Laudy Agnieszka E, Róg Patrycja, Smolińska-Król Katarzyna, Ćmiel Milena, Słoczyńska Alicja, Patzer Jan, Dzierżanowska Danuta, Wolinowska Renata, Starościak Bohdan, Tyski Stefan
Department of Pharmaceutical Microbiology, Medical University of Warsaw, Warsaw, Poland.
Department of Clinical Microbiology and Immunology, The Children's Memorial Health Institute, Warsaw, Poland.
PLoS One. 2017 Jun 28;12(6):e0180121. doi: 10.1371/journal.pone.0180121. eCollection 2017.
Knowledge of the prevalence of ESBL enzymes among P. aeruginosa strains compared to the Enterobacteraiceae family is limited. The phenotypic tests recommended by EUCAST for the detection of ESBL-producing Enterobacteriaceae are not always suited for P. aeruginosa strains. This is mainly due to the presence of other families of ESBLs in P. aeruginosa isolates more often than in Enterobacteriaceae, production of natural AmpC cephalosporinase and its overexpression, and co-production of metallo-β-lactamases. The aim of this study was to determine the occurrence of ESBLs in P. aeruginosa isolated from patients from hospitals in Warsaw, to evaluate the ESBL production of these isolates using currently available phenotypic tests, their modifications, multiplex PCR and molecular typing of ESBL-positive isolates by PFGE. Clinical isolates of P. aeruginosa were collected in 2000-2014 from four Warsaw hospitals. Based on the data obtained in this study, we suggest using three DDST methods with inhibitors, such as clavulanic acid, sulbactam and imipenem, to detect ESBL-producing P. aeruginosa strains. Depending on the appearance of the plates, we suggest a reduction in the distance between discs with antibiotics to 15 mm and the addition of boronic acid at 0.4 mg per disc. The analysed isolates carried genes encoding ESBL from the families VEB (69 isolates with VEB-9), GES (6 with GES-1, 1 GES-5, 5 GES-13 and 2 with GES-15), OXA-2 (12 with OXA-15, 1 OXA-141, 1 OXA-210, 1 OXA-543 and 1 with OXA-544) and OXA-10 (5 isolates with OXA-74 and one with OXA-142). The most important result of this study was the discovery of three new genes, blaGES-15, blaOXA-141 and blaOXA-142; their nucleotide sequences have been submitted to the NCBI GenBank. It is also very important to note that this is the first report on the epidemiological problem of VEB-9-producing bacterial strains, not only in Poland but also worldwide.
与肠杆菌科相比，铜绿假单胞菌菌株中ESBL酶的流行情况相关知识有限。欧盟CAST推荐的用于检测产ESBL肠杆菌科细菌的表型试验并不总是适用于铜绿假单胞菌菌株。这主要是因为铜绿假单胞菌分离株中ESBL的其他家族比肠杆菌科中更常见，存在天然AmpC头孢菌素酶及其过表达，以及金属β-内酰胺酶的共同产生。本研究的目的是确定从华沙医院患者中分离出的铜绿假单胞菌中ESBL的发生情况，使用目前可用的表型试验、其改良方法、多重PCR评估这些分离株的ESBL产生情况，并通过PFGE对ESBL阳性分离株进行分子分型。2000年至2014年期间，从华沙的四家医院收集了铜绿假单胞菌的临床分离株。基于本研究获得的数据，我们建议使用三种带有抑制剂（如克拉维酸、舒巴坦和亚胺培南）的双纸片协同试验方法来检测产ESBL的铜绿假单胞菌菌株。根据平板的外观，我们建议将含抗生素纸片之间的距离缩短至15毫米，并在每张纸片中添加0.4毫克硼酸。分析的分离株携带了来自VEB家族（69株携带VEB-9）、GES家族（6株携带GES-1、1株携带GES-5、5株携带GES-13和2株携带GES-15）、OXA-2家族（12株携带OXA-15、1株携带OXA-141、1株携带OXA-210、1株携带OXA-543和1株携带OXA-544）和OXA-10家族（5株携带OXA-74和1株携带OXA-142）的编码ESBL的基因。本研究最重要的结果是发现了三个新基因，blaGES-15、blaOXA-141和blaOXA-142；它们的核苷酸序列已提交至NCBI基因库。同样非常重要的是要注意，这不仅是波兰，也是全球范围内关于产VEB-9细菌菌株流行病学问题的首份报告。
