一株ST477型单增李斯特菌的全基因组测序及inlA基因遗传多样性
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摘要
【背景】单增李斯特菌是一种重要的条件致病菌,不同型别菌株在宿主范围和毒力等方面存在差异。内化素基因inlA在入侵宿主上皮细胞中具有重要作用。【目的】研究单增李斯特菌序列型(Sequence type,ST)为477菌株的基因组特征及内化素基因inlA的遗传多样性。【方法】使用相关软件对测序数据进行多位点序列分型(Mutilocussequencetyping,MLST)、单核苷酸多态性(Single nucleotide polymorphism,SNP)及基因inlA遗传多样性分析。【结果】MLST进化分析结果显示,分离自不同国家的菌株具有较近亲缘关系。以分离自中国食品的ST477型菌株为参考菌株,通过SNP分析表明,加拿大食品中的ST9型菌株发生的突变位点最少(91-93个)。7株复合克隆系(Clonal complex,CC)为9的菌株其inlA基因序列间核苷酸相似性为29.8%-100%。【结论】初步分析了ST477型别菌株的进化及基因组特征,同时研究了部分CC9克隆系菌株inlA基因突变情况,为研究ST477型别菌株的进化及单增李斯特菌的毒力提供基础数据。
[Background] Listeria monocytogenes is an important opportunistic pathogen. There are some differences in host range and virulence among different strains. Internalin gene inlA play an important role in host epithelial cells invasion. [Objective] Analyze the genomic characteristics of the Listeria monocytogenes STS477 isolate and investigate the genetic diversity of gene inlA. [Methods] Mutilocus sequence typing(MLST), single nucleotide polymorphism(SNP) and inlA genetic diversity were analyzed based on the sequencing data with software. [Results] MLST analysis showed that isolates from different countries have close relationship. Based on the SNP analysis, ST9 strains isolated from Canadian food owned the fewest mutation sites(91-93) by aligning six CC9 isolates against the chromosome of the ST477. The nucleotide homology of seven inlA gene sequences was 29.8%-100%. [Conclusion] We preliminarily analyzed the evolution of ST477 isolate and investigate the mutation type of inlA gene belonging to CC9. This study provides basic data for analysis of the evolution of ST477 strains and the virulence of L. monocytogenes.
[Background] Listeria monocytogenes is an important opportunistic pathogen. There are some differences in host range and virulence among different strains. Internalin gene inlA play an important role in host epithelial cells invasion. [Objective] Analyze the genomic characteristics of the Listeria monocytogenes STS477 isolate and investigate the genetic diversity of gene inlA. [Methods] Mutilocus sequence typing(MLST), single nucleotide polymorphism(SNP) and inlA genetic diversity were analyzed based on the sequencing data with software. [Results] MLST analysis showed that isolates from different countries have close relationship. Based on the SNP analysis, ST9 strains isolated from Canadian food owned the fewest mutation sites(91-93) by aligning six CC9 isolates against the chromosome of the ST477. The nucleotide homology of seven inlA gene sequences was 29.8%-100%. [Conclusion] We preliminarily analyzed the evolution of ST477 isolate and investigate the mutation type of inlA gene belonging to CC9. This study provides basic data for analysis of the evolution of ST477 strains and the virulence of L. monocytogenes.
引文
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[17]Kuenne C,Billion A,Mraheil MA,et al.Reassessment of the Listeria monocytogenes pan-genome reveals dynamic integration hotspots and mobile genetic elements as major components of the accessory genome[J].BMC Genomics,2013,14:47
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[19]Liu EL,Yuan MY,Deng JY,et al.Multilocus sequence typing of Listeria monocytogenes isolated from imported and exported animal-derived foods[J].Food Science,2016,37(12):212-216(in Chinese)刘二龙,袁慕云,邓建英,等.出入境动物源性食品中单增李斯特菌的多位点序列分型分析[J].食品科学,2016,37(12):212-216
[20]Moura A,Criscuolo A,Pouseele H,et al.Whole genome-based population biology and epidemiological surveillance of Listeria monocytogenes[J].Nature Microbiology,2016,2(2):16185
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[22]Zhang JM,Cao GJ,Xu XB,et al.Evolution and diversity of Listeria monocytogenes from clinical and food samples in Shanghai,China[J].Frontiers in Microbiology,2016,7:1138
[23]Halbedel S,Prager R,Fuchs S,et al.Whole-genome sequencing of recent Listeria monocytogenes isolates from Germany reveals population structure and disease clusters[J].Journal of Clinical Microbiology,2018,56(6):e00119-18
[24]Toledo V,den Bakker HC,Hormazábal JC,et al.Genomic diversity of Listeria monocytogenes isolated from clinical and non-clinical samples in Chile[J].Genes,2018,9(8):396
[25]Gelbí?ováT,Kolá?kováI,Pant??ek R,et al.A novel mutation leading to a premature stop codon in inlA of Listeria monocytogenes isolated from neonatal listeriosis[J].New Microbiologica,2015,38(2):293-296
[26]van Stelten A,Simpson JM,Ward TJ,et al.Revelation by single-nucleotide polymorphism genotyping that mutations leading to a premature stop codon in inlA are common among Listeria monocytogenes isolates from ready-to-eat foods but not human listeriosis cases[J].Applied and Environmental Microbiology,2010,76(9):2783-2790
[27]Manuel CS,van Stelten A,Wiedmann M,et al.Prevalence and distribution of Listeria monocytogenes inlA alleles prone to phase variation and inlA alleles with premature stop codon mutations among human,food,animal,and environmental isolates[J].Applied and Environmental Microbiology,2015,81(24):8339-8345
[28]Palma F,Pasquali F,Lucchi A,et al.Whole genome sequencing for typing and characterisation of Listeria monocytogenes isolated in a rabbit meat processing plant[J].Italian Journal of Food Safety,2017,6(3):6879
[2]Angelo KM,Conrad AR,Saupe A,et al.Multistate outbreak of Listeria monocytogenes infections linked to whole apples used in commercially produced,prepackaged caramel apples:United States,2014-2015[J].Epidemiology&Infection,2017,145(5):848-856
[3]Jackson BR,Tarr C,Strain E,et al.Implementation of nationwide real-time whole-genome sequencing to enhance listeriosis outbreak detection and investigation[J].Clinical Infectious Diseases,2016,63(3):380-386
[4]Jiao Y,Zhang W,Ma JR,et al.Early onset of neonatal listeriosis[J].Pediatrics International,2011,53(6):1034-1037
[5]Wang H,Wang Y,Zhang ZD,et al.Clinical and epidemiological characteristics of three cases of Listeria monocytogenes infection[J].Chinese Journal of Zoonoses,2015,31(7):678-680(in Chinese)王红,王艳,张正东,等.3例单增李斯特菌感染的病原学、临床及流行病学特征分析[J].中国人兽共患病学报,2015,31(7):678-680
[6]Wu S,Wu QP,Zhang JM,et al.Listeria monocytogenes prevalence and characteristics in retail raw foods in China[J].PLoS One,2015,10(8):e0136682
[7]Ma AJ,Wang Y,Wang Y,et al.Molecular epidemiological characteristics of Listeria monocytogenes isolated from raw meat samples in some regions of Beijing,China[J].Chinese Journal of Zoonoses,2015,31(5):403-407(in Chinese)马爱静,王艳,王毅,等.北京市一些地区生肉标本中单增李斯特菌的分离及其分子流行病学特征分析[J].中国人兽共患病学报,2015,31(5):403-407
[8]Li ZY,Jia JT,Zeng J,et al.Identification of Listeria monocytogenes from imported Trachurus japonicas and prokaryotic expression of flagellin[J].Microbiology China,2014,41(6):1160-1167(in Chinese)李正义,贾俊涛,曾静,等.进口竹荚鱼中单核细胞增生李斯特菌的鉴定及其鞭毛蛋白的原核表达[J].微生物学通报,2014,41(6):1160-1167
[9]Ragon M,Wirth T,Hollandt F,et al.A new perspective on Listeria monocytogenes evolution[J].PLoS Pathogen,2008,4(9):e1000146
[10]Ward TJ,Ducey TF,Usgaard T,et al.Multilocus genotyping assays for single nucleotide polymorphism-based subtyping of Listeria monocytogenes isolates[J].Applied and Environmental Microbiology,2008,74(24):7629-7642
[11]Nightingale KK,Windham K,Martin KE,et al.Select Listeria monocytogenes subtypes commonly found in foods carry distinct nonsense mutations in inlA,leading to expression of truncated and secreted internalin A,and are associated with a reduced invasion phenotype for human intestinal epithelial cells[J].Applied and Environmental Microbiology,2005,71(12):8764-8772
[12]Wang Y,Zhao AL,Zhu RF,et al.Genetic diversity and molecular typing of Listeria monocytogenes in China[J].BMCMicrobiology,2012,12:119
[13]Wu S,Wu QP,Zhang JM,et al.Analysis of multilocus sequence typing and virulence characterization of Listeria monocytogenes isolates from Chinese retail ready-to-eat food[J].Frontiers in Microbiology,2016,7:168
[14]Yan H,Neogi SB,Mo ZY,et al.Prevalence and characterization of antimicrobial resistance of foodborne Listeria monocytogenes isolates in Hebei province of Northern China,2005-2007[J].International Journal of Food Microbiology,2010,144(2):310-316
[15]Rabinovich L,Sigal N,Borovok I,et al.Prophage excision activates Listeria competence genes that promote phagosomal escape and virulence[J].Cell,2012,150(4):792-802
[16]Wang X,Li J,Zhang ZH,et al.Comparative genomic analysis of one Listeria monocytogenes containing five prophages[J].Genomics and Applied Biology,2017,36(2):670-685(in Chinese)王旭,李骏,张昭寰,等.一株含有5个原噬菌体的单增李斯特菌比较基因组分析[J].基因组学与应用生物学,2017,36(2):670-685
[17]Kuenne C,Billion A,Mraheil MA,et al.Reassessment of the Listeria monocytogenes pan-genome reveals dynamic integration hotspots and mobile genetic elements as major components of the accessory genome[J].BMC Genomics,2013,14:47
[18]Chenal-Francisque V,Lopez J,Cantinelli T,et al.Worldwide distribution of major clones of Listeria monocytogenes[J].Emerging Infectious Diseases,2011,17(6):1110-1112
[19]Liu EL,Yuan MY,Deng JY,et al.Multilocus sequence typing of Listeria monocytogenes isolated from imported and exported animal-derived foods[J].Food Science,2016,37(12):212-216(in Chinese)刘二龙,袁慕云,邓建英,等.出入境动物源性食品中单增李斯特菌的多位点序列分型分析[J].食品科学,2016,37(12):212-216
[20]Moura A,Criscuolo A,Pouseele H,et al.Whole genome-based population biology and epidemiological surveillance of Listeria monocytogenes[J].Nature Microbiology,2016,2(2):16185
[21]Fretz R,Pichler J,Sagel U,et al.Update:multinational listeriosis outbreak due to‘Quargel’,a sour milk curd cheese,caused by two different L.monocytogenes serotype 1/2a strains,2009-2010[J].EuroSurveillance,2010,15(16):19543
[22]Zhang JM,Cao GJ,Xu XB,et al.Evolution and diversity of Listeria monocytogenes from clinical and food samples in Shanghai,China[J].Frontiers in Microbiology,2016,7:1138
[23]Halbedel S,Prager R,Fuchs S,et al.Whole-genome sequencing of recent Listeria monocytogenes isolates from Germany reveals population structure and disease clusters[J].Journal of Clinical Microbiology,2018,56(6):e00119-18
[24]Toledo V,den Bakker HC,Hormazábal JC,et al.Genomic diversity of Listeria monocytogenes isolated from clinical and non-clinical samples in Chile[J].Genes,2018,9(8):396
[25]Gelbí?ováT,Kolá?kováI,Pant??ek R,et al.A novel mutation leading to a premature stop codon in inlA of Listeria monocytogenes isolated from neonatal listeriosis[J].New Microbiologica,2015,38(2):293-296
[26]van Stelten A,Simpson JM,Ward TJ,et al.Revelation by single-nucleotide polymorphism genotyping that mutations leading to a premature stop codon in inlA are common among Listeria monocytogenes isolates from ready-to-eat foods but not human listeriosis cases[J].Applied and Environmental Microbiology,2010,76(9):2783-2790
[27]Manuel CS,van Stelten A,Wiedmann M,et al.Prevalence and distribution of Listeria monocytogenes inlA alleles prone to phase variation and inlA alleles with premature stop codon mutations among human,food,animal,and environmental isolates[J].Applied and Environmental Microbiology,2015,81(24):8339-8345
[28]Palma F,Pasquali F,Lucchi A,et al.Whole genome sequencing for typing and characterisation of Listeria monocytogenes isolated in a rabbit meat processing plant[J].Italian Journal of Food Safety,2017,6(3):6879