ArticleJournal of clinical microbiology2004
Rapid quantification of Yersinia enterocolitica in pork samples by a novel sample preparation method, flotation, prior to real-time PCR.
Article in Journal of clinical microbiology, 2004. 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, 85 citations in OpenAlex.
- Non-continuous Percoll density gradient: a method for purifying Mycobacterium tuberculosis from dust.BMC infectious diseases · 2026Article
- Biochemical indexes and gut microbiota testing as diagnostic methods forFood science & nutrition · 2022Article
- Detection of Salmonella enterica in Meat in Less than 5 Hours by a Low-Cost and Noncomplex Sample Preparation Method.Applied and environmental microbiology · 2017Article
- Article
- Detection of Yersinia enterocolitica in food: an overview.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2015Review
- Quantification of Gram-positive bacteria: adaptation and evaluation of a preparation strategy using high amounts of clinical tissue.BMC veterinary research · 2014Article
- Enteroaggregative Escherichia coli quantification in children stool samples using quantitative PCR.APMIS : acta pathologica, microbiologica, et immunologica Scandinavica · 2013Article
- Vibrio vulnificus DNA load and mortality.Journal of clinical microbiology · 2011Article
- A comparative analysis of coprologic diagnostic methods for detection of Toxoplama gondii in cats.The American journal of tropical medicine and hygiene · 2010Article
- Enumeration of salmonella bacteria in food and feed samples by real-time PCR for quantitative microbial risk assessment.Applied and environmental microbiology · 2008Review
- Rapid separation and concentration of food-borne pathogens in food samples prior to quantification by viable-cell counting and real-time PCR.Applied and environmental microbiology · 2007Article
- Direct quantitation and detection of salmonellae in biological samples without enrichment, using two-step filtration and real-time PCR.Applied and environmental microbiology · 2006Article
- Quantification of Campylobacter spp. in chicken rinse samples by using flotation prior to real-time PCR.Applied and environmental microbiology · 2005Article
- Current and developing technologies for monitoring agents of bioterrorism and biowarfare.Clinical microbiology reviews · 2005Review
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The development of real-time PCR thermal cycles in the late 1990s has opened up the possibility of accurate quantification of microorganisms in clinical, environmental, and food samples. However, a lack of suitable sample preparation methods that allow rapid quantification of the nucleic acids, remove PCR inhibitors, and prevent false-positive results due to DNA originating from dead cells has limited the use of quantitative PCR. We have used for the first time a new variant of density gradient centrifugation, called flotation, as a user-friendly sample preparation method prior to PCR. This paper describes the use of this sample preparation method, without DNA purification, for direct detection and quantification of Yersinia enterocolitica in PCR-inhibitory meat juice from pork. Flotation combined with qPCR could overcome PCR interference in juice from pork, as was shown by amplification efficiencies of 1.006 +/- 0.021 and 1.007 +/- 0.025, which are comparable to the amplification efficiency obtained for purified DNA samples (1.005 +/- 0.059). Applying flotation to meat juice samples containing natural background flora and spiked with different levels of Y. enterocolitica showed that direct quantification of Y. enterocolitica was possible down to a level of at least 4.2 x 10(3) CFU per ml of meat juice, even in the presence of 10(6) CFU of background flora per ml. Finally, the results showed that samples containing large amounts of Y. enterocolitica DNA did not result in a positive PCR signal. This indicates that the risk of false-positive results due to detection of DNA originating from dead cells can be greatly reduced by using flotation prior to PCR.
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