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Article

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

by
Peter Braun
,
Immanuel Wolfschläger
,
Leonie Reetz
,
Lilia Bachstein
,
Ana Clara Jacinto
,
Carolina Tocantins
,
Johannes Poppe
and
Gregor Grass
*
Department of Bacteriology and Toxinology, Bundeswehr Institute of Microbiology (IMB), 80937 Munich, Germany
*
Author to whom correspondence should be addressed.
Microorganisms 2020, 8(6), 934; https://doi.org/10.3390/microorganisms8060934
Submission received: 29 May 2020 / Revised: 18 June 2020 / Accepted: 19 June 2020 / Published: 21 June 2020
(This article belongs to the Special Issue An Update on Anthrax)
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Abstract

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.
Keywords: anthrax; Bacillus anthracis; bacteriophage; receptor binding protein; reporter fusions; detection assay anthrax; Bacillus anthracis; bacteriophage; receptor binding protein; reporter fusions; detection assay
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MDPI and ACS Style

Braun, P.; Wolfschläger, I.; Reetz, L.; Bachstein, L.; Jacinto, A.C.; Tocantins, C.; Poppe, J.; Grass, G. Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages. Microorganisms 2020, 8, 934. https://doi.org/10.3390/microorganisms8060934

AMA Style

Braun P, Wolfschläger I, Reetz L, Bachstein L, Jacinto AC, Tocantins C, Poppe J, Grass G. Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages. Microorganisms. 2020; 8(6):934. https://doi.org/10.3390/microorganisms8060934

Chicago/Turabian Style

Braun, Peter, Immanuel Wolfschläger, Leonie Reetz, Lilia Bachstein, Ana Clara Jacinto, Carolina Tocantins, Johannes Poppe, and Gregor Grass. 2020. "Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages" Microorganisms 8, no. 6: 934. https://doi.org/10.3390/microorganisms8060934

APA Style

Braun, P., Wolfschläger, I., Reetz, L., Bachstein, L., Jacinto, A. C., Tocantins, C., Poppe, J., & Grass, G. (2020). Rapid Microscopic Detection of Bacillus anthracis by Fluorescent Receptor Binding Proteins of Bacteriophages. Microorganisms, 8(6), 934. https://doi.org/10.3390/microorganisms8060934

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