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

doi:10.3969/cjz.j.issn.1002-2694.2013.12.005

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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、Lineage Ⅲ、Lineage Ⅳ)及2个亚系(Lineage ⅢA、Lineage ⅢC)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、Lineage ⅢA、Lineage ⅢC和 Lineage Ⅳ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、Lineage ⅢA 4株、Lineage ⅢC 13株和 Lineage Ⅳ 1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。

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	王毅
	王艳
	叶正兴
	马爱静
	代航
	许华清
	李东迅
	叶长芸

关键词 ：单增李斯特菌, 进化家系, 多重 PCR

Abstract：Listeria monocytogenes, an import food-borne pathogen in human and livestocks, encompasses four phylogenetic lineages. Lineage I and II are responsible for most listeriosis cases, 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 reaction 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.

Key words： Listeria monocytogenes Lineages Multiplex PCR 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)

收稿日期: 2013-09-16

PACS:

R378.99+4

基金资助:国家重大传染病防治科技重大专项（No.2011ZX10004-001，No.2013ZX10004-101）资助

通讯作者: 叶长芸, Email:yechangyun@icdc.cn E-mail: yechangyun@icdc.cn

引用本文:

王毅, 王艳, 叶正兴, 马爱静, 代航, 许华清, 李东迅, 叶长芸. 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[J]. 中国人兽共患病学报, 2013, 29(12): 1151-1156. WANG Yi, WANG Yan, YE Zheng-xing, MA Ai-jing, DAI Hang, XU Hua-qing, LI Dong-xun, YE Chang-yun. Differentiation of 4 Listeria monocytogene phylogenetic lineages and 2 sublineages by a novel multiplex PCR. Chinese Journal of Zoonoses, 2013, 29(12): 1151-1156.

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http://www.rsghb.cn/CN/10.3969/cjz.j.issn.1002-2694.2013.12.005 或 http://www.rsghb.cn/CN/Y2013/V29/I12/1151

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