[18] The enzyme-linked immunosorbent assay (ELISA) for the detection and determination of Clostridium botulinum toxins A, B, and E

Notermans, S.; Hagenaars, A. M.; Kozaki, Shunji

doi:10.1016/0076-6879(82)84020-2

Cited by 34 publications

(15 citation statements)

References 20 publications

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“…Briefly, Falcon microtest assay plates (BD Biosciences) were coated with an optimal concentration of purified BoNT/A (100 l of 2 g/ml) in PBS overnight at 4°C (35). Two-fold serial dilutions of samples were added after blocking with 1% BSA.…”

Section: Ab Assaysmentioning

confidence: 99%

A Novel Neurotoxoid Vaccine Prevents Mucosal Botulism

Kobayashi

Kohda

Kataoka

et al. 2005

The Journal of Immunology

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The threat posed by botulism, classically a food- and waterborne disease with a high morbidity and mortality, has increased exponentially in an age of bioterrorism. Because botulinum neurotoxin (BoNT) could be easily disseminated by terrorists using an aerosol or could be used to contaminate the food or water supply, the Centers for Disease Control and Prevention and the National Institute of Allergy and Infectious Diseases has classified it as a category A agent. Although clearly the development of a safe and effective mucosal vaccine against this toxin should be a high priority, essentially no studies to date have assessed mucosal immune responses to this disease. To bridge this gap in our knowledge, we immunized mice weekly for 4 wk with nasal doses of BoNT type A toxoid and a mutant of cholera toxin termed E112K. We found elevated levels of BoNT-specific IgG Abs in plasma and of secretory IgA Abs in external secretions (nasal washes, saliva, and fecal extracts). When mice given nasal BoNT vaccine were challenged with 4 × 103 LD50 of BoNT type A (BoNT/A) via the i.p. route, complete protection was seen, while naive mice given the same dosage died within 2 h. To further confirm the efficacy of this nasal BoNT vaccine, an oral LD50 was determined. When mice were given an oral challenge of 5 μg (2 × oral LD50) of progenitor BoNT/A, all immunized mice survived beyond 5 days, while nonimmunized mice did not. The fecal extract samples from nasally vaccinated mice were found to contain neutralizing secretory IgA Abs. Taken together, these results show that nasal BoNT/A vaccine effectively prevents mucosal BoNT intoxication.

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Section: Ab Assaysmentioning

confidence: 99%

A Novel Neurotoxoid Vaccine Prevents Mucosal Botulism

Kobayashi

Kohda

Kataoka

et al. 2005

The Journal of Immunology

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“…A number of in vitro assays have been developed; these included enzyme-linked immunosorbent assays, radioimmunoassays, and PCR probe identification assays. At this time, none of these is sensitive enough to replace the mouse bioassay (1,8,15,16,23,25).…”

mentioning

confidence: 99%

Quantification of Clostridium botulinum Toxin Gene Expression by Competitive Reverse Transcription-PCR

McGrath¹,

Dooley²,

Haylock³

2000

Appl Environ Microbiol

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Clostridium botulinum produces a characteristic botulinum neurotoxin which can cause an often fatal neuroparalytic condition known as botulism. Although food-borne botulism is rare, critical screening by food companies is necessary to ensure that food products are safe. At present, the food industry assesses the risks of botulinum neurotoxin production by challenge testing to check any new food products and to check the efficacy of new storage regimes. Challenge testing involves artificial introduction of defined strains of microorganisms into food, and microbial growth and possible toxin production are then monitored. Botulinum toxin is normally analyzed by using the mouse bioassay. However, the mouse bioassay is expensive, slow, and politically sensitive because of animal rights issues. In this paper we describe adaptation of a new assay, competitive reverse transcription-PCR (RT-PCR), to monitor botulinum neurotoxin production. This method accurately measures the level of toxin-encoding mRNA in C. botulinum cells. Measurement of mRNA should provide a good indication of gene expression as mRNA is turned over rapidly in bacterial cells. In addition, the method is rapid, specific, and sensitive. 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 broth. The results which we obtained with the competitive RT-PCR method demonstrated that this method is more rapid and more sensitive than the mouse bioassay.

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“…ELISAs have been developed for C. botulinum toxin type A (185,186), type B (187,188), and type E (188). These assays, however, are generally insufficiently sensitive to fully substitute mouse tests.…”

Section: Determination Of Toxinsmentioning

confidence: 98%

Changing perspectives in food microbiology: Implementation of rapid microbiological analyses in modern food processing

Veld

Hartog

Hofstra

1988

Food Reviews International

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This review article summarizes the perspective for the introduction of novel microbiological techniques into modern food manufacturing and processing. Changes are discussed that have occurred recently in the microbiological control of food operations (process microbiology)-for example, the "hazard analysis critical control point" (HACCP) concept, which can be elaborated to a longitudinal integrated chain assessment by monitoring critical control points. In addition, we describe the recent developments in rapid analytical methodology that can be implemented under these concepts. These techniques are based on two major approaches; that is, they are either aimed at estimating the total microbial load as a hygiene parameter, or at detecting and enumerating specific pathogenic or toxigenic microorganisms and their metabolites. We conclude that, at this moment, several of these methods are significant improvements compared with the conventional techniques. However their use in preventive quality assurance by integrated chain assessment, as well as in routine food (end product) control, has been limited to certain areas of the food industries as yet.

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[18] The enzyme-linked immunosorbent assay (ELISA) for the detection and determination of Clostridium botulinum toxins A, B, and E

Cited by 34 publications

References 20 publications

A Novel Neurotoxoid Vaccine Prevents Mucosal Botulism

A Novel Neurotoxoid Vaccine Prevents Mucosal Botulism

Quantification of Clostridium botulinum Toxin Gene Expression by Competitive Reverse Transcription-PCR

Changing perspectives in food microbiology: Implementation of rapid microbiological analyses in modern food processing

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