Fernández-Cuenca Felipe, Delgado-Valverde Mercedes, Guridi-Fernández Pablo, Gimeno-Cardona Concepción, Hidalgo-Díaz Carmen, Pascual Álvaro
Unidad de Gestión Clínica de Enfermedades Infecciosas y Microbiología, Hospital Universitario Virgen Macarena, Sevilla, Spain.
Instituto de Biomedicina de Sevilla (IBIs), Hospital Universitario Virgen Macarena/CSIC/Universidad de Sevilla, Sevilla, Spain.
Front Microbiol. 2025 Jun 10;16:1605590. doi: 10.3389/fmicb.2025.1605590. eCollection 2025.
The objective was to assess the ability of Spanish clinical microbiology laboratories to report reliable carbapenem susceptibility test results and to detect carbapenemase production (CP) and/or carbapenemase genes in double (DCP) and single carbapenemase-producing (SCP) isolates.
Twelve isolates (8 SCP and 4 DCP) selected from the Andalusian Reference Laboratory were sent to 83 laboratories with requests for MICs and phenotypic and genotypic tests used for CP.
Overall, there was lower essential agreement and a higher number of clinical errors in DCP than in SCP isolates. Phenotypic tests showed higher sensitivity for DCP isolates than for SCP isolates: lateral flow immunoassay (99.0% vs. 95.1%), carbapenem inactivation method (100% vs. 93%) and chromogenic media (100% vs. 83.3%); conversely, sensitivities for DCP versus SCP isolates was lower using disk diffusion with inhibitors (83.3% vs. 90.4%) and hydrolysis-based assays (81.3% vs. 86.1%). Molecular methods showed higher sensitivity for DCP isolates than phenotypic methods, and higher sensitivity for DCP isolates than for SCP isolates. In addition, concordance between genes detected and the reference was higher in DCP than in SCP isolates (98.9% vs. 83%). However, VIM-1 and IMP-8 were not detected in 77.5% and 42.2% of the determinations, respectively, for DCP isolates.
The main differences between DCP versus SCP isolates were the lower reliability of gradient strips, higher overall sensitivity of phenotypic methods for DCP isolates, but lower sensitivity with disk diffusion inhibitors and hydrolysis-based assays. Molecular assays were generally more sensitive for carbapenemase gene detection in DCP than in SCP isolates, although concordance was lower.
本研究旨在评估西班牙临床微生物实验室报告可靠的碳青霉烯类药敏试验结果,以及检测双碳青霉烯酶产生菌(DCP)和单碳青霉烯酶产生菌(SCP)中碳青霉烯酶产生情况(CP)和/或碳青霉烯酶基因的能力。
从安达卢西亚参考实验室选取12株分离菌(8株SCP和4株DCP),送至83家实验室,要求进行最低抑菌浓度(MIC)测定以及用于检测CP的表型和基因型试验。
总体而言,与SCP分离菌相比,DCP分离菌的基本一致性较低,临床错误数量较多。表型试验显示,DCP分离菌的敏感性高于SCP分离菌:侧向流动免疫测定法(99.0%对95.1%)、碳青霉烯灭活法(100%对93%)和显色培养基(100%对83.3%);相反,使用含抑制剂的纸片扩散法(83.3%对90.4%)和基于水解的检测方法时,DCP分离菌的敏感性低于SCP分离菌(81.3%对86.1%)。分子方法显示,DCP分离菌的敏感性高于表型方法,且DCP分离菌的敏感性高于SCP分离菌。此外,DCP分离菌中检测到的基因与参考基因的一致性高于SCP分离菌(98.9%对83%)。然而,分别有77.5%和42.2%的DCP分离菌测定未检测到VIM-1和IMP-8。
DCP与SCP分离菌之间的主要差异在于梯度条带的可靠性较低,DCP分离菌表型方法的总体敏感性较高,但含抑制剂的纸片扩散法和基于水解的检测方法的敏感性较低。分子检测对DCP碳青霉烯酶基因的检测通常比SCP分离菌更敏感,尽管一致性较低。