Field-Based Multiplex Detection of Biothreat Agents

Pöhlmann, Christopher; Elßner, Thomas

doi:10.1007/978-3-319-91791-7_5

Cited by 3 publications

(5 citation statements)

References 14 publications

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“…In the present study, electrochemical readable anti-idiotypic antibody based competitive immunoassays allowing the detection of the phycotoxin STX and the mycotoxins T-2 as well as AFM1 and corresponding congeners, respectively, using a portable and fully automated detection platform (pBDi, portable BioDetector integrated) (Pöhlmann and Elßner, 2018) were developed. This electrochemical detection platform with corresponding biochips based on a non-competitive immunoassay format has already been tested for detection of high molecular weight toxins such as ricin (Worbs et al, 2015) and staphylococcal enterotoxin B (Nia et al, 2016) in the field of biosecurity.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical Biochip Assays Based on Anti-idiotypic Antibodies for Rapid and Automated On-Site Detection of Low Molecular Weight Toxins

et al. 2019

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Phycotoxins and mycotoxins, such as paralytic shellfish poisoning toxins, type A trichothecenes, and aflatoxins are among the most toxic low molecular weight toxins associated with human poisoning incidents through the consumption of naturally contaminated food. Therefore, there is an utmost need for rapid and sensitive on-site detection systems. Herein, an electrochemical biochip for fast detection of saxitoxin, T-2 toxin as well as aflatoxin M1 and their corresponding congeners, respectively, using a portable and fully automated detection platform (pBDi, portable BioDetector integrated) was developed. Toxin analysis is facilitated upon the biochip via an indirect competitive immunoassay using toxin-specific antibodies combined with anti-idiotypic antibodies. The developed biochips enable detection in the low ng/mL-range within 17 min. Moreover, the assays cover a wide linear working range of 2–3 orders of magnitude above the limit of detection with an inter-chip coefficient of variation lower than 15%. The broad specificity of the employed antibodies which react with a large number of congeners within the respective toxin group allows efficient screening of contaminated samples for the presence of these low molecular weight toxins. With respect to the analysis of human urine samples, we focused here on the detection of saxitoxin, HT-2 toxin, and aflatoxin M1, all known as biomarkers of acute toxin exposure. Overall, it was proved that the developed biochip assays can be used to rapidly and reliably identify severe intoxications caused by these low molecular weight toxins.

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Section: Introductionmentioning

confidence: 99%

Electrochemical Biochip Assays Based on Anti-idiotypic Antibodies for Rapid and Automated On-Site Detection of Low Molecular Weight Toxins

et al. 2019

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“…The toxin group specificity of the antibodies enables that as many congeners as possible can be detected in a single analysis. In combination with the existing electrochemical biochip assays for high molecular weight toxins (e.g., botulinum neurotoxins, ricin) as well as for bioterrorism relevant microorganisms (e.g., Bacillus anthracis, Yersinia pestis) [36], the pBDi platform is a valuable tool for rapid and straightforward on-site detection of various kinds of biothreat agents. Moreover, the detection system has the flexibility to combine these assays for different kinds of biological agents on one biochip without modifying the general workflow, thus, offering a new level of biothreat detection.…”

Section: Discussionmentioning

confidence: 99%

“…All pumping and incubation steps as well as electrochemical detection are performed by the fully automated on-site biochip processing platform [36]. Prior to the assay, mAb1 (T-2, AFB1) or mAb2 (STX, MC-LR, RoA) were immobilized on specific positions by physisorption on IDA gold electrodes of the biochip.…”

Section: Fiveplex Biochip Assay Designmentioning

confidence: 99%

“…In this work, we present for the first time a fiveplex electrochemical biochip assay for on-site detection of low molecular weight toxins relying solely on anti-idiotypic antibodies as epitope-mimicking molecules. Moreover, due to the generation of reporter enzyme-labeled monoclonal detection antibodies used as tracer molecules, we demonstrate a straightforward assay allowing simultaneous detection of bioterrorism relevant STX, MC-LR, T-2, RoA and AFB1 as well as corresponding congeners with minimal hands-on time using the automated on-site biochip processing platform pBDi (portable BioDetector integrated) [36]. Furthermore, we demonstrated the applicability of the robust multiplex detection system to screen human serum-beside urine another relevant body fluid-for the presence of STX, MC-LR, HT-2, RoA and AFB1 after intentional or natural occurring poisoning incidents.…”

Section: Introductionmentioning

confidence: 99%

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An Electrochemical Fiveplex Biochip Assay Based on Anti-Idiotypic Antibodies for Fast On-Site Detection of Bioterrorism Relevant Low Molecular Weight Toxins

Schulz

Pöhlmann

Dietrich

et al. 2019

Toxins

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Modern threats of bioterrorism force the need for multiple detection of biothreat agents to determine the presence or absence of such agents in suspicious samples. Here, we present a rapid electrochemical fiveplex biochip screening assay for detection of the bioterrorism relevant low molecular weight toxins saxitoxin, microcystin-LR, T-2 toxin, roridin A and aflatoxin B1 relying on anti-idiotypic antibodies as epitope-mimicking reagents. The proposed method avoids the use of potentially harmful toxin-protein conjugates usually mandatory for competitive immunoassays. The biochip is processed and analyzed on the automated and portable detection platform pBDi within 13.4 min. The fiveplex biochip assay revealed toxin group specificity to multiple congeners. Limits of detection were 1.2 ng/mL, 1.5 ng/mL, 0.4 ng/mL, 0.5 ng/mL and 0.6 ng/mL for saxitoxin, microcystin-LR, T-2 toxin, roridin A or aflatoxin B1, respectively. The robustness of the fiveplex biochip for real samples was demonstrated by detecting saxitoxin, microcystin-LR, HT-2 toxin, roridin A and aflatoxin B1 in contaminated human blood serum without elaborate sample preparation. Recovery rates were between 52–115% covering a wide concentration range. Thus, the developed robust fiveplex biochip assay can be used on-site to quickly detect one or multiple low molecular weight toxins in a single run.

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“…Quite early during development, the first applications based on this electrochemical biochip technology focused on multiplex detection of security sensitive toxins were developed using a prototype reader platform (ePaTox system) [ 142 , 143 ]. Based on these promising results, Bruker developed a portable and ruggedized platform for on-site usage called pBDi as well as several commercially available detection kits for the detection of security sensitive proteotoxins, bacteria and orthopox viruses [ 92 , 93 ]. The pBDi employs electrochemical biochip technology for multiplex fully automated ELISA-based detection of five to six agents simultaneously (for commercially available pBDi Test Kits; for customer specific assays in development, 12-plex immunoassays were already demonstrated).…”

Section: Portable Immunoassay Techniques With Multiplexing Capabilmentioning

confidence: 99%

Multiplex Immunoassay Techniques for On-Site Detection of Security Sensitive Toxins

Pöhlmann

Elßner

2020

Toxins

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Biological toxins are a heterogeneous group of high molecular as well as low molecular weight toxins produced by living organisms. Due to their physical and logistical properties, biological toxins are very attractive to terrorists for use in acts of bioterrorism. Therefore, among the group of biological toxins, several are categorized as security relevant, e.g., botulinum neurotoxins, staphylococcal enterotoxins, abrin, ricin or saxitoxin. Additionally, several security sensitive toxins also play a major role in natural food poisoning outbreaks. For a prompt response to a potential bioterrorist attack using biological toxins, first responders need reliable, easy-to-use and highly sensitive methodologies for on-site detection of the causative agent. Therefore, the aim of this review is to present on-site immunoassay platforms for multiplex detection of biological toxins. Furthermore, we introduce several commercially available detection technologies specialized for mobile or on-site identification of security sensitive toxins.

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Field-Based Multiplex Detection of Biothreat Agents

Cited by 3 publications

References 14 publications

Electrochemical Biochip Assays Based on Anti-idiotypic Antibodies for Rapid and Automated On-Site Detection of Low Molecular Weight Toxins

Electrochemical Biochip Assays Based on Anti-idiotypic Antibodies for Rapid and Automated On-Site Detection of Low Molecular Weight Toxins

An Electrochemical Fiveplex Biochip Assay Based on Anti-Idiotypic Antibodies for Fast On-Site Detection of Bioterrorism Relevant Low Molecular Weight Toxins

Multiplex Immunoassay Techniques for On-Site Detection of Security Sensitive Toxins

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