A new single-tube platform of melting temperature curve analysis based on multiplex real-time PCR using EvaGreen for simultaneous screening detection of Shiga toxin-producing Escherichia coli, Salmonella spp. and Listeria monocytogenes in food

Bundidamorn, Damkerng; Supawasit, Wannakarn; Trevanich, Sudsai

doi:10.1016/j.foodcont.2018.07.001

Cited by 17 publications

(5 citation statements)

References 38 publications

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“…Most multiplex qPCR developed for the identification of different foodborne pathogens use hydrolysis probes [ 38 , 39 ] or high resolution melting analysis [ 15 ], which need appropriate reagents and a more complex downstream analysis, increasing the price in the first case or the time of the analysis in the second. On other hand, studies for multiplex real-time PCR without these approach, using only intercalating dyes have reported only the amplification of one or two target when an internal control is used [ 40 , 41 ], or when more pathogens are reported no internal control is employed [ 42 , 43 ]. The implementation of an internal control have been highly recommended by different authors, and the International Organization for Standardization (ISO) established this component as requirement for routine qPCR testing in food samples [ 44 ].…”

Section: Discussionmentioning

confidence: 99%

Multiplex Detection of Salmonella spp., E. coli O157 and L. monocytogenes by qPCR Melt Curve Analysis in Spiked Infant Formula

et al. 2020

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Food poisoning continue to be a threat in the food industry showing a need to improve the detection of the pathogen responsible for the hospitalization cases and death. DNA-based techniques represent a real advantage and allow the detection of several targets at the same time, reducing cost and time of analysis. The development of new methodology using SYBR Green qPCR for the detection of L. monocytogenes, Salmonella spp. and E. coli O157 simultaneously was developed and a non-competitive internal amplification control (NC-IAC) was implemented to detect reaction inhibition. The formulation and supplementation of the enrichment medium was also optimized to allow the growth of all pathogens. The limit of detection (LoD) 95% obtained was <1 CFU/25 g for E. coli O157, and 2 CFU/25 g for Salmonella spp. and L. monocytogenes and regarding the multiplex detection a LoD 95% of 1.7 CFU/25 g was observed. The specificity, relative sensitivity and accuracy of full methodology were 100% and the use of the NC-IAC allowed the reliability of the results without interfering with the sensitivity of the methodology. The described study proved to obtain results comparable to those of probe-based qPCR, and more economically than classical high resolution melting qPCR, being both important aspects for its implementation in the food industry.

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Section: Discussionmentioning

confidence: 99%

Multiplex Detection of Salmonella spp., E. coli O157 and L. monocytogenes by qPCR Melt Curve Analysis in Spiked Infant Formula

et al. 2020

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“…The sensitivity of the platform developed in this study was compared to that of the colloidal gold-labeled lateral flow assay, traditional enzyme-linked immunosorbent assay, gold magnetic bifunctional nanobeads-labeled immunochromatographic test strip, multiplex real-time polymerase chain reaction (PCR), − propidium monoazide–multiplex PCR, and multiplex magnetic capture hybridization with multiplex real-time PCR, as shown in Table . In all of the assays, immunological methods, such as traditional ELISA and lateral flow assay, were simple to operate and tolerant to the sample matrix.…”

Section: Results and Discussionmentioning

confidence: 99%

Multicolor and Ultrasensitive Enzyme-Linked Immunosorbent Assay Based on the Fluorescence Hybrid Chain Reaction for Simultaneous Detection of Pathogens

Huang²,

Liu

et al. 2019

J. Agric. Food Chem.

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Various pathogens may coexist in one sample; however, detection methods that rely on traditional selective culture media or immune agents designed specifically for a certain target are unsuitable for multiple targets. It is important to develop a simultaneous and sensitive detection method for multiple pathogens. Here, a multicolor and ultrasensitive enzymelinked immunosorbent assay (ELISA) platform based on the fluorescence hybridization chain reaction (HCR) was developed. In the assay, multicolor fluorescence concatemers formed as signal amplifiers and signal reporters in the presence of target pathogens. When HCR occurred, Escherichia coli O157:H7, Salmonella serotype Choleraesuis, and Listeria monocytogenes were detected simultaneously with three different fluorescences. Additionally, the limits of detection for E. coli O157:H7, Salmonella Choleraesuis, and L. monocytogenes were 3.4 × 10 1 , 6.4 × 10 0 , and 7.0 × 10 1 CFU/mL, respectively. The assay achieved ultrasensitive, specific, and simultaneous detection of three pathogens and can be applied to the detection of pathogens in milk samples. Therefore, this multicolor and ultrasensitive ELISA platform has great potential in the application of simultaneous detection of pathogens.

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“…Based on a Badan Pengawas Obat dan Makanan (BPOM) report (2019), approximately 61% of the microbes detected in several food products were pathogenic. Two bacterial species, Escherichia coli and Staphylococcus aureus have been confirmed as the dominant pathogenic microorganisms in food products (Bundidamorn et al, 2018;Incili et al, 2019;Qi et al, 2021;Wang et al, 2021b). These infections cause several poisoning symptoms, such as diarrhea, fever, nausea, vomiting and occasionally even death.…”

Section: Introductionmentioning

confidence: 99%

Antimicrobial and physicochemical characterization of Lactobacillus brevis biofilms as biopreservative agents

Solichah¹,

Sapalina²,

Retnaningrum³

2022

MJM

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Aims: Lactic acid bacteria (LAB) biofilms constitute one of the most remarkable breakthroughs in the field of food biopreservatives and can be employed to prevent foodborne disease. The purposes of this study were to investigate the efficacy of inhibitory LAB biofilms against foodborne pathogens and evaluate their tolerance to acidic pH and bile salts, as well as their physicochemical properties. Methodology and results: Four strains of Lactobacillus brevis biofilms isolated from kimchi showed antipathogenic activity to the bacteria Staphylococcus aureus FNCC 0049 and Escherichia coli FNCC 0091. These biofilms were also tolerant to pH 2.5, 0.3% bile salt and strong adhesion. Two of the four L. brevis biofilms (L. brevis biofilm KA2 and KB1) produced the highest inhibitory activity against both pathogenic bacterial indicators, tolerance to acidic pH and bile salts, and the strongest adhesion. In addition, based on Scanning Electron Microscope-Energy Dispersion X-ray Spectroscopy (SEM-EDS) analysis, both biofilm strains had a smooth surface texture; the cell morphology was rod-shaped and consisted of several elements such as carbon, oxygen and nitrogen, which was built up of extracellular polymeric substances (EPS). Conclusion, significance and impact of study:The presence of EPS as a constituent of LAB biofilms influenced their survival abilities in an acidic pH and bile salt environment. As a result, the characteristics of L. brevis biofilm KA2 and KB1 made them excellent candidates for use as antimicrobial packaging systems in food biopreservative applications.

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A new single-tube platform of melting temperature curve analysis based on multiplex real-time PCR using EvaGreen for simultaneous screening detection of Shiga toxin-producing Escherichia coli, Salmonella spp. and Listeria monocytogenes in food

Cited by 17 publications

References 38 publications

Multiplex Detection of Salmonella spp., E. coli O157 and L. monocytogenes by qPCR Melt Curve Analysis in Spiked Infant Formula

Multiplex Detection of Salmonella spp., E. coli O157 and L. monocytogenes by qPCR Melt Curve Analysis in Spiked Infant Formula

Multicolor and Ultrasensitive Enzyme-Linked Immunosorbent Assay Based on the Fluorescence Hybrid Chain Reaction for Simultaneous Detection of Pathogens

Antimicrobial and physicochemical characterization of Lactobacillus brevis biofilms as biopreservative agents

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