Phage-based detection of bacterial pathogens

Merwe, R. G. van der; Helden, Paul D. van; Warren, Robin M.; Sampson, Samantha L.; Pittius, Nicolaas C. Gey van

doi:10.1039/c4an00208c

Cited by 57 publications

(39 citation statements)

References 92 publications

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“…Several phage-based detection systems have been developed for human and animal pathogens (van der Merwe et al, 2014). Recently however, work has been published on the promising application of these methods for the detection of plant pathogens.…”

Section: Other Phage Applications Of the Past And Possible Future Witmentioning

confidence: 99%

Bacteriophages and Bacterial Plant Diseases

et al. 2017

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Losses in crop yields due to disease need to be reduced in order to meet increasing global food demands associated with growth in the human population. There is a well-recognized need to develop new environmentally friendly control strategies to combat bacterial crop disease. Current control measures involving the use of traditional chemicals or antibiotics are losing their efficacy due to the natural development of bacterial resistance to these agents. In addition, there is an increasing awareness that their use is environmentally unfriendly. Bacteriophages, the viruses of bacteria, have received increased research interest in recent years as a realistic environmentally friendly means of controlling bacterial diseases. Their use presents a viable control measure for a number of destructive bacterial crop diseases, with some phage-based products already becoming available on the market. Phage biocontrol possesses advantages over chemical controls in that tailor-made phage cocktails can be adapted to target specific disease-causing bacteria. Unlike chemical control measures, phage mixtures can be easily adapted for bacterial resistance which may develop over time. In this review, we will examine the progress and challenges for phage-based disease biocontrol in food crops.

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Section: Other Phage Applications Of the Past And Possible Future Witmentioning

confidence: 99%

Bacteriophages and Bacterial Plant Diseases

et al. 2017

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“…While the bioengineering of reporter phage is not new (Schmelcher and Loessner, 2014, Singh, et al, 2012, van der Merwe, et al, 2014), to our knowledge we are the first to demonstrate the use of a protease as a reporter with application in multiple detection platforms. While our proof-of-principle is specific for E. coli , we believe our scheme can be readily modified for the detection of other bacterial species in combination with other specific proteases.…”

Section: Discussionmentioning

confidence: 99%

“…While our proof-of-principle is specific for E. coli , we believe our scheme can be readily modified for the detection of other bacterial species in combination with other specific proteases. Successful bioengineering of phage specific for other species, like Salmonella and Listeria , has been well demonstrated (van der Merwe, et al, 2014). The successful expression of viral proteases in gram negative bacteria, like E. coli , has also been demonstrated (Graves, et al, 1988, Liu, et al, 1999, Raran-Kurussi, et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Phage-protease-peptide: a novel trifecta enabling multiplex detection of viable bacterial pathogens

Alcaine

Tilton

Serrano

et al. 2015

Appl Microbiol Biotechnol

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Bacteriophages represent rapid, readily targeted, and easily produced molecular probes for the detection of bacterial pathogens. Molecular biology techniques have allowed researchers to make significant advances in the bioengineering of bacteriophage to further improve speed and sensitivity of detection. Despite their host-specificity, bacteriophages have not been meaningfully leveraged in multiplex detection of bacterial pathogens. We propose a proof-of-principal phage-based scheme to enable multiplex detection. Our scheme involves bioengineering bacteriophage to carry a gene for a specific protease, which is expressed during infection of the target cell. Upon lysis, the protease is released to cleave a reporter peptide, and the signal detected. Here we demonstrate the successful: i) modification of T7 bacteriophage to carry TEV protease; ii) expression of TEV protease by E. coli following infection by our modified T7, an average of 2000 units of protease per phage are produced during infection; and iii) proof-of-principle detection of E. coli in 3 hours after a primary enrichment via TEV protease activity using a fluorescent peptide and using a designed target peptide for MALDI-TOF MS analysis. This proof-of-principle can be translated to other phage-protease-peptide combinations to enable multiplex bacterial detection, and readily adopted on multiple platforms, like MALDI-TOF MS or fluorescent readers, commonly found in labs.

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“…In addition to their target specificity, another key feature of phages is their ability to distinguish viable versus inactive host bacterial cells, because phages can only replicate and express enzymes within viable bacterial cells. 28 The specificity of a phage-based detection system requires careful host-range screening of phage libraries. Also, it is relatively easy and inexpensive to synthesize and purify phage, providing a new platform to detect bacteria.…”

Section: Bacteriophage-based Detection Of Bacteriamentioning

confidence: 99%