An Engineered Reporter Phage for the Fluorometric Detection of Escherichia coli in Ground Beef

Chen, Anqi; Wang, Danhui; Nugen, Sam R.; Chen, Juhong

doi:10.3390/microorganisms9020436

Cited by 13 publications

(5 citation statements)

References 33 publications

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“…The chicken was processed according to the previous method [ 25 ]. Skim milk powder (10 g) was dissolved in 100 mL of distilled water and sterilized at 65 °C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Phage-Based Biosensing for Rapid and Specific Detection of Staphylococcus aureus

Li,

Huang

et al. 2023

Microorganisms

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Staphylococcus aureus (S. aureus) is a major foodborne pathogen. Rapid and specific detection is crucial for controlling staphylococcal food poisoning. This study reported a Staphylococcus phage named LSA2302 showing great potential for applications in the rapid detection of S. aureus. Its biological characteristics were identified, including growth properties and stability under different pH and temperature conditions. The genomic analysis revealed that the phage has no genes associated with pathogenicity or drug resistance. Then, the phage-functionalized magnetic beads (pMB), serving as a biological recognition element, were integrated with ATP bioluminescence assays to establish a biosensing method for S. aureus detection. The pMB enrichment brought high specificity and a tenfold increase in analytical sensitivity during detection. The whole detection process could be completed within 30 min, with a broad linear range of 1 × 104 to 1 × 108 CFU/mL and a limit of detection (LOD) of 2.43 × 103 CFU/mL. After a 2 h pre-cultivation, this method is capable of detecting bacteria as low as 1 CFU/mL. The recoveries of S. aureus in spiked skim milk and chicken samples were 81.07% to 99.17% and 86.98% to 104.62%, respectively. Our results indicated that phage-based biosensing can contribute to the detection of target pathogens in foods.

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“…The chicken was processed according to the previous method [ 25 ]. Skim milk powder (10 g) was dissolved in 100 mL of distilled water and sterilized at 65 °C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Phage-Based Biosensing for Rapid and Specific Detection of Staphylococcus aureus

Li,

Huang

et al. 2023

Microorganisms

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“…In recently, Chen et al used a genetically engineered reporter T7 phage to detect a food-borne pathogen E. coli via fluorometric detection [ 28 ]. This study suggests that a fluorogenic substrate to detect β-gal activity caused by the genetically modified phage infection is a unique and very promising way to detect bacterial pathogens.…”

Section: Fluorescent Signal Generated By Genetically Engineered Phagesmentioning

confidence: 99%

Advances in the Development of Phage-Based Probes for Detection of Bio-Species

et al. 2022

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Bacteriophages, abbreviated as “phages”, have been developed as emerging nanoprobes for the detection of a wide variety of biological species, such as biomarker molecules and pathogens. Nanosized phages can display a certain length of exogenous peptides of arbitrary sequence or single-chain variable fragments (scFv) of antibodies that specifically bind to the targets of interest, such as animal cells, bacteria, viruses, and protein molecules. Metal nanoparticles generally have unique plasmon resonance effects. Metal nanoparticles such as gold, silver, and magnetism are widely used in the field of visual detection. A phage can be assembled with metal nanoparticles to form an organic–inorganic hybrid probe due to its nanometer-scale size and excellent modifiability. Due to the unique plasmon resonance effect of this composite probe, this technology can be used to visually detect objects of interest under a dark-field microscope. In summary, this review summarizes the recent advances in the development of phage-based probes for ultra-sensitive detection of various bio-species, outlining the advantages and limitations of detection technology of phage-based assays, and highlighting the commonly used editing technologies of phage genomes such as homologous recombination and clustered regularly interspaced palindromic repeats/CRISPR-associated proteins system (CRISPR-Cas). Finally, we discuss the possible scenarios for clinical application of phage-probe-based detection methods.

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“…Several reporter phages have been developed for the detection of foodborne bacteria. For instance, T7-ALP [ 54 ], Φ V10 lux [ 55 ], ΦV10 NanoLuc luciferase (Nluc) [ 56 ], T7-NRGp5 [ 57 ], and T4-NRGp17 [ 58 ] have been developed for the detection of different E. coli strains from various food matrices [ 59 ].…”

Section: Phage-mediated Bacterial Detection Approachesmentioning

confidence: 99%

Bacteriophage-Based Biosensors: A Platform for Detection of Foodborne Bacterial Pathogens from Food and Environment

et al. 2022

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Foodborne microorganisms are an important cause of human illness worldwide. Two-thirds of human foodborne diseases are caused by bacterial pathogens throughout the globe, especially in developing nations. Despite enormous developments in conventional foodborne pathogen detection methods, progress is limited by the assay complexity and a prolonged time-to-result. The specificity and sensitivity of assays for live pathogen detection may also depend on the nature of the samples being analyzed and the immunological or molecular reagents used. Bacteriophage-based biosensors offer several benefits, including specificity to their host organism, the detection of only live pathogens, and resistance to extreme environmental factors such as organic solvents, high temperatures, and a wide pH range. Phage-based biosensors are receiving increasing attention owing to their high degree of accuracy, specificity, and reduced assay times. These characteristics, coupled with their abundant supply, make phages a novel bio-recognition molecule in assay development, including biosensors for the detection of foodborne bacterial pathogens to ensure food safety. This review provides comprehensive information about the different types of phage-based biosensor platforms, such as magnetoelastic sensors, quartz crystal microbalance, and electrochemical and surface plasmon resonance for the detection of several foodborne bacterial pathogens from various representative food matrices and environmental samples.

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An Engineered Reporter Phage for the Fluorometric Detection of Escherichia coli in Ground Beef

Cited by 13 publications

References 33 publications

Phage-Based Biosensing for Rapid and Specific Detection of Staphylococcus aureus

Phage-Based Biosensing for Rapid and Specific Detection of Staphylococcus aureus

Advances in the Development of Phage-Based Probes for Detection of Bio-Species

Bacteriophage-Based Biosensors: A Platform for Detection of Foodborne Bacterial Pathogens from Food and Environment

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