Chen Jianshun, Jiang Lingli, Chen Qiaomiao, Zhao Huancan, Luo Xiaokai, Chen Xueyan, Fang Weihuan
Institute of Preventive Veterinary Medicine, Zhejiang University , Hangzhou, P.R. China.
Foodborne Pathog Dis. 2009 Apr;6(3):365-76. doi: 10.1089/fpd.2008.0207.
The genus Listeria comprises two pathogenic species, L. monocytogenes and L. ivanovii, as well as four nonpathogenic species, L. innocua, L. weishimeri, L. seeligeri, and L. grayi. Within L. monocytogenes, lineages I and II are responsible for most listeriosis cases, while lineage III strains are rarely associated with human morbidity but providing important clues for Listeria evolution. The gene lmo0038, belonging to the peptidylarginine deiminase family, was involved in the optimal growth under stress conditions, including low pH and heat shock (52 degrees C), and virulence potential. Further, this gene was specific to L. monocytogenes lineages I and II and L. ivanovii with significant similarities at nucleotide and amino acid levels. A novel multiplex PCR, based on lmo0038 in combination with optimized iap migration profiles, was developed for simultaneous identification of Listeria species and discrimination of L. monocytogenes lineage III, with a detection limit down to 1.0-9.0 x 10(2) CFU/mL. This assay was evaluated by 119 suspected Listeria food-related isolates and corrected 4 and 5 misidentifications by Listeria selective agar plate screening and API system, respectively. Therefore, this one-step molecular assay provides a rapid, reliable, and inexpensive screening test to detect Listeria species-particularly, the pathogenic species in surveillance programs concerning food safety and foodborne disease cases.
李斯特菌属包括两种致病菌种,即单核细胞增生李斯特菌和伊氏李斯特菌,以及四种非致病菌种,即无害李斯特菌、威氏李斯特菌、斯氏李斯特菌和格氏李斯特菌。在单核细胞增生李斯特菌中,I 系和 II 系导致了大多数李斯特菌病病例,而 III 系菌株很少与人类发病相关,但为李斯特菌的进化提供了重要线索。基因 lmo0038 属于肽基精氨酸脱氨酶家族,参与了在包括低 pH 和热休克(52 摄氏度)等应激条件下的最佳生长以及毒力潜能。此外,该基因是单核细胞增生李斯特菌 I 系和 II 系以及伊氏李斯特菌所特有的,在核苷酸和氨基酸水平上具有显著相似性。基于 lmo0038 并结合优化的 iap 迁移图谱,开发了一种新型多重 PCR,用于同时鉴定李斯特菌属菌种并区分单核细胞增生李斯特菌 III 系,检测限低至 1.0 - 9.0×10² CFU/mL。该检测方法通过 119 株疑似与李斯特菌相关的食品分离株进行评估,分别纠正了李斯特菌选择性琼脂平板筛选和 API 系统的 4 例和 5 例错误鉴定。因此,这种一步式分子检测方法为检测李斯特菌属菌种,特别是在食品安全和食源性疾病病例监测项目中的致病菌种,提供了一种快速、可靠且廉价地筛选检测方法。