Next-day detection of viable Listeria monocytogenes by multiplex reverse transcriptase real-time PCR

Azinheiro, Sarah; Ghimire, Dipak; Carvalho, Joana; Prado, Marta; Garrido‐Maestu, Alejandro

doi:10.1016/j.foodcont.2021.108593

Cited by 8 publications

(2 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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

View full text Add to dashboard Cite

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.

show abstract

Section: Cellular Components As Molecular Targetsmentioning

confidence: 99%

Molecular Targets for Foodborne Pathogenic Bacteria Detection

Paramithiotis

2023

Pathogens

View full text Add to dashboard Cite

show abstract

“…for marine species should follow the footsteps of its human-pathogen counterparts (such as V. parahaemolyticus [ 39 , 40 ], V. cholerae [ 41 ], or V. vulnificus [ 42 ]) and embark on the molecular era. Despite all of the advantages of qPCR, one major drawback is the fact that this technique is unable to distinguish live and dead bacteria compared to the conventional culturing methods [ 43 ].…”

Section: Introductionmentioning

confidence: 99%

Real-Time PCR Protocol for Detection and Quantification of Three Pathogenic Members of the Vibrionaceae Family

Costa¹,

Ferreira

Simões³

et al. 2022

Microorganisms

View full text Add to dashboard Cite

Vibriosis, an often-fatal disease induced by pathogenic members of the Vibrionaceae family, causes severe economic losses in aquacultures. To mitigate/avoid vibriosis outbursts, it is vital to detect and quantify these pathogens as early as possible. However, standard microbiological methods are time-consuming and often underestimate cell counts, which calls for the development of valid alternatives. In this study, real-time polymerase chain reaction (qPCR) was employed to detect the pathogenic species Vibrio alginolyticus, Listonella anguillara, and Vibrio harveyi using a new primer pair targeting the groEL gene. In addition, the DNA extraction efficiency of three methods, two commercial kits and the boiling method, was compared. The most efficient method was the DNeasy Blood and Tissue kit, with a detection limit ranging between 154 and 600 CFU mL−1 in the case of V. alginolyticus and L. anguillara, and 48 CFU mL−1 for V. harveyi. Thus, this study presents the development and evaluation of a method for the early quantification of all three species in saline suspensions. However, the results obtained by spiking a microalgae sample with V. harveyi emphasize the importance of adjusting the DNA control’s standard curve to the relevant extraction matrices, as it affects the DNA extraction efficiency and may hamper an accurate quantification with qPCR.

show abstract