Gato Eva, Anantharajah Ahalieyah, Arroyo Manuel J, Artacho María José, Caballero Juan de Dios, Candela Ana, Chudějová Kateřina, Constanso Ignacio Pedro, Elías Cristina, Fernández Javier, Jiménez Jesús, Lumbreras Pilar, Méndez Gema, Mulet Xavier, Pérez-Palacios Patricia, Rodríguez-Sánchez Belén, Cantón Rafael, Hrabák Jaroslav, Mancera Luis, Martínez-Martínez Luis, Oliver Antonio, Pascual Álvaro, Verroken Alexia, Bou Germán, Oviaño Marina
Servicio de Microbiología, Red Española de Investigación en Patología Infecciosa, Instituto de Investigación Biomédica da Coruña, CIBER de Enfermedades Infecciosas (CIBERIFEC), Instituto de Salud Carlos III, Complejo Hospitalario Universitario A Coruña, A Coruña, Spain.
Service de Microbiologie, Cliniques Universitaires Saint-Luc, Brussels, Belgium.
Front Microbiol. 2022 Jan 27;12:789731. doi: 10.3389/fmicb.2021.789731. eCollection 2021.
In this study, we evaluate the performance of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for rapid detection of carbapenemase activity in Enterobacterales in clinical microbiology laboratories during a multicenter networking validation study. The study was divided into three different stages: "software design," "intercenter evaluation," and "clinical validation." First, a standardized procedure with an online software for data analysis was designed. Carbapenem resistance was detected by measuring imipenem hydrolysis and the results were automatically interpreted using the Clover MS data analysis software (Clover BioSoft, Spain). Second, a series of 74 genotypically characterized Enterobacterales (46 carbapenemase-producers and 28 non carbapenemase-producers) were analyzed in 8 international centers to ensure the reproducibility of the method. Finally, the methodology was evaluated independently in all centers during a 2-month period and results were compared with the reference standard for carbapenemase detection used in each center. The overall agreement rate relative to the reference method for carbapenemase resistance detection in clinical samples was 92.5%. The sensitivity was 93.9% and the specificity, 100%. Results were obtained within 60 min and accuracy ranged from 83.3 to 100% among the different centers. Further, our results demonstrate that MALDI-TOF MS is an outstanding tool for rapid detection of carbapenemase activity in Enterobacterales in clinical microbiology laboratories. The use of a simple in-house procedure with online software allows routine screening of carbapenemases in diagnostics, thereby facilitating early and appropriate antimicrobial therapy.
在这项研究中，我们在一项多中心网络验证研究中评估了基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）在临床微生物实验室中快速检测肠杆菌科细菌碳青霉烯酶活性的性能。该研究分为三个不同阶段：“软件设计”、“中心间评估”和“临床验证”。首先，设计了一种带有在线数据分析软件的标准化程序。通过测量亚胺培南水解来检测碳青霉烯耐药性，并使用三叶草MS数据分析软件（西班牙三叶草生物软件公司）自动解释结果。其次，在8个国际中心对一系列74株基因特征明确的肠杆菌科细菌（46株碳青霉烯酶产生菌和28株非碳青霉烯酶产生菌）进行了分析，以确保该方法的可重复性。最后，在两个月的时间里，所有中心独立评估了该方法，并将结果与每个中心用于碳青霉烯酶检测的参考标准进行了比较。临床样本中碳青霉烯耐药性检测相对于参考方法的总体符合率为92.5%。敏感性为93.9%，特异性为100%。在60分钟内获得结果，不同中心的准确性范围为83.3%至100%。此外，我们的结果表明，MALDI-TOF MS是临床微生物实验室中快速检测肠杆菌科细菌碳青霉烯酶活性的出色工具。使用简单的内部程序和在线软件可在诊断中对碳青霉烯酶进行常规筛查，从而有助于早期和适当的抗菌治疗。
