上海市一例肠出血性大肠埃希菌O157∶H7感染病例的病原学分析
|
摘要
目的通过流行病学调查与病原菌鉴定,并结合既往相关菌株资料,对2016年6月上海市腹泻病综合监测中报告的1例肠出血性大肠埃希菌(EHEC)O157∶H7感染病例进行病原学分析。方法对患者开展流行病学调查。采集标本后进行菌株分离培养、生化鉴定并确定血清型;利用聚合酶链式反应(PCR)检测所携带的毒力基因;采用微量肉汤稀释法进行药敏试验;应用脉冲场凝胶电泳(PFGE)技术进行分子分型;通过全基因组测序获取序列信息进行多位点序列分型(MLST)及单核苷酸多态性分析。结果流行病学调查未发现可疑传染源。患者样本分离出EHEC O157∶H7;携带eae、stx2基因;对进行试验的16种抗生素均敏感;菌株图谱带型与以往监测中分离菌株带型均不相同;MLST分型为ST11型,与以往分离菌株核心基因组差异较大。结论上海市较长时间无检出EHEC O157∶H7菌株的病例记录,通过对此次分离的菌株的全面检测,确定了其毒力基因、耐药性和MLST型别,并发现新的PFGE带型,其核心基因组与以往菌株呈现出一定差异,值得进一步关注。
Objective To understand the etiologic characteristics of an infection case of enterohemorrhagic Escherichia coli(EHEC) O157∶H7 in June, 2016 in Shanghai. Methods The patient information was collected by epidemiological investigation. Two strains were isolated from case's stool samples and identified by biochemical analysis and serologic tests.Polymerase chain reaction was conducted to detect the virulence related genes and microdilution broth method was used to analyze the antibiotic resistance. Pulsed-field gel electrophoresis(PFGE), multi-locus sequence typing(MLST) and single nucleotide polymorphisms(SNP) were conducted for further analysis on the strain's characteristics. Results No suspected sources of infection were found by epidemiological investigation. The strains were confirmed to be EHEC O157∶H7 carrying eae and stx2, and they were sensitive to 16 kinds of antibiotics tested. Two strains were classified to ST11 while showed a new PFGE pattern. Compared with the reference strain, they showed more SNPs than the previously identified strains by core genome alignment. Conclusion No EHEC O157∶H7 has been isolated from patient for about ten years in Shanghai since2006. The virulence related genes, the antibiotic resistance and the ST type of the strains were confirmed in this study. They were more different from the previously identified strains according to the genome sequence alignment and PFGE results,further attention should be paid to it.
Objective To understand the etiologic characteristics of an infection case of enterohemorrhagic Escherichia coli(EHEC) O157∶H7 in June, 2016 in Shanghai. Methods The patient information was collected by epidemiological investigation. Two strains were isolated from case's stool samples and identified by biochemical analysis and serologic tests.Polymerase chain reaction was conducted to detect the virulence related genes and microdilution broth method was used to analyze the antibiotic resistance. Pulsed-field gel electrophoresis(PFGE), multi-locus sequence typing(MLST) and single nucleotide polymorphisms(SNP) were conducted for further analysis on the strain's characteristics. Results No suspected sources of infection were found by epidemiological investigation. The strains were confirmed to be EHEC O157∶H7 carrying eae and stx2, and they were sensitive to 16 kinds of antibiotics tested. Two strains were classified to ST11 while showed a new PFGE pattern. Compared with the reference strain, they showed more SNPs than the previously identified strains by core genome alignment. Conclusion No EHEC O157∶H7 has been isolated from patient for about ten years in Shanghai since2006. The virulence related genes, the antibiotic resistance and the ST type of the strains were confirmed in this study. They were more different from the previously identified strains according to the genome sequence alignment and PFGE results,further attention should be paid to it.
引文
[1]Nataro JP,Kaper JB.Diarrheagenic Escherichia coli[J].Clin Microbiol Rev,1998,11(1):142-201.DOI:10.1128/CMR.11.1.142.
[2]Xiong YW,Wang P,Lan RT,et al.A novel Escherichia coli O157∶H7 clone causing a major hemolytic uremic syndrome outbreak in China[J].PLoS One,2012,7(4).DOI:10.1371/journal.pone.0036144.
[3]顾宝柯,许学斌,金汇明,等.上海地区家畜、家禽及肉制品大肠杆菌O157∶H7监测[J].疾病监测,2003,18(1):5-7.DOI:10.3784/j.issn.1003-9961.2003.01.005.Gu BK,Xu XB,Jin HM,et al.Surveillance of E.coli O157∶H7in meat food of Shanghai district[J].Dis Surveill,2003,18(1):5-7.DOI:10.3784/j.issn.1003-9961.2003.01.005.
[4]Wang SH,Zhang SX,Liu Z,et al.Molecular characterization of enterohemorrhagic E.coli O157 isolated from animal fecal and food samples in Eastern China[J].Scientific World Journal,2014,2014:946394.DOI:10.1155/2014/946394.
[5]沈小明,赵冰,胡培玉,等.上海市浦东新区肠出血性大肠埃希菌O157∶H7的监测研究[J].中国病原生物学杂志,2008,3(8):570-573.DOI:10.3969/j.issn.1673-5234.2008.08.004.Shen XM,Zhao B,Hu PY,et al.Surveillance on Escherichia coli O157∶H7 in Shanghai Pudong new area[J].J Pathog Biol,2008,3(8):570-573.DOI:10.3969/j.issn.1673-5234.2008.08.004.
[6]孔令娜,黎健,马飞飞,等.上海市由腹泻综合监测系统发现的1例O157∶H7感染性腹泻病例的确认及流行病学调查[J].职业与健康,2018,34(1):120-122.Kong LN,Li J,Ma FF,et al.Identification and epidemiological investigation of an infectious diarrhea case of O157∶H7 owing to the diarrhea surveillance system in Shanghai[J].Occup Health,2018,34(1):120-122.
[7]Ribot EM,Fair MA,Gautom R,et al.Standardization of pulsedfield gel electrophoresis protocols for the subtyping of Escherichia coli O157∶H7,Salmonella,and Shigella for PulseNet[J].Foodborne Pathog Dis,2006,3(1):59-67.DOI:10.1089/fpd.2006.3.59.
[8]Parks DH,Imelfort M,Skennerton CT,et al.CheckM:assessing the quality of microbial genomes recovered from isolates,single cells,and metagenomes[J].Genome Res,2015,25(7):1043-1055.DOI:10.1101/gr.186072.114.
[9]Wirth T,Falush D,Lan RT,et al.Sex and virulence in Escherichia coli:an evolutionary perspective[J].Mol Microbiol,2006,60(5):1136-1151.DOI:10.1111/j.1365-2958.2006.05172.x.
[10]Treangen TJ,Ondov BD,Koren S,et al.The Harvest suite for rapid core-genome alignment and visualization of thousands of intraspecific microbial genomes[J].Genome Biol,2014,15(11):524.DOI:10.1186/s13059-014-0524-x.
[11]盛跃颖,庄源,陈洪友,等.上海地区分离的大肠埃希菌O157的分子分型研究[J].中国卫生检验杂志,2016,26(7):917-920.Sheng YY,Zhuang Y,Chen HY,et al.Study on the molecular typing of Eschierichia coli O157 isolated in Shanghai[J].Chin JHealth Lab Tec,2016,26(7):917-920.
[12]Eklund M,Leino K,Siitonen A.Clinical Escherichia coli strains carrying stx genes:stx variants and stx-positive virulence profiles[J].J Clin Microbiol,2002,40(12):4585-4593.DOI:10.1128/JCM.40.12.4585-4593.2002.
[13]Ji XW,Liao YL,Zhu YF,et al.Multilocus sequence typing and virulence factors analysis of Escherichia coli O157 strains in China[J].J Microbiol,2010,48(6):849-855.DOI:10.1007/s12275-010-0132-8.
[14]Noller AC,McEllistrem MC,Stine OC,et al.Multilocus sequence typing reveals a lack of diversity among Escherichia coli O157∶H7 isolates that are distinct by pulsed-field gel electrophoresis[J].J Clin Microbiol,2003,41(2):675-679.DOI:10.1128/JCM.41.2.675-679.2003.
[2]Xiong YW,Wang P,Lan RT,et al.A novel Escherichia coli O157∶H7 clone causing a major hemolytic uremic syndrome outbreak in China[J].PLoS One,2012,7(4).DOI:10.1371/journal.pone.0036144.
[3]顾宝柯,许学斌,金汇明,等.上海地区家畜、家禽及肉制品大肠杆菌O157∶H7监测[J].疾病监测,2003,18(1):5-7.DOI:10.3784/j.issn.1003-9961.2003.01.005.Gu BK,Xu XB,Jin HM,et al.Surveillance of E.coli O157∶H7in meat food of Shanghai district[J].Dis Surveill,2003,18(1):5-7.DOI:10.3784/j.issn.1003-9961.2003.01.005.
[4]Wang SH,Zhang SX,Liu Z,et al.Molecular characterization of enterohemorrhagic E.coli O157 isolated from animal fecal and food samples in Eastern China[J].Scientific World Journal,2014,2014:946394.DOI:10.1155/2014/946394.
[5]沈小明,赵冰,胡培玉,等.上海市浦东新区肠出血性大肠埃希菌O157∶H7的监测研究[J].中国病原生物学杂志,2008,3(8):570-573.DOI:10.3969/j.issn.1673-5234.2008.08.004.Shen XM,Zhao B,Hu PY,et al.Surveillance on Escherichia coli O157∶H7 in Shanghai Pudong new area[J].J Pathog Biol,2008,3(8):570-573.DOI:10.3969/j.issn.1673-5234.2008.08.004.
[6]孔令娜,黎健,马飞飞,等.上海市由腹泻综合监测系统发现的1例O157∶H7感染性腹泻病例的确认及流行病学调查[J].职业与健康,2018,34(1):120-122.Kong LN,Li J,Ma FF,et al.Identification and epidemiological investigation of an infectious diarrhea case of O157∶H7 owing to the diarrhea surveillance system in Shanghai[J].Occup Health,2018,34(1):120-122.
[7]Ribot EM,Fair MA,Gautom R,et al.Standardization of pulsedfield gel electrophoresis protocols for the subtyping of Escherichia coli O157∶H7,Salmonella,and Shigella for PulseNet[J].Foodborne Pathog Dis,2006,3(1):59-67.DOI:10.1089/fpd.2006.3.59.
[8]Parks DH,Imelfort M,Skennerton CT,et al.CheckM:assessing the quality of microbial genomes recovered from isolates,single cells,and metagenomes[J].Genome Res,2015,25(7):1043-1055.DOI:10.1101/gr.186072.114.
[9]Wirth T,Falush D,Lan RT,et al.Sex and virulence in Escherichia coli:an evolutionary perspective[J].Mol Microbiol,2006,60(5):1136-1151.DOI:10.1111/j.1365-2958.2006.05172.x.
[10]Treangen TJ,Ondov BD,Koren S,et al.The Harvest suite for rapid core-genome alignment and visualization of thousands of intraspecific microbial genomes[J].Genome Biol,2014,15(11):524.DOI:10.1186/s13059-014-0524-x.
[11]盛跃颖,庄源,陈洪友,等.上海地区分离的大肠埃希菌O157的分子分型研究[J].中国卫生检验杂志,2016,26(7):917-920.Sheng YY,Zhuang Y,Chen HY,et al.Study on the molecular typing of Eschierichia coli O157 isolated in Shanghai[J].Chin JHealth Lab Tec,2016,26(7):917-920.
[12]Eklund M,Leino K,Siitonen A.Clinical Escherichia coli strains carrying stx genes:stx variants and stx-positive virulence profiles[J].J Clin Microbiol,2002,40(12):4585-4593.DOI:10.1128/JCM.40.12.4585-4593.2002.
[13]Ji XW,Liao YL,Zhu YF,et al.Multilocus sequence typing and virulence factors analysis of Escherichia coli O157 strains in China[J].J Microbiol,2010,48(6):849-855.DOI:10.1007/s12275-010-0132-8.
[14]Noller AC,McEllistrem MC,Stine OC,et al.Multilocus sequence typing reveals a lack of diversity among Escherichia coli O157∶H7 isolates that are distinct by pulsed-field gel electrophoresis[J].J Clin Microbiol,2003,41(2):675-679.DOI:10.1128/JCM.41.2.675-679.2003.