MALDI-TOF MS applied to indirect carbapenemase detection: a validated procedure to clearly distinguish between carbapenemase-positive and carbapenemase-negative bacterial strains
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- 作者：Lijun Wang (1)
Chao Han (2)
Wenjun Sui (1)
Mei Wang (1)
Xinxin Lu (1) - 关键词：MALDI ; Carbapenem ; Carbapenemases ; Resistance ; Classification
- 刊名：Analytical and Bioanalytical Chemistry
- 出版年：2013
- 出版时间：June 2013
- 年：2013
- 卷：405
- 期：15
- 页码：5259-5266
- 全文大小：356KB
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Chao Han (2)
Wenjun Sui (1)
Mei Wang (1)
Xinxin Lu (1)
1. Department of Laboratory Medicine, Beijing Tongren Hospital, No. 1. Dongjiaominxiang Road, Dongcheng District, Beijing, 100730, China
2. Department of Laboratory Medicine, Xi’an No. 1 Hospital, Xi’an City 029, Shanxi Province, China - ISSN：1618-2650
文摘
Laboratory identification of carbapenemase-producing clinical isolates is crucial to limit the spread of the bacteria. In this study, we shall first develop the matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS) assay in automatic identification of carbapenemase producers. A total of 143 well-characterized isolates were studied. After an incubation of bacteria with meropenem trihydrate, the mixture was centrifuged and the supernatant analyzed by MALDI-TOF MS. A genetic algorithm model with ClinProTools software was built using spectra of 43 carbapenemase-positive isolates and 40 carbapenemase-negative isolates after 2?h of incubation. This model was externally validated using 60 test isolates. All spectra of supernatants of the carbapenemase-negative isolates showed peak profiles comparable to that of pure meropenem (m/z 384.159, 406.140, and 428.122 of its two sodium salt variants) regardless of the incubation time tested. For the carbapenemase-positive isolates, the specific peak for meropenem at m/z 384.159 disappeared during the incubation time, two products of meropenem degradation were identified with m/z 358.18 (the decarboxylated product) and 380.161 (sodium salt of the decarboxylated product), and other degradation products were observed (native molecule with disrupted amide bond with m/z 402.169, three sodium salt variants with m/z 424.151, 446.133, and 468.115). Sixty test isolates were 100?% correctly classified as carbapenemase positive and carbapenemase negative with the genetic algorithm model. MALDI-TOF MS coupled with ClinProTools is capable of rapidly, accurately, and automatically identifying carbapenemase producers. Figure The average spectra of the carbapenemase-positive (red) and carbapenemasenegative isolates (green) were shown. Nine peaks differentiating the two classes are highlighted by arrows. x axis, mass per charge [m/z (in daltons)]; y axis, intensity(arbitrary units [arb.u.]).