A phage-based magnetic relaxation switching biosensor using bioorthogonal reaction signal amplification for Salmonella detection in foods

Huang, Chenxi; Zhao, Junpeng; Lu, Rongsheng; Wang, Jia; Nugen, Sam R.; Chen, Yiping; Wang, Xiaohong

doi:10.1016/j.foodchem.2022.134035

Cited by 39 publications

(16 citation statements)

References 37 publications

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“…typhimurium, the MNP 1μm -phage captures and binds specifically to bacteria surfaces, thereby trapping them. The phage LPST10, which was referenced from our previous work, can distinguish between viable and nonviable bacteria, which greatly reduces the likelihood of false positive results . After the MNP 1μm -phage has trapped the bacteria, the phage multiplies within the bacteria and releases lytic enzymes that break down the bacterial cell walls, causing them to disintegrate and release their internal DNA, including the target DNA.…”

Section: Resultsmentioning

confidence: 99%

“…The phage LPST10, which was referenced from our previous work, 27 can distinguish between viable and nonviable bacteria, which greatly reduces the likelihood of false positive results. 28 After the MNP 1μm -phage has trapped the bacteria, the phage multiplies within the bacteria and releases lytic enzymes that break down the bacterial cell walls, causing them to disintegrate and release their internal DNA, including the target DNA. Following magnetic separation, the target DNA is retained for subsequent analysis without the need for purification or amplification.…”

Section: Resultsmentioning

confidence: 99%

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DNA Extraction- and Amplification-Free Nucleic Acid Biosensor for the Detection of Foodborne Pathogens Based on CRISPR/Cas12a and Argonaute Protein-Mediated Cascade Signal Amplification

Chen,

Zhao,

Han

et al. 2023

J. Agric. Food Chem.

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A novel method for detecting low levels of viable foodborne pathogens, specifically Salmonella typhimurium (S. typhimurium), has been developed. Traditional nucleic acid assay, such as polymerase chain reaction (PCR), often requires complex DNA extraction and amplification, making it challenging to differentiate between viable and nonviable pathogens. This assay employed a phage as the recognition element to precisely identify and lyse viable S. typhimurium that can undergo DNA extraction. It combined the efficient trans-cleavage activities of CRISPR/Cas12a with the specific cleavage advantages of Argonaute proteins, enabling ultrasensitive detection. This double-enzyme-mediated nucleic acid test can accurately distinguish viable and nonviable S. typhimurium with a detection limit of 23 CFU/mL without DNA amplification. The method was successfully applied to common food samples, producing results consistent with quantitative PCR tests. This work provides a promising platform for easily detecting viable foodborne pathogens with high sensitivity without the need for DNA extraction and amplification.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

DNA Extraction- and Amplification-Free Nucleic Acid Biosensor for the Detection of Foodborne Pathogens Based on CRISPR/Cas12a and Argonaute Protein-Mediated Cascade Signal Amplification

Chen,

Zhao,

Han

et al. 2023

J. Agric. Food Chem.

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“… The biomolecule–probe conjugate can be imaged provided that it gives a detectable signal or affinity captured for downstream analysis if it binds solid support. Consequently, the technique has implications for bioimaging, biosensing, chemical immunology, drug development, targeted drug delivery, and omics analysis of biomolecules. , Motivated by the interdisciplinary flavor of this technique and its far-reaching implications for emerging disciplines such as synthetic biology, chemical immunology, biotechnology, and biomedicine, we have designed a BML experiment. The experiment is well-suited for an undergraduate research laboratory course to prepare students lacking prior experience in a chemical biology laboratory to work with minimal supervision in a research setting or motivate them to pursue a career in chemical biology and these emerging disciplines.…”

Section: Introductionmentioning

confidence: 99%

“…20 The biomolecule−probe conjugate can be imaged provided that it gives a detectable signal or affinity captured for downstream analysis if it binds solid support. Consequently, the technique has implications for bioimaging, 21 biosensing, 22 chemical immunology, 23 drug development, 24 targeted drug delivery, 25 and omics analysis of biomolecules. 26,27 Motivated by the interdisciplinary flavor of this technique and its far-reaching implications for emerging disciplines such as synthetic biology, chemical immunology, biotechnology, and biomedicine, we have designed a BML experiment.…”

Section: ■ Introductionmentioning

confidence: 99%

Bioorthogonal Metabolic Labeling Experiments to Introduce Undergraduate Students to Interdisciplinarity at the Interface of Chemistry and Biology

Fura,

Oluwole,

Hakim

et al. 2023

J. Chem. Educ.

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Chemists have always designed and deployed crossdisciplinary approaches that interface chemistry with other disciplines to proffer solutions to many human problems. We designed a laboratory experiment that implements chemistry concepts in a biological model to prime undergraduate students to appreciate interdisciplinarity as an approach to solving complex problems in contemporary society. The experiments can also be implemented with graduate students transitioning to chemical biology research. The experiment introduces students to classical and emerging chemistry and molecular biology techniques�mammalian cell culture, bioorthogonal metabolic labeling, azide−alkyne cycloaddition reaction ("click" chemistry), microplate photometry, gel electrophoresis, and far-western blot analysis�that are routinely used in research at the interface of chemistry and biology. The students cultured a Chinese hamster ovary cell line (CHO-K1) and metabolically labeled the cellular sialoglycans with azide groups. Next, they used a click chemistry reaction to bioconjugate a fluorogenic molecule to the labeled sialoglycans, enabling the detection of the glycans using microplate photometry or far-western blot analysis. The experiment created a context for a first-hand demonstration of bioorthogonal chemical reaction, a concept novel to most undergraduates with traditional chemistry (organic, inorganic, physical chemistry, and analytical) background. The experience allows the students to apply and connect organic chemistry, bioanalytical chemistry, and molecular biology concepts while learning techniques such as microplate reading, SDS PAGE, and western blot imaging that are indispensable to the growing fields of drug discovery, chemical toxicology, chemical biology, and molecular biology. This experiment is ideal for preparing and motivating students to pursue future research opportunities at the interface of chemistry and biology.

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“…Many-tailed phages use their receptor-binding proteins at the distal end of the tail to interact with receptors such as lipopolysaccharides, teichoic acids, and porins on the surface of host bacteria, allowing phages to capture their host bacteria from complex matrixes . Recently, some detection techniques employing single phages as bacterial recognition agents have been reported to detect Salmonella , Escherichia coli, Staphylococcus aureus, Mycobacterium smegmatis, and so on. It is worth noting that these single-phage-based techniques can distinguish dead/living bacteria as phages only infect living ones .…”

Section: Introductionmentioning

confidence: 99%

Employment of the Phage Cocktail as a Species-Specific Recognition Agent for Wide-Spectrum Detection of Bacterial Strains

Yang,

Gou,

Yuan

et al. 2023

Anal. Chem.

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Phages have already been employed to detect bacteria because of their specific recognition capability and strong infectious activity toward their host. However, the reported single-phage-based techniques are inevitably restricted by false negative results that arose from extremely high strain specificity of phages. In this study, a cocktail composed of three Klebsiella pneumoniae (K. pneumoniae) phages was prepared as a recognition agent to broaden the recognition spectrum for detecting this bacterial species. A total of 155 clinically isolated strains of K. pneumoniae collected from four hospitals were adopted to test its recognition spectrum. A superior recognition rate of 91.6% for the strains was achieved due to the complementarity of the recognition spectra of the three phages composed of the cocktail. However, the recognition rate is as low as 42.3−62.2% if a single phage is employed. Based on the wide-spectrum recognition capability of the phage cocktail, a fluorescence resonance energy transfer method was established for detecting K. pneumoniae strains by employing fluorescein isothiocyanate labeled to the phage cocktail and Au nanoparticles labeled to p-mercaptophenylboronic acid as energy donors and acceptors, respectively. The detection process can be completed within 35 min, with a wide dynamic range of 5.0 × 10 2 −1.0 × 10 7 CFU/mL. The application potential was verified by applying it to quantitate K. pneumoniae in different sample matrixes. This pioneer work opens an avenue for achieving wide-spectrum detection of different strains belonging to the same bacterial species with the phage cocktail.

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A phage-based magnetic relaxation switching biosensor using bioorthogonal reaction signal amplification for Salmonella detection in foods

Cited by 39 publications

References 37 publications

DNA Extraction- and Amplification-Free Nucleic Acid Biosensor for the Detection of Foodborne Pathogens Based on CRISPR/Cas12a and Argonaute Protein-Mediated Cascade Signal Amplification

DNA Extraction- and Amplification-Free Nucleic Acid Biosensor for the Detection of Foodborne Pathogens Based on CRISPR/Cas12a and Argonaute Protein-Mediated Cascade Signal Amplification

Bioorthogonal Metabolic Labeling Experiments to Introduce Undergraduate Students to Interdisciplinarity at the Interface of Chemistry and Biology

Employment of the Phage Cocktail as a Species-Specific Recognition Agent for Wide-Spectrum Detection of Bacterial Strains

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