Moreira Natália Kehl, Wilhelm Camila Mörschbächer, Echevarria Aymê Duarte, Volpato Fabiana Caroline Zempulski, Wink Priscila Lamb, Barth Afonso Luís, Caierão Juliana
Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre 90610000, Rio Grande do Sul, Brazil.
Laboratório de Pesquisa em Bacteriologia Clínica, Universidade Federal do Rio Grande do Sul, Porto Alegre 90610000, Rio Grande do Sul, Brazil.
Antibiotics (Basel). 2023 Mar 17;12(3):601. doi: 10.3390/antibiotics12030601.
Detecting carbapenemase-associated carbapenem resistance is a subject of major clinical and epidemiological concern as it influences therapeutic choice. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed as a means to assess bacterial resistance mechanisms. We aimed to detect the KPC enzyme directly from positive blood cultures using MALDI-TOF MS. To do so, 102 clinical were evaluated, including 59 positives. Proteins were extracted using formic acid, isopropyl alcohol, and water (17:33:50) and spotted onto a steel target plate using the double-layer sinapinic acid technique. Two parameters were considered: (i) the visual detection of a clear peak with the expected KPC / and (ii) the evaluation of the relative intensity of the ions in the peak. A peak was observed in 56/59 -positive isolates (94.9% sensitivity), with no false-positive results (100% specificity). When considering intensity, with a cut-off ≥120 (a.u.), sensitivity was 94.9% and specificity was 95.3%. We proposed a "buffer" zone, with intermediate values of intensity (115 to 125) reaching 100% sensitivity and specificity. The detection of KPC peaks directly from positive blood cultures using MALDI-TOF MS is feasible and rapid, which may improve appropriate patient therapy and antimicrobial stewardship.
检测与碳青霉烯酶相关的碳青霉烯耐药性是临床和流行病学主要关注的问题,因为它会影响治疗选择。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)已被提议作为评估细菌耐药机制的一种手段。我们旨在使用MALDI-TOF MS直接从阳性血培养物中检测KPC酶。为此,对102份临床样本进行了评估,其中包括59份阳性样本。使用甲酸、异丙醇和水(17:33:50)提取蛋白质,并采用双层芥子酸技术将其点样到钢靶板上。考虑了两个参数:(i)目视检测到预期的KPC / 的清晰峰;(ii)评估峰中离子的相对强度。在56/59份阳性分离株中观察到一个峰(灵敏度为94.9%),无假阳性结果(特异性为100%)。当考虑强度时,截断值≥120(a.u.)时,灵敏度为94.9%,特异性为95.3%。我们提出了一个“缓冲”区,强度中间值(115至125)时灵敏度和特异性均达到100%。使用MALDI-TOF MS直接从阳性血培养物中检测KPC峰是可行且快速的,这可能会改善患者的适当治疗和抗菌药物管理。