ArticleApplied and environmental microbiology1999
Detection of viable Listeria monocytogenes with a 5' nuclease PCR assay.
Article in Applied and environmental microbiology, 1999. 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.
- Label-free technique for universal and sequence independent detection of oligonucleotides and nuclease activity.Nucleic acids research · 2025Article
- Exploring specific primers targeted against different genes for a multiplex PCR for detection of Listeria monocytogenes.3 Biotech · 2015Article
- Diagnosis of Listeria monocytogenes meningoencephalitis by real-time PCR for the hly gene.Journal of clinical microbiology · 2011Article
- Stress- and growth rate-related differences between plate count and real-time PCR data during growth of Listeria monocytogenes.Applied and environmental microbiology · 2009Article
- Development of a quantitative PCR method to differentiate between viable and nonviable bacteria in environmental water samples.Applied microbiology and biotechnology · 2009Article
- Detection of Vibrio cholerae by real-time nucleic acid sequence-based amplification.Applied and environmental microbiology · 2007Article
- Use of ethidium monoazide and PCR in combination for quantification of viable and dead cells in complex samples.Applied and environmental microbiology · 2005Article
- Real-time PCR analysis of Vibrio vulnificus from oysters.Applied and environmental microbiology · 2003Article
- Detection of Listeria monocytogenes from a model food by fluorescence resonance energy transfer-based PCR with an asymmetric fluorogenic probe set.Applied and environmental microbiology · 2003Article
- TaqMan PCR for detection of Vibrio cholerae O1, O139, non-O1, and non-O139 in pure cultures, raw oysters, and synthetic seawater.Applied and environmental microbiology · 2001Article
- Application of the 5' fluorogenic exonuclease assay (TaqMan) for quantitative ribosomal DNA and rRNA analysis in sediments.Applied and environmental microbiology · 2001Article
- Rapid detection and quantification of members of the archaeal community by quantitative PCR using fluorogenic probes.Applied and environmental microbiology · 2000Article
- Application of 5'-nuclease PCR for quantitative detection of Listeria monocytogenes in pure cultures, water, skim milk, and unpasteurized whole milk.Applied and environmental microbiology · 2000Article
- Application of the 5'-nuclease PCR assay in evaluation and development of methods for quantitative detection of Campylobacter jejuni.Applied and environmental microbiology · 2000Article
Corrections and comments
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A 5' nuclease assay has been developed to detect viable Listeria monocytogenes. The assay targets the hemolysin A (hlyA) transcript, which is found only in L. monocytogenes. The single-tube, reverse transcriptase (RT), fluorogenic probe-based assay was formatted by using Tth DNA polymerase whose activity was modulated by using the manganese-chelating morpholinepropanesulfonic acid (MOPS) buffer. This assay was quantitative over a 3-log-unit range of template concentrations when tested with an in vitro-transcribed hlyA mRNA. The viability of L. monocytogenes was reduced by heating at various temperatures and times up to a maximum of a 9-D inactivation. The location of the primer had a pronounced effect on the utility of the assay, and primers located in the most distal regions of the hlyA transcript appeared to correlate with the number of CFU while primers located more internal on the amplicon overestimated the cell viability. The assay with primers that included the 3' end of the transcript was an accurate indicator of viability as measured by CFU determination or staining with 5-sulfofluorescein diacetate.
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