鉴定单增李斯特菌四个进化家系及两个亚系的多PCR方法的建立(英文)  

Differentiation of 4 Listeria monocytogenes phylogenetic lineages and 2sublineages by a novel multiplex PCR

作　　者：王毅[1] 王艳[1] 叶正兴[1] 马爱静[1] 代航[1] 许华清[2] 李东迅[2] 叶长芸[1] 

机构地区：[1]传染病预防控制国家重点实验室,中国疾病预防控制中心传染病预防控制所,北京102206 [2]贵阳医学院微生物学教研室,贵阳550004

出　　处：《中国人兽共患病学报》2013年第12期1151-1156,共6页Chinese Journal of Zoonoses

基　　金：Supported by the Mega Project of Research on the Prevention and Control of HIV/AIDS and Viral Hepatitis Infectious Diseases(No.2011ZX10004-001,No.2013ZX10004-101)~~

摘　　要：目的利用多重PCR(multiplex PCR)技术,建立针对单增李斯特菌4个进化家系(LineageⅠ、LineageⅡ、LineageⅢ、LineageⅣ)及2个亚系(LineageⅢA、LineageⅢC)的鉴定方法。方法以InlA、Lmo0733、Lmo0038、Lmo0735和LmaA作为靶基因设计引物,建立同时鉴别LineageⅠ、LineageⅡ、LineageⅢA、LineageⅢC和LineageⅣ菌株的多重PCR反应体系,并对387株单增李斯特菌进行了所属进化家系及亚系的鉴定。结果 387株单增李斯特菌可用本方法进行所属家系和亚系的分型,包括LineageⅠ246株、LineageⅡ123株、LineageⅢA 4株、LineageⅢC 13株和LineageⅣ1株。结论本方法能准确鉴定单增李斯特菌的4个进化家系及2个亚系的菌株,可用于食品、临床标本中单增李斯特菌的病原学诊断和疫情暴发时的溯源调查。Listeria monocytogenes, an import food-borne pathogen in human and livestocks, encompasses four phyloge- netie lineages. Lineage I and II are responsible for most listeriosis eases, while lineages III （including sublineages IIIA and IIIC） and IV strains are rarely associated with human morbidity but providing significant clues for Listeria evolution. In this study, we have developed a multiplex PCR for rapid and simultaneous detection of four lineages and two sublineages in only one reac- tion step with easily interpretable results. This multiplex PCR strategy was successfully evaluated with 392 Listeria strains, and the classification of lineages and sublineages was consistent with other phenotypic and genotypic methods. This established multiplex PCR assay provides rapid and reliable results and will be useful for the detection of L. monocytogenes lineages and sublineages in contaminated food products and clinical samples.

关 键 词：单增李斯特菌 进化家系 多重PCR 

分 类 号：R378.994[医药卫生—病原生物学]