Evaluation of a modified meropenem hydrolysis assay on a large cohort of KPC and VIM carbapenemase-producing Enterobacteriaceae.
Evaluation of a modified meropenem hydrolysis assay on a large cohort of KPC and VIM carbapenemase-producing Enterobacteriaceae.
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DOI：
10.1371/journal.pone.0174908
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
De Conto F
中科院分区：
文献类型：
--
作者：
Calderaro A;Buttrini M;Piergianni M;Montecchini S;Martinelli M;Covan S;Piccolo G;Medici MC;Arcangeletti MC;Chezzi C;De Conto F
耐碳青霉烯类肠杆菌科（CRE）已在全球范围内蔓延，并对公共卫生构成严重且日益严重的威胁。采用快速、灵敏的方法检测产碳青霉烯酶细菌越来越重要。碳青霉烯酶的产生可以使用非分子方法（如改良的Hodge试验、协同试验、Carba NP试验和抗生素水解试验）和基于DNA的方法进行检测。在这项研究中，我们提出了一个修改后的版本，以前描述的美罗培南水解试验（MHA）的MALDI-TOF MS的表型检测在2小时的碳青霉烯酶生产肠杆菌。MHA成功应用于检测981株产KPC或Vim碳青霉烯酶的肠杆菌科菌株和146株碳青霉烯完全敏感菌株的碳青霉烯酶活性。该检测方法也适用于NDM和OXA-48产生菌株以及具有碳青霉烯酶产生以外耐药机制的CRE，已证明能够区分产碳青霉烯酶和不产碳青霉烯酶的肠杆菌科。如前所述和我们手中观察到的，通过MALDI-TOF MS分析的MHA与所涉及的碳青霉烯酶类型无关，它比基于培养的方法更快，更容易执行/解释。另一方面，它不能检测其他碳青霉烯耐药机制，如孔蛋白改变和外排机制。
Carbapenem-resistant Enterobacteriaceae (CRE) have spread globally and represent a serious and growing threat to public health. The introduction of rapid and sensitive methods for the detection of carbapenemase-producing bacteria is of increasing importance. The carbapenemase production can be detected using non-molecular methods (such as the modified Hodge test, the synergy test, the Carba NP test and the antibiotic hydrolysis assays) and DNA-based methods. In this study, we propose a modified version of a previously described meropenem hydrolysis assay (MHA) by MALDI-TOF MS for the phenotypic detection in 2h of carbapenemase-producing Enterobacteriaceae. The MHA was successfully applied to detect carbapenemase activity in 981 well-characterized Enterobacteriaceae strains producing KPC or VIM carbapenemases, and in 146 carbapenem fully susceptible strains. This assay, applied also to NDM and OXA-48-producing strains and to CRE with resistance mechanisms other than carbapenemase production, has proved to be able to distinguish between carbapenemase-producing and -nonproducing Enterobacteriaceae. As already stated and as observed in our hands, MHA by MALDI-TOF MS analysis is independent from the type of carbapenemases involved, it is faster and easier to perform/interpret than culture-based methods. On the other hand, it cannot detect other carbapenem resistance mechanisms, such as porin alterations and efflux mechanisms.