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[产超广谱β-内酰胺酶(ESBL)和AmpCβ-内酰胺酶的肺炎克雷伯菌、奇异变形杆菌和大肠埃希菌氟喹诺酮耐药临床分离株的特征]

[Characteristic of fluoroquinolone resistant clinical isolates of K. pneumoniae, P. mirabilis and E. coli producing ESBL and AmpC beta-lactamases].

作者信息

Rzeczkowska Magdalena, Piekarska Katarzyna, Gierczyński Rafał

机构信息

Zakład Bakteriologii Narodowego Instytutu Zdrowia Publicznego - Państwowego Zakładu Higieny w Warszawie.

出版信息

Med Dosw Mikrobiol. 2012;64(4):285-95.

Abstract

INTRODUCTION

Extended-spectrum beta-lactamases (ESBLs) are the predominant mechanism for acquired antibiotic resistance in Enterobacteriaceae. During the past few years, increasing occurrence of plasmid-mediated AmpC beta-lactamases (pAmpCs) particularly in K. pneumonia, P. mirabilis and E. coli was reported. Therefore, the aim of our study was to analyse of the diversity of plasmid-mediated beta-lactamases such as pAmpCs and ESBLs among clinical K. pneumonia, P. mirabilis and E. coli strains in Poland.

METHODS

A total of 19 clinical Enterobacteriaceae strains (E. coli, n = 9; K. pneumoniae, n = 7; P. mirabilis, n = 3) resistant to third-generation cephalosporin were selected from collection of fluoroquinolone resistant isolates recovered during a 6-months period in regular hospital in Warsaw, Poland. ESBLs and AmpCs were detected by using phenotypic methods: double-disc tests (DDSTs), MAST ID D68C test, sensitivity to cefoxitin, disk potentiation test (DPT) and Tris-EDTA test. Polymerase chain reaction (PCR) amplification of the bla(AmpC), bla(CTM-M), bla(TEM), and bla(SHV), genes. PCR-products for these genes were sequenced. To determine the possible clonality of the tested isolates PFGE with the XbaI was performed.

RESULTS

Nine of 19 fluoroquinolone-resistant strains tested produced extended-spectrum beta-lactamases of TEM, SHV and CTX-M families. These ESBLs were most commonly detected in E. coli. AmpC beta-lactamases were produced by 6 tested strains, including 3 isolates of P. mirabilis. The AmpC found in our study belonged to CMY and DHA families, Furthermore, 4 isolates of K. pneumoniae were found to co-produce both ESBL and AmpC beta-lactamases. XbaI-PFGE profiles pointed significant differences of tested strains.

CONCLUSION

Horizontal transfer of genes encoding for acquired beta-lactamases such ESBL and AmpC seem to play primary role in dissemination of these resistance traits among fluoroquinolone-resistant clinical strains of Enterobacteriaceae in Poland.

摘要

引言

超广谱β-内酰胺酶(ESBLs)是肠杆菌科细菌获得性抗生素耐药的主要机制。在过去几年中,有报道称质粒介导的AmpCβ-内酰胺酶(pAmpCs)的出现频率不断增加,尤其是在肺炎克雷伯菌、奇异变形杆菌和大肠杆菌中。因此,我们研究的目的是分析波兰临床分离的肺炎克雷伯菌、奇异变形杆菌和大肠杆菌菌株中质粒介导的β-内酰胺酶如pAmpCs和ESBLs的多样性。

方法

从波兰华沙一家普通医院6个月期间收集的耐氟喹诺酮类菌株中,选取19株对第三代头孢菌素耐药的临床肠杆菌科菌株(大肠杆菌9株、肺炎克雷伯菌7株、奇异变形杆菌3株)。采用表型方法检测ESBLs和AmpCs:双纸片协同试验(DDSTs)、MAST ID D68C试验、对头孢西丁的敏感性、纸片增效试验(DPT)和Tris-EDTA试验。对bla(AmpC)、bla(CTM-M)、bla(TEM)和bla(SHV)基因进行聚合酶链反应(PCR)扩增。对这些基因的PCR产物进行测序。为了确定受试菌株可能的克隆性,进行了XbaI酶切脉冲场凝胶电泳(PFGE)分析。

结果

19株耐氟喹诺酮类菌株中有9株产生了TEM、SHV和CTX-M家族的超广谱β-内酰胺酶。这些ESBLs最常见于大肠杆菌中。6株受试菌株产生了AmpCβ-内酰胺酶,其中包括3株奇异变形杆菌分离株。我们研究中发现的AmpC属于CMY和DHA家族。此外,发现4株肺炎克雷伯菌分离株同时产生ESBL和AmpCβ-内酰胺酶。XbaI-PFGE图谱显示受试菌株存在显著差异。

结论

编码获得性β-内酰胺酶如ESBL和AmpC的基因水平转移似乎在波兰耐氟喹诺酮类临床肠杆菌科菌株中这些耐药性状的传播中起主要作用。

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