血流感染大肠埃希菌的耐药性分析
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摘要
目的对血流感染中分离的大肠埃希菌进行筛查并检测相关的耐药基因,为合理用药提供实验室依据。方法选取2015年6月至2016年5月温州医科大学附属第一医院门诊及住院患者血液中首次分离的216株大肠埃希菌,采用VITEK-2全自动微生物分析仪进行药敏试验;采用双纸片扩散法进行ESBLs表型检测;采用PCR方法筛选ESBLs基因,并测序确定其基因型。结果 216株大肠埃希菌对氨基糖苷类(除庆大霉素外)、头霉素类、呋喃妥因和碳青霉烯类等药物具有较好的敏感性,但对青霉素类、喹诺酮类和磺胺类药物耐药率较高;检出产ESBLs菌株56.94%(123/216);在130株第三代头孢菌素耐药株中,22株bla_(TEM)基因检测阳性,59株bla_(CTX-M)基因检测阳性,经测序比对显示:29株携带bla_(CTX-M-14)基因,23株携带bla_(CTX-M-15)基因。结论引起血流感染的大肠埃希菌多为对第三代头孢菌素耐药的产ESBLs菌株,携带的耐药基因主要为bla_(CTX-M)和bla_(TEM)。
Objective To analyze the drug resistance of Escherichia coli(E.coli) isolated from patients with bloodstream infections, and to provide evidence for the use of antibiotics. Methods A total of 216 E.coli strains isolated from patients with BSI in the First Affiliated Hospital of Wenzhou Medical University from June 2015 to May 2016 were selected.The identification and antimicrobial susceptibility testing of E.coli isolates were determined by VITEK-2 full automated microbiology analyzer.The double disk diffusion test was used for the determination of extended-spectrum β-lactamases(ESBLs) phenotype of E.coli isolates.ESBLs genes were screened by PCR and were sequenced for genotype. Results 216 E.coli isolates were all sensitive to aminoglycosides(except gentamicin), cephamycins, nitrofurantion and carbapenems, but were highly resistant to penicillins, quinolones and sulfonamides.123 of 216 E.coli isolates(56.94%) were found to produce ESBLs.Out of 130 strains resistant to the third-generation cephalosporins, 22 and 59 were positive for bla_(TEM) and bla_(CTX-M) determined by PCR, while 29 and 23 strains harbored bla_(CTX-M-14) and bla_(CTX-M-15) genes, respectively. Conclusions Majority of E.coli strains isolated from patients with BSI are ESBLs positive and highly resistant to the third-generation cephalosporins.bla_(CTX-M-14) and bla_(CTX-M-15) genes are approved to be closely related to the drug resistance of E.coli isolates.
Objective To analyze the drug resistance of Escherichia coli(E.coli) isolated from patients with bloodstream infections, and to provide evidence for the use of antibiotics. Methods A total of 216 E.coli strains isolated from patients with BSI in the First Affiliated Hospital of Wenzhou Medical University from June 2015 to May 2016 were selected.The identification and antimicrobial susceptibility testing of E.coli isolates were determined by VITEK-2 full automated microbiology analyzer.The double disk diffusion test was used for the determination of extended-spectrum β-lactamases(ESBLs) phenotype of E.coli isolates.ESBLs genes were screened by PCR and were sequenced for genotype. Results 216 E.coli isolates were all sensitive to aminoglycosides(except gentamicin), cephamycins, nitrofurantion and carbapenems, but were highly resistant to penicillins, quinolones and sulfonamides.123 of 216 E.coli isolates(56.94%) were found to produce ESBLs.Out of 130 strains resistant to the third-generation cephalosporins, 22 and 59 were positive for bla_(TEM) and bla_(CTX-M) determined by PCR, while 29 and 23 strains harbored bla_(CTX-M-14) and bla_(CTX-M-15) genes, respectively. Conclusions Majority of E.coli strains isolated from patients with BSI are ESBLs positive and highly resistant to the third-generation cephalosporins.bla_(CTX-M-14) and bla_(CTX-M-15) genes are approved to be closely related to the drug resistance of E.coli isolates.
引文
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[16] 王莉洁,梁志欣,喻航,等.大肠埃希菌与肺炎克雷伯菌血流感染的临床特征及耐药性比较[J].解放军医学院学报,2017,38(4):306-308,316.
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[20] Shin J,Kim DH,Ko KS.Comparison of CTX-M-14-and CTX-M-15 producing Escherichia coli and Klebsiella pneumoniae isolates from patients with bacteremia[J].J Infect,2011,63(1):39-47.
[2] Shi H,Sun F,Chen J,et al.Epidemiology of CTX-M-type extended-spectrum beta-lactamase (ESBL)-producing nosocomial-Escherichia coli infection in China[J].Ann Clin Microbiol Antimicrob,2015,14:4.
[3] Hayakawa K,Gattu S,Marchaim D,et al.Epidemiology and risk factors for isolation of Escherichia coli producing CTX-M-type extended-spectrum β-lactamase in a large U.S.Medical Center[J].Antimicrob Agents Chemother,2013,57(8):4010-4018.
[4] Shams S,Hashemi A,Esmkhani M,et al.Imipenem resistance in clinical Escherichia coli from Qom,Iran[J].BMC Res Notes,2018,11(1):314.
[5] Mokady D,Gophna U,Ron EZ.Virulence factors of septicemic Escherichia coli strains[J].Int J Med Microbiol,2005,295(6-7):455-462.
[6] Concei??o RA,Ludovico MS,Andrade CG,et al.Human sepsis-associated Escherichia coli (SEPEC) is able to adhere to and invade kidney epithelial cells in culture[J].Braz J Med Biol,Res,2012,45(5):417-424.
[7] 王小龙,栾旭波.某院血流感染大肠埃希菌耐药表型及耐药率研究[J].检验医学与临床,2018,15(4):451-453.
[8] Ananias M,Yano T.Serogroups and virulence genotypes of Escherichia coli isolated from patients with sepsis[J].Braz J Med Biol Res,2008,41(10):877-883.
[9] 冯琳涵,徐修礼,刘家云,等.63229份血培养标本中病原菌群分布及耐药性分析[J].国际检验医学杂志,2015,36(19):2800-2802.
[10] Laupland KB,Church DL.Population-based epidemiology and microbiology of community-onset bloodstream infections[J].Clin Microbiol Rev,2014,27(4):647-664.
[11] 郭小兵,饶玉婷,贺小红,等.1061株血标本分离菌的分布及耐药性[J].中国感染控制杂志,2018,17(4):304-309.
[12] 吕媛,李耘,薛峰,等.卫生部全国细菌耐药监测网(Mohnarin)2011-2012年度血流感染细菌耐药监测报告[J].中国临床药理学杂志,2014,30(3):278-288.
[13] 艾根伟.血流感染大肠埃希菌的耐药性及耐药性变迁[J].中国现代医药杂志,2016,18(5):98-99.
[14] 田俊华,陈蕾.血流感染非产超广谱β-内酰胺酶大肠埃希菌和产超广谱β-内酰胺酶大肠埃希菌的耐药性分析[J].中国卫生检验杂志,2017,27(20):3023-3026.
[15] Li XZ,Plésiat P,Nikaido H.The challenge of efflux-mediated antibiotic resistance in Gram-negative bacteria[J].Clin Microbiol Rev,2015,28(2):337-418.
[16] 王莉洁,梁志欣,喻航,等.大肠埃希菌与肺炎克雷伯菌血流感染的临床特征及耐药性比较[J].解放军医学院学报,2017,38(4):306-308,316.
[17] Livermore DM,Canton R,Gniadkowski M,et al.CTX-M:changing the face of ESBLs in Europe[J].J Antimicrob Chemother,2007,59(2):165-174.
[18] Pitout JD,Church DL,Gregson DB,et al.Molecular epidemiology of CTX-M-producing escherichia coli in the calgary health region:emergence of CTX-M-15-producing Isolates[J].Antimicrob Agents Chemother,2007,51(4):1281-1286.
[19] Brolund A,Edquist PJ,M?kitalo B,et al.Epidemiology of extended-spectrum β-lactamase-producing Escherichia coli in Sweden 2007-2011[J].Clin Microbiol Infect,2014,20(6):344-352.
[20] Shin J,Kim DH,Ko KS.Comparison of CTX-M-14-and CTX-M-15 producing Escherichia coli and Klebsiella pneumoniae isolates from patients with bacteremia[J].J Infect,2011,63(1):39-47.