Centre for Modern Interdisciplinary Technologies, Nicolaus Copernicus University, Toruń, Poland.
Bruker Daltonik GmbH, Bremen, Germany.
PLoS One. 2021 Mar 18;16(3):e0247369. doi: 10.1371/journal.pone.0247369. eCollection 2021.
The development of new techniques for the detection of carbapenemase activity is of great importance since the increased incident of resistance against carbapenems represents a serious threat to global public health. In this context, the matrix-assisted laser desorption/ionization approach already demonstrated to be a reliable tool for rapid carbapenemase detection. As a newly developed test, there is still a lack of in-depth analysis of its robustness and possible wider application. The main goal of this study was to evaluate the potential for using the design MBT STAR-Carba assay as the pre-characterization method for Enterobacterales and P. aeruginosa strains in terms of the produced classes of carbapenemases using modified procedure parameters-various suspension densities and incubation times. Moreover, its usefulness for the in-depth analysis and characterization of metallo-β-lactamases (MBL) was tested by applying inhibition assays. In this study, the designed assay proved to be a sensitive tool for the detection of carbapenemase hydrolytic activity, which can be successfully used to partially classify the class of carbapenemase present. Additionally, the use of defined high concentration suspensions would allow to shorten the incubation time to 1 minute for certain strains. Considering that the assay was also suitable to investigate the effect of different inhibitors on the MBL activity, it demonstrates far higher discriminatory potential than only a rapid routine carbapenemase detection tool and could be used as a susceptibility assay.
由于碳青霉烯类耐药性的增加对全球公共健康构成了严重威胁,因此开发新的技术来检测碳青霉烯酶活性非常重要。在这种情况下,基质辅助激光解吸/电离方法已被证明是一种快速检测碳青霉烯酶的可靠工具。作为一种新开发的检测方法,其稳健性和可能更广泛的应用仍缺乏深入分析。本研究的主要目的是评估使用设计的 MBT STAR-Carba 测定法作为肠杆菌科和铜绿假单胞菌菌株的预特征化方法的潜力,使用修改后的程序参数-各种悬浮密度和孵育时间来检测产生的碳青霉烯酶类别。此外,通过应用抑制试验测试了其对金属β-内酰胺酶 (MBL) 的深入分析和特征描述的有用性。在本研究中,设计的测定法被证明是一种检测碳青霉烯酶水解活性的敏感工具,可成功用于对存在的碳青霉烯酶类别进行部分分类。此外,使用定义的高浓度悬浮液可以将某些菌株的孵育时间缩短至 1 分钟。考虑到该测定法也适用于研究不同抑制剂对 MBL 活性的影响,它表明其鉴别潜力远高于仅作为快速常规碳青霉烯酶检测工具,可用于药敏测定。