A Rapid, Sensitive, and Portable Biosensor Assay for the Detection of Botulinum Neurotoxin Serotype A in Complex Food Matrices

Tam, Christina; Flannery, Andrew R.; Cheng, Luisa W.

doi:10.3390/toxins10110476

Cited by 12 publications

(11 citation statements)

References 37 publications

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“…Recent developments of cell-based assays provide detection methods that are more sensitive than the MBA, and reflect all BoNT steps of activity; however, they have to be adapted for each BoNT type, whereas the MBA is sensitive to all BoNT types [ 107 , 109 ].…”

Section: Discussion: Suitability Of Each Methods For Their Applicationsmentioning

confidence: 99%

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Recent Developments in Botulinum Neurotoxins Detection

Rasetti-Escargueil

Popoff

2022

Microorganisms

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Botulinum neurotoxins (BoNTs) are produced as protein complexes by bacteria of the genus Clostridium that are Gram-positive, anaerobic and spore forming (Clostridium botulinum, C. butyricum, C. baratii and C. argentinense spp.). BoNTs show a high immunological and genetic diversity. Therefore, fast, precise, and more reliable detection methods are still required to monitor outbreaks and ensure surveillance of botulism. The botulinum toxin field also comprises therapeutic uses, basic research studies and biodefense issues. This review presents currently available detection methods, and new methods offering the potential of enhanced precision and reproducibility. While the immunological methods offer a range of benefits, such as rapid analysis time, reproducibility and high sensitivity, their implementation is subject to the availability of suitable tools and reagents, such as specific antibodies. Currently, the mass spectrometry approach is the most sensitive in vitro method for a rapid detection of active or inactive forms of BoNTs. However, these methods require inter-laboratory validation before they can be more widely implemented in reference laboratories. In addition, these surrogate in vitro models also require full validation before they can be used as replacement bioassays of potency. Cell-based assays using neuronal cells in culture recapitulate all functional steps of toxin activity, but are still at various stages of development; they are not yet sufficiently robust, due to high batch-to-batch cell variability. Cell-based assays have a strong potential to replace the mouse bioassay (MBA) in terms of BoNT potency determination in pharmaceutical formulations; they can also help to identify suitable inhibitors while reducing the number of animals used. However, the development of safe countermeasures still requires the use of in vivo studies to complement in vitro immunological or cell-based approaches.

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Section: Discussion: Suitability Of Each Methods For Their Applicationsmentioning

confidence: 99%

“…Luminescence is measured with a luminometer. The sensitivities of this assay are 10 ng/mL with BoNT/A in buffer; 7.4 to 62.5 ng/mL according to various food matrices (milk, vegetables, meat, fish); and 171 ng/mL in egg matrix [ 107 ].…”

Section: Bont Detectionmentioning

confidence: 99%

Recent Developments in Botulinum Neurotoxins Detection

Rasetti-Escargueil

Popoff

2022

Microorganisms

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“…Christina CT et al tried to detect botulinum toxin in undiluted liquid eggs but failed. Therefore, it is proposed that before adding BoNT/A, liquid eggs must be diluted 1:10 with detection buffer ( 5 ).…”

Section: Discussionmentioning

confidence: 99%

“…Besides neurotoxicity, BoNTs also kills animals by entering the digestive system and ultimately weakening the vital organs. The exceptionally low lethal dose of BoNTs is estimated from primate studies to be 1 µg/kg body weight when received orally, 10 ng/kg by inhalation, and 1 ng/kg when received intravenously or intramuscularly (2)(3)(4)(5).…”

Section: Introductionmentioning

confidence: 99%

Rapid and sensitive detection of botulinum toxin type A in complex sample matrices by AlphaLISA

Zhang

Lv²,

Zheng³

et al. 2022

Front. Public Health

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Botulinum toxin A(BoNT/A) is a neurotoxin produced by the bacteria Clostridium botulinum, which can cause serious food poisoning and is recognized as a potential biological warfare agent. BoNT/A is does not degrade easily and can remain in the complex matrix for a long time. Meanwhile, the poisonous dose of botulinum toxin exceptionally low and intravenous human lethal doses estimated at 1-3 ng/kg. Therefore, sensitive and accurate detection methods suitable for testing a wide range of complex samples are urgently needed. To this end, the “amplified luminescent proximity homogeneous assay linked immunosorbent assay” (AlphaLISA) was established for the detection of BoNT/A and its detection efficacy in plasma, beverage, food, and other complex samples was evaluated. The results showed that this method can very effectively resist matrix interference. The detection time is rapid, reaching a detection limit for all samples of up to 0.1 ng/mL in only 30 min. BoNT/A can also be accurately detected in vomit samples of patients with clinical food poisoning. This study demonstrates that AlphaLISA is an effective tool for the detection of BoNT/A in complex samples and can potentially be developed for commercial use in the future.

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“…BoNT binds irreversibly to the presynaptic membranes of peripheral neuromuscular and autonomic nerve junctions, causing a symmetric, descending flaccid paralysis. [2,3] Clostridium botulinum is found in the environment as dormant spores but germinates and produces toxins in foods under permissive conditions of pH, water activity, anaerobic environment, and temperature. Bacterial growth is inhibited by refrigeration below 4°C or by heating above 121°C.…”

Section: Introductionmentioning

confidence: 99%

Foodborne botulism in eastern Poland: a hospital-based retrospective study and epidemiological data review

Krzowska-Firych

Mikłaszewska

Tomasiewicz

2020

International Journal of Food Properties

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Foodborne botulism is a potentially fatal illness caused by consuming food with neurotropic exotoxins produced by Clostridium botulinum and other species capable of producing botulinum toxins. Its incidence in Poland has decreased but new cases are still noted. The purpose of this paper is to provide a detailed information on epidemiological data and clinical features of patients with botulism hospitalized at the Department of Infectious Diseases in Lublin Province (Eastern Poland). We have retrospectively analyzed medical records from the years 2010-2018 of all patients with foodborne botulism that have been hospitalized at the Department of Infectious Diseases, Medical University of Lublin. A standard case report form was completed for each patient. This form included case demographics, clinical features, botulinum testing results, and type of exposure. During the period 2010-2018, a total of 35 patients with foodborne botulism were hospitalized. Of these, 24 cases were laboratory confirmed. Males accounted for 80.0 %, and 71.4% of all cases were residents from rural areas. All patients presented with typical signs and symptoms of botulism, and in were given trivalent ABE antitoxin on admission. Toxin type B was implicated in 58.3% of cases. The most common food source was home-preserved pork meat. Stronger efforts should be made toward raising public awareness of the risk of foodborne botulism, especially with respect to home-preserved foods.

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A Rapid, Sensitive, and Portable Biosensor Assay for the Detection of Botulinum Neurotoxin Serotype A in Complex Food Matrices

Cited by 12 publications

References 37 publications

Recent Developments in Botulinum Neurotoxins Detection

Recent Developments in Botulinum Neurotoxins Detection

Rapid and sensitive detection of botulinum toxin type A in complex sample matrices by AlphaLISA

Foodborne botulism in eastern Poland: a hospital-based retrospective study and epidemiological data review

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