Combination of filtration and immunomagnetic separation based on real-time PCR to detect foodborne pathogens in fresh-cut apple

Lee, So‐Young; Kim, Jinhee; Oh, Se‐Wook

doi:10.1016/j.mimet.2022.106577

Cited by 9 publications

(4 citation statements)

References 58 publications

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“…This result might be due to the existence of strains in fresh milk that have a greater growth advantage than P. fragi, and there was a phenomenon of competitive inhibition. Previous studies had shown that E.coli was able to coexist with spoilage Pseudomonas, which would lead to meat food spoilage and has a clear leading role than other microorganisms [44,45]. We guess microorganisms in fresh milk may have more dominant strains， which may affect the detection of the P. fragi.…”

Section: Analysis Of Anti-interference Abilitymentioning

confidence: 89%

Evaluation of Sensitivity and Applicability of a Droplet Digital Pcr Assay for Simultaneous Detecting Pseudomonas aeruginosa and Pseudomonas fragi in Foods

Huang,

Zhai,

Wang

et al. 2024

Preprint

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Achieving effective control over microbial contamination necessitates the precise and concurrent identification of numerous pathogens. In this research, we have devised a remarkably sensitive duplex droplet digital PCR (dddPCR) reaction system to simultaneously detect Pseudomonas aeruginosa (P. aeruginosa) and Pseudomonas fragi (P. fragi). Employing comparative genomics, we identified four genes of P. fragi. By specific analysis, RS22680 gene was selected as the detection target of P. fragi and LasR gene was chosed as P. aeruginosa, which were applied to construct a dddPCR reaction. In terms of specificity, sensitivity and anti-interference ability, the constructed dddPCR detection system was verified and analyzed. The assay showed excellent sensitivity and applicability, as evidenced by a limit of detection of 100 CFU/mL. When the concentration of natural background bacteria in milk or fresh meat was 100 times that of the target detection bacteria, the method was still capable of completing the absolute quantification. In the simulation of actual sample contamination, P. aeruginosa could be detected after 3 h of enrichment culture, and P. fragi could be detected after 6 h. The established ddPCR detection system exhibits exceptional performance, serving as a foundation for the simultaneous detection of various pathogenic bacteria in food products.

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Section: Analysis Of Anti-interference Abilitymentioning

confidence: 89%

Evaluation of Sensitivity and Applicability of a Droplet Digital Pcr Assay for Simultaneous Detecting Pseudomonas aeruginosa and Pseudomonas fragi in Foods

Huang,

Zhai,

Wang

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…This result might be due to the existence of strains in fresh milk that have a greater growth advantage than P. fragi, and there was a phenomenon of competitive inhibition. Previous studies have shown that E.coli was able to coexist with spoilage Pseudomonas, which could lead to meat food spoilage and has a clear leading role compared to other microorganisms [48,49]. We guess that microorganisms in fresh milk may have more dominant strains, which may affect the detection of P. fragi.…”

Section: Analysis Of Anti-interference Abilitymentioning

confidence: 92%

An Evaluation of the Sensitivity and Applicability of a Droplet Digital Polymerase Chain Reaction Assay to Simultaneously Detect Pseudomonas aeruginosa and Pseudomonas fragi in Foods

Huang,

Zhai,

Wang

et al. 2024

Foods

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Achieving effective control over microbial contamination necessitates the precise and concurrent identification of numerous pathogens. As a common bacterium in the environment, Pseudomonas is rich in variety. It not only has pathogenic strains, but also spoilage bacteria that cause food spoilage. In this research, we devised a remarkably sensitive duplex droplet digital PCR (dddPCR) reaction system to simultaneously detect pathogenic Pseudomonas aeruginosa (P. aeruginosa) and spoilage Pseudomonas fragi (P. fragi). By employing comparative genomics, we identified four genes of P. fragi. Through a specific analysis, the RS22680 gene was selected as the detection target for P. fragi, and the lasR gene was chosen for P. aeruginosa, which were applied to construct a dddPCR reaction. In terms of specificity, sensitivity and anti-interference ability, the constructed dddPCR detection system was verified and analyzed. The assay showed excellent sensitivity and applicability, as evidenced by a limit of detection of 100 cfu/mL. When the concentration of natural background bacteria in milk or fresh meat was 100 times that of the target detection bacteria, the method was still capable of completing the absolute quantification. In the simulation of actual sample contamination, P. aeruginosa could be detected after 3 h of enrichment culture, and P. fragi could be detected after 6 h. The established dddPCR detection system exhibits exceptional performance, serving as a foundation for the simultaneous detection of various pathogenic bacteria in food products.

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“…By employing this approach, the authors achieved successful detection of E. coli O157:H7 in milk, ground beef, and cabbage samples at concentrations ranging from 10 to 10 2 cfu/mL. Interestingly, their method required a short enrichment time of only 1 h, and the entire assay was completed within 3 h. Similarly, another noteworthy study by Lee et al (2022b) , who also employed IMS-qPCR detect E. coli O157:H7 and Salmonella Typhimurium in fresh-cut apples, reported a successful detection of these pathogens at concentrations of 2.7 × 10 cfu/mL and 1.8 × 10 2 cfu/mL, respectively, without the need for sample enrichment. However, it's important to note that IMS-qPCR's performance may vary depending on the specific food sample being analyzed.…”

Section: Recent Advances In Naa-based Methodsmentioning

confidence: 98%

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

Ndraha,

Lin,

Wang

et al. 2023

Food Chemistry: Molecular Sciences

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Combination of filtration and immunomagnetic separation based on real-time PCR to detect foodborne pathogens in fresh-cut apple

Cited by 9 publications

References 58 publications

Evaluation of Sensitivity and Applicability of a Droplet Digital Pcr Assay for Simultaneous Detecting Pseudomonas aeruginosa and Pseudomonas fragi in Foods

Evaluation of Sensitivity and Applicability of a Droplet Digital Pcr Assay for Simultaneous Detecting Pseudomonas aeruginosa and Pseudomonas fragi in Foods

An Evaluation of the Sensitivity and Applicability of a Droplet Digital Polymerase Chain Reaction Assay to Simultaneously Detect Pseudomonas aeruginosa and Pseudomonas fragi in Foods

Rapid detection methods for foodborne pathogens based on nucleic acid amplification: Recent advances, remaining challenges, and possible opportunities

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