急性腹泻患者致泻性大肠埃希菌流行情况及耐药性分析
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摘要
目的了解致泻性大肠埃希菌(DEC)在急性腹泻患者中的分布及对常用抗菌药物的敏感性,为疾病的防控和治疗提供依据。方法收集2014年1月至2015年12月期间就诊于浙江大学医学院附属第一医院急性腹泻患者粪便标本,采用常规病原菌检验流程对常见肠道致病菌进行分离培养鉴定,分离到的疑似大肠埃希菌采用多重PCR和单重PCR进行鉴定、分型,采用纸片扩散法(K-B法)对各型DEC进行18种常用药物敏感试验。结果从1 019例急性腹泻患者标本中分离到396株病原菌,其中DEC230株,分离率为22.6%,占所有分离菌株的58.1%,居病原菌首位。230株DEC中,ETEC占56.5%,EAEC占30.4%,EPEC占8.7%,STEC占0.4%,混合型DEC占0.9%,未发现EIEC。各型DEC全年均有检出,其中夏季(6~8月)分离率最高,达24.0%,患者主要分布在19~45岁,男女检出率分别为21.8%和23.2%,差异无统计学意义。DEC对氨苄西林的耐药率最高,达49.3%,对头孢唑林、复方新诺明、氨苄西林/舒巴坦的耐药率分别为46.4%、35.7%、30.9%,其余抗菌药物耐药率均低于15.0%,未出现亚胺培南耐药株。207株DEC中,产ESBLs菌株占41.1%,多重耐药菌株占44.9%。结论 DEC是本地区急性腹泻最常见的细菌性病原,是临床肠道感染中不可忽视的病原之一,尤其是同时携带多型别毒力基因菌株及产ESBLs菌株和多重耐药菌。长期持续监测其分布及耐药趋势,对疾病的预防、治疗具有重要意义。
Objective To learn the distribution and antimicrobial susceptibility of diarrheagenic Escherichia coli(DEC)isolated from patients with acute diarrhea,and provide evidence for the prevention and treatment.Methods The suspicious strains were isolated from acute diarrhea patients in Hangzhou area from January 2014 to December 2015 by using conventional methods.Multiplex PCR and monoplex PCR were applied to detect 12 virulence genes,and K-B method was used to carry out 18 commonly used drug susceptibility tests for all types of DEC.Results A total of 396 enteric pathogens were isolated from 1 019 specimens.DEC was found in 230cases(22.6%,230/1019)and accounted for 58.1% of all isolates,ranked 1st among the pathogenic bacteria.Among the 230 DECs,ETEC accounted for 56.5%,EAEC 30.4%,EPEC 8.7%,and STEC 0.4%.Two different pathotypes of DEC strains were isolated from 9specimens of patients with mixed infections.No EIEC was found.All types of DEC were isolated throughout the year,mainly from June to August.DEC infections occurred mainly in young adults of 18-45 years old.There was no significant difference in the detection rate between male(21.8%)and female(23.2%).Of the 207 isolates,49.3% were resistant to Ampicillin.The resistance rate to Cefazolin,Trimethoprim/Sulfamethoxazole and Ampicillin/Sulbactam were 46.4%,35.7% and 30.9%respectively,while those to the rest antibiotics were all less than 15.0%.All the isolates were sensitive to Imipenem.ESBLs-producing strains accounted for 41.1%and multidrug-resistant strains accounted for44.9%.Conclusion DEC,especially ESBLs-producing strains,multidrug-resistant strains and some pathotypes with multiple virulence genes,were the unneglectable pathogens in intestinal infections,which were the most common pathogens that cause diarrhea in Hangzhou area.Long-term monitoring on epidemic trend and drug resistance is of great significance for disease prevention and treatment.
Objective To learn the distribution and antimicrobial susceptibility of diarrheagenic Escherichia coli(DEC)isolated from patients with acute diarrhea,and provide evidence for the prevention and treatment.Methods The suspicious strains were isolated from acute diarrhea patients in Hangzhou area from January 2014 to December 2015 by using conventional methods.Multiplex PCR and monoplex PCR were applied to detect 12 virulence genes,and K-B method was used to carry out 18 commonly used drug susceptibility tests for all types of DEC.Results A total of 396 enteric pathogens were isolated from 1 019 specimens.DEC was found in 230cases(22.6%,230/1019)and accounted for 58.1% of all isolates,ranked 1st among the pathogenic bacteria.Among the 230 DECs,ETEC accounted for 56.5%,EAEC 30.4%,EPEC 8.7%,and STEC 0.4%.Two different pathotypes of DEC strains were isolated from 9specimens of patients with mixed infections.No EIEC was found.All types of DEC were isolated throughout the year,mainly from June to August.DEC infections occurred mainly in young adults of 18-45 years old.There was no significant difference in the detection rate between male(21.8%)and female(23.2%).Of the 207 isolates,49.3% were resistant to Ampicillin.The resistance rate to Cefazolin,Trimethoprim/Sulfamethoxazole and Ampicillin/Sulbactam were 46.4%,35.7% and 30.9%respectively,while those to the rest antibiotics were all less than 15.0%.All the isolates were sensitive to Imipenem.ESBLs-producing strains accounted for 41.1%and multidrug-resistant strains accounted for44.9%.Conclusion DEC,especially ESBLs-producing strains,multidrug-resistant strains and some pathotypes with multiple virulence genes,were the unneglectable pathogens in intestinal infections,which were the most common pathogens that cause diarrhea in Hangzhou area.Long-term monitoring on epidemic trend and drug resistance is of great significance for disease prevention and treatment.
引文
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[9]QU Mei,ZHANG Xin,QIAN Haikun,et al.Study on the genotype and epidemic characteristics of diarrheagenic Escherichia coli isolated from diarrheal patients in Beijing[J].Chin J Epidemiol,2014,35(10):1123-1126.(in Chinese)曲梅,张新,钱海坤,等.北京地区腹泻病患者致泻性大肠埃希菌感染类型及其流行特征[J].中华流行病学杂志,2014,35(10):1123-1126.
[10]Frank C,Werber D,Cramer JP,et al.Epidemic profile of Shiga-toxin-producing Escherichia coli O104:H4outbreak in Germany[J].NEJM,2011.365(19):1771-80.
[11]Trabulsi LR,Keller R,Tardelli Gomes TA.Typical and atypical enteropathogenic Escherichia coil[J].Emerg Infect Dis,2002,8(5):508-513.
[12]Hazen TH,Kaper JB,Nataro JP,et al.Comparative genomics provides insight into the diversity of the attaching and effacing Escherichia coli virulence plasmids[J].Infect Immun,2015,83(10):4103-4117.
[13]Grif K,Dierich MP,Karch H,et al.Strain-specific differences in the amount of Shiga toxin released from enterohemorrhagic Escherichia coli O157following exposure to subinhibitory concentrations of antimicrobial agents[J].Eur J Clin Microbiol Infect Dis,1998,17(11):761-766.
[14]ZHOU Fangmei,CHEN Xiao,YU Fei,et al.Characterization and antimicrobial resistance analysis of diarrheagenic Escherichia coli in Hangzhou area from 2011to 2013[J].Chin J Health Lab Tec,2016,26(11):1655-1657.(in Chinese)周芳美,陈晓,余斐,等.2011—2013年杭州地区致泻性大肠埃希菌的流行特征及耐药性分析[J].中国卫生检验杂志,2016,26(11):1655-1657.
[15]YU Miao,WANG Zhuo,FENG Li,et al.Surveillance and analysis of antimicrobial resistance of diagrrheagenic Escherichia coli in Shenyang during 2012-2014[J].Chin J Health Lab Tec,2015,25(24):4301-4303.(in Chinese)于淼,王卓,冯立,等.沈阳地区2012年-2014年致泻性大肠埃希菌耐药性监测分析[J].中国卫生检验杂志,2015,25(24):4301-4303.
[16]Randrianirina F,Ratsima EH,Ramparany L,et al.Antimicrobial resistance of bacterial enteropathogens isolated from stools in Madagascar[J].BMC Infect Dis,2014,14:104.
[17]Saviolli JY,Cunha MP,Guerra MF,et al.Free-Ranging Frigates(Fregaa magnificens)of the southeast coast of brazil harbor extraintestinal pathogenic Escherichia coli resistant to antimicrobials[J].PLoS One,2016,11(2):e0148624.
[2]Chen Y,Chen X,Zheng S,et al.Serotypes,genotypes and antimicrobial resistance patterns of human diarrhoeagenic Escherichia coli isolates circulating in southeastern China[J].Clin Microbiol Infect,2014,20(1):52-58.
[3]Muller D,Greune L,Heusipp G,et al.Identification of unconventional intestinal pathogenic Escherichia coli isolates expressing intermediate virulence factor profiles by using a novel single-step multiplex PCR[J].Appl Environ Microbiol,2007,73(10):3380-3390.
[4]Clinical and Laboratory Standards Institute(CLSI).Performance standards for antimicrobial susceptibility testing[S].Pennsylvania:Clinical and Laboratory Standards Institute,2016.
[5]Magiorakos AP,Srinivasan A,Carey RB,et al.Multidrug-resistant,extensively drug-resistant and pandrug-resistant bacteria:An international expert proposal for interim standard definitions for acquired resistance[J].Clin Microbiol Infect,2012,18(3):268-281.
[6]WANG Ruonan,LI Yi,CHEN Xiao,et al.Prevalence and antimicrobial susceptibility of Plesiomonas shigelloides in patients with acute diarrhea[J].Chin J Microecol,2015,27(12):1413-1416.(in Chinese)王若南,李医,陈晓,等,急性腹泻患者类志贺邻单胞菌流行及药物敏感性分析[J].中国微生态学杂志,2015,27(12):1413-1416.
[7]ZHAO Xuetao,GAO Kun,ZHANG Chunhua.The etiological survey and epidemiology research of diagrrheagenic Escherichia coli in Xuhui district from 2011to 2012[J].Lab Med,2013,28(8):671-675.(in Chinese)赵雪涛,高昆,张春华.2011至2012年徐汇区致泻性大肠埃希菌病原学监测与流行病学研究[J].检验医学,2013,28(8):671-675.
[8]LI Yinghui,QIU Yaqun,XIAN Huixia,et al.Epidemiologic and etiologic characteristics of diarrheagenic Escherichia coli infection in population in Shenzhen[J].Chin J Epidemiol,2016,37(1):115-118.(in Chinese)李迎慧,邱亚群,冼慧霞,等.深圳市腹泻人群致泻性大肠埃希菌流行及病原特征研究[J].中华流行病学杂志,2016,37(1):115-118.
[9]QU Mei,ZHANG Xin,QIAN Haikun,et al.Study on the genotype and epidemic characteristics of diarrheagenic Escherichia coli isolated from diarrheal patients in Beijing[J].Chin J Epidemiol,2014,35(10):1123-1126.(in Chinese)曲梅,张新,钱海坤,等.北京地区腹泻病患者致泻性大肠埃希菌感染类型及其流行特征[J].中华流行病学杂志,2014,35(10):1123-1126.
[10]Frank C,Werber D,Cramer JP,et al.Epidemic profile of Shiga-toxin-producing Escherichia coli O104:H4outbreak in Germany[J].NEJM,2011.365(19):1771-80.
[11]Trabulsi LR,Keller R,Tardelli Gomes TA.Typical and atypical enteropathogenic Escherichia coil[J].Emerg Infect Dis,2002,8(5):508-513.
[12]Hazen TH,Kaper JB,Nataro JP,et al.Comparative genomics provides insight into the diversity of the attaching and effacing Escherichia coli virulence plasmids[J].Infect Immun,2015,83(10):4103-4117.
[13]Grif K,Dierich MP,Karch H,et al.Strain-specific differences in the amount of Shiga toxin released from enterohemorrhagic Escherichia coli O157following exposure to subinhibitory concentrations of antimicrobial agents[J].Eur J Clin Microbiol Infect Dis,1998,17(11):761-766.
[14]ZHOU Fangmei,CHEN Xiao,YU Fei,et al.Characterization and antimicrobial resistance analysis of diarrheagenic Escherichia coli in Hangzhou area from 2011to 2013[J].Chin J Health Lab Tec,2016,26(11):1655-1657.(in Chinese)周芳美,陈晓,余斐,等.2011—2013年杭州地区致泻性大肠埃希菌的流行特征及耐药性分析[J].中国卫生检验杂志,2016,26(11):1655-1657.
[15]YU Miao,WANG Zhuo,FENG Li,et al.Surveillance and analysis of antimicrobial resistance of diagrrheagenic Escherichia coli in Shenyang during 2012-2014[J].Chin J Health Lab Tec,2015,25(24):4301-4303.(in Chinese)于淼,王卓,冯立,等.沈阳地区2012年-2014年致泻性大肠埃希菌耐药性监测分析[J].中国卫生检验杂志,2015,25(24):4301-4303.
[16]Randrianirina F,Ratsima EH,Ramparany L,et al.Antimicrobial resistance of bacterial enteropathogens isolated from stools in Madagascar[J].BMC Infect Dis,2014,14:104.
[17]Saviolli JY,Cunha MP,Guerra MF,et al.Free-Ranging Frigates(Fregaa magnificens)of the southeast coast of brazil harbor extraintestinal pathogenic Escherichia coli resistant to antimicrobials[J].PLoS One,2016,11(2):e0148624.