Simultaneous Detection of Four Main Foodborne Pathogens in Ready-to-Eat Food by Using a Simple and Rapid Multiplex PCR (mPCR) Assay

Boukharouba, Aya; González, Ana; García-Ferrús, Miguel; Ferrús, María Antonia; Botella, S.

doi:10.3390/ijerph19031031

Cited by 25 publications

(4 citation statements)

References 56 publications

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“…Detection aims to find the pathogens and provides information and direction for the downstream process of intervention. This can range from traditional culturing (gold standard) to more recent polymerase chain reaction (PCR) methods [ 4 , 5 , 6 ], biochemical methods [ 7 ], and novel instrumentations such as the elastic mass spectrometry (MALDI-TOF) light-scattering (ELS) technology [ 8 , 9 , 10 ] and laser-induced breakdown spectroscopy (LIBS) [ 11 ], as well as others. Most current detection methods rely on sampling-based techniques that require a significant length of time-to-result.…”

Section: Introductionmentioning

confidence: 99%

Surface Environment and Energy Density Effects on the Detection and Disinfection of Microorganisms Using a Portable Instrument

Shin

Dowden

Doh

et al. 2023

Sensors

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Real-time detection and disinfection of foodborne pathogens are important for preventing foodborne outbreaks and for maintaining a safe environment for consumers. There are numerous methods for the disinfection of hazardous organisms, including heat treatment, chemical reaction, filtration, and irradiation. This report evaluated a portable instrument to validate its simultaneous detection and disinfection capability in typical laboratory situations. In this challenging study, three gram-negative and two gram-positive microorganisms were used. For the detection of contamination, inoculations of various concentrations were dispensed on three different surface types to estimate the performance for minimum-detectable cell concentration. Inoculations higher than 103~104 CFU/mm2 and 0.15 mm of detectable contaminant size were estimated to generate a sufficient level of fluorescence signal. The evaluation of disinfection efficacy was conducted on three distinct types of surfaces, with the energy density of UVC light (275-nm) ranging from 4.5 to 22.5 mJ/cm2 and the exposure time varying from 1 to 5 s. The study determined the optimal energy dose for each of the microorganisms species. In addition, surface characteristics may also be an important factor that results in different inactivation efficacy. These results demonstrate that the proposed portable device could serve as an in-field detection and disinfection unit in various environments, and provide a more efficient and user-friendly way of performing disinfection on large surface areas.

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Section: Introductionmentioning

confidence: 99%

Surface Environment and Energy Density Effects on the Detection and Disinfection of Microorganisms Using a Portable Instrument

Shin

Dowden

Doh

et al. 2023

Sensors

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show abstract

“…With the rapid development of molecular diagnostic technologies, PCR is widely used for the detection of various microorganisms ( Takahashi et al., 2021 ; Boukharouba et al., 2022 ; Serigstad et al., 2022 ). A PCR method was developed for the detection of H. influenzae based on the 16S rRNA gene.…”

Section: Introductionmentioning

confidence: 99%

Establishment and Clinical Application of a RPA-LFS Assay for Detection of Capsulated and Non-Capsulated Haemophilus influenzae

Wang

Liu

et al. 2022

Front. Cell. Infect. Microbiol.

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A recombinase polymerase amplification-lateral flow strip assay was established for detection of the outer membrane protein P6 (omp6) and the capsule encoding gene bexA of Haemophilus influenzae and the detection limit, sensitivity, and specificity were determined. Specific primers and probes were designed based on the published nucleotide sequences of omp6 and bexA. The minimum detection limit was determined with standard strains and the practical applicability of the RPA-LFS assay was assessed by detection of 209 clinical samples. The results confirmed that the RPA-LFS assay was both specific and sensitive for the detection of capsulated and non-capsulated H. influenzae with a detection limit of 1 CFU/µL. The detection rate of the 209 clinical samples was 97.1%, while the detection rate of capsulated H. influenzae was 63.2%. The detection results were consistent with the traditional culture method and dual polymerase chain reaction (PCR), confirming the applicability of the RPA-LFS assay.

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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 ].…”

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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Simultaneous Detection of Four Main Foodborne Pathogens in Ready-to-Eat Food by Using a Simple and Rapid Multiplex PCR (mPCR) Assay

Cited by 25 publications

References 56 publications

Surface Environment and Energy Density Effects on the Detection and Disinfection of Microorganisms Using a Portable Instrument

Surface Environment and Energy Density Effects on the Detection and Disinfection of Microorganisms Using a Portable Instrument

Establishment and Clinical Application of a RPA-LFS Assay for Detection of Capsulated and Non-Capsulated Haemophilus influenzae

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

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