P3-27 Detection of Cronobacter sakazakii in Milk Powder Using Loop-mediated Isothermal Amplification

Wednesday, July 31, 2013
Exhibit Hall (Charlotte Convention Center)
Young-Ju Kim, Chung-Ang University, Ansung, South Korea
Sheungwoo Seo, 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
Xiaoyu Wang, 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: Loop-mediated isothermal amplification (LAMP) is an emerging detection technology for the amplifying DNA under isothermal condition.

Purpose: The aim of this study was to develop a rapid and reliable LAMP technique for the detection of Cronobacter sakazakii in milk powder or powdered food.

Methods: Thirty-five C. sakazakii strains and 14 pathogenic microorganisms were used to examine the specificity of LAMP on C. sakazakii. LAMP products were submitted to restriction enzyme fragment polymorphism (RFLP) to prove the specificity of C. sakazakii. The sensitivity of polymerase chain reaction (PCR) and Real-time PCR were compared with that of LAMP in this study.

Results: LAMP for C. sakazakii ompA was successfully developed. RFLP pattern of LAMP product with HhaI and NruI confirmed the specificity. The LAMP for C. sakazakii did not show the cross reactivity with other foodborne pathogens. Based on the sensitivity of C. sakazakii, LAMP had 1,000-fold and 100-fold higher sensitivity than PCR and real-time PCR, respectively.

Significance: According to this study, diagnosis time is shortened and early diagnosis is performed by LAMP when food poisoning caused by pathogenic microorganisms like C. sakazakii in the future.