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Strategies for capturing Bacillus thuringiensis spores on surfaces of (001) GaAs-based biosensors.
Moteshareie, Houman; Hassen, Walid M; Vermette, Jonathan; Dubowski, Jan J; Tayabali, Azam F.
Afiliação
  • Moteshareie H; Biotechnology Laboratory, Environmental Health Science and Research Bureau, Healthy Environments and Consumer Safety Branch, Health Canada, Ottawa, Ontario, Canada; Interdisciplinary Institute for Technological Innovation (3IT), CNRS UMI-3463, Laboratory for Quantum Semiconductors and Photon-based B
  • Hassen WM; Interdisciplinary Institute for Technological Innovation (3IT), CNRS UMI-3463, Laboratory for Quantum Semiconductors and Photon-based BioNanotechnology, Department of Electrical and Computer Engineering, Sherbrooke, Québec, Canada.
  • Vermette J; Interdisciplinary Institute for Technological Innovation (3IT), CNRS UMI-3463, Laboratory for Quantum Semiconductors and Photon-based BioNanotechnology, Department of Electrical and Computer Engineering, Sherbrooke, Québec, Canada.
  • Dubowski JJ; Interdisciplinary Institute for Technological Innovation (3IT), CNRS UMI-3463, Laboratory for Quantum Semiconductors and Photon-based BioNanotechnology, Department of Electrical and Computer Engineering, Sherbrooke, Québec, Canada. Electronic address: Jan.J.Dubowski@USherbrooke.ca.
  • Tayabali AF; Biotechnology Laboratory, Environmental Health Science and Research Bureau, Healthy Environments and Consumer Safety Branch, Health Canada, Ottawa, Ontario, Canada; Interdisciplinary Institute for Technological Innovation (3IT), CNRS UMI-3463, Laboratory for Quantum Semiconductors and Photon-based B
Talanta ; 236: 122813, 2022 Jan 01.
Article em En | MEDLINE | ID: mdl-34635209
ABSTRACT
Bacillus thuringiensis (Bt) is used as a bioinsecticide since it effectively kills insect larvae. Bt is also genetically similar to Bacillus cereus (Bc), a well recognized foodborne human pathogen; they are both members of the Bacillus cereus group (BC group). Although approved Bt bioinsecticide products have been confirmed to be non-pathogenic to humans, close monitoring of Bt during dissemination is important for cost considerations and to limit impact on biodiversity towards nontarget organisms. As such, developing rapid, sensitive, and specific tools for quantitative detection of Bt spores during and following spray operations is highly desirable. The goals of this study were to investigate commercially available detection reagents for sensitivity and selectivity in detecting Bt spores, and then functionalize a surface of (001) GaAs used in photonic biosensing. To achieve these goals, we (1) screened commercial antibodies for their capacity to bind recombinant proteins from Bt spores, (2) screened antibodies and aptamers for their sensitivity and selectivity against Bt spores, and (3) tested the efficiency of selected antibodies and aptamers in capturing Bt spores on the surface of functionalized GaAs biochips. Seven genes encoding Bt spore proteins were cloned and expressed in Escherichia coli. The binding of each purified spore antigen was tested by commercially available polyclonal and monoclonal antibodies claimed to exclusively target spores. Of the seven targets, Bacillus collagen-like protein A, was the most abundant protein on Bt spores and demonstrated the strongest binding affinity to all test antibodies. The commercial antibodies (Abs) were also tested for specificity to BC Group versus non-BC Group spores. Three of six commercial antibodies showed selectivity to Bt spores, with recombinant Abs providing the most robust lower range of detection (102 to 6 × 103 spores/mL). The sensitivity and selectivity of three published DNA aptamer sequences demonstrated a wide range of detection sensitivity for Bt spores. Two of the three test aptamers also showed reasonable selectivity towards Bt spores while the third demonstrated reactivity to non-BC Group B. megaterium and B. subtilis. Of the reagents tested, a thiolated aptamer and llama recombinant Ab showed highest Bt spore capture efficiency as measured by spore coverage of the GaAs surface. These results confirm that the selected aptamer and llama rAb can be considered strong candidates for the development of GaAs-based biosensing devices.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bacillus anthracis / Bacillus thuringiensis / Técnicas Biossensoriais Limite: Humans Idioma: En Revista: Talanta Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bacillus anthracis / Bacillus thuringiensis / Técnicas Biossensoriais Limite: Humans Idioma: En Revista: Talanta Ano de publicação: 2022 Tipo de documento: Article