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利比亚某社区中产超广谱β-内酰胺酶和产AmpC酶大肠埃希菌的粪便携带情况

Fecal carriage of extended-spectrum β-lactamases and AmpC-producing Escherichia coli in a Libyan community.

作者信息

Ahmed Salwa Fouad, Ali Mostafa Mohamed M, Mohamed Zienat Kamel, Moussa Tarek A, Klena John D

机构信息

United States Naval Medícal Research Unit No,3, Cairo, Egypt.

出版信息

Ann Clin Microbiol Antimicrob. 2014 Jun 16;13:22. doi: 10.1186/1476-0711-13-22.

Abstract

BACKGROUND

Extended-spectrum β-lactamases (ESBLs), including the AmpC type, are important mechanisms of resistance among Enterobacteriaeceae. CTX-M type extended-spectrum β- lactamases, of which there are now over 90 variants, are distributed globally, yet appear to vary in regional distribution. AmpC β-lactamases hydrolyze third generation cephalosporins, but are resistant to inhibition by clavulanate or other β-lactamase inhibitors in vitro. Fecal carriage and rates of colonization by bacteria harboring these resistance mechanisms have been reported in patients with community-acquired infections and in healthy members of their households. Expression of these ESBLs compromises the efficacy of current antibacterial therapies, potentially increasing the seriousness of hospital- and community-acquired Escherichia coli (E. coli) infections.To investigate the occurrence of ESBL-producing E. coli in human fecal flora isolated from two pediatric populations residing in the Libyan cities Zleiten and Abou El Khoms. Isolates were further studied to characterize genes encoding β-lactam resistance, and establish genetic relationships.

METHODS

Antibiotic resistance profiles of phenotypically characterized E. coli isolates recovered from the stools of 243 Libyan children during two surveillance periods in 2001 and 2007 were determined by the disk diffusion method. ESBL-screening was performed using the cephalosporin/clavulanate double synergy disc method, and the AmpC-phenotype was confirmed by the aminophenyl-boronic acid test. ESBL genes were molecularly characterized. Phylogenetic group and multilocus sequence typing (MLST) were determined for ESBL-producing isolates and PFGE was performed to compare banding profiles of some dominant strains.

RESULTS

ESBLs were identified in 13.4% (18/134) of E. coli isolates, and nine isolates (6.7%) demonstrated AmpC activity; all 18 isolates contained a CTX-M gene. Three CTX-M gene families (CTX-M-1, n=9; CTX-M-15, n=8 and CTX-M-3, n=1) were distributed in diverse E. coli backgrounds (phylogenetic group D, 39%; B2, 28%; B1, 22% and A, 11%). MLST analysis revealed 14 sequence type (ST) with six new sequence types. The gene encoding the CMY-2 enzyme was detected in five AmpC-positive E. coli.

CONCLUSIONS

These results identified heterogeneous clones of CTX-M-producing E. coli in the fecal isolates, indicating that the intestinal tract acts as a reservoir for ESBL-producing organisms, and a trafficker of antibiotic resistance genes.

摘要

背景

超广谱β-内酰胺酶(ESBLs),包括AmpC型,是肠杆菌科细菌重要的耐药机制。CTX-M型超广谱β-内酰胺酶目前有90多种变体,在全球范围内均有分布,但区域分布似乎有所不同。AmpCβ-内酰胺酶可水解第三代头孢菌素,但在体外对克拉维酸或其他β-内酰胺酶抑制剂的抑制作用具有抗性。据报道,在社区获得性感染患者及其家庭健康成员中,携带这些耐药机制的细菌存在粪便携带和定植率。这些ESBLs的表达会影响当前抗菌治疗的效果,可能会增加医院获得性和社区获得性大肠杆菌(E. coli)感染的严重性。为了调查从利比亚兹利坦和阿布胡姆斯市的两个儿科人群中分离出的人类粪便菌群中产ESBLs大肠杆菌的发生情况。对分离株进行进一步研究,以表征编码β-内酰胺抗性的基因,并建立遗传关系。

方法

采用纸片扩散法测定了2001年和2007年两个监测期从243名利比亚儿童粪便中分离出的表型特征明确的大肠杆菌分离株的抗生素耐药谱。采用头孢菌素/克拉维酸双协同纸片法进行ESBL筛查,通过氨基苯基硼酸试验确认AmpC表型。对ESBL基因进行分子表征。对产ESBLs分离株进行系统发育群和多位点序列分型(MLST),并进行脉冲场凝胶电泳(PFGE)以比较一些优势菌株的条带图谱。

结果

在13.4%(18/134)的大肠杆菌分离株中鉴定出ESBLs,9株(6.7%)表现出AmpC活性;所有18株分离株均含有CTX-M基因。三个CTX-M基因家族(CTX-M-1,n = 9;CTX-M-15,n = 8和CTX-M-3,n = 1)分布在不同的大肠杆菌背景中(系统发育群D,39%;B2,28%;B1,22%和A,11%)。MLST分析揭示了14种序列类型(ST),其中有6种新的序列类型。在5株AmpC阳性大肠杆菌中检测到编码CMY-2酶的基因。

结论

这些结果在粪便分离株中鉴定出了产CTX-M大肠杆菌的异质克隆,表明肠道是产ESBLs生物体的储存库,也是抗生素耐药基因的传播者。

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