P3-63 Development of Loop-mediated Isothermal Amplification for the Rapid Detection of Arcobacter Species in Chicken

Wednesday, July 31, 2013
Exhibit Hall (Charlotte Convention Center)
Xiaoyu Wang, Chung-Ang University, Ansung-Si, South Korea
Min Hwa Lee, Chung-Ang University, Ansung-Si, South Korea
Dong Joo Seo, Chung-Ang University, Ansung-Si, South Korea
Sheungwoo Seo, Chung-Ang University, Ansung-Si, South Korea
Na Ry Son, Chung-Ang University, Ansung-Si, South Korea
Changsun Choi, Chung-Ang University, Ansung-Si, South Korea
Introduction: Arcobacter is one of foodborne pathogenic bacteria which may cause gastroenteritis in human and abortion in animal.

Purpose: The aim of this study was to develop a more sensitive, specific and rapid technique of loop-mediated isothermal amplification (LAMP) for detecting of Arcobacter species.

Methods: The LAMP reaction was optimized under different conditions. Specificity and sensitivity of LAMP were compared with multiplex polymerase chain reaction. Bacterial isolation, multiplex PCR, and LAMP were performed in chicken samples collected from retailed markets.

Results: The optimization reaction condition for LAMP was achieved at 61°C for 50 min and ladder-like DNA products were produced with reference strains and field isolates of Arcobacter species. Because multiplex PCR showed the cross-reactivity with Campylobacter species, the LAMP assay was specific rather than multiplex PCR which was used in previous studies. The detection limit of LAMP was 2-20 CFU/reaction in vitro and 200 CFU/reaction in chicken samples. The sensitivity of LAMP assay showed a tenfold to thousand-fold times more than that of multiplex PCR assay.

Significance: The LAMP assay developed in this research is rapid and reliable detection technique for Arcobacter species which can cause food poisoning in human.