Detection of live<i>Salmonella enterica</i>in fresh-cut vegetables by a TaqMan-based one-step reverse transcription real-time PCR

Miao, Yingjie; Xiong, Guotong; Bai, Meirong; Ge, Yunsheng; Wu, Zufang

doi:10.1111/lam.12871

Cited by 10 publications

(2 citation statements)

References 23 publications

(27 reference statements)

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“…The usefulness of the latter is getting increased recognition, and its ability to reach strain level has been exhibited [ 148 ]. Therefore, RT-qPCR protocols combining the aforementioned adjustments are the method of choice for the detection and quantification of foodborne pathogenic bacteria in food samples in many studies [ 149 , 150 , 151 , 152 ]. By combining the utilization of RNA as the target molecule with suitably adjusted PCR conditions and the use of hybridization probes or melting curve analysis, the detection and quantification of metabolically active bacterial foodborne pathogens with adequate sensitivity can be achieved.…”

Section: Cellular Components As 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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Section: Cellular Components As Molecular Targetsmentioning

confidence: 99%

Molecular Targets for Foodborne Pathogenic Bacteria Detection

Paramithiotis

2023

Pathogens

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“…Most of the foodborne outbreaks come from animal origin food, including beef meat, poultry, eggs, and milk products, which may be contaminated by multiple pathogens including Salmonella enterica [ 14 ]. Fresh-cut produce is at great risk of Salmonella contamination, and a one-step quantitative real-time polymerase chain reaction (qRT-PCR) assay has been used to detect Salmonella in fresh-cut vegetables [ 15 ]. A fluorescent biosensor with multiple fluorescent signal amplification based on a streptavidin biotin system was established to detect Salmonella in milk.…”

Section: Introductionmentioning

confidence: 99%

Development of a Duplex TaqMan Real-Time Polymerase Chain Reaction for Accurate Identification and Quantification of Salmonella Enteritidis from Laboratory Samples and Contaminated Chicken Eggs

Xiong

Zhou

Song

et al. 2022

Foods

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Salmonella enteritidis is a major causative agent of foodborne illnesses worldwide. As the traditional serotyping and quantification methods are labor-intensive, time-consuming, and expensive, faster and more convenient molecular diagnostic methods are needed. In this study, we developed and validated a rapid duplex TaqMan real-time polymerase chain reaction (PCR) for the accurate identification and quantification of S. enteritidis. The primers and TaqMan probes were designed based on the S. enteritidis-specific gene lygD and the Salmonella genus-specific gene invA. The melt curve and gel electrophoresis analysis showed that the designed primers had potent specificity for the amplification of lygD and invA. The duplex real-time PCR specifically identified S. enteritidis from a panel of 40 Salmonella strains that represented 29 serovars and 12 non-Salmonella organisms. The duplex real-time PCR assay detected four copies of S. enteritidis DNA per reaction. The intra- and inter- assays indicated a high degree of reproducibility. The real-time PCR could accurately detect and quantify S. enteritidis in chicken organs after Salmonella infection. Furthermore, the assay identified 100% of the S. enteritidis and Salmonella genus isolates from chicken egg samples with superior sensitivity after 6 h of pre-enrichment compared to the traditional culture method. Additionally, the most-probable-number (MPN) combined with qPCR and a shortened incubation time (MPN-qPCR-SIT) method was developed for the population determination of S. enteritidis and compared with various enumeration methods. Thus, we have established and validated a new duplex real-time PCR assay and MPN-qPCR-SIT method for the accurate detection and quantification of S. enteritidis, which could contribute to meeting the need for fast detection and identification in prevention and control measures for food safety.

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Detection and Characterization of Salmonella enterica Serotypes by Simple PCR Technologies

Kariuki

Kiiru

2020

Methods in Molecular Biology

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Detection of liveSalmonella entericain fresh-cut vegetables by a TaqMan-based one-step reverse transcription real-time PCR

Cited by 10 publications

References 23 publications

Molecular Targets for Foodborne Pathogenic Bacteria Detection

Molecular Targets for Foodborne Pathogenic Bacteria Detection

Development of a Duplex TaqMan Real-Time Polymerase Chain Reaction for Accurate Identification and Quantification of Salmonella Enteritidis from Laboratory Samples and Contaminated Chicken Eggs

Detection and Characterization of Salmonella enterica Serotypes by Simple PCR Technologies

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