Zhang Jianmin, Cao Guojie, Xu Xuebin, Allard Marc, Li Peng, Brown Eric, Yang Xiaowei, Pan Haijian, Meng Jianghong
National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, College of Veterinary Medicine, South China Agricultural University Guangzhou, China.
Department of Nutrition and Food Science and Joint Institute for Food Safety and Applied Nutrition, University of Maryland, College Park College Park, MD, USA.
Front Microbiol. 2016 Jul 22;7:1138. doi: 10.3389/fmicb.2016.01138. eCollection 2016.
Listeria monocytogenes is a significant foodborne pathogen causing severe systemic infections in humans with high mortality rates. The objectives of this work were to establish a phylogenetic framework of L. monocytogenes from China and to investigate sequence diversity among different serotypes. We selected 17 L. monocytogenes strains recovered from patients and foods in China representing serotypes 1/2a, 1/2b, and 1/2c. Draft genome sequences were determined using Illumina MiSeq technique and associated protocols. Open reading frames were assigned using prokaryotic genome annotation pipeline by NCBI. Twenty-four published genomes were included for comparative genomic and phylogenetic analysis. More than 154,000 single nucleotide polymorphisms (SNPs) were identified from multiple genome alignment and used to reconstruct maximum likelihood phylogenetic tree. The 41 genomes were differentiated into lineages I and II, which consisted of 4 and 11 subgroups, respectively. A clinical strain from China (SHL009) contained significant SNP differences compared to the rest genomes, whereas clinical strain SHL001 shared most recent common ancestor with strain SHL017 from food. Moreover, clinical strains SHL004 and SHL015 clustered together with two strains (08-5578 and 08-5923) recovered from an outbreak in Canada. Partial sequences of a plasmid found in the Canadian strain were also present in SHL004. We investigated the presence of various genes and gene clusters associated with virulence and subgroup-specific genes, including internalins, L. monocytogenes pathogenicity islands (LIPIs), L. monocytogenes genomic islands (LGIs), stress survival islet 1 (SSI-1), and clustered regularly interspaced short palindromic repeats (CRISPR)/cas system. A novel genomic island, denoted as LGI-2 was identified. Comparative sequence analysis revealed differences among the L. monocytogenes strains related to virulence, survival abilities, and attributes against foreign genetic elements. L. monocytogenes from China were genetically diverse. Strains from clinical specimens and food related closely suggesting foodborne transmission of human listeriosis.
单核细胞增生李斯特菌是一种重要的食源性病原体,可导致人类严重的全身感染,死亡率很高。这项工作的目的是建立来自中国的单核细胞增生李斯特菌的系统发育框架,并研究不同血清型之间的序列多样性。我们选择了从中国患者和食品中分离出的17株单核细胞增生李斯特菌菌株,分别代表血清型1/2a、1/2b和1/2c。使用Illumina MiSeq技术和相关方案测定了基因组草图序列。使用NCBI的原核生物基因组注释管道分配开放阅读框。纳入了24个已发表的基因组进行比较基因组和系统发育分析。通过多个基因组比对鉴定出超过154,000个单核苷酸多态性(SNP),并用于重建最大似然系统发育树。这41个基因组被分为I和II两个谱系,分别由4个和11个亚组组成。来自中国的一株临床菌株(SHL009)与其余基因组相比存在显著的SNP差异,而临床菌株SHL001与来自食品的菌株SHL017有最近的共同祖先。此外,临床菌株SHL004和SHL015与从加拿大一次暴发中分离出的两株菌株(08 - 5578和08 - 5923)聚集在一起。在加拿大菌株中发现的一个质粒的部分序列也存在于SHL004中。我们研究了与毒力和亚组特异性基因相关的各种基因和基因簇的存在情况,包括内化素、单核细胞增生李斯特菌致病岛(LIPIs)、单核细胞增生李斯特菌基因组岛(LGIs)、应激存活岛1(SSI - 1)以及成簇规律间隔短回文重复序列(CRISPR)/cas系统。鉴定出了一个新的基因组岛,命名为LGI - 2。比较序列分析揭示了单核细胞增生李斯特菌菌株在毒力、存活能力以及对外源遗传元件的抗性方面的差异。来自中国的单核细胞增生李斯特菌具有遗传多样性。临床标本和食品中的菌株关系密切,提示人类李斯特菌病通过食物传播。