Abstract
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.
Highlights
The development of quick and reliable methods for pathogen detection and identification is critical to improve the prevention and treatment of bacterial diseases in various fields from food production to health care
This review focuses on the design principles that govern the engineering of heterologous into phages (Section 3) and how these phages can be used to detect a wide variety of clinically and reporters into phages (Section 3) and how these phages can be used to detect a wide variety of industrially important bacterial pathogens
The rapid and sensitive nature of reporter phage-based systems has made them ideal tools for detecting low levels of viable, contaminating bacteria in many matrices, including foods, water and clinical samples
Summary
The development of quick and reliable methods for pathogen detection and identification is critical to improve the prevention and treatment of bacterial diseases in various fields from food production to health care. Cell wall-binding domains (CBDs) of phage endolysins have been successfully used for detecting various Gram-positive bacteria, Viruses 2020, 12, x FOR PEER REVIEW e.g., Listeria [32,33], Bacillus cereus [34], and Clostridium tyrobutyricum [35] Phages recognize their bacterial using specialized receptor-binding proteins (RBPs),[35]. The rapid and sensitive nature of reporter phage-based systems has made them ideal tools for detecting low levels of viable, contaminating bacteria in many matrices, including foods, water and clinical samples These phages express the reporter proteins during host infection to produce an amplifying fluorescent or bioluminescent signal upon the addition of a substrate.
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