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Original Research ARTICLE Provisionally accepted The full-text will be published soon. Notify me

Front. Microbiol. | doi: 10.3389/fmicb.2019.02443

A Sensitive, Specific and Simple Loop Mediated Isothermal Amplification Method for Rapid Detection of Campylobacter Spp. in Broiler Production

  • 1DTU Bioengineering, Technical University of Denmark, Denmark
  • 2Novo Nordisk (Denmark), Denmark
  • 3National Food Institute, Technical University of Denmark, Denmark
  • 4Technical University of Denmark, Denmark

Campylobacteriosis is one of the most common foodborne diseases worldwide. Two Campylobacter species - Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) in poultry and poultry products are considered to be the main source of human campylobacteriosis. Therefore, studying Campylobacter status in poultry flock is needed in order to prevent transmission of disease and reduce human risk, health cost, and economic losses. In this study, we adapted and used a Loop-Mediated Isothermal Amplification (LAMP) assay for specific, sensitive, simple and cost-effective rapid detection of C. jejuni and C. coli in the poultry production chain. By using optimized conditions for amplification and a small, low-cost commercial blue LED transilluminator for detection, a limit of detection (LOD) of 1 CFU/ reaction was achieved for testing of C. jejuni and C. coli in spiked chicken feces. The method took less than 70 min from receiving the samples to the final results (including 30 min for amplification). Comparison results of the LAMP and real-time PCR methods showed a relative accuracy, specificity, and sensitivity of 98.43%, 97.87%, and 100%, respectively. Cohen's kappa index also showed an excellent agreement between the two methods. The LAMP was used to study Campylobacter status in broiler flocks during the production of five weeks. The results showed that the method was specific, sensitive and was suitable to develop for online rapid detection of Campylobacter spp. at poultry production.

Keywords: Campylobacteriosis, Loop-mediated isothermal amplification (LAMP), Broiler faecal sample, Rapid detection, Broiler chicken production, Campylobacter spp.

Received: 20 Jun 2019; Accepted: 10 Oct 2019.

Copyright: © 2019 Linh Quyen, Nordentoft, Chidambara Vinayaka, Ngo, Engelsmenn, Sun, Madsen, Duong Bang and Wolff. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence:
Prof. Dang Duong Bang, National Food Institute, Technical University of Denmark, Kgs Lyngby, DK-2800, Denmark, ddba@food.dtu.dk
Prof. Anders Wolff, Technical University of Denmark, DTU Bioengineering, Kongens Lyngby, Denmark, awol@dtu.dk