Reporter Phage-Based Detection of Bacterial Pathogens: Design Guidelines and Recent Developments

Viruses. 2020 Aug 26;12(9):944. doi: 10.3390/v12090944.


Fast and reliable detection of bacterial pathogens in clinical samples, contaminated food products, and water supplies can drastically improve clinical outcomes and reduce the socio-economic impact of disease. As natural predators of bacteria, bacteriophages (phages) have evolved to bind their hosts with unparalleled specificity and to rapidly deliver and replicate their viral genome. Not surprisingly, phages and phage-encoded proteins have been used to develop a vast repertoire of diagnostic assays, many of which outperform conventional culture-based and molecular detection methods. While intact phages or phage-encoded affinity proteins can be used to capture bacteria, most phage-inspired detection systems harness viral genome delivery and amplification: to this end, suitable phages are genetically reprogrammed to deliver heterologous reporter genes, whose activity is typically detected through enzymatic substrate conversion to indicate the presence of a viable host cell. Infection with such engineered reporter phages typically leads to a rapid burst of reporter protein production that enables highly sensitive detection. In this review, we highlight recent advances in infection-based detection methods, present guidelines for reporter phage construction, outline technical aspects of reporter phage engineering, and discuss some of the advantages and pitfalls of phage-based pathogen detection. Recent improvements in reporter phage construction and engineering further substantiate the potential of these highly evolved nanomachines as rapid and inexpensive detection systems to replace or complement traditional diagnostic approaches.

Keywords: CRISPR-Cas editing; bacterial detection; bacteriophage; genetic engineering; luciferase; reporter phage.

Publication types

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

MeSH terms

  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • Bacteriolysis
  • Bacteriophages* / genetics
  • Bacteriophages* / physiology
  • Biosensing Techniques
  • CRISPR-Cas Systems
  • Cloning, Molecular
  • Colorimetry
  • Genes, Reporter
  • Genetic Engineering
  • Genome, Viral
  • Luminescent Measurements
  • Microscopy, Fluorescence