ArticleApplied and environmental microbiology2000
Application of the 5'-nuclease PCR assay in evaluation and development of methods for quantitative detection of Campylobacter jejuni.
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 31 papers.
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Who cites it
31 citing papers in PubMed.
- Developments in Rapid Detection Methods for the Detection of FoodborneFrontiers in microbiology · 2018Review
- Comparison of analytic methods for quantitative real-time polymerase chain reaction data.Journal of computational biology : a journal of computational molecular cell biology · 2015Article
- Detection, identification and quantification of Campylobacter jejuni, coli and lari in food matrices all at once using multiplex qPCR.Gut pathogens · 2014Article
- Survival of host-associated bacteroidales cells and their relationship with Enterococcus spp., Campylobacter jejuni, Salmonella enterica serovar Typhimurium, and adenovirus in freshwater microcosms as measured by propidium monoazide-quantitative PCR.Applied and environmental microbiology · 2012Article
- Highly specific and quick detection of Mycobacterium avium subsp. paratuberculosis in feces and gut tissue of cattle and humans by multiple real-time PCR assays.Journal of clinical microbiology · 2011Article
- Evaluation of Salmonella-vectored Campylobacter peptide epitopes for reduction of Campylobacter jejuni in broiler chickens.Clinical and vaccine immunology : CVI · 2011Article
- Improved detection of five major gastrointestinal pathogens by use of a molecular screening approach.Journal of clinical microbiology · 2010Article
- Susceptibility of Campylobacter jejuni to organic acids and monoacylglycerols.Folia microbiologica · 2010Article
- Simultaneous Screening of 24 Target Genes of Foodborne Pathogens in 35 Foodborne Outbreaks Using Multiplex Real-Time SYBR Green PCR Analysis.International journal of microbiology · 2010Article
- Comparison of the EntericBio multiplex PCR system with routine culture for detection of bacterial enteric pathogens.Journal of clinical microbiology · 2009Article
- Article
- Risk factors for campylobacteriosis: an epidemiological surveillance study of patients and retail poultry.Journal of clinical microbiology · 2009Article
- Development of cpn60-based real-time quantitative PCR assays for the detection of 14 Campylobacter species and application to screening of canine fecal samples.Applied and environmental microbiology · 2009Article
- Comprehensive and rapid real-time PCR analysis of 21 foodborne outbreaks.International journal of microbiology · 2009Article
- Method To Detect Only Live Bacteria during PCR Amplification.Journal of clinical microbiology · 2008Article
- Co-infection dynamics of a major food-borne zoonotic pathogen in chicken.PLoS pathogens · 2007Article
- Feasibility of a molecular screening method for detection of Salmonella enterica and Campylobacter jejuni in a routine community-based clinical microbiology laboratory.Journal of clinical microbiology · 2007Article
- Distribution and ecology of campylobacters in coastal plain streams (Georgia, United States of America).Applied and environmental microbiology · 2007Article
- Comparison of culture and a novel 5' Taq nuclease assay for direct detection of Campylobacter fetus subsp. venerealis in clinical specimens from cattle.Journal of clinical microbiology · 2006Article
- Quantification of Campylobacter spp. in chicken rinse samples by using flotation prior to real-time PCR.Applied and environmental microbiology · 2005Article
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Campylobacter jejuni is recognized as a leading human food-borne pathogen. Traditional diagnostic testing for C. jejuni is not reliable due to special growth requirements and the possibility that this bacterium can enter a viable but nonculturable state. Nucleic acid-based tests have emerged as a useful alternative to traditional enrichment testing. In this article, we present a 5'-nuclease PCR assay for quantitative detection of C. jejuni and describe its evaluation. A probe including positions 381121 to 381206 of the published C. jejuni strain NCTC 11168 genome sequence was identified. When this probe was applied, the assay was positive for all of the isolates of C. jejuni tested (32 isolates, including the type strain) and negative for all other Campylobacter spp. (11 species tested) and several other bacteria (41 species tested). The total assay could be completed in 3 h with a detection limit of approximately 1 CFU. Quantification was linear over at least 6 log units. Quantitative detection methods are important for both research purposes and further development of C. jejuni detection methods. In this study, we used the assay to investigate to what extent the PCR signals generated by heat-killed bacteria interfere with the detection of viable C. jejuni after exposure at elevated temperatures for up to 5 days. An approach to the reduction of the PCR signal generated by dead bacteria was also investigated by employing externally added DNases to selectively inactivate free DNA and exposed DNA in heat-killed bacteria. The results indicated relatively good discrimination between exposed DNA from dead C. jejuni and protected DNA in living bacteria.
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