Korean Journal of Veterinary Research 2003;43(2):231-237.
Development of Differential Media and Multiplex PCR Assays for the Rapid Detection of Listeria monocytogenes
Byeong-yeal Jung1, Hyun-sook Lim2, Suk-chan Jung1
1National Veterinary Research and Quarantine Service, MAF
2Research Institute of Public Health & Environment
Listeria monocytogenes의 신속검출을 위한 선택배지 및 multiplex PCR 기법 개발
정병열1, 임현숙2, 정석찬1
1국립수의과학검역원
2대구광역시 보건환경연구원
Abstract
Listeria (L.) monocytogenes in samples could not be detected occasioally by faster growth of other Listeria spp. especially L. innocua. The aim of this study was to develop the differential media and multiplex polymerase chain reaction (PCR) assays for the rapid detection of L. monocytogenes. L. monocytogenes colonies were characterized by their ${eta}$-hemolysis with fluorescence under 366 nm UV light on the Listeria hemolysis agar (LHA). L. innocua, a species commonly present in foods, did not produce ${eta}$-hemolysis on LHA. Therefore, one or more colonies of L. monocytogenes were easily distinguished from large populations of L. innocua. The multiplex PCR assays were developed to distinguish from L. monocytogenes and other Listeria spp. with two pairs of primers. The primers were designed in 16S rRNA and listeriolysin O gene for specific amplification of all members of the genus Listeria and L. monocytogenes, respectively. The multiplex PCR assays produced 560 and 938 bp products in L. monocytogenes; only 938 bp products in the genus Listeria. The multiplex PCR assays could detect as little as 50 pg of L monocytogenes DNA. These results indicated that the differential media and multiplex PCR assays might be useful diagnostic tools for the rapid detection of L. monocytogenes.
Key Words: Listeria monocytogenes, differential media, multiplex PCR


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