A single thiolated-phage displayed nanobody-based biosensor for label-free detection of foodborne pathogen

Wang, Peng; Yu, Gege; Wei, Juan; Liao, Xingrui; Zhang, Yao; Ren, Yarong; Zhang, Cui; Wang, Yueqi; Zhang, Daohong; Wang, Jianlong; Wang, Yanru

doi:10.1016/j.jhazmat.2022.130157

Cited by 28 publications

(18 citation statements)

References 31 publications

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“…These results demonstrate that microbes can be detected after being converted from unstable bioparticles to solid polymer microspheres using various size analysis strategies after “dramatic growth”. The “dramatic growth” strategy has the advantages of physical monodispersity, chemical stability, biological specificity, sensitivity, and reproducibility, especially compared with methods requiring exogenous signaling probes …”

Section: Resultsmentioning

confidence: 99%

“…The "dramatic growth" strategy has the advantages of physical monodispersity, chemical stability, biological specificity, sensitivity, and reproducibility, especially compared with methods requiring exogenous signaling probes. 35 Simulated and Actual Environmental Detection of Bacterial Aerosols. To demonstrate the application of the "dramatic growth" strategy for bacterial aerosols in the air environment, a simulation experiment was carried out to probe the possible interference of impurities in polymerization.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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“Dramatic Growth” of Microbial Aerosols for Visualization and Accurate Counting of Bioaerosols

Pan,

Zhao,

Zhang

et al. 2023

Anal. Chem.

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While the global COVID-19 pandemic has subsided, microbial aerosol detection has become of high concern. Timely, accurate, and highly sensitive monitoring of microbial aerosols in indoor air is the basis for effective prevention and control of infectious diseases. At present, no commercial equipment or reliable technology can simultaneously control the detection time and limit at 6 h and 10 2 CFU/mL, respectively. Based on the "safety size range" of particulate matter in the air, we propose a new method of microbial dilation detection, which enables the pathogen to grow rapidly and dramatically into a polymeric microsphere, larger in size than the coexisting aerosol particles. "Like a crane standing among chickens", the microorganism can be easily visualized and counted. Different from routine chemical and biological sensing technologies, this method can achieve absolute counting of microbial particles, and the simple principles can be developed into devices for different life scenarios.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

“Dramatic Growth” of Microbial Aerosols for Visualization and Accurate Counting of Bioaerosols

Pan,

Zhao,

Zhang

et al. 2023

Anal. Chem.

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“…Some of the latter, especially the ones referring to the adsorbent surface and colorimetric detection, have been effectively addressed through the introduction of nanomaterials [ 21 ]. However, only marginal improvement has been achieved regarding the sensitivity and specificity of the approach, which was only feasible through the substitution of antibodies as recognition elements with aptamers, nanobodies or the use of haptens [ 22 , 23 , 24 , 25 ].…”

Section: Surface-residing Molecular Targetsmentioning

confidence: 99%

Molecular Targets for Foodborne Pathogenic Bacteria Detection

Paramithiotis

2023

Pathogens

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The detection of foodborne pathogenic bacteria currently relies on their ability to grow on chemically defined liquid and solid media, which is the essence of the classical microbiological approach. Such procedures are time-consuming and the quality of the result is affected by the selectivity of the media employed. Several alternative strategies based on the detection of molecular markers have been proposed. These markers may be cell constituents, may reside on the cell envelope or may be specific metabolites. Each marker provides specific advantages and, at the same time, suffers from specific limitations. The food matrix and chemical composition, as well as the accompanying microbiota, may also severely compromise detection. The aim of the present review article is to present and critically discuss all available information regarding the molecular targets that have been employed as markers for the detection of foodborne pathogens. Their strengths and limitations, as well as the proposed alleviation strategies, are presented, with particular emphasis on their applicability in real food systems and the challenges that are yet to be effectively addressed.

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“…nanobody NbS22 gene 13 was cloned into the pET25b-AP plasmid with Sfi I restriction sites, while anti-V. parahaemolyticus nanobody NbV20 gene sequence was cloned using Hind III and Xho II as restriction sites. 4 After the plasmid vector transformed into E. coli DH5α, positive colonies were selected on an LB-ampicillin agar plate and validated by polymerase chain reaction (PCR). The inset xed DNA was further confirmed by DNA sequencing.…”

Section: Materials and Reagentsmentioning

confidence: 99%

“…3 Thus, the establishment of a sensitive and reliable detection method is imperative to safeguard the food supply, ensure public health, and mitigate financial costs. 4,5 Among the various bacteria detection assays, immunoassays have garnered considerable attention due to their unique advantages, including good selectivity, simplicity, high throughput, low cost, safety, and universal applicability. 6−9 However, traditional immunoassays still suffer from limitations such as relatively low sensitivity and stability.…”

Section: ■ Introductionmentioning

confidence: 99%

Enhancing Oriented Immobilization Efficiency: A One-for-Two Organism-Bispecific Nanobody Scaffold for Highly Sensitive Detection of Foodborne Pathogens

Wang,

Zhang,

Wang

et al. 2023

Anal. Chem.

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Nanobodies have gained widespread application in immunoassays. However, their small size presents a significant challenge in achieving effective immobilization and optimal sensitivity. Here, we present a novel “one-for-two”-oriented immobilization platform based on an organism-bispecific nanobody (O-BsNb) scaffold, enabling highly sensitive detection of two bacterial pathogens. Through genetic engineering, a bispecific nanobody (BsNb) was engineered, targeting Salmonella spp. and Vibrio parahaemolyticus. The O-BsNb scaffold allowed one nanobody to bind specifically to inactivated bacteria, forming an organism-oriented immobilization platform, while the other served as the capture antibody. Consequently, the O-BsNb bioscaffold-based ELISA (O-ELISA) for individual detection of S. enteritidis and V. parahaemolyticus was established. When compared to the sandwich ELISA utilizing passive immobilization of monovalent nanobodies, the O-ELISA exhibited a remarkable 13.4- and 13.7-fold improvement in LOD for S. enteritidis and V. parahaemolyticus, respectively, highlighting the enhanced immobilization efficacy of the O-ELISA. Furthermore, the feasibility and reproducibility of the assay in practical samples were meticulously evaluated, revealing exemplary performance in terms of recovery precision and assay stability. These findings demonstrate the significant potential of the O-ELISA platform for the sensitive detection of macromolecules, opening new avenues for efficient pathogen identification in foodborne safety and clinical diagnostics.

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A single thiolated-phage displayed nanobody-based biosensor for label-free detection of foodborne pathogen

Cited by 28 publications

References 31 publications

“Dramatic Growth” of Microbial Aerosols for Visualization and Accurate Counting of Bioaerosols

“Dramatic Growth” of Microbial Aerosols for Visualization and Accurate Counting of Bioaerosols

Molecular Targets for Foodborne Pathogenic Bacteria Detection

Enhancing Oriented Immobilization Efficiency: A One-for-Two Organism-Bispecific Nanobody Scaffold for Highly Sensitive Detection of Foodborne Pathogens

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