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Open AccessJournal ArticleDOI

Development of a sensitive and specific qPCR assay in conjunction with propidium monoazide for enhanced detection of live Salmonella spp. in food

Baoguang Li, +1 more
- 01 Dec 2013 - 
- Vol. 13, Iss: 1, pp 273-273
TLDR
A 130-bp amplicon in invA gene was demonstrated to be optimal for PMA treatment for selective detection of live Salmonella cells by PCR and provided excellent inhibition of DNA amplification from dead cells while only a slight DNA amplification difference was noted between the PMA-treated and untreated live cells.
Abstract
Background Although a variety of methodologies are available for detection of Salmonella, sensitive, specific, and efficient methods are urgently needed for differentiation of live Salmonella cells from dead cells in food and environmental samples. Propidium monoazide (PMA) can preferentially penetrate the compromised membranes of dead cells and inhibit their DNA amplification, however, such inhibition has been reported to be incomplete by some studies. In the present study, we report an efficient qPCR assay targeting a conserved region of the invA gene of Salmonella in conjunction with PMA treatment for detection of DNA from live Salmonella cells in food samples.

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Citations
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Culture‐Independent Rapid Detection Methods for Bacterial Pathogens and Toxins in Food Matrices

TL;DR: An overview of detection methods for various foodborne pathogens, including Listeria monocytogenes, Salmonella enterica, and shiga toxin-producing Escherichia coli, and bacterial toxins in food matrices, with emphasis on those methods which do not require cultural enrichment.
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Recent and emerging innovations in Salmonella detection: a food and environmental perspective.

TL;DR: This review will focus on current approaches and state‐of‐the‐art next‐generation advances in the detection, identification and subtyping of Salmonella from food and environmental sources.
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Presence and Persistence of Salmonella in Water: The Impact on Microbial Quality of Water and Food Safety.

TL;DR: The presence and persistence ofSalmonella in water and the mechanisms Salmonella uses to persist in the aquatic environment, particularly irrigation water are discussed to better understand the impact on the microbial quality of water and food safety due to the presence of Salmoneella in the water environment.
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The microbiome and antibiotic resistance in integrated fishfarm water: Implications of environmental public health.

TL;DR: The presence of intersecting health risk factors in aquaculture facilities is demonstrated and can lay the foundation for addressing these risks in Aquaculture management in rural China, with potential applicability in other developing regions dependent on aquacculture.
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Advances and Challenges in Viability Detection of Foodborne Pathogens.

TL;DR: The incorporation of PMA into PCR-based assays for viability detection of pathogens in foods has increased significantly in the last decade, particularly to achieve complete suppression of signal of DNA from the dead cells present in some particular food matrix.
References
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Journal ArticleDOI

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TL;DR: Foodborne outbreaks associated with fresh produce in the United States have increased in absolute numbers and as a proportion of all reported foodborne outbreaks, and Cyclospora and Escherichia coli O157:H7 were newly recognized as causes of foodborne illness.
Journal ArticleDOI

Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella.

TL;DR: The detection of 99.4% of Salmonella strains tested and the failure to specifically amplify DNA from non-Salmonellastrains confirm that the invA gene contains sequences unique to Salmoneella and demonstrate that this gene is a suitable PCR target, with potential diagnostic applications.
Journal ArticleDOI

Comparison of propidium monoazide with ethidium monoazide for differentiation of live vs. dead bacteria by selective removal of DNA from dead cells

TL;DR: A novel chemical, propidium monoazide (PMA), that (like propidium iodide) is highly selective in penetrating only into 'dead' bacterial cells with compromised membrane integrity but not into live cells with intact cell membranes/cell walls.
Journal ArticleDOI

Foodborne illness acquired in the United States--unspecified agents.

TL;DR: Each year, unspecified agents caused an estimated 38.4 million episodes of illness, resulting in 71,878 hospitalizations and 1,686 deaths, according to the United States Environmental Protection Agency.
Journal ArticleDOI

Fresh fruit and vegetables as vehicles for the transmission of human pathogens

TL;DR: A better understanding of plant, microbiological, environmental, processing and food handling factors that facilitate contamination will allow development of evidence-based policies, procedures and technologies aimed at reducing the risk of contamination of fresh produce.
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