Bacteriophage tailspike proteins as molecular probes for sensitive and selective bacterial detection

Singh, Amit; Arya, Sunil K.; Glass, Nick; Hanifi-Moghaddam, Pejman; Naidoo, Ravendra; Szymanski, Christine M.; Tanha, Jamshid; Evoy, Stéphane

doi:10.1016/j.bios.2010.05.024

Cited by 119 publications

(111 citation statements)

References 27 publications

(24 reference statements)

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“…Moreover, single proteins or even protein complexes (such as gp37-gp38) are still much smaller than whole phage particles, less complicated to produce as a single entity, and easier to modify in order to alter or optimize their recognition properties (47,64,65). This was elegantly demonstrated by Singh et al, who observed a 6-fold increase in Salmonella immobilization in an SPR-based biosensor by removing the inherent endorhamnosidase activity of the phage P22 tail spike probe (39). We here demonstrate the ideal properties of phage S16 LTF-MBs for rapid detection of low-level Salmonella contaminations in foods.…”

Section: Discussionmentioning

confidence: 69%

“…This "self-sandwich"-based detection assay exploits the Salmonella selectivity of the phage S16 LTF to provide dual levels of Salmonella-specific interaction. The ELLTA assay only takes 2 h to complete, including all wash steps, and without enrichment or amplification is able to reliably detect S. Typhimurium DB7155 down to 10 2 CFU · ml Ϫ1 , equivalent to the limits documented for other phage-based detection tests (17), including SPR biosensors based on other RBPs for detection of Salmonella (39) and Campylobacter jejuni (41). Similar to the phage S16 LTF, these RBP-based sensors also showed no cross-reactivity, a frequent feature of phage proteinbased recognition.…”

Section: Discussionmentioning

confidence: 99%

“…The use of affinity-based magnetic separation can simultaneously tackle two critical parameters to reduce detection assay time and increase sensitivity, by (i) enriching viable cell numbers to a detectable level and (ii) isolating cells from contaminants and debris found in food samples for more accurate downstream identification and characterization (51). Toward our aim to develop a sensitive and specific assay for bacterial enrichment that can outcompete the conventional application of antibodies for biomolecular recognition, the bacteriophage S16 LTF joins other approaches employing either whole phages (25,32,33,58,59) or phage-encoded proteins, such as endolysin cell wall-binding domains (CBDs) (36)(37)(38) and tail spike RBPs (39)(40)(41).…”

Section: Discussionmentioning

confidence: 99%

“…Previously, the tail spike RBP from phage P22 has been developed as a probe for detecting Salmonella in real-time biosensors (39) and, due to its inherent enzymatic activity, as a treatment against Salmonella colonization in chickens (40). Campylobacter phage RBPs have also been developed as probes for detecting the organism via surface plasmon resonance (SPR) biosensors (41), as well as for a cell agglutination assay (42).…”

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confidence: 99%

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Modified Bacteriophage S16 Long Tail Fiber Proteins for Rapid and Specific Immobilization and Detection of Salmonella Cells

Denyes

Dunne

Steiner

et al. 2017

Appl Environ Microbiol

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Bacteriophage-based assays and biosensors rival traditional antibodybased immunoassays for detection of low-level Salmonella contaminations. In this study, we harnessed the binding specificity of the long tail fiber (LTF) from bacteriophage S16 as an affinity molecule for the immobilization, enrichment, and detection of Salmonella. We demonstrate that paramagnetic beads (MBs) coated with recombinant gp37-gp38 LTF complexes (LTF-MBs) are highly effective tools for rapid affinity magnetic separation and enrichment of Salmonella. Within 45 min, the LTF-MBs consistently captured over 95% of Salmonella enterica serovar Typhimurium cells from suspensions containing from 10 to 10 5 CFU · ml Ϫ1 , and they yielded equivalent recovery rates (93% Ϯ 5%, n ϭ 10) for other Salmonella strains tested. LTF-MBs also captured Salmonella cells from various food sample preenrichments, allowing the detection of initial contaminations of 1 to 10 CFU per 25 g or ml. While plating of bead-captured cells allowed ultrasensitive but time-consuming detection, the integration of LTF-based enrichment into a sandwich assay with horseradish peroxidaseconjugated LTF (HRP-LTF) as a detection probe produced a rapid and easy-to-use Salmonella detection assay. The novel enzyme-linked LTF assay (ELLTA) uses HRP-LTF to label bead-captured Salmonella cells for subsequent identification by HRP-catalyzed conversion of chromogenic 3,3=,5,5=-tetramethylbenzidine substrate. The color development was proportional for Salmonella concentrations between 10 2 and 10 7 CFU · ml Ϫ1 as determined by spectrophotometric quantification. The ELLTA assay took 2 h to complete and detected as few as 10 2 CFU · ml Ϫ1 S. Typhimurium cells. It positively identified 21 different Salmonella strains, with no cross-reactivity for other bacteria. In conclusion, the phage-based ELLTA represents a rapid, sensitive, and specific diagnostic assay that appears to be superior to other currently available tests. IMPORTANCEThe incidence of foodborne diseases has increased over the years, resulting in major global public health issues. Conventional methods for pathogen detection can be laborious and expensive, and they require lengthy preenrichment steps. Rapid enrichment-based diagnostic assays, such as immunomagnetic separation, can reduce detection times while also remaining sensitive and specific. A critical component in these tests is implementing affinity molecules that retain the ability to specifically capture target pathogens over a wide range of in situ applications. The protein complex that forms the distal tip of the bacteriophage S16 long tail fiber is shown here to represent a highly sensitive affinity molecule for the specific enrichment and detection of Salmonella. Phage-encoded long tail fibers have huge potential for development as novel affinity molecules for robust and specific diagnostics of a vast spectrum of bacteria.

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Section: Discussionmentioning

confidence: 69%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Modified Bacteriophage S16 Long Tail Fiber Proteins for Rapid and Specific Immobilization and Detection of Salmonella Cells

Denyes

Dunne

Steiner

et al. 2017

Appl Environ Microbiol

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“…They include susceptibility testing with phage amplification assays to detect multidrugresistant pathogens (31)(32)(33). Tailspike proteins, which are used by many phages to bind specifically to the cell surface, have recently been suggested for use as molecular probes for bacterial detection (34). Our study investigated a novel application of engineered bacteriophage host-binding protein as a rapid diagnostic tool in the form of a LAT.…”

Section: Discussionmentioning

confidence: 99%

Bacteriophage-Based Latex Agglutination Test for Rapid Identification of Staphylococcus aureus

Idelevich

Walther²,

Molinaro³

et al. 2014

J Clin Microbiol

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e Rapid diagnosis is essential for the management of Staphylococcus aureus infections. A host recognition protein from S. aureus bacteriophage phiSLT was recombinantly produced and used to coat streptavidin latex beads to develop a latex agglutination test (LAT). The diagnostic accuracy of this bacteriophage-based test was compared with that of a conventional LAT, Pastorex StaphPlus, by investigating a clinical collection of 86 S. aureus isolates and 128 coagulase-negative staphylococci (CoNS) from deep tissue infections. All of the clinical S. aureus isolates were correctly identified by the bacteriophage-based test. While most of the CoNS were correctly identified as non-S. aureus isolates, 7.9% of the CoNS caused agglutination. Thus, the sensitivity of the bacteriophage-based LAT for identification of S. aureus among clinical isolates was 100%, its specificity was 92.1%, its positive predictive value (PPV) was 89.6%, and its negative predictive value (NPV) was 100%. The sensitivity, specificity, PPV, and NPV of the Pastorex LAT for the identification of S. aureus were 100%, 99.2%, 98.9%, and 100%, respectively. Among the additionally tested 35 S. aureus and 91 non-S. aureus staphylococcal reference and type strains, 1 isolate was false negative by each system; 13 and 8 isolates were false positive by the bacteriophage-based and Pastorex LATs, respectively. The ability of the phiSLT protein to detect S. aureus was dependent on the presence of wall teichoic acid (WTA) and corresponded to the production of ribitol phosphate WTA, which is found in most S. aureus clones but only a small minority of CoNS. Bacteriophage-based LAT identification is a promising strategy for rapid pathogen identification. Finding more specific bacteriophage proteins would increase the specificity of this novel diagnostic approach.

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The Role of Phages in Food‐Borne Pathogen Detection

Nevin

Coffey

O’Mahony

2015

Advances in Food Biotechnology

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Bacteriophage tailspike proteins as molecular probes for sensitive and selective bacterial detection

Cited by 119 publications

References 27 publications

Modified Bacteriophage S16 Long Tail Fiber Proteins for Rapid and Specific Immobilization and Detection of Salmonella Cells

Modified Bacteriophage S16 Long Tail Fiber Proteins for Rapid and Specific Immobilization and Detection of Salmonella Cells

Bacteriophage-Based Latex Agglutination Test for Rapid Identification of Staphylococcus aureus

The Role of Phages in Food‐Borne Pathogen Detection

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