens  beta2-toxin and alpha-toxin but is free of a  Clostridium perfringens  beta-toxin,  Clostridium perfringens  epsilon-toxin and  Clostridium perfringens  iota-toxin. 
     
     
         34 . The method of  claim 1 , wherein the beta2-toxin-gene-positive  Clostridium perfringens  in the gut of an autistic subject produces a  Clostridium perfringens  beta2-toxin but is free of a  Clostridium perfringens  alpha-toxin,  Clostridium perfringens  beta-toxin,  Clostridium perfringens  epsilon-toxin and  Clostridium perfringens  iota-toxin. 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . The method of  claim 1 , wherein the one or more agent(s) is an antimicrobial, a bacteriophage, a probiotic, a probiotic group, a prebiotic or a vaccine which would lead to production of an anti-toxin against a toxin produced by the beta2-toxin-gene-positive  Clostridium perfringens  in the gut of the autistic subject, or a combination thereof. 
     
     
         38 . The method of  claim 1 , wherein the agent to reduce or eliminate beta2-toxin-gene-positive  Clostridium perfringens  is selected from the group consisting of ABT-773, ampicillin, sulbactam, amphomycin, azithromycin, bacitracin, carboxmycin, cephlosporins, clarithromycin, erythromycin, furazolidone, nitrofuran, fusidic acid, sodium fusidate, gramicidin, a penem, imipenem, josamycin, linezolid, oxazolidinone, a macrolide, metronidazole, nitroimidazole, mikamycin, minocycline, novobiocin, oleandomycin, triacetyloleandomycin, ostreogrycin, piperacillin, tazobactam, pristinamycin, ramoplanin, ristocetin, rosamicin, rosaramicin, spectinomycin, spiramycin, streptogramin, synergistin, teicoplanin, telithromycin, ticarcillin, clavulanic acid, tyrocidin, tyrothricin, vancomycin, vernamycin, and virginiamycin. 
     
     
         39 . The method of  claim 37 , wherein the antimicrobial is selected from the group consisting of ABT-773, ampicillin, sulbactam, amphomycin, azithromycin, bacitracin, carboxmycin, cephlosporins, clarithromycin, erythromycin, furazolidone, nitrofuran, fusidic acid, sodium fusidate, gramicidin, a penem, imipenem, josamycin, linezolid, oxazolidinone, a macrolide, metronidazole, nitroimidazole, mikamycin, minocycline, novobiocin, oleandomycin, triacetyloleandomycin, ostreogrycin, piperacillin, tazobactam, pristinamycin, ramoplanin, ristocetin, rosamicin, rosaramicin, spectinomycin, spiramycin, streptogramin, synergistin, teicoplanin, telithromycin, ticarcillin, clavulanic acid, tyrocidin, tyrothricin, vancomycin, vernamycin, and virginiamycin. 
     
     
         40 . The method of  claim 37 , wherein the probiotic or the probiotic group is selected from the group consisting of  Bacteroides thetaiotaomicron, Bacteroides vulgatus, Bacteroides distasonis, Bacteroides fragilis, Bifidobacterium adolescentis  group,  Eubacterium aerofaciens, Clostridium ramosum, Escherichia coli, Streptococcus faecalis  group,  Lactobacillus  spp.,  L. acidophilus , gram-negative anaerobes, enterococci,  Bacteroides  sp.,  Parabacteroides, Prevotella, Porphyromonas , gram-positive anaerobic cocci,  Clostridium  sp., Enterobacteriaceae,  E. coli, L. bulgaricus, S. thermophilus, Collinsella  genus,  Bifidobacterium  genus,  Bifidobacterium longum, Bifidobacterium angulatum, Dialister invisus, Clostridium leptum, Firmicutes, Actinobacteria, Faecalibacterium, Ruminococcus, Eubacterium, Alistipes, Roseburia, Anaerofilum, Streptococcus, Turicibacter, Parabacteroides, Dorea, Veillonella, Akkermansia, Sporobacter, Ethanoligenens, Papillibacter, Holdemania, Weissella, Dialister, Pseudoramibacter, Streptococcus, Anaerovorax, Lactococcus, Leuconostoc, Ethanoligenens, Helcococcus, Alkaliphilus, Clostridium bolteae, Clostridium methylpentosum, Eubacterium ruminantium, Phascolarctobacterium faecium, Alistipes  species,  Bifidobacterium  species,  Lactobacillus  species, and  Bacteroides caccae.    
     
     
         41 .- 45 . (canceled) 
     
     
         46 . The method of  claim 37 , wherein the toxin is  Clostridium perfringens  beta2-toxin. 
     
     
         47 . (canceled) 
     
     
         48 . A method of treating autism associated with an overgrowth of beta2-toxin-gene-positive  Clostridium perfringens  in the gut of an autistic subject comprising: a) determining existence of an overgrowth of  Clostridium perfringens  positive for beta2-toxin gene in a sample from the subject; and (b) administering to the subject one or more agents to reduce or eliminate beta2-toxin-gene positive  Clostridium perfringens  in the subject so as to relieve one or more symptoms of autism; thereby, treating autism associated with an overgrowth of beta2-toxin-gene-positive  Clostridium perfringens  in the autistic subject. 
     
     
         49 .- 56 . (canceled) 
     
     
         57 . A method of identifying an autistic subject whose autism may be alleviated by a course of treatment directed against an overgrowth of  Clostridium perfringens  so as to reduce or eliminate the overgrowth comprising (a) determining existence of said overgrowth in the autistic subject, thereby, identifying the autistic subject whose autism may be alleviated by a course of treatment directed against the overgrowth of  Clostridium perfringens.    
     
     
         58 .- 65 . (canceled) 
     
     
         66 . A method for monitoring the course of autism in an autistic subject by identifying the subject by the method of  claim 57  or  58  and quantitatively determining a level of overgrowth of  Clostridium perfringens , overgrowth of beta2-toxin-gene-positive  Clostridium perfringens , activity of  Clostridium perfringens  beta2-toxin from a biological sample from the subject at one time point and comparing a value so determined with the value determined from a second sample from the subject, such samples being taken at different points in time, a difference in the values determined being indicative of the course of the autistic condition. 
     
     
         67 . (canceled) 
     
     
         68 . (canceled) 
     
     
         69 . The method of  claim 66 , wherein a decrease in the overgrowth of  Clostridium perfringens , overgrowth of beta2-toxin-gene-positive  Clostridium perfringens , activity of  Clostridium perfringens  beta2-toxin is indicative or predictive of reducing severity or duration of one or more symptoms associated with autism. 
     
     
         70 . The method of  claim 66 , wherein an increase in the overgrowth of  Clostridium perfringens , overgrowth of beta2-toxin-gene-positive  Clostridium perfringens , activity of  Clostridium perfringens  beta2-toxin is indicative or predictive of increasing severity or duration of one or more symptoms associated with autism. 
     
     
         71 . (canceled)