Hashem Hany, Hanora Amro, Abdalla Salah, Shaeky Alaa, Saad Alaa
Department of Microbiology and Immunology, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.
Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt.
APMIS. 2017 May;125(5):499-505. doi: 10.1111/apm.12669. Epub 2017 Mar 15.
This study was designed to investigate the prevalence of metallo-β-lactamase (MBL) in Pseudomonas aeruginosa isolates collected from Suez Canal University Hospital in Ismailia, Egypt. Antibiotic susceptibility testing and phenotypic and genotypic screening for MBLs were performed on 147 isolates of P. aeruginosa. MICs were determined by agar dilution method for carbapenem that was ≥2 μg/mL for meropenem. MBL genes were detected by multiplex and monoplex PCR for P. aeruginosa-harbored plasmids. Mutation profile of sequenced MBL genes was screened using online software Clustal Omega. Out of 147 P. aeruginosa, 39 (26.5%) were carbapenem-resistant isolates and 25 (64%) were confirmed to be positive for MBLs. The susceptibility rate of P. aeruginosa toward polymyxin B and norfloxacin was 99% and 88%, respectively. Identification of collected isolates by API analysis and constructed phylogenetic tree of 16S rRNA showed that the isolates were related to P. aeruginosa species. The frequency of blaGIM-1, blaSIM-1, and blaSPM-1 was 52%, 48%, and 24%, respectively. BlaVIM and blaIMP-like genes were 20% and 4% and the sequences confirm the isolate to be blaVIM-1, blaVIM-2, blaVIM-4, and blaIMP-1. Three mutations were identified in blaVIM-4 gene. Our study emphasizes the high occurrence of multidrug-resistant P. aeruginosa-producing MBL enzymes.
本研究旨在调查从埃及伊斯梅利亚苏伊士运河大学医院收集的铜绿假单胞菌分离株中金属β-内酰胺酶(MBL)的流行情况。对147株铜绿假单胞菌进行了抗生素敏感性测试以及MBL的表型和基因型筛选。通过琼脂稀释法测定碳青霉烯类抗生素的最低抑菌浓度(MIC),美罗培南的MIC≥2μg/mL。通过多重PCR和单重PCR检测铜绿假单胞菌携带的质粒中的MBL基因。使用在线软件Clustal Omega筛选测序的MBL基因的突变谱。在147株铜绿假单胞菌中,39株(26.5%)为耐碳青霉烯类分离株,25株(64%)被确认为MBL阳性。铜绿假单胞菌对多粘菌素B和诺氟沙星的敏感率分别为99%和88%。通过API分析鉴定收集的分离株,并构建16S rRNA系统发育树,结果表明这些分离株与铜绿假单胞菌属相关。blaGIM-1、blaSIM-1和blaSPM-1的频率分别为52%、48%和24%。BlaVIM和blaIMP样基因分别为20%和4%,序列证实分离株为blaVIM-1、blaVIM-2、blaVIM-4和blaIMP-1。在blaVIM-4基因中鉴定出三个突变。我们的研究强调了产生MBL酶的多重耐药铜绿假单胞菌的高发生率。