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

Frontiers in Microbiology

Mogens Madsen¹  Anders Wolff¹  Than Linh Quyen¹  Yi Sun²  Steen Nordentoft³  Aaydha Chidambara Vinayaka³  Dang Duong Bang³  Pia Engelsmenn³  Tien Anh Ngo³ 

[1] Department of Biotechnology and Biomedicine, Technical University of Denmark (DTU-Bioengineering), Lyngby, Denmark;Department of Health Technology, Technical University of Denmark (DTU-Health Tech), Lyngby, Denmark;National Food Institute, Technical University of Denmark (DTU-Food), Lyngby, Denmark;

关键词: campylobacteriosis;    Campylobacter spp.;    loop mediated isothermal amplification;    broiler fecal sample;    broiler chicken production;    rapid detection;   

DOI  :  10.3389/fmicb.2019.02443

来源: DOAJ

【 摘 要 】

Campylobacteriosis is one of the most common foodborne diseases worldwide. Two Campylobacter species – C. jejuni and C. coli in poultry and poultry products are considered to be the main source of human campylobacteriosis. Therefore, studying Campylobacter status in poultry flocks is needed 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. Amplified LAMP products were detected using a small, low-cost portable commercial blue LED transilluminator and a direct visual detection strategy was demonstrated. By using optimized conditions for amplification a limit of detection (LOD) of 50 CFU/ml was achieved for testing of C. jejuni and C. coli in spiked chicken feces without enrichment. The method took 60–70 min from receiving the samples to the final results (including 30 min for amplification). The optimized LAMP showed a relative accuracy of 98.4%, a specificity of 97.9%, and a sensitivity of 100% in comparison to real-time PCR method. Cohen’s kappa index also showed an excellent agreement (0.94) between the two methods. The results showed that the method is specific, sensitive and is suitable to develop for rapid detection of Campylobacter spp. at poultry production.

【 授权许可】

Unknown