Barbarini D, Russello G, Brovarone F, Capatti C, Colla R, Perilli M, Moro M L, Carretto E
Clinical Virology and Microbiology Laboratory - Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
Clinical Microbiology Laboratory - IRCCS Arcispedale Santa Maria Nuova, Reggio Emilia, Italy.
Infect Genet Evol. 2015 Mar;30:8-14. doi: 10.1016/j.meegid.2014.11.025. Epub 2014 Nov 29.
The spread of carbapenem resistant Enterobacteriaceae (CRE) has recently become a matter of concern in public health, mainly due to the wide distribution of carbapenemase genes. Italy is a country considered endemic for the spread of blaKPC Klebsiella pneumoniae (KP). The aim of this study was to depict the epidemiological trend of CRE in one Italian hospital over a long period (3 years surveillance, from May 2011 to April 2014). Based on defined MIC cut-off for specific carbapenems, 164 strains isolated from 146 different patients were analyzed both phenotypically and genotypically to establish the resistance genes. Molecular typing was performed using the RAPD technique. 77 strains were demonstrated to harbor the blaKPC gene (73 KP, 4 Escherichia coli - EC), 51 strains the blaVIM gene (44 KP, 3 EC, 2 Enterobacter cloacae and 2 Klebsiella oxytoca), 8 the blaNDM gene (3 KP, 4 EC and one Providencia stuartii), 3 the blaOXA-48 gene (2 KP, 1 EC), whereas 25 out of the 164 isolates (of different genera and species) had a negative multiplex-PCR amplification for all the targets tested. 39 out of the 164 strains analyzed (23.8%) revealed discrepancies between the MICs obtained with automated instrument and gradient MICs of more than two logs of difference; the broth microdilution provided a better agreement with the results obtained with the gradient MIC. The use of RAPD allowed to distinguish different clusters, closely related, both for blaKPC and for blaVIM KP.
耐碳青霉烯类肠杆菌科细菌（CRE）的传播最近已成为公共卫生领域关注的问题，主要原因是碳青霉烯酶基因广泛分布。意大利是一个被认为blaKPC肺炎克雷伯菌（KP）传播为地方病的国家。本研究的目的是描述一家意大利医院在很长一段时间内（2011年5月至2014年4月的3年监测期）CRE的流行病学趋势。根据特定碳青霉烯类药物定义的最低抑菌浓度（MIC）临界值，对从146名不同患者中分离出的164株菌株进行了表型和基因型分析，以确定耐药基因。使用随机扩增多态性DNA（RAPD）技术进行分子分型。77株菌株被证明携带blaKPC基因（73株KP、4株大肠杆菌 - EC），51株携带blaVIM基因（44株KP、3株EC、2株阴沟肠杆菌和2株产酸克雷伯菌），8株携带blaNDM基因（3株KP、4株EC和1株斯氏普罗威登斯菌），3株携带blaOXA - 48基因（2株KP、1株EC），而164株分离株（不同属和种）中有25株对所有测试靶点的多重PCR扩增均为阴性。在分析的164株菌株中，有39株（23.8%）显示自动仪器获得的MIC与梯度MIC之间的差异超过两个对数；肉汤微量稀释法与梯度MIC获得的结果一致性更好。使用RAPD可以区分blaKPC和blaVIM KP的不同且密切相关的簇。
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