Early, Real-Time Medical Diagnosis of Botulism by Endopeptidase-Mass Spectrometry

Rosen, Osnat; Feldberg, Liron; Gura, Sigalit; Brosh‐Nissimov, Tal; Guri, Alex; Zimhony, Oren; Shapiro, Eli; Beth-Din, Adi; Stein, Dana; Ozeri, Eyal; Barnea, Ada; Turgeman, Amram; David, Alon Ben; Schwartz, Arieh; Elhanany, Eytan; Diamant, Eran; Yitzhaki, Shmuel; Zichel, Ran

doi:10.1093/cid/civ861

Cited by 12 publications

(7 citation statements)

References 12 publications

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“…The Endopep-MS is an in-vitro activity assay that can be used to detect botulinum toxins 10 , 15 , 16 ; this assay only requires a few hours to complete, is highly specific and is more sensitive than the gold standard in-vivo mouse bioassay 10 , 15 , 16 . The superior sensitivity, specificity and speed of the Endopep-MS assay have the potential to facilitate both diagnosis and treatment.…”

Section: Discussionmentioning

confidence: 99%

“…The assay only requires a few hours to complete, is highly specific and is more sensitive than the gold standard in-vivo mouse bioassay. Recently, we demonstrated the utility of Endopep-MS in diagnosing infant botulism in a clinically relevant timeframe 10 . The detection of toxin A was reported to clinicians in an intervention-relevant timeframe of 24 hours, days before results could be obtained from the mouse bioassay, which in any case failed to detect the low levels of toxin.…”

Section: Introductionmentioning

confidence: 99%

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Development of a multiplex Endopep-MS assay for simultaneous detection of botulinum toxins A, B and E

Rosen

Feldberg

Yamin

et al. 2017

Sci Rep

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Botulinum neurotoxins (BoNTs) are bacterial proteins that cause botulism, a life-threatening disease. The Endopep-MS assay permits rapid detection and serotypic differential diagnosis of BoNTs. The serotype-specific nature of this assay requires that each serum sample be aliquoted and individually tested, which in addition to the limited volume of clinical samples, especially in infants, points to the need for a multiplex assay. However, previous attempts to develop such an assay have been challenging, mainly due to inhibition of BoNT/A activity by the BoNT/E peptide substrate. BoNT/A and BoNT/E share the same native target protein as their substrate. We hypothesized that the steric interference between the BoNT/A and BoNT/E substrate peptides is responsible for the difficulty in simultaneously assaying these two toxins. To explore the basis for steric interference, we used the reported structure of BoNT/A in complex with SNAP-25 and modelled the structure of BoNT/E with SNAP-25. Following this thorough structural analysis, we designed a new peptide substrate for BoNT/A that maintained the assay sensitivity and allowed, for the first time, simultaneous detection of the three most abundant human botulinum serotypes. Adopting the multiplex assay will minimize the required sample volume and assay time for botulinum detection while maintaining the superior Endopep-MS assay performance.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of a multiplex Endopep-MS assay for simultaneous detection of botulinum toxins A, B and E

Rosen

Feldberg

Yamin

et al. 2017

Sci Rep

Self Cite

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“…The main disadvantage of this test is its 96-h duration and lack of sufficient sensitivity. A new method of endopeptidase-mass spectrometry is gaining credibility as a promising method that addresses the limitations of the mouse assay [ 15 – 19 ]. Electromyography can strongly support the clinical diagnosis showing a typical pattern for botulism.…”

Section: Botulinum Neurotoxinmentioning

confidence: 99%

Toxins as biological weapons for terror—characteristics, challenges and medical countermeasures: a mini-review

Berger

Eisenkraft

Bar-Haim

et al. 2016

Disaster and Mil Med

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Toxins are hazardous biochemical compounds derived from bacteria, fungi, or plants. Some have mechanisms of action and physical properties that make them amenable for use as potential warfare agents. Currently, some toxins are classified as potential biological weapons, although they have several differences from classic living bio-terror pathogens and some similarities to manmade chemical warfare agents. This review focuses on category A and B bio-terror toxins recognized by the Centers for Disease Control and Prevention: Botulinum neurotoxin, staphylococcal enterotoxin B, Clostridium perfringens epsilon toxin, and ricin. Their derivation, pathogenesis, mechanism of action, associated clinical signs and symptoms, diagnosis, and treatment are discussed in detail. Given their expected covert use, the primary diagnostic challenge in toxin exposure is the early detection of morbidity clusters, apart from background morbidity, after a relatively short incubation period. For this reason, it is important that clinicians be familiar with the clinical manifestations of toxins and the appropriate methods of management and countermeasures.

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“…This method has found utilization in various matrices [ 151 ]. In the study, in which all conventional tests were used, botulinum toxin was successfully detected in patient serum using the endopeptidase-MS assay, although all conventional tests gave negative results [ 152 ].…”

Section: Detection Of Selected Protein Toxinsmentioning

confidence: 99%

Proteomic Methods of Detection and Quantification of Protein Toxins

Duracova

Klimentová

Fučíková

et al. 2018

Toxins

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Biological toxins are a heterogeneous group of compounds that share commonalities with biological and chemical agents. Among them, protein toxins represent a considerable, diverse set. They cover a broad range of molecular weights from less than 1000 Da to more than 150 kDa. This review aims to compare conventional detection methods of protein toxins such as in vitro bioassays with proteomic methods, including immunoassays and mass spectrometry-based techniques and their combination. Special emphasis is given to toxins falling into a group of selected agents, according to the Centers for Disease Control and Prevention, such as Staphylococcal enterotoxins, Bacillus anthracis toxins, Clostridium botulinum toxins, Clostridium perfringens epsilon toxin, ricin from Ricinus communis, Abrin from Abrus precatorius or control of trade in dual-use items in the European Union, including lesser known protein toxins such as Viscumin from Viscum album. The analysis of protein toxins and monitoring for biological threats, i.e., the deliberate spread of infectious microorganisms or toxins through water, food, or the air, requires rapid and reliable methods for the early identification of these agents.

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Early, Real-Time Medical Diagnosis of Botulism by Endopeptidase-Mass Spectrometry

Cited by 12 publications

References 12 publications

Development of a multiplex Endopep-MS assay for simultaneous detection of botulinum toxins A, B and E

Development of a multiplex Endopep-MS assay for simultaneous detection of botulinum toxins A, B and E

Toxins as biological weapons for terror—characteristics, challenges and medical countermeasures: a mini-review

Proteomic Methods of Detection and Quantification of Protein Toxins

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