ArticleApplied and environmental microbiology2005
A novel real-time PCR for Listeria monocytogenes that monitors analytical performance via an internal amplification control.
Article in Applied and environmental microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- Real-Time PCR Method Combined with a Matrix Lysis Procedure for the Quantification ofFoods (Basel, Switzerland) · 2021Article
- Evaluation and implementation of commercial antibodies for improved nanoparticle-based immunomagnetic separation and real-time PCR for faster detection ofJournal of food science and technology · 2020Article
- Colorimetric Detection ofSensors (Basel, Switzerland) · 2020Article
- The Response to Oxidative Stress inMicroorganisms · 2020Article
- Rapid sample preparation with Lyse-It® for Listeria monocytogenes and Vibrio cholerae.PloS one · 2018Article
- Article
- 3D-printed microfluidic device for the detection of pathogenic bacteria using size-based separation in helical channel with trapezoid cross-section.Scientific reports · 2015Article
- Molecular Epidemiology of Invasive Listeriosis due to Listeria monocytogenes in a Spanish Hospital over a Nine-Year Study Period, 2006-2014.BioMed research international · 2015Article
- Development of a real time PCR assay for rapid detection of Vibrio parahaemolyticus from seafood.Protein & cell · 2012Article
- Design and Application of Nucleic Acid Standards for Quantitative Detection of Enteric Viruses by Real-Time PCR.Food and environmental virology · 2011Article
- Mycoplasma agalactiae p40 Gene, a novel marker for diagnosis of contagious agalactia in sheep by real-time PCR: assessment of analytical performance and in-house validation using naturally contaminated milk samples.Journal of clinical microbiology · 2009Article
- Enumeration of viable Listeria monocytogenes cells by real-time PCR with propidium monoazide and ethidium monoazide in the presence of dead cells.Applied and environmental microbiology · 2007Article
- Quantitative detection of Clostridium tyrobutyricum in milk by real-time PCR.Applied and environmental microbiology · 2007Article
- Internally controlled real-time PCR method for quantitative species-specific detection and vapA genotyping of Rhodococcus equi.Applied and environmental microbiology · 2006Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe a novel quantitative real-time (Q)-PCR assay for Listeria monocytogenes based on the coamplification of a target hly gene fragment and an internal amplification control (IAC). The IAC is a chimeric double-stranded DNA containing a fragment of the rapeseed BnACCg8 gene flanked by the hly-specific target sequences. This IAC is detected using a second TaqMan probe labeled with a different fluorophore, enabling the simultaneous monitoring of the hly and IAC signals. The hly-IAC assay had a specificity and sensitivity of 100%, as assessed using 49 L. monocytogenes isolates of different serotypes and 96 strains of nontarget bacteria, including 51 Listeria isolates. The detection and quantification limits were 8 and 30 genome equivalents, and the coefficients for PCR linearity (R2) and efficiency (E) were 0.997 and 0.80, respectively. We tested the performance of the hly-IAC Q-PCR assay using various broth media and food matrices. Fraser and half-Fraser media, raw pork, and raw or cold-smoked salmon were strongly PCR-inhibitory. This Q-PCR assay for L. monocytogenes, the first incorporating an IAC to be described for quantitative detection of a food-borne pathogen, is a simple and robust tool facilitating the identification of false negatives or underestimations of contamination loads due to PCR failure.
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