Detection and Identification of Bacillus anthracis: From Conventional to Molecular Microbiology Methods

Zasada, Aleksandra Anna

doi:10.3390/microorganisms8010125

Cited by 41 publications

(48 citation statements)

References 101 publications

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“…The confirmatory specific identification of B. anthracis is often achieved by means of antigen–antibody interaction, be it in the form of enzyme-linked immunosorbent assays (ELISA) [ 38 ], lateral flow assays [ 39 , 40 ] or by the use of fluorescently labeled antibodies in microscopy [ 41 ] (further alternative detection techniques for B. anthracis are reviewed in a contribution to the special issue “An Update on Anthrax” of Microorganisms [ 42 ]). The direct fluorescent-antibody (DFA) assay [ 41 ] may be seen as being related to the study at hand insofar as both methods take advantage of fluorescence reporters for detecting presumptive B. anthracis cultures and thus helping confirming the identity of this notorious biothreat agent.…”

Section: Discussionmentioning

confidence: 99%

Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages

Braun¹,

Wolfschläger²,

Reetz³

et al. 2020

Microorganisms

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Bacillus anthracis, the etiological agent of anthrax disease, is typically diagnosed by immunological and molecular methods such as polymerase chain reaction (PCR). Alternatively, mass spectrometry techniques may aid in confirming the presence of the pathogen or its toxins. However, because of the close genetic relationship between B. anthracis and other members of the Bacillus cereus sensu lato group (such as Bacillus cereus or Bacillus thuringiensis) mis- or questionable identification occurs frequently. Also, bacteriophages such as phage gamma (which is highly specific for B. anthracis) have been in use for anthrax diagnostics for many decades. Here we employed host cell-specific receptor binding proteins (RBP) of (pro)-phages, also known as tail or head fibers, to develop a microscopy-based approach for the facile, rapid and unambiguous detection of B. anthracis cells. For this, the genes of (putative) RBP from Bacillus phages gamma, Wip1, AP50c and from lambdoid prophage 03 located on the chromosome of B. anthracis were selected. Respective phage genes were heterologously expressed in Escherichia coli and purified as fusions with fluorescent proteins. B. anthracis cells incubated with either of the reporter fusion proteins were successfully surface-labeled. Binding specificity was confirmed as RBP fusion proteins did not bind to most isolates of a panel of other B. cereus s.l. species or to more distantly related bacteria. Remarkably, RBP fusions detected encapsulated B. anthracis cells, thus RBP were able to penetrate the poly-γ-d-glutamate capsule of B. anthracis. From these results we anticipate this RBP-reporter assay may be useful for rapid confirmative identification of B. anthracis.

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Section: Discussionmentioning

confidence: 99%

Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages

Braun¹,

Wolfschläger²,

Reetz³

et al. 2020

Microorganisms

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“…Conserved regions within the 16S rRNA gene of the microbes’ DNA are usually utilized, which enhance sequence comparison. These provide an essential tool for the study evolutionary phylogenetic and molecular diversity [ 55 ]. Therefore, the antagonists isolates were subjected to molecular analysis for identification.…”

Section: Discussionmentioning

confidence: 99%

In vitro antibacterial activities and molecular characterization of bacterial species isolated from farmlands against selected pathogens

Onajobi

Idowu

Adeyemi

et al. 2020

Biotechnology Reports

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Highlights Screening bacterial isolates for antagonistic potential. Molecular methods for identification of isolates. Six potent antibacterial isolates against the test pathogenic species. Pseudomonas was susceptible to antagonizing strains. Necessity for new effective antibacterial formulation.

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“…Another paper describes an agar plate medium, named PLET, which contains the following ingredients added to Difco Heart Infusion Agar (HIA): polymyxin, lysozyme, disodium ethylenediaminetetraacetate, (EDTA), and thallous [ 57 ]. These plates seem to be superior to other plates for the isolation of B. anthracis from other spore-forming bacilli [ 58 ]. There are some limitations for this medium such as stricter safety and environmental regulations due to the presence of thallium acetate [ 59 ], the ability of other Bacillus strains to grow on the PLET agar [ 60 ], and the reduction in the germination of B. anthracis spores on this media [ 61 ].…”

Section: Bacterial Bioterror Agents— B Anthracis Y Pestis and F Tularensismentioning

confidence: 99%

Beating the Bio-Terror Threat with Rapid Antimicrobial Susceptibility Testing

et al. 2021

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A bioterror event using an infectious bacterium may lead to catastrophic outcomes involving morbidity and mortality as well as social and psychological stress. Moreover, a bioterror event using an antibiotic resistance engineered bacterial agent may raise additional concerns. Thus, preparedness is essential to preclude and control the dissemination of the bacterial agent as well as to appropriately and promptly treat potentially exposed individuals or patients. Rates of morbidity, death, and social anxiety can be drastically reduced if the rapid delivery of antimicrobial agents for post-exposure prophylaxis and treatment is initiated as soon as possible. Availability of rapid antibiotic susceptibility tests that may provide key recommendations to targeted antibiotic treatment is mandatory, yet, such tests are only at the development stage. In this review, we describe the recently published rapid antibiotic susceptibility tests implemented on bioterror bacterial agents and discuss their assimilation in clinical and environmental samples.

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Detection and Identification of Bacillus anthracis: From Conventional to Molecular Microbiology Methods

Cited by 41 publications

References 101 publications

Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages

Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages

In vitro antibacterial activities and molecular characterization of bacterial species isolated from farmlands against selected pathogens

Beating the Bio-Terror Threat with Rapid Antimicrobial Susceptibility Testing

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