분유에 오염된 Cronobacter sakazakii 검출을 위한 중합효소연쇄반응, 실시간중합효소연쇄반응, 등온검출법의 비교

• 발행기관: 한국축산식품학회
• 저자명: 김영주 서승우 왕효우 서동주 이민화 손나리 이복희 최창순
• 간행물 정보: 『Food Science of Animal Resources』Korean Journal for Food Science of Animal Resources 제33권 제5호, 610~616쪽, 전체 7쪽
• 주제분류: 농수해양 > 식품과학
• 파일형태: PDF
• 발행일자: 2013.10.31

영문 초록

Loop-mediated isothermal amplification (LAMP) is an emerging detection technology for the amplification of DNA under isothermal conditions. The aim of this study was to develop a rapid and reliable LAMP technique for the detection of Cronobacter sakazakii in milk powder. In order to enhance the sensitivity and specificity, LAMP primers targeting outer membrane protein A (ompA) gene of C. sakazakii were designed using Explorer V4 software. Thirty seven C. sakazakii strains and 13 pathogenic microorganisms were used for comparative detection of C. sakazakii using polymerase chain reaction (PCR), real-time PCR, and LAMP. LAMP developed in this study could specifically detect C. sakazakii strains without cross-reactivity with other foodborne pathogens. LAMP products amplified from ompA gene of C. sakazakii were digested with with HhaI and NruI enzyme. The specificity of LAMP was confirmed by restriction fragment length polymorphism (RFLP) analysis. LAMP could detect C. sakazakii within 1 h without bacterial culture and its detection limit was as low as 1 CFU/mL C. sakazakii in milk. In the comparison of the sensitivity, LAMP showed 10,000- and 100-times higher detection limit than PCR or real-time PCR, respectively. Therefore, this study can conclude that LAMP is a rapid and reliable detection technique for C. sakazakii contaminated in powdered milk.

키워드

loop-mediated isothermal amplification LAMP Cronobacter sakazakii ompA specificity sensitivity