Clinical Microbiology, Laboratory Medicine, Royal Infirmary of Edinburgh, Edinburgh, UK.
University of Edinburgh Centre for Inflammation Research, Queen's Medical Research Institute, Edinburgh BioQuarter, UK.
J Med Microbiol. 2019 Apr;68(4):560-565. doi: 10.1099/jmm.0.000931. Epub 2019 Mar 14.
This study evaluated in-house PCR testing for local identification of bacteria carrying the major carbapenemase genes (blaOXA-48-like, blaVIM, blaNDM, blaKPC and blaIMP).
Carbapenemase-producing organisms (CPOs) isolated from patients managed in two tertiary care hospitals in Scotland from September 2014-January 2017 were investigated. A combination of chromogenic screening agar (ChromID CARBA SMART), a carbapenem hydrolysis test (Rapidec Carba NP) and in-house real-time PCR for the blaOXA-48-like, blaVIM, blaNDM, blaKPC and blaIMP genes were utilized. All isolates were sent to the AMRHAI reference unit for confirmatory testing.
During the 29-month study period 39 CPO were isolated from 34 patients. The average turnaround time for a workflow involving phenotypic and molecular testing was 4.2 days. PCR had a sensitivity and specificity of 100 %. The most common carbapenemase genes were blaOXA-48-like (31%), blaVIM (23%) and blaNDM (20%). Resistance to antimicrobials other than beta-lactams was common; the most active agents were colistin, amikacin and fosfomycin. Twenty-seven patients were considered to be colonized (although CPO detection influenced empiric antimicrobials in five) and a CPO was implicated in infection in seven patients (bacteraemia in immunocompromised patients, n=2; surgical site infections, n=2; osteomyelitis in a patient with diabetes mellitus, n=1; and urinary tract infections, n=2). All patients survived infection.
In a lowincidence setting we demonstrate the efficacy of a combined local laboratory workflow for rapid detection of CPOs, incorporating phenotypic and molecular testing. In 7/34 patients the CPO was implicated as a pathogen and detection influenced antimicrobial decision-making in five colonized patients.
本研究评估了用于本地鉴定携带主要碳青霉烯酶基因(blaOXA-48 样、blaVIM、blaNDM、blaKPC 和 blaIMP)的细菌的内部 PCR 检测。
从 2014 年 9 月至 2017 年 1 月在苏格兰的两家三级保健医院管理的患者中分离出产碳青霉烯酶的生物体(CPO),并对其进行调查。使用组合显色筛选琼脂(ChromID CARBA SMART)、碳青霉烯水解试验(Rapidec Carba NP)和blaOXA-48 样、blaVIM、blaNDM、blaKPC 和 blaIMP 基因的内部实时 PCR。所有分离物均被送往 AMRHAI 参考单位进行确认测试。
在 29 个月的研究期间,从 34 名患者中分离出 39 株 CPO。涉及表型和分子检测的工作流程的平均周转时间为 4.2 天。PCR 的灵敏度和特异性均为 100%。最常见的碳青霉烯酶基因是 blaOXA-48 样(31%)、blaVIM(23%)和 blaNDM(20%)。除了β-内酰胺类抗生素外,对其他抗生素的耐药性很常见;最有效的药物是粘菌素、阿米卡星和磷霉素。27 名患者被认为是定植(尽管有 5 名患者的 CPO 检测影响了经验性抗生素),7 名患者的 CPO 被认为与感染有关(免疫功能低下患者的菌血症,n=2;手术部位感染,n=2;1 名糖尿病患者的骨髓炎,n=1;尿路感染,n=2)。所有患者均存活感染。
在低发生率环境中,我们证明了快速检测 CPO 的综合本地实验室工作流程的功效,该流程结合了表型和分子检测。在 34 名患者中的 7 名患者中,CPO 被认为是病原体,在 5 名定植患者中,检测影响了抗生素的决策。