大肠埃希菌质粒介导的bla_(NDM-1)基因耐药及传播性研究
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摘要
目的探讨某院携带bla_(NDM-1)基因大肠埃希菌的流行现况及传播机制。方法收集2016年1—12月南昌大学某附属医院对碳青霉烯类抗生素不敏感的大肠埃希菌92株,对其进行bla_(NDM-1)基因筛查,并对bla_(NDM-1)基因阳性的大肠埃希菌进行其他常见碳青霉烯酶基因检测。将bla_(NDM-1)基因阳性的大肠埃希菌(供体菌)与大肠埃希菌J53(受体菌)进行接合试验,采用药敏试验及mCIM试验对接合子进行表型验证,提取供体菌及接合子的质粒采用Southern blot技术对bla_(NDM-1)基因进行基因定位,并验证其水平转移的能力。结果在对碳青霉类抗生素不敏感的92株大肠埃希菌中发现2株携带bla_(NDM-1)基因,携带率2.2%。此2株大肠埃希菌未发现同时携带其他常见碳青霉烯酶基因。接合试验及Southern blot发现此2株大肠埃希菌的bla_(NDM-1)基因均位于菌株的质粒上,且有1株大肠埃希菌的bla_(NDM-1)基因可以通过质粒向大肠埃希菌J53转移。药敏试验结果显示,此2株大肠埃希菌对临床使用的多种抗菌药物耐药,且接合子获得了与供体菌相似的药敏结果。mCIM试验表明,此2株大肠埃希菌及接合子均产NDM-1型碳青霉烯酶。结论该院存在携带bla_(NDM-1)基因而产NDM-1型碳青霉烯酶的大肠埃希菌,携带bla_(NDM-1)基因的大肠埃希菌对临床使用的大多数抗菌药物耐药,质粒可介导bla_(NDM-1)基因的水平传播。
Objective To investigate the epidemic status and transmission mechanism of bla_(NDM-1) gene-carrying Escherichia coli(E.coli)in a hospital.Methods 92 strains of carbapenem-non-susceptible E.coli were collected from a hospital affiliated to Nanchang University between January and December 2016,bla_(NDM-1) gene was screened and other common carbapenemase genes in bla_(NDM-1) positive E.coli were also detected.bla_(NDM-1) positive E.coli(donor)and E.coli J53(receptor)was performed conjugation testing,phenotyping of conjugants was validated by antimicrobial susceptibility testing and modified carbapenem inactivation method(mCIM),plasmids of donor strain and conjugator were extracted,bla_(NDM-1) gene was located by Southern blot,and horizontally transfer ability of bla_(NDM-1) gene was validated.Results Among 92 strains of carbapenem-non-susceptible E.coli,2 strains were found to carry bla_(NDM-1) gene,carrying rate was 2.2%.Other common carbapenemase genes were also found in these two strains of E.coli.Conjugation testing and Southern blot found that bla_(NDM-1) gene of these two E.coli strains was located on the plasmid of E.coli strain,and bla_(NDM-1) gene of one E.coli strain could be transferred to E.coli J53 through plasmid.Antimicrobial susceptibility testing showed that these two E.coli strains were both resistant to multiple clinically used antimicrobial agents,conjugants obtained similar susceptibility results to the donor bacteria.mCIM showed that these two E.coli strains and their conjugants all produced NDM-1 carbapenemase.Conclusion E.coli carrying bla_(NDM-1) gene and producing NDM-1 carbapenemase is found in this hospital,E.coli carrying bla_(NDM-1) gene is resistant to most clinically used antimicrobial agents,plasmids can mediate the horizontal transfer of bla_(NDM-1) gene.
Objective To investigate the epidemic status and transmission mechanism of bla_(NDM-1) gene-carrying Escherichia coli(E.coli)in a hospital.Methods 92 strains of carbapenem-non-susceptible E.coli were collected from a hospital affiliated to Nanchang University between January and December 2016,bla_(NDM-1) gene was screened and other common carbapenemase genes in bla_(NDM-1) positive E.coli were also detected.bla_(NDM-1) positive E.coli(donor)and E.coli J53(receptor)was performed conjugation testing,phenotyping of conjugants was validated by antimicrobial susceptibility testing and modified carbapenem inactivation method(mCIM),plasmids of donor strain and conjugator were extracted,bla_(NDM-1) gene was located by Southern blot,and horizontally transfer ability of bla_(NDM-1) gene was validated.Results Among 92 strains of carbapenem-non-susceptible E.coli,2 strains were found to carry bla_(NDM-1) gene,carrying rate was 2.2%.Other common carbapenemase genes were also found in these two strains of E.coli.Conjugation testing and Southern blot found that bla_(NDM-1) gene of these two E.coli strains was located on the plasmid of E.coli strain,and bla_(NDM-1) gene of one E.coli strain could be transferred to E.coli J53 through plasmid.Antimicrobial susceptibility testing showed that these two E.coli strains were both resistant to multiple clinically used antimicrobial agents,conjugants obtained similar susceptibility results to the donor bacteria.mCIM showed that these two E.coli strains and their conjugants all produced NDM-1 carbapenemase.Conclusion E.coli carrying bla_(NDM-1) gene and producing NDM-1 carbapenemase is found in this hospital,E.coli carrying bla_(NDM-1) gene is resistant to most clinically used antimicrobial agents,plasmids can mediate the horizontal transfer of bla_(NDM-1) gene.
引文
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[11]宁长秀,汪红,钟桥石,等.耐碳青霉烯类抗菌药物肠杆菌科细菌耐药特性的分析[J].中国抗生素杂志,2013,38(10):788-790.
[12]Bocanegra-Ibarias P,Garza-González E,Morfin-Otero R,et al.Molecular and microbiological report of a hospital outbreak of NDM-1-carrying Enterobacteriaceae in Mexico[J].PLoSOne,2017,12(6):e0179651.
[13]Kocsis E,Gu6vinec M,Buti'c I,et al.blaNDM-1carriage on IncR plasmid in Enterobacteriaceae strains[J].Microb Drug Resist,2016,22(2):123-128.
[14]Pedersen T,Sekyere JO,Govinden U,et al.Spread of plasmid-encoded NDM-1 and GES-5 carbapenemases among extensively drug-resistant and pandrug-resistant clinical Enterobacteriaceae in Durban,South Africa[J].Antimicrob Agents Chemother,2018,62(5),pii:e02178-17.
[15]贾园春,袁敏,陈东科,等.临床来源碳青霉烯耐药大肠埃希菌药物敏感性及分子流行病学特征分析[J].疾病监测,2017,32(4):346-350.
[16]俞凤,邓林强,钟桥石,等.发现1株产NDM-1型碳青霉烯酶非脱羧勒克菌[J].临床检验杂志,2018,36(1):25-28.
[17]Doyle D,Peirano G,Lascols C,et al.Laboratory detection of Enterobacteriaceae that produce carbapenemases[J].J Clin Microbiol,2012,50(12):3877-3880.
[18]Butler-Wu SM,Abbott AN.Is this the carbapenemase test we’ve been waiting for?A multicenter evaluation of the modified carbapenem inactivation method[J].J Clin Microbiol,2017,55(8):2309-2312.
[2]Ozbek B,Mataraci-Kara E,Er S,et al.In vitro activities of colistin,tigecycline and tobramycin alone or in combination,against carbapenem-resistant Enterobacteriaceae strains[J].J Glob Antimicrob Resist,2015,3(4):278-282.
[3]王峰,孙景勇,张芳芳,等.碳青霉烯类耐药的大肠埃希菌中blaNDM基因型检测及流行病学分析[J].中国感染与化疗杂志,2017,17(1):56-60.
[4]邹大阳.NDM-1耐药菌在中国的流行及特性研究[D].北京:中国人民解放军军事医学科学院,2015.
[5]Wei DD,Wan L G,Yu Y,et al.Characterization of extendedspectrum beta-lactamase,carbapenemase,and plasmid quinolone determinants in Klebsiella pneumoniae isolates carrying distinct types of 16S rRNA methylase genes,and their association with mobile genetic elements[J].Microb Drug Resist,2015,21(2):186-193.
[6]Pierce VM,Simner PJ,Lonsway DR,et al.Modified carbapenem inactivation method for phenotypic detection of carbapenemase production among Enterobacteriaceae[J].J Clin Microbiol,2017,55(8):2321-2333.
[7]Jeon JH,Lee J H,Lee JJ,et al.Structural basis for carbapenem-hydrolyzing mechanisms of carbapenemases conferring antibiotic resistance[J].Int J Mol Sci,2015,16(5):9654-9692.
[8]Mathers A.Mobilization of carbapenemase-mediated resistance in Enterobacteriaceae[J].Microbiol Spectr,2016,4(3):1-10.
[9]Kumarasamy KK,Toleman MA,Walsh TR,et al.Emergence of a new antibiotic resistance mechanism in India,Pakistan,and the UK:a molecular,biological,and epidemiological study[J].Lancet Infect Dis,2010,10(9):597-602.
[10]Liu Z,Li W,Wang J,et al.Identification and characterization of the first Escherichia coli strain carrying NDM-1 gene in China[J].PLoS One,2013,8(6):e66666.
[11]宁长秀,汪红,钟桥石,等.耐碳青霉烯类抗菌药物肠杆菌科细菌耐药特性的分析[J].中国抗生素杂志,2013,38(10):788-790.
[12]Bocanegra-Ibarias P,Garza-González E,Morfin-Otero R,et al.Molecular and microbiological report of a hospital outbreak of NDM-1-carrying Enterobacteriaceae in Mexico[J].PLoSOne,2017,12(6):e0179651.
[13]Kocsis E,Gu6vinec M,Buti'c I,et al.blaNDM-1carriage on IncR plasmid in Enterobacteriaceae strains[J].Microb Drug Resist,2016,22(2):123-128.
[14]Pedersen T,Sekyere JO,Govinden U,et al.Spread of plasmid-encoded NDM-1 and GES-5 carbapenemases among extensively drug-resistant and pandrug-resistant clinical Enterobacteriaceae in Durban,South Africa[J].Antimicrob Agents Chemother,2018,62(5),pii:e02178-17.
[15]贾园春,袁敏,陈东科,等.临床来源碳青霉烯耐药大肠埃希菌药物敏感性及分子流行病学特征分析[J].疾病监测,2017,32(4):346-350.
[16]俞凤,邓林强,钟桥石,等.发现1株产NDM-1型碳青霉烯酶非脱羧勒克菌[J].临床检验杂志,2018,36(1):25-28.
[17]Doyle D,Peirano G,Lascols C,et al.Laboratory detection of Enterobacteriaceae that produce carbapenemases[J].J Clin Microbiol,2012,50(12):3877-3880.
[18]Butler-Wu SM,Abbott AN.Is this the carbapenemase test we’ve been waiting for?A multicenter evaluation of the modified carbapenem inactivation method[J].J Clin Microbiol,2017,55(8):2309-2312.