Resistance to third-generation cephalosporins and carbapenems in Gram-negative bacteria is chiefly mediated by beta-lactamases including extended-spectrum beta-lactamase (ESBL), AmpC, and carbapenemase enzymes. Routine phenotypic detection methods do not provide timely results, and there is a lack of comprehensive molecular panels covering all important markers. An ESBL/carbapenemase high-resolution melt analysis (HRM) assay (SHV, TEM, CTX-M ESBL families, and NDM, IMP, KPC, VIM and OXA-48–like carbapenemases) and an AmpC HRM assay (16S rDNA control, FOX, MOX, ACC, EBC, CIT, and DHA) were designed and evaluated on 111 Gram-negative isolates with mixed resistance patterns. The sensitivity for carbapenemase, ESBL, and AmpC genes was 96.7% (95% confidence interval [CI]: 82.8–99.9%), 93.6% (95% CI: 85.7–97.9%), and 93.8% (95% CI: 82.8–98.7%), respectively, with a specificity of 100% (95% CI: 95.6–100%), 93.9% (95% CI: 79.8–99.3%), and 93.7% (95% CI: 84.5–98.2%). The HRM assays enable the simultaneous detection of the 14 most important ESBL, carbapenemase, and AmpC genes and could be used as a molecular surveillance tool or to hasten detection of antimicrobial resistance for treatment management.

革兰氏阴性菌对第三代头孢菌素和碳青霉烯类药物的耐药性主要由β-内酰胺酶介导，包括超广谱β-内酰胺酶(ESBL)、AmpC和碳青霉烯酶。常规表型检测方法不能提供及时的结果，并且缺乏涵盖所有重要标记的综合分子组。 ESBL/碳青霉烯酶高分辨率熔解分析 (HRM) 测定（SHV、TEM、CTX-M ESBL 家族以及 NDM、IMP、KPC、VIM 和 OXA-48 类碳青霉烯酶）和 AmpC HRM 测定（16S rDNA 对照、 FOX、MOX、ACC、EBC、CIT 和 DHA）在 111 种具有混合耐药模式的革兰氏阴性分离株上进行设计和评估。碳青霉烯酶、ESBL 和 AmpC 基因的敏感性分别为 96.7%（95% 置信区间 [CI]：82.8–99.9%）、93.6%（95% CI：85.7–97.9%）和 93.8%（95% CI：82.8） –98.7%），特异性分别为 100%（95% CI：95.6–100%）、93.9%（95% CI：79.8–99.3%）和 93.7%（95% CI：84.5–98.2%） 。 HRM 检测能够同时检测 14 个最重要的 ESBL、碳青霉烯酶和 AmpC 基因，并可用作分子监测工具或加速检测抗菌药物耐药性以进行治疗管理。