Bacillus anthracis diagnostic detection and rapid antibiotic susceptibility determination using ‘bioluminescent’ reporter phage

• Published in: Journal of microbiological methods 2013-11, Vol.95 (2), p.156-161
• Main Authors: Schofield, David A., Sharp, Natasha J., Vandamm, Joshua, Molineux, Ian J., Spreng, Krista A., Rajanna, Chythanya, Westwater, Caroline, Stewart, George C.
• Format: Article
• Language: English
• Subjects: Anthrax; antibiotic resistance; antibiotics; Bacillus anthracis; Bacillus anthracis - drug effects; Bacillus anthracis - isolation & purification; Bacillus cereus; Bacillus cereus - isolation & purification; bacteria; bacteriophages; Bacteriophages - genetics; Bioluminescence; Detection; Diagnosis; Drug Resistance, Bacterial; Genes, Reporter; Humans; Luminescent Measurements - methods; pathogens; patients; phenotype; prognosis; rapid methods; Reporter phage; Species Specificity; therapeutics
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2013.08.013

Genetically modified phages have the potential to detect pathogenic bacteria from clinical, environmental, or food-related sources. Herein we assess an engineered ‘bioluminescent’ reporter phage (Wß::luxAB) as a clinical diagnostic tool for Bacillus anthracis, the etiological agent of anthrax. Wß::luxAB is able to rapidly (within minutes) detect a panel of B. anthracis strains by transducing a bioluminescent phenotype. The reporter phage displays species specificity by its inability, or significantly reduced ability, to detect members of the closely related Bacillus cereus group and other common bacterial pathogens. Using spiked clinical specimens, Wß::luxAB detects B. anthracis within 5h at clinically relevant concentrations, and provides antibiotic susceptibility information that mirrors the CLSI method, except that data are obtained at least 5-fold faster. Although anthrax is a treatable disease, a positive patient prognosis is dependent on timely diagnosis and appropriate therapy. Wß::luxAB rapidly detects B. anthracis and determines antibiotic efficacy, properties that will help patient outcome. •Diagnostic that displays 100% inclusivity with species specificity.•Detection of B. anthracis at clinically relevant concentrations.•Detection of B. anthracis directly in clinical matrices without culture isolation.•Determine antibiotic susceptibility 5 times faster than conventional methods.•Potentially improve outcome of patients infected with anthrax.