Detection of Extended-Spectrum β-Lactamase and Carbapenemase Activity in Gram-Negative Bacilli Using Liquid Chromatography - Tandem Mass Spectrometry

使用液相色谱-串联质谱法检测革兰氏阴性杆菌中的超广谱 β-内酰胺酶和碳青霉烯酶活性

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作者:Vlad Serafim, Ajit J Shah, Monica Licker, Florin George Horhat, Silvana Vulpie, Corina Musuroi, Delia Muntean

Conclusion

By using a sensitive strain for results interpretation and simultaneous use of multiple antibiotics, the proposed protocol offers improved robustness and multiplexing over previously described methods for antibiotic sensitivity testing. Nevertheless, further research is needed before routine assimilation of the method, especially for strains with intermediate resistance.

Methods

Four species of Gram-negative bacilli (Klebsiella pneumoniae, Escherichia coli, Providencia stuartii and Acinetobacter baumannii) were tested against six antibiotics from three different classes: ampicillin, meropenem, imipenem, ceftazidime, ceftriaxone and cefepime. Bacterial suspensions from each species were incubated with a mixture of the six antibiotics. Any remaining antibiotic following incubation was measured using LC-MS/MS. The

Purpose

Several mass spectrometry-based

Results

Overall, LC-MS/MS assay and commercial antimicrobial susceptibility platform results showed good agreement in terms of an organism being resistant/sensitive to an antibiotic. The time required to complete the LC-MS/MS-based hydrolysis test was under 5 h, significantly shorter that commercially available susceptibility testing platforms.

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