Enzyme-Linked Phage Receptor Binding Protein Assays (ELPRA) Enable Identification of Bacillus anthracis Colonies

Viruses. 2021 Jul 27;13(8):1462. doi: 10.3390/v13081462.

Abstract

Bacteriophage receptor binding proteins (RBPs) are employed by viruses to recognize specific surface structures on bacterial host cells. Recombinant RBPs have been utilized for detection of several pathogens, typically as fusions with reporter enzymes or fluorescent proteins. Identification of Bacillus anthracis, the etiological agent of anthrax, can be difficult because of the bacterium's close relationship with other species of the Bacillus cereussensu lato group. Here, we facilitated the identification of B. anthracis using two implementations of enzyme-linked phage receptor binding protein assays (ELPRA). We developed a single-tube centrifugation assay simplifying the rapid analysis of suspect colonies. A second assay enables identification of suspect colonies from mixed overgrown solid (agar) media derived from the complex matrix soil. Thus, these tests identified vegetative cells of B. anthracis with little processing time and may support or confirm pathogen detection by molecular methods such as polymerase chain reaction.

Keywords: Bacillus anthracis; ELPRA; anthrax; bacteriophage; enzyme-linked phage protein assay; receptor binding protein; reporter fusions.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anthrax / microbiology*
  • Bacillus Phages / genetics
  • Bacillus Phages / physiology
  • Bacillus anthracis / genetics
  • Bacillus anthracis / isolation & purification*
  • Bacillus anthracis / metabolism
  • Bacillus anthracis / virology
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Bacteriological Techniques / instrumentation
  • Bacteriological Techniques / methods*
  • Bacteriophage Receptors / chemistry*
  • Bacteriophage Receptors / genetics
  • Bacteriophage Receptors / metabolism
  • Genes, Reporter
  • Humans
  • Luciferases / chemistry
  • Luciferases / genetics
  • Luciferases / metabolism
  • Luminescent Measurements / methods*
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Red Fluorescent Protein
  • Soil Microbiology

Substances

  • Bacterial Proteins
  • Bacteriophage Receptors
  • Luminescent Proteins
  • Luciferases