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2 edition of development of an RNA assay for Clostridium Botulinum toxin gene expression. found in the catalog.

development of an RNA assay for Clostridium Botulinum toxin gene expression.

Susan McGrath

development of an RNA assay for Clostridium Botulinum toxin gene expression.

by Susan McGrath

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Published by The Author] in [S.l .
Written in English


Edition Notes

Thesis (D. Phil. ) - University of Ulster, 1999.

ID Numbers
Open LibraryOL18341507M

Clostridium botulinum is an anaerobic bacterium that produces a neurotoxin thought to be the most lethal substance known (on a per molecule or per weight basis). Strains of C. botulinum have been identified that produce 7 different types of this neurotoxin, designated as types A through G. The toxin is synthesized as a single polypeptide of approximately kD. The Clostridia: Molecular Biology and Pathogenesis is a unique work, comprising the most complete reference on the clostridia for over 20 years, bringing together the results from some of the most innovative and exciting research in the past decade. Using a principle-oriented rather than taxonomic approach, the results from molecular biology.

The toxin gene clusters of proteolytic C. botulinum (toxin types A, B, F and H) contain the regulatory gene botR in both the ha and orfX toxin gene clusters that codes for a σ factor that positively controls expression of the structural gene for botulinum toxin as well as of its accessory genes, –. Botulism can be spread in several ways. The bacterial spores which cause it are common in both soil and water. They produce the botulinum toxin when exposed to low oxygen levels and certain temperatures. Foodborne botulism happens when food containing the toxin is eaten. Infant botulism happens when the bacteria develops in the intestines and releases the toxin. This typically only occurs in children fewer than six months old, as protective mechanisms develop Complications: Respiratory failure.

Clostridium botulinum is a Gram-positive, rod-shaped, anaerobic, spore-forming, motile bacterium with the ability to produce the neurotoxin botulinum.. The botulinum toxin can cause a severe flaccid paralytic disease in humans and other animals and is the most potent toxin known to humankind, natural or synthetic, with a lethal dose of – ng/kg in : Clostridia.   Eating an amount of toxin just th the weight of a grain of salt can be fatal, which is why so much effort has been put into keeping Clostridium botulinum, which produces the toxin, out of .


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Development of an RNA assay for Clostridium Botulinum toxin gene expression by Susan McGrath Download PDF EPUB FB2

Cordivari C, Misra VP, Vincent A, et al. Secondary nonresponsiveness to botulinum toxin A in cervical dystonia: the role of electromyogram-guided injections, botulinum toxin A antibody assay, and the extensor digitorum brevis by: Detection of C. botulinum E VH toxin gene expression by RT-PCR following DNase I treatment.

A total cellular RNA extract was used in an RT-PCR assay to detect C. botulinum E VH toxin-specific mRNA (Fig. (Fig.2). An analysis of the gel showed that RT-PCR amplified a bp fragment from C. botulinum E VH target mRNA, as by: sion of the type B botulinum neurotoxin (BoNT/B) gene (cntB)inClostridium botulinum.

The levels ofcntB mRNA in five type B strains were accurately monitored by using primers specific for cntB and for the reference gene encoding the 16S rRNA.

The patterns and relative expression of cntB were different in the different by:   The competitive RT-PCR method was developed to examine C. botulinum E VH toxin gene expression and was used to investigate the level of toxin production by C. botulinum E VH when the organism was grown in two different types of by: Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E Uma Basavanna, 1 Tim Muruvanda, 1 Eric W.

Brown, 1 and Shashi K. Sharma 1 1 Division of Microbiology, Office of Regulatory Science, Center for Food Safety and Applied Nutrition, Food and Drug Administration, CPK1, HFS, Paint Branch Parkway, College Park, Cited by: A specific and sensitive combined selection and enrichment PCR procedure was developed for the detection of Clostridium botulinum types B, E, and F in fecal samples from slaughtered pigs.

Botulism outbreak due to consumption of food contaminated with botulinum neurotoxins (BoNTs) is a public health emergency. The threat of bioterrorism through deliberate distribution in food sources and/or aerosolization of BoNTs raises global public health and security concerns due to the potential for high mortality and morbidity.

Rapid and reliable detection methods are necessary to Cited by: 5. A quantitative reverse transcription-PCR (qRT-PCR) method was developed to monitor the relative expression of the type B botulinum neurotoxin (BoNT/B) gene (cntB) in Clostridium botulinum.

the C. botulinum bacterium that will later release the botulism toxin. Adult intestinal toxemia botulism-Rare but follows the same route of infection as infant botulism. Inhalation botulism-Results from breathing in botulinum toxin as an aerosol, but is rare. Wound botulism-Occurs when C.

botulinum spores contaminate a wound. Size: KB. dose of botulinum toxin compared to that of an international standard antitoxin with known potency.

However, the mouse assay is time consuming, labor intensive, costly, necessitates a large number of. A quantitative reverse transcription-PCR (qRT-PCR) method was developed to monitor the relative expression of the type B botulinum neurotoxin (BoNT/B) gene (cntB) in Clostridium botulinum.

The levels of cntB mRNA in five type B strains were accurately monitored by using primers specific for cntB and for the reference gene encoding the 16S rRNA. The patterns and relative expression Cited by: The competitive RT-PCR method was developed to examineC.

botulinum E VH toxin gene expression and was used to investigate the level of toxin production by C. botulinum E VH. Clostridium botulinum, the causative agent of botulism, is an example of a non-monophyletic bacterial species where horizontal gene transfer events have unlinked taxonomy from phylogeny.A strain is classified as C.

botulinum based on the presence of a gene cluster encoding a functional botulinum neurotoxin (BoNT) and other associated components. However, the bont gene cluster has a history. @article{osti_, title = {Arrangement of the Clostridium baratii F7 Toxin Gene Cluster with Identification of a σ Factor That Recognizes the Botulinum Toxin Gene Cluster Promoters}, author = {Dover, Nir and Barash, Jason R.

and Burke, Julianne N. and Hill, Karen K. and Detter, John C. and Arnon, Stephen S.}, abstractNote = {Botulinum neurotoxin (BoNT) is the most poisonous substances. BotR/A and TetR are alternative RNA sigma factors controlling the expression of the neurotoxin and associated protein genes in Clostridium botulinum type A and Clostridium tetani.

Mol. Microbiol. –/jxCited by: Clostridia are the bacteria which produce the most numerous and most potent toxins. • Genetic characteristics have been mainly unraveled in Clostridium perfringens, Clostridium difficile and Clostridium botulinum. Horizontal toxin gene transfer and subsequent evolution are the main mechanisms of toxin by: C2 toxin produced from Clostridium botulinum serotypes C and D has a potential role in many pathophysiological mechanisms in birds and animals.

It has encompassed an ADP ribosyltransferase subunit (C2I) and a translocation/binding subunit (C2II). In the present study, we intended to produce C2I mutant proteins as recombinant subunit vaccines by using glutathione-S-transferase-gene fusion Cited by: 2.

Chapter Clostridium botulinum Toxin Formation (A Biological Hazard) Continued Hazard Analysis Worksheet STEP # UNDERSTAND THE POTENTIAL HAZARD. Clostridium botulinum toxin formation can result in consumer illness and death. This chapter covers the potential for C.

botulinum growth and toxin forma- tion as a result of time/temperature abuse duringFile Size: KB. Positive Regulation of Botulinum Neurotoxin Gene Expression by CodY in Clostridium botulinum ATCC Article (PDF Available) in Applied and Environmental Microbiology 80(24).

A rapid competitive RT/PCR assay was developed to determine the effects of nutrients on Clostridium botulinum type E toxin gene expression. The type E strain (EVH) was grown in a nutrient-rich broth containing 1% glucose (base medium).

Toxin gene expression was quantified at both mid and late exponential phases of by: 6. Avian botulism is a serious neuroparalytic disease mainly caused by a type C/D botulinum neurotoxin produced by Clostridium botulinum group III, one of the entwined bacterial species from the Clostridium novyi sensu lato genospecies.

Its isolation is very challenging due to the absence of selective media and the instability of the phage carrying the gene encoding for the neurotoxin.Genetic analysis focused on BoNT genes and flanking genes, which elucidated the botulinum locus encompassing the genes encoding BoNT, associated nontoxic proteins, and the regulator Bot/R, and provided evidence of its variation among the different toxinotypes.

Whole genomes sequencing is now available for eight Clostridium botulinum strains and is in progress for several other strains.Competitive reverse transcription polymerase chain reaction (cRT-PCR) was used to quantify the toxin-encoding mRNA production of a Clostridium botulinum type E strain in media containing either sorbic acid or sodium nitrite.

A fold reduction in toxin mRNA production and a fold reduction in the proportion of toxin mRNA to total RNA, was estimated when either 1 mg ml −1 sorbic acid or Cited by: