Laboratory of Physics of Living Matter, BSP, EPFL, Lausanne, Switzerland.
Institute of Microbiology, University of Lausanne and University Hospital Centre, Lausanne, Switzerland.
Clin Microbiol Infect. 2017 Jun;23(6):400-405. doi: 10.1016/j.cmi.2016.12.028. Epub 2017 Jan 3.
The management of bloodstream infection, a life-threatening disease, largely relies on early detection of infecting microorganisms and accurate determination of their antibiotic susceptibility to reduce both mortality and morbidity. Recently we developed a new technique based on atomic force microscopy capable of detecting movements of biologic samples at the nanoscale. Such sensor is able to monitor the response of bacteria to antibiotic's pressure, allowing a fast and versatile susceptibility test. Furthermore, rapid preparation of a bacterial pellet from a positive blood culture can improve downstream characterization of the recovered pathogen as a result of the increased bacterial concentration obtained.
Using artificially inoculated blood cultures, we combined these two innovative procedures and validated them in double-blind experiments to determine the susceptibility and resistance of Escherichia coli strains (ATCC 25933 as susceptible and a characterized clinical isolate as resistant strain) towards a selection of antibiotics commonly used in clinical settings.
On the basis of the variance of the sensor movements, we were able to positively discriminate the resistant from the susceptible E. coli strains in 16 of 17 blindly investigated cases. Furthermore, we defined a variance change threshold of 60% that discriminates susceptible from resistant strains.
By combining the nanomotion sensor with the rapid preparation method of blood culture pellets, we obtained an innovative, rapid and relatively accurate method for antibiotic susceptibility test directly from positive blood culture bottles, without the need for bacterial subculture.
血流感染是一种危及生命的疾病,其治疗主要依赖于早期发现感染微生物,并准确测定其对抗生素的敏感性,以降低死亡率和发病率。最近,我们开发了一种基于原子力显微镜的新技术,能够在纳米尺度上检测生物样本的运动。这种传感器能够监测细菌对抗生素压力的反应,从而实现快速、多功能的药敏试验。此外,从阳性血培养物中快速制备细菌沉淀,可提高下游对回收病原体的特征描述,因为获得了更高的细菌浓度。
我们使用人工接种的血培养物,将这两种创新方法结合起来,并在双盲实验中对其进行验证,以确定大肠杆菌菌株(ATCC 25933 为敏感株,经鉴定的临床分离株为耐药株)对临床常用抗生素的敏感性和耐药性。
基于传感器运动的方差,我们能够在 17 个盲检案例中的 16 个案例中,阳性区分敏感和耐药的大肠杆菌菌株。此外,我们定义了一个 60%的方差变化阈值来区分敏感和耐药菌株。
通过将纳米运动传感器与快速制备血培养物沉淀的方法相结合,我们获得了一种创新的、快速且相对准确的方法,可直接从阳性血培养瓶中进行抗生素药敏试验,而无需进行细菌传代培养。