Luminescence based enzyme-labeled phage (Phazyme) assays for rapid detection of Shiga toxin producing<i>Escherichia coli</i>serogroups

Willford, John; Bisha, Bledar; Bolenbaugh, Kyle E.; Goodridge, Lawrence

doi:10.4161/bact.1.2.15666

Cited by 26 publications

(15 citation statements)

References 29 publications

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“…Instead of a lacZ carrying reporter phage, this device involves so-called Phazymes, i.e., phage particles specific for the target pathogen that are chemically labeled with horse radish peroxidase (HRP), and are contained within the cap of the SnapValve tube. 93 As for the PhastSwab, the bottom of the tube contains a swab for sample collection, suitable enrichment broth, and beads for IMS of STEC cells. Following IMS, the Phazymes are added to the immobilized cells by snapping the cap, and after several washing steps, STEC can be detected by addition of either a colorimetric or luminescent HRP substrate.…”

Section: Labeled Phage Particlesmentioning

confidence: 99%

Application of bacteriophages for detection of foodborne pathogens

2014

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Bacterial contamination of food products presents a challenge for the food industry and poses a high risk for the consumer. Despite increasing awareness and improved hygiene measures, foodborne pathogens remain a threat for public health, and novel methods for detection of these organisms are needed. Bacteriophages represent ideal tools for diagnostic assays because of their high target cell specificity, inherent signal-amplifying properties, easy and inexpensive production, and robustness. Every stage of the phage lytic multiplication cycle, from the initial recognition of the host cell to the final lysis event, may be harnessed in several ways for the purpose of bacterial detection. Besides intact phage particles, phage-derived affinity molecules such as cell wall binding domains and receptor binding proteins can serve for this purpose. This review provides an overview of existing phage-based technologies for detection of foodborne pathogens, and highlights the most recent developments in this field, with particular emphasis on phage-based biosensors.

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Section: Labeled Phage Particlesmentioning

confidence: 99%

Application of bacteriophages for detection of foodborne pathogens

2014

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“…Bacteriophages have been intensively used as molecular biology tools since their discovery [ 9 ]. In the last decade, several phage-based bioassays have been developed using a variety of output signals such as fluorescence and luminescence and based on direct (phage protein fusions or phage DNA staining) [ 19 – 21 ] or indirect (phages labeled with antibodies or quantum dots) detection [ 18 ]. Most techniques require a high level of technicality as well as dedicated detection device that are rarely transportable on the sampling site.…”

Section: Discussionmentioning

confidence: 99%

“…Most phage-based biosensors use capsid gene fusions that allow bacterial detection upon binding of the labeled phage to its bacterial receptor [ 18 , 19 , 21 ]. This is a source of false positive, since metabolically inactive bacteria are detected together with metabolically active ones.…”

Section: Discussionmentioning

confidence: 99%

“…Currently, several based-phage methods are developed to detect pathogenic bacteria using engineered phages expressing and coding for fluorescent [ 13 , 14 ], luminescent [ 15 , 16 ] or colorimetric [ 17 ] markers that require host cell infection for detection. Alternatively, labeled phage protein [ 18 , 19 ] and phage DNA staining [ 20 , 21 ] allow direct detection of phages bound to host bacterial cells. Several strategies have been developed for phage engineering such as in vitro packaging of modified DNA, transposition and post-infection homologous recombination [ 22 – 24 ].…”

Section: Introductionmentioning

confidence: 99%

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Phage-Based Fluorescent Biosensor Prototypes to Specifically Detect Enteric Bacteria Such as E. coli and Salmonella enterica Typhimurium

Vinay

Franche²,

Grégori³

et al. 2015

PLoS ONE

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Water safety is a major concern for public health and for natural environment preservation. We propose to use bacteriophages to develop biosensor tools able to detect human and animal pathogens present in water. For this purpose, we take advantage of the highly discriminating properties of the bacteriophages, which specifically infect their bacterial hosts. The challenge is to use a fluorescent reporter protein that will be synthesized, and thus detected, only once the specific recognition step between a genetically modified temperate bacteriophage and its bacterial host has occurred. To ensure the accuracy and the execution speed of our system, we developed a test that does not require bacterial growth, since a simple 1-hour infection step is required. To ensure a high sensitivity of our tool and in order to detect up to a single bacterium, fluorescence is measured using a portable flow cytometer, also allowing on-site detection. In this study, we have constructed and characterized several "phagosensor" prototypes using the HK620 bacteriophage and its host Escherichia coli TD2158 and we successfully adapted this method to Salmonella detection. We show that the method is fast, robust and sensitive, allowing the detection of as few as 10 bacteria per ml with no concentration nor enrichment step. Moreover, the test is functional in sea water and allows the detection of alive bacteria. Further development will aim to develop phagosensors adapted on demand to the detection of any human or animal pathogen that may be present in water.

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“…ml À1 . 46 A similar method combined immuno-separation of E. coli O157 with ow-cytometry-based detection of bacteriophage-host binding. 47 The study made use of bacteriophage LG1 stained with the nucleic acid dye YOYO-1 (Molecular probes, Inc., Eugene, Oreg.).…”

Section: Labeled Phagementioning

confidence: 99%

Phage-based detection of bacterial pathogens

Merwe

Helden

Warren

et al. 2014

Analyst

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Bacterial pathogens cause significant morbidity and mortality annually to both humans and animals. With the rampant spread of drug resistance and the diminishing effectiveness of current antibiotics, there is a pressing need for effective diagnostics for detection of bacterial pathogens and their drug resistances. Bacteriophages offer several unique opportunities for bacterial detection. This review highlights the means by which bacteriophages have been utilized to achieve and facilitate specific bacterial detection.

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Luminescence based enzyme-labeled phage (Phazyme) assays for rapid detection of Shiga toxin producingEscherichia coliserogroups

Cited by 26 publications

References 29 publications

Application of bacteriophages for detection of foodborne pathogens

Application of bacteriophages for detection of foodborne pathogens

Phage-Based Fluorescent Biosensor Prototypes to Specifically Detect Enteric Bacteria Such as E. coli and Salmonella enterica Typhimurium

Phage-based detection of bacterial pathogens

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