ArticleApplied and environmental microbiology2000
Application of 5'-nuclease PCR for quantitative detection of Listeria monocytogenes in pure cultures, water, skim milk, and unpasteurized whole milk.
Article in Applied and environmental microbiology, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
47 citing papers in PubMed, 254 citations in OpenAlex.
- Development of an IMSA assay targeting the hly gene for detection of Listeria monocytogenes in food samples.World journal of microbiology & biotechnology · 2026Article
- Viability Detection of Foodborne Bacterial Pathogens in Food Environment by PMA-qPCR and by Microscopy Observation.Methods in molecular biology (Clifton, N.J.) · 2025Article
- The Detection of Foodborne Pathogenic Bacteria in Seafood Using a Multiplex Polymerase Chain Reaction System.Foods (Basel, Switzerland) · 2022Article
- Distribution and Characterization of Antimicrobial Resistant Pathogens in a Pig Farm, Slaughterhouse, Meat Processing Plant, and in Retail Stores.Microorganisms · 2022Article
- Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification ofFoods (Basel, Switzerland) · 2021Article
- Spiking a Silty-Sand Reference Soil with Bacterial DNA: Limits and Pitfalls in the Discrimination of Live and Dead Cells When Applying Ethidium Monoazide (EMA) Treatment.Current microbiology · 2019Article
- Controlled release of nisin from Neusilin particles to enhance food safety of sour curd cheese.Journal of food science and technology · 2019Article
- Risk factors associated with fecal shedding of Listeria monocytogenes by dairy cows and calves.Journal of veterinary internal medicine · 2018Article
- Pathogen Identification by Multiplex LightMix Real-Time PCR Assay in Patients with Meningitis and Culture-Negative Cerebrospinal Fluid Specimens.Journal of clinical microbiology · 2018Article
- Article
- Detection ofJournal of veterinary research · 2017Article
- An assessment of the microbiological quality of liver-based pâté in England 2012-13: comparison of samples collected at retail and from catering businesses.Epidemiology and infection · 2017Article
- An assessment of the microbiological quality of lightly cooked food (including sous-vide) at the point of consumption in England.Epidemiology and infection · 2017Article
- Persistence and Potential Viable but Non-culturable State of Pathogenic Bacteria during Storage of Digestates from Agricultural Biogas Plants.Frontiers in microbiology · 2016Article
- Fate of Viable but Non-culturable Listeria monocytogenes in Pig Manure Microcosms.Frontiers in microbiology · 2016Article
- Experimental design for the optimization of propidium monoazide treatment to quantify viable and non-viable bacteria in piggery effluents.BMC microbiology · 2015Article
- Diagnostics method for the rapid quantitative detection and identification of low-level contamination of high-purity water with pathogenic bacteria.Journal of industrial microbiology & biotechnology · 2013Article
- Development and validation of qualitative SYBR®Green real-time PCR for detection and discrimination of Listeria spp. and Listeria monocytogenes.Applied microbiology and biotechnology · 2013Article
- Highly specific fiber optic immunosensor coupled with immunomagnetic separation for detection of low levels of Listeria monocytogenes and L. ivanovii.BMC microbiology · 2012Article
- Quantification of bacterial indicators and zoonotic pathogens in dairy wastewater ponds.Applied and environmental microbiology · 2012Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
PCR techniques have significantly improved the detection and identification of bacterial pathogens. Countless adaptations and applications have been described, including quantitative PCR and the latest innovation, real-time PCR. In real-time PCR, e.g., the 5'-nuclease chemistry renders the automated and direct detection and quantification of PCR products possible (P. M. Holland et al., Proc. Natl. Acad. Sci. USA 88:7276-7280, 1991). We present an assay for the quantitative detection of Listeria monocytogenes based on the 5'-nuclease PCR using a 113-bp amplicon from the listeriolysin O gene (hlyA) as the target. The assay was positive for all isolates of L. monocytogenes tested (65 isolates including the type strain) and negative for all other Listeria strains (16 isolates from five species tested) and several other bacteria (18 species tested). The application of 5'-nuclease PCR in diagnostics requires a quantitative sample preparation step. Several magnetic bead-based strategies were evaluated, since these systems are simple and relatively easy to automate. The combination of nonspecific binding of bacteria to paramagnetic beads, with subsequent DNA purification by use of the same beads, gave the most satisfactory result. The detection limit was approximately 6 to 60 CFU, quantification was linear over at least 7 log units, and the method could be completed within 3 h. In conclusion, a complete quantitative method for L. monocytogenes in water and in skimmed and raw milk was developed.
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