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埃及开罗肺炎克雷伯菌临床分离株中OmpK35、OmpK36孔蛋白及β-内酰胺酶产生对亚胺培南敏感性的作用

The Role of OmpK35, OmpK36 Porins, and Production of β-Lactamases on Imipenem Susceptibility in Klebsiella pneumoniae Clinical Isolates, Cairo, Egypt.

作者信息

Wassef Mona, Abdelhaleim Mona, AbdulRahman Eiman, Ghaith Doaa

机构信息

Department of Clinical and Chemical Pathology, Faculty of Medicine, Cairo University , Cairo, Egypt .

出版信息

Microb Drug Resist. 2015 Dec;21(6):577-80. doi: 10.1089/mdr.2014.0226. Epub 2015 May 6.

DOI:10.1089/mdr.2014.0226
PMID:25945596
Abstract

BACKGROUND

OmpK35 and OmpK36 are the major outer membrane porins of Klebsiella pneumoniae. We aimed to study the effect of combined porin loss and production of extended-spectrum β-lactamases (ESBLs) on imipenem susceptibility among K. pneumoniae clinical isolates.

MATERIALS AND METHODS

This study included 91 suspected ESBL-producing K. pneumoniae clinical isolates, isolated from different patient specimens at the Cairo University hospital from January to June 2010. All isolates were subjected to genotypic analysis of the outer membrane protein gene expression using reverse transcription-PCR (RT-PCR) and analysis of OmpK35/36 of 38 isolates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

RESULTS

By RT-PCR, loss of Omp35 was detected in 78 (85.7%) isolates, loss of Omp36 was detected in 64 (70.32%), and loss of both porins was detected in 62 (68.1%). Out of 91 isolates, 45 (49.5%) were resistant to cefoxitin, and 17 (18.7%) were confirmed as derepressed AmpC producers. Omp35 was lost in all FOX-resistant isolates, whereas Omp36 was lost in 42 (93.3%) (p-value 0.002). The mean of ceftazidime inhibition zone diameter was significantly decreased among ESBL-producing isolates with loss of Omp35/36 (p-value 0.041 and 0.006), respectively. The mean of imipenem minimal inhibitory concentration (MIC) was markedly increased to 8.55 μg/ml among AmpC-producing isolates with Omp35/36 loss, while the mean of imipenem MIC among the 66 confirmed ESBL producers was 0.32 μg/ml.

CONCLUSION

Imipenem MIC was markedly increased among K. pneumoniae isolates showing AmpC production with loss of both porins OmpK35/36. Meanwhile, the association of porin OmpK35/36 loss with ESBL production was not a direct cause of resistance to imipenem.

摘要

背景

外膜孔蛋白OmpK35和OmpK36是肺炎克雷伯菌的主要外膜孔蛋白。我们旨在研究孔蛋白缺失与超广谱β-内酰胺酶(ESBLs)产生共同作用对肺炎克雷伯菌临床分离株亚胺培南敏感性的影响。

材料与方法

本研究纳入了91株疑似产ESBL的肺炎克雷伯菌临床分离株,于2010年1月至6月从开罗大学医院不同患者标本中分离获得。所有分离株均采用逆转录聚合酶链反应(RT-PCR)对外膜蛋白基因表达进行基因分型分析,并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)对38株分离株的OmpK35/36进行分析。

结果

通过RT-PCR检测到,78株(85.7%)分离株存在Omp35缺失,64株(70.32%)存在Omp36缺失,62株(68.1%)两种孔蛋白均缺失。在91株分离株中,45株(49.5%)对头孢西丁耐药,17株(18.7%)被确认为去阻遏AmpC产生菌。所有对FOX耐药的分离株均存在Omp35缺失,而42株(93.3%)存在Omp36缺失(p值为0.002)。在Omp35/36缺失的产ESBL分离株中,头孢他啶抑菌圈直径均值显著降低(p值分别为0.041和0.006)。在Omp35/36缺失的产AmpC分离株中,亚胺培南最低抑菌浓度(MIC)均值显著升高至8.55μg/ml,而在66株确诊的产ESBL分离株中,亚胺培南MIC均值为0.32μg/ml。

结论

在同时产AmpC且OmpK35/36两种孔蛋白均缺失的肺炎克雷伯菌分离株中,亚胺培南MIC显著升高。同时,孔蛋白OmpK35/36缺失与ESBL产生之间的关联并非对亚胺培南耐药的直接原因。

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