Amplified Immunoassay ELISA-ELCA for Measuring Clostridium botulinum Type E Neurotoxin in Fish Fillets

Roman, Michael; Humber, John Y.; Hall, Paul A.; Reddy, N. R.; Solomon, Haim M.; Triscott, Mark; Beard, G. Alan; Bottoms, J D; Cheng, Therese M.; Doellgast, George J.

doi:10.4315/0362-028x-57.11.985

Cited by 24 publications

(20 citation statements)

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“…The sensitivity of the conventional ELISAs for botulinum neurotoxin detection is some 10-to 100-fold lower than that of the mouse bioassay (47,164,174,177) (Table 2). Signal amplification using a chromogenic diaphorase system (189), biotinylated antibodies and avidin-enzyme conjugate (66,68), or enzyme-linked coagulation assay (ELCA) (53,54,178) has enabled increased assay sensitivity.…”

Section: Detection Of Botulinum Neurotoxinmentioning

confidence: 99%

Laboratory Diagnostics of Botulism

2006

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SUMMARY Botulism is a potentially lethal paralytic disease caused by botulinum neurotoxin. Human pathogenic neurotoxins of types A, B, E, and F are produced by a diverse group of anaerobic spore-forming bacteria, including Clostridium botulinum groups I and II, Clostridium butyricum, and Clostridium baratii. The routine laboratory diagnostics of botulism is based on the detection of botulinum neurotoxin in the patient. Detection of toxin-producing clostridia in the patient and/or the vehicle confirms the diagnosis. The neurotoxin detection is based on the mouse lethality assay. Sensitive and rapid in vitro assays have been developed, but they have not yet been appropriately validated on clinical and food matrices. Culture methods for C. botulinum are poorly developed, and efficient isolation and identification tools are lacking. Molecular techniques targeted to the neurotoxin genes are ideal for the detection and identification of C. botulinum, but they do not detect biologically active neurotoxin and should not be used alone. Apart from rapid diagnosis, the laboratory diagnostics of botulism should aim at increasing our understanding of the epidemiology and prevention of the disease. Therefore, the toxin-producing organisms should be routinely isolated from the patient and the vehicle. The physiological group and genetic traits of the isolates should be determined.

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Section: Detection Of Botulinum Neurotoxinmentioning

confidence: 99%

Laboratory Diagnostics of Botulism

2006

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“…ELISA-ELCA detected toxin in fillets packaged in all atmospheres on the same sampling day as the mouse bioassay (Roman et al, 1994).…”

Section: Relationship Of Sensory Spoilage To Toxin Productionmentioning

confidence: 99%

Toxin Development by Clostridium botulinum in Modified Atmosphere‐Packaged Fresh Tilapia Fillets During Storage

Reddy

Paradis

Roman

et al. 1996

Journal of Food Science

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Potential for toxin development by Clostridium botulinum type E was investigated in retail-type packages of fresh tilapia fillets packaged in high barrier film under selected atmospheres [100% air, a modified atmosphere (MA) containing 75%CO 2 :25%N 2 , and vacuum] and stored under refrigeration (4ЊC) and abuse temperatures (8 and 16ЊC). Toxin development coincided with sensory spoilage at 16ЊC storage for fillets packaged in either MA, or vacuum. At 8ЊC, toxin development in fillets packaged in either of the atmospheres occurred 7-23 days after sensory spoilage. At 4ЊC, none of MA-packaged fillets became toxic until 10 days after onset of sensory spoilage. Toxin development did not precede sensory spoilage in any treatments or temperatures studied.

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“…The clear filtrate was tested for the presence of C. botulinum toxin by standard mouse bioassay (FDA, 1992) with minor modifications to reduce trypsin solution concentration (5% trypsin solution used instead of 10%). It was also tested by amplified enzymelinked immunosorbent assay -enzyme-linked coagulation assay (ELISA-ELCA) (Roman et al, 1994). The filtrate was divided into three aliquots.…”

Section: Determination Of the Presence Of C Botulinum Toxinmentioning

confidence: 99%

Shelf Life and Clostridium botulinum Toxin Development during Storage of Modified Atmosphere‐packaged Fresh Catfish Fillets

Reddy¹,

Roman²,

Villanueva³

et al. 1997

Journal of Food Science

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Shelf life (onset of sensory spoilage) and potential for toxin production by Clostridium botulinum type E in retail type packages of fresh catfish fillets in high barrier film were investigated under selected atmospheres when stored under refrigeration and temperature-abuse conditions. Shelf life of fillets in all atmospheres decreased with increase of storage temperature from 4ЊC to 16ЊC. Trimethylamine content associated with onset of spoilage was different for each storage temperature and atmosphere. Surface pH and K-values were not good indicators of onset of sensory spoilage. Toxin development coincided with sensory spoilage at 16ЊC storage for fillets packaged in either atmosphere. At 4ЊC, none of the MA-packaged fillets became toxic, even after 37 days of sensory spoilage.

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Amplified Immunoassay ELISA-ELCA for Measuring Clostridium botulinum Type E Neurotoxin in Fish Fillets

Cited by 24 publications

References 0 publications

Laboratory Diagnostics of Botulism

Laboratory Diagnostics of Botulism

Toxin Development by Clostridium botulinum in Modified Atmosphere‐Packaged Fresh Tilapia Fillets During Storage

Shelf Life and Clostridium botulinum Toxin Development during Storage of Modified Atmosphere‐packaged Fresh Catfish Fillets

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