ArticleBMC microbiology2013
Development of a sensitive and specific qPCR assay in conjunction with propidium monoazide for enhanced detection of live Salmonella spp. in food.
Article in BMC microbiology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 63 citations in OpenAlex.
- Viability Detection of Foodborne Bacterial Pathogens in Food Environment by PMA-qPCR and by Microscopy Observation.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Optimization of the viability PCR for accurate detection of Staphylococcus aureus in food samples.PloS one · 2025Article
- Viability discrimination of bacterial microbiomes in home kitchen dish sponges using propidium monoazide treatment.Environmental microbiology reports · 2024Article
- Identification and quantification of viableFrontiers in microbiology · 2024Article
- Advances in detection methods for viable Salmonella spp.: current applications and challenges.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2023Review
- Detection ofFrontiers in veterinary science · 2022Article
- Article
- How to Evaluate Non-Growing Cells-Current Strategies for Determining Antimicrobial Resistance of VBNC Bacteria.Antibiotics (Basel, Switzerland) · 2021Review
- Development of PMAxxFrontiers in microbiology · 2020Article
- Microcin J25 Exhibits Inhibitory Activity AgainstFrontiers in microbiology · 2020Article
- Detection of Cell-Dissociated Non-TypeableInternational journal of chronic obstructive pulmonary disease · 2020Observational
- Evaluating the fate of bacterial indicators, viral indicators, and viruses in water resource recovery facilities.Water environment research : a research publication of the Water Environment Federation · 2019Article
- Quantitative Polymerase Chain Reaction Coupled With Sodium Dodecyl Sulfate and Propidium Monoazide for Detection of ViableFrontiers in microbiology · 2019Article
- Molecular viability testing of viable but non-culturable bacteria induced by antibiotic exposure.Microbial biotechnology · 2018Article
- Minimal Influence of Extracellular DNA on Molecular Surveys of Marine Sedimentary Communities.Frontiers in microbiology · 2018Article
- Presence and Persistence ofFrontiers in public health · 2018Review
- A Novel qPCR Method for Simultaneous Detection and Quantification of Viable Pathogenic and Non-pathogenicFrontiers in microbiology · 2018Article
- Designing primers and evaluation of the efficiency of propidium monoazide - Quantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius.Journal of food and drug analysis · 2017Article
- False-Positive Viability PCR Results: An Association with Microtubes.Current microbiology · 2017Article
- Improved sample treatment protocol for accurate detection of live Salmonella spp. in food samples by viability PCR.PloS one · 2017Article
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundAlthough 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.
resultsWe investigated the relationship between amplicon length and inhibitory effect of PMA treatment to prevent DNA amplification from dead cells while allowing for DNA amplification from live cells, and found that the two factors are well correlated with each other. An amplicon that is 130 bp in length was determined to be optimal for PMA treatment and was selected for further PMA-qPCR assay development. A PMA-qPCR assay was established by utilizing this amplicon and adopting a modified PMA-treatment procedure. The PMA-qPCR assay provided excellent inhibition of DNA amplification from dead cells (a 17-CT-value, or 128,000-fold reduction) while only a slight DNA amplification difference (0.5 CT value) was noted between the PMA-treated and untreated live cells. This assay has been validated through stringent inclusivity and exclusivity studies using a large number of (n = 167) Salmonella, including all strains of SARA and SARB collections, and non-Salmonella strains (n = 36). This PMA-qPCR assay is capable of detecting live Salmonella cells in live/dead cell mixtures, or 30 CFU/g live Salmonella cells from enriched spiked spinach samples as early as 4 h.
conclusionsA 130-bp amplicon in invA gene was demonstrated to be optimal for PMA treatment for selective detection of live Salmonella cells by PCR. This PMA-qPCR assay provides a sensitive, specific, and efficient method for detecting live Salmonella cells in foods and environmental samples and may have an impact on the accurate microbiological monitoring of Salmonella in foods and environment samples.
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