Detection of Salmonella typhimurium in fat free milk using a phage immobilized magnetoelastic sensor

Lakshmanan, Ramji S.; Guntupalli, Rajesh; Hu, Jing; Petrenko, Valery A.; Barbaree, James M.; Chin, Bryan A.

doi:10.1016/j.snb.2007.04.003

Cited by 135 publications

(89 citation statements)

References 39 publications

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“…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%

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: 99%

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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“…Favrin et al [40] also used phage amplification but with phage SJ2 to detect Salmonella at the low detection limit of three cells per 25 g -1 ground beef or per 25 mL -1 milk powder or chicken rinses in approximately 20 h. De Siqueira et al [71] used phage amplification with phage P22 to detect Salmonella in artificially contaminated chicken breasts at a limit of 10 4 CFU g -1 after 3-4 h of incubation. Using a magnetoelastic biosensor interface with immobilized phage E2, Lakshmanan et al [72] and Li et al [73] detected Salmonella Typhimurium in fat-free milk at 10 3 CFU mL -1 in 20 min and directly on the surface of whole tomato at 10 2 CFU mL -1 in 30 min.…”

Section: Detection Of Foodborne Pathogensmentioning

confidence: 99%

Pathogen detection using engineered bacteriophages

Smartt

Jegier

et al. 2011

Anal Bioanal Chem

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Bacteriophages, or phages, are bacterial viruses that can infect a broad or narrow range of host organisms. Knowing the host range of a phage allows it to be exploited in targeting various pathogens. Applying phages for the identification of microorganisms related to food and waterborne pathogens and pathogens of clinical significance to humans and animals has a long history, and there has to some extent been a recent revival in these applications as phages have become more extensively integrated into novel detection, identification, and monitoring technologies. Biotechnological and genetic engineering strategies applied to phages are responsible for some of these new methods, but even natural unmodified phages are widely applicable when paired with appropriate innovative detector platforms. This review highlights the use of phages as pathogen detector interfaces to provide the reader with an up-to-date inventory of phage-based biodetection strategies.

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“…Other systems based on the same principle have been developed before for detecting other biological agents (Lakshmanan et al, 2007;Wan et al, 2007) The system consists of the following elements ( Fig.1):…”

Section: In Situ Measurement Of the Mass Evolution Of Cell Culturementioning

confidence: 99%

Magnetic Sensors for Biomedical Applications

Rivero¹,

Multigner²,

Spottorno³

2012

Magnetic Sensors - Principles and Applications

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Detection of Salmonella typhimurium in fat free milk using a phage immobilized magnetoelastic sensor

Cited by 135 publications

References 39 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

Pathogen detection using engineered bacteriophages

Magnetic Sensors for Biomedical Applications

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