我国施万菌分布及其分子特征分析

黄振洲; 于可艺; 汪永禄; 李颖; 高鹤; 白雪梅; 肖悦; 赖玖连; 李坤; 王多春

doi:10.3784/jbjc.202212300556

我国施万菌分布及其分子特征分析

doi: 10.3784/jbjc.202212300556

黄振洲^1,,
于可艺¹,
汪永禄²,
李颖³,
高鹤¹,
白雪梅¹,
肖悦¹,
赖玖连^{1, 4},
李坤^{1, 4},
王多春^1, ,

1.
中国疾病预防控制中心传染病预防控制所, 北京 102206
2.
马鞍山市疾病预防控制中心, 安徽马鞍山 243000
3.
北京市顺义区疾病预防控制中心, 北京 101300
4.
兰州大学公共卫生学院, 甘肃兰州 730000

基金项目: 国家科技基础资源调查专项（No. 2021FY100904）

详细信息

作者简介:
黄振洲，男，浙江省绍兴市人，博士，主要从事病原生物学相关工作，Email：ronnie_0414@163.com

通讯作者:
王多春，Tel：010−58900773，Email：wangduochun@icdc.cn

中图分类号: R211; R378
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出版历程
- 收稿日期: 2022-12-30
- 网络出版日期: 2023-03-14
- 刊出日期: 2023-04-30

Distribution and molecular characterization of Shewanella in China

Huang Zhenzhou^1
,,
Yu Keyi¹,
Wang Yonglu²,
Li Ying³,
Gao He¹,
Bai Xuemei¹,
Xiao Yue¹,
Lai Jiulian^{1, 4},
Li Kun^{1, 4},
Wang Duochun^{1
, ,}

1.
National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
2.
Ma'anshan Prefectural Center for Disease Control and Prevention, Ma'anshan 243000, Anhui, China
3.
Shunyi District Center for Disease Control and Prevention, Beijing 101300, China
4.
School of Public Health, Lanzhou University, Lanzhou 730000, Gansu, China

Funds: This study was supported by the National Science and Technology Fundamental Resources Investigation Program of China (No. 2021FY100904)

More Information

Corresponding author: Wang Duochun, Email: wangduochun@icdc.cn

摘要

摘要: 目的探索中国施万菌的种群分布、分子流行病学特征及遗传进化关系，为施万菌的流行病学监测和进化研究提供依据。方法利用基质辅助激光解吸电离–飞行时间质谱（MALDI-TOF MS）联合gyrB基因测序进行施万菌种水平鉴定；利用7个管家基因（16S rRNA、gyrA、gyrB、infB、recN、rpoA和topA）进行多位点序列分型（MLST），利用BioNumerics 7.1软件对各个序列类型（ST）构建最小生成树。结果本研究共收集全国201株施万菌，鉴定到10个不同的菌种，其中海藻施万菌所占数量最多。不同分离来源中的施万菌种分布有所差异，临床和食品来源的施万菌种分布相似。 MLST将施万菌实验室分离株划分成136个ST型和15 个克隆复合体（CCs），其中CC12为海藻施万菌优势克隆群。结论我国分布的施万菌病原谱丰富多样，提示这些菌株具有高度的遗传多样性。
- 施万菌 /
- 种群分布 /
- 分子分型 /
- 遗传多样性
Abstract: Objective To explore the distribution and molecular epidemiological characteristics genetic evolution of Shewanella in China and provide evidence for epidemiological surveillance and evolution research of Shewanella. Methods The identification of Shewanella species was performed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) combined with gyrB gene sequencing. Multilocus sequence typing (MLST) was performed using seven housekeeping genes (16S rRNA, gyrA, gyrB, infB, recN, rpoA and topA). A minimal spanning tree was constructed for each sequence type (ST) by using software BioNumerics 7.1. Results The 201 Shewanella isolates collected in China were identified as 10 different species, with S. algae accounting for the largest proportion. There were differences in the distribution of Shewanella among the isolated from different sources, the distribution of S. algae from clinical and food sources was similar. MLST can classified the Shewanella isolates into 136 STs, clonal complex 12 (CC12) was the dominant clonal complexes of S. algae. Conclusion The rich pathogenic spectrum of Shewanella distributed in China suggests that the genetic diversity of these strains is high.
- Shewanella /
- Population distribution /
- Molecular typing /
- Genetic diversity

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图 1 施万菌种鉴定及构成分布

注：A. 施万菌gyrB基因Neighbor-Joining （NJ）进化树。该进化树共纳入10株模式菌株和201株实验室分离株，采用邻接法构建，bootstrap参数设为1 000。施万菌模式菌种用粗体标记，括号里数字分别代表模式菌株和实验室分离株数量。B. 施万菌种构成比

Figure 1. Identification and composition distribution of Shewanella spp. collected in this study

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图 2 施万菌分离来源分布

注：A. 按照临床和环境划分不同来源的施万菌。B. 不同分离来源中的施万菌种分布

Figure 2. Distribution of Shewanella spp. isolation sources

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图 3 施万菌分离时间分布

Figure 3. Folding Line Chart of isolation year of Shewanella spp.

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图 4 施万菌多位点序列分析最小生成树

注：A. 按照种水平划分不同菌种的施万菌。B. 按照分离地区划分施万菌。C. 按照分离来源划分施万菌。圆圈内数字代表所属序列类型（sequence type，ST），圆圈大小代表菌株数量多少，CC1～CC15代表本研究鉴定的15个克隆复合体（clonal complexes，CCs）

Figure 4. Minimum spanning tree of Shewanella spp. in the multilocus sequence typing analysis

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表 1 多位点序列分析管家基因引物序列信息表

Table 1. Summary of housekeeping genes primer sequences in multilocus sequence typing

基因名称	引物名称	引物序列（5'~3'）	退火温度（℃）	扩增长度（bp）
16S rRNA	8F	AGAGTTTGATCCTGGCTCAG	50	1414
16S rRNA	1492R	GGTTACCTTGTTACGACTT	50	1414
gyrA	gyrA-F	TGAAGAACGATTGGAACAARCCNTAYAARAARTC	56	664
gyrA	gyrA-R	TTTTCAATCAAACGAGCTTTGTTHACYTGRTAHGG	56	664
gyrB	gyrB-F	GAAGTGGCKATGCAGTGGAA	56	627
gyrB	gyrB-R	CGRCRAATACCACAGCCRAG	56	627
infB	infB-F	ATGCCACAGACTATTGAAGCDATYCARCAYGC	56	830
infB	infB-R	GCATCAGCACGAACGTTAAARCCNAYMAKRATNGC	56	830
recN	recN-F	AGTGAGCATCAACTGACCYTRYTNGAYAGYTAYGC	54	863
recN	recN-R	GGTTGTAAAGGTTGCCCTGGGTTDGTNSWNAC	54	863
rpoA	rpoA-F	TGGAGCCGCTTGAGCGTGGTTTYGGHCAYAC	56	751
rpoA	rpoA-R	ATGTAATGAATCGCTTCGGCYTTYARRCAGTT	56	751
topA	topA-F	GAATTCATCGTTAAGTCGAGYGTDGGBCAYRT	60	860
topA	topA-R	CGCTGGGCCATCATCATGGTYTTYTTNACNCC	60	860

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