Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Rapid quantification of viable Campylobacter bacteria on chicken carcasses, using real-time pcr and propidium monoazide treatment, as a tool for quantitative risk assessment
DTU Technical University of Denmark, Denmark.ORCID iD: 0000-0001-6492-1245
Show others and affiliations
2010 (English)In: Applied and Environmental Microbiology, ISSN 0099-2240, E-ISSN 1098-5336, Vol. 76, no 15, p. 5097-5104Article in journal (Refereed) Published
Abstract [en]

A number of intervention strategies against Campylobacter-contaminated poultry focus on postslaughter reduction of the number of cells, emphasizing the need for rapid and reliable quantitative detection of only viable Campylobacter bacteria. We present a new and rapid quantitative approach to the enumeration of food-borne Campylobacter bacteria that combines real-time quantitative PCR (Q-PCR) with simple propidium monoazide (PMA) sample treatment. In less than 3 h, this method generates a signal from only viable and viable but nonculturable (VBNC) Campylobacter bacteria with an intact membrane. The method's performance was evaluated by assessing the contributions to variability by individual chicken carcass rinse matrices, species of Campylobacter, and differences in efficiency of DNA extraction with differing cell inputs. The method was compared with culture-based enumeration on 50 naturally infected chickens. The cell contents correlated with cycle threshold (CT.) values (R2 = 0.993), with a quantification range of 1 × 102 to 1 × 107 CFU/ml. The correlation between the Campylobacter counts obtained by PMA-PCR and culture on naturally contaminated chickens was high (R 2 = 0.844). The amplification efficiency of the Q-PCR method was not affected by the chicken rinse matrix or by the species of Campylobacter. No Q-PCR signals were obtained from artificially inoculated chicken rinse when PMA sample treatment was applied. In conclusion, this study presents a rapid tool for producing reliable quantitative data on viable Campylobacter bacteria in chicken carcass rinse. The proposed method does not detect DNA from dead Campylobacter bacteria but recognizes the infectious potential of the VBNC state and is thereby able to assess the effect of control strategies and provide trustworthy data for risk assessment. Copyright © 2010, American Society tor Microbiology. All Rights Reserved.

Place, publisher, year, edition, pages
2010. Vol. 76, no 15, p. 5097-5104
Keywords [en]
Campylobacters, Control strategies, DNA extraction, Intervention strategy, matrix, PCR method, Propidium, Quantitative approach, Quantitative data, Quantitative detection, Quantitative risk assessment, Real-time PCR, Real-time quantitative PCR, Sample treatment, Viable but non-culturable, Amplification, Bacteriology, Cell culture, DNA, DNA sequences, Extraction, Genes, Risk assessment, Risk management, Signal detection, Bacteria, azide, drug derivative, propidium iodide, propidium monoazide, bacterium, bioassay, membrane, polymerase chain reaction, poultry, quantitative analysis, real time, animal, article, bacterial count, Campylobacter, chicken, comparative study, evaluation, isolation and purification, metabolism, methodology, microbial viability, microbiology, time, analogs and derivatives, evaluation study, procedures, Animals, Azides, Chickens, Colony Count, Microbial, Time Factors, Bacteria (microorganisms), Gallus gallus
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:ri:diva-39028DOI: 10.1128/AEM.00411-10Scopus ID: 2-s2.0-77955580586OAI: oai:DiVA.org:ri-39028DiVA, id: diva2:1324732
Available from: 2019-06-14 Created: 2019-06-14 Last updated: 2019-06-14Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textScopus

Authority records BETA

Löfström, Charlotta

Search in DiVA

By author/editor
Löfström, Charlotta
In the same journal
Applied and Environmental Microbiology
Natural Sciences

Search outside of DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 1 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
v. 2.35.8