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基于 MALDI-TOF 质谱的方法快速检测血培养中第三代肟基头孢菌素耐药的大肠埃希菌和克雷伯菌属。

Performance of a MALDI-TOF mass spectrometry-based method for rapid detection of third-generation oxymino-cephalosporin-resistant Escherichia coli and Klebsiella spp. from blood cultures.

机构信息

Microbiology Service, Hospital Clínico Universitario, Instituto de Investigación INCLIVA, Valencia, Spain.

Infectious Diseases Unit, Hospital Clínico Universitario, Instituto de Investigación INCLIVA, Valencia, Spain.

出版信息

Eur J Clin Microbiol Infect Dis. 2021 Sep;40(9):1925-1932. doi: 10.1007/s10096-021-04251-0. Epub 2021 Apr 19.

Abstract

We optimized and prospectively evaluated a simple MALDI-TOF MS-based method for direct detection of third-generation oxymino-cephalosporin resistance (3rd CephR) in Escherichia coli and Klebsiella spp. from blood cultures (BC). In addition, we assessed the performance of a lateral flow immunochromatographic assay (LFIC) for detecting extended-spectrum β-lactamases (ESBL) (NG-Test CTX-M MULTI assay) using bacterial pellets from BC. A total of 168 BCs from unique patients were included. A pre-established volume of BC flagged as positive was transferred in brain heart infusion with or without ceftriaxone (2 mg/ml). After 2-h incubation, intact bacterial pellets were used for MALDI-TOF MS testing. Identification of bacterial species (index score > 2) in the presence of CRO was considered marker of 3rd CephR. The LFIC assay was evaluated in 141 BC. Bacteremia episodes were caused by E. coli (n = 115) or Klebsiella spp. (n = 53). A total of 49 strains were 3rd CephR by broth microdilution, of which 41 were ESBL producers, seven expressed ESBL and OXA-48 type D carbapenemase, and one harbored a plasmid-mediated AmpC. The MALDI-TOF MS method yielded four very major errors (false susceptibility) and two major errors (false resistance). The overall sensitivity of the assay was 91.8% and the specificity 98.3%. Concordance between the LFIC assay and the MALDI-TOF MS method for detection of ESBL-mediated 3rd CephR was 100%. Both evaluated methods may prove useful for early adjustment of empirical therapy in patients with E. coli and Klebsiella spp. bloodstream infections. Whether their use has a beneficial impact on patient outcomes is currently under investigation.

摘要

我们优化并前瞻性评估了一种基于 MALDI-TOF MS 的简单方法,用于直接检测血培养物(BC)中的第三代肟基头孢菌素耐药性(3rd CephR)的大肠埃希菌和克雷伯菌属。此外,我们评估了使用 BC 中的细菌沉淀检测扩展谱β-内酰胺酶(ESBL)(NG-Test CTX-M MULTI 测定法)的侧向流动免疫层析测定法(LFIC)的性能。共纳入了 168 例来自独特患者的 BC。将预定量的 flagged 为阳性的 BC 转移到含有或不含有头孢曲松(2mg/ml)的脑心浸液中。孵育 2 小时后,使用完整的细菌沉淀进行 MALDI-TOF MS 检测。在存在 CRO 的情况下鉴定细菌物种(指数得分>2)被认为是 3rd CephR 的标志物。LFIC 测定法在 141 例 BC 中进行了评估。菌血症发作由大肠埃希菌(n = 115)或克雷伯菌属(n = 53)引起。通过肉汤微量稀释法,共有 49 株为 3rd CephR,其中 41 株为 ESBL 产生菌,7 株表达 ESBL 和 OXA-48 型 D 碳青霉烯酶,1 株携带质粒介导的 AmpC。MALDI-TOF MS 方法产生了 4 个非常大的错误(假敏感性)和 2 个主要错误(假耐药性)。该测定法的总灵敏度为 91.8%,特异性为 98.3%。LFIC 测定法和 MALDI-TOF MS 法用于检测 ESBL 介导的 3rd CephR 的一致性为 100%。两种评估方法都可能有助于早期调整大肠埃希菌和克雷伯菌属血流感染患者的经验性治疗。它们的使用是否对患者结局有有益影响,目前正在研究中。

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