基质辅助激光解吸电离飞行时间质谱法直接检测肠杆菌科中的 KPC。
MALDI-TOF mass spectrometry for direct KPC detection among Enterobacterales.
机构信息
Programa de Pós-Graduação Em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal Do Rio Grande Do Sul, Avenida Ipiranga, 2752, Porto Alegre, Rio Grande do Sul, 90610-000, Brazil.
Laboratório de Pesquisa Em Resistência Bacteriana, Hospital de Clínicas de Porto Alegre, Porto Alegre, Rio Grande do Sul, Brazil.
出版信息
Braz J Microbiol. 2022 Dec;53(4):1907-1913. doi: 10.1007/s42770-022-00798-y. Epub 2022 Jul 18.
In this study, we evaluate a method for the KPC enzyme detection, using MALDI-TOF MS, for Enterobacterales. A total of 300 clinical Enterobacterales isolates were selected. The collection included 259 carbapenemase-producing (157 KPC and 102 non-KPC) and 41 carbapenemase non-producing isolates. Bacterial proteins were extracted from Mueller-Hinton agar plates using formic acid, isopropyl alcohol, and water (17:33:50). Samples were prepared with a double layer of synapinic acid. Analyses were performed using a Microflex LT mass spectrometer (Bruker Daltonics) and flexAnalysis 4.0 software (Bruker Daltonics). Statistical analyses were performed using SPSS Software. A distinctive peak at m/z 28,643-28,731 was found in all 157 KPC-producing isolates, and it was consistently absent in the 143 KPC non-producing group. KPC-producing peak intensities ranged from 77 to 3893. Considering an intensity cutoff value ≥ 120 for the presence of KPC, this methodology presented 98.09% and 97.90% of sensitivity and specificity, respectively.
在这项研究中，我们评估了一种使用 MALDI-TOF MS 检测肠杆菌科中的 KPC 酶的方法。共选择了 300 株临床肠杆菌科分离株。该集合包括 259 株产碳青霉烯酶（157 株 KPC 和 102 株非 KPC）和 41 株产碳青霉烯酶非产株。使用甲酸、异丙醇和水（17:33:50）从 Mueller-Hinton 琼脂平板中提取细菌蛋白。使用双层 synapinic 酸制备样品。使用 Microflex LT 质谱仪（布鲁克·道尔顿）和 flexAnalysis 4.0 软件（布鲁克·道尔顿）进行分析。使用 SPSS 软件进行统计分析。在所有 157 株产 KPC 的分离株中均发现了 m/z 28,643-28,731 的特征峰，而在 143 株非产 KPC 组中则一致缺失。KPC 产生的峰强度范围为 77 至 3893。考虑到存在 KPC 的强度截断值≥120，该方法的灵敏度和特异性分别为 98.09%和 97.90%。
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