Molecular determinants of ß-lactamase producing Klebsiella pneumoniae in Mansoura University Neonatal Intensive Care Unit

E. Hammad, Hamdia Askar, M. Saleh, B. Shouman
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引用次数: 1

Abstract

Background: The emergence of s-lactamase- producing Klebsiella pneumonia (K. pneumoniae), represents a significant diagnostic and therapeutic challenge to the management of infections caused by this organism. This prospective study aimed at studying the frequency of s-lactamase production by K. pneumoniae in neonatal intensive care unit (NICU) of Mansoura University Children’s Hospital.Methods: This prospective study was conducted over a period of thirty six months from September 2010 to August 2013, where 684 samples were collected from different body sites of neonates in the NICU. Microbial isolation, identification and antimicrobial susceptibility testing were carried out.  s-lactamase production by K. pneumoniae isolates was tested by phenotypic methods and PCR amplification of related genes using a six-gene panel for the amplification of the blaCMY-2, blaDHA, blaACC, blaSHV, blaTEM and blaCTX-M genes. In vitro transformation and conjugation were carried out to test for plasmid mediated AmpC s- lactamase resistance transmission to E. coli. Results: K. pneumoniae was isolated at a percentage of 12.6% and s-lactamase production was detected in 62.8% of the isolates.  The most commonly detected s-lactamase gene was blaSHV (51.9%), followed by BlaCMY-2 (16.67), blaDHA (12.96%), blaTEM (9.2%), blaCTX- M(7.4%) and lastly blaACC (1.85%). Additionally, some strains carried combinations of two or three genes. The plasmid carrying blaCMY-2 was 100% successfully transformed into the competent E. coli LE392 while conjugation with the E. coli ATCC 25922 was (77.8%) successful.Conclusion: K. pneumoniae is a common multidrug resistant isolate; the production of s-lactamases is the mechanism of resistance in a significant number of cases and represents a real risk for failure of many therapeutic options. This problem is highly complicated by the horizontal spread of resistance plasmids among microbial population.
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曼苏拉大学新生儿重症监护室产ß-内酰胺酶肺炎克雷伯菌的分子决定因素
背景:产生s-内酰胺酶的肺炎克雷伯菌(肺炎克雷伯菌)的出现,对由该菌引起的感染的管理提出了重大的诊断和治疗挑战。本前瞻性研究旨在研究曼苏拉大学儿童医院新生儿重症监护病房(NICU)肺炎克雷伯菌s-内酰胺酶产生频率。方法:本前瞻性研究于2010年9月至2013年8月为期36个月,在NICU新生儿不同身体部位采集样本684份。进行微生物分离鉴定和药敏试验。利用blaCMY-2、blacha、blaACC、blaSHV、blaTEM和blaCTX-M基因6基因板扩增相关基因,采用表型法和PCR扩增方法检测肺炎克雷伯菌分离株s-内酰胺酶产量。采用体外转化和偶联法检测质粒介导的AmpC s-内酰胺酶耐药性在大肠杆菌中的传播。结果:肺炎克雷伯菌分离率为12.6%,s-内酰胺酶产率为62.8%。s-内酰胺酶基因检出率最高的是blaSHV(51.9%),其次是BlaCMY-2(16.67%)、blaDHA(12.96%)、blaTEM(9.2%)、blaCTX- M(7.4%)和blaACC(1.85%)。此外,一些菌株携带两个或三个基因的组合。携带blaCMY-2的质粒100%成功转化到大肠杆菌LE392,与大肠杆菌ATCC 25922的结合成功率为77.8%。结论:肺炎克雷伯菌是常见的多药耐药分离株;在许多病例中,s-内酰胺酶的产生是耐药性的机制,并代表着许多治疗方案失败的真正风险。由于耐药质粒在微生物群体中的水平传播,这一问题变得高度复杂。
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