Development of a rapid screening method for Carbapenem-Resistant Enterobacteriaceae using high resolution mass spectrometry

利用高分辨率质谱法开发耐碳青霉烯类肠杆菌科细菌的快速筛查方法

• 批准号: 2621037
• 金额: --
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2621037
• 关键词: Development; rapid; screening; method; Carbapenem

Carbapenem-resistant Enterobacteriaceae are categorised by the World Health Organisation (WHO) as a critical priority for the development of novel drugs and treatment surveillance strategies. Species include Escherichia coli and Klebsiella pneumoniae both of which have developed multi-drug resistant strains. Carbapenems are referred to as the last resort antimicrobial by the WHO, and without immediate action antimicrobial resistance will be the leading cause of death per year by 2050 costing 10 million lives and causing economic damage in excess of $100 trillion. This project will contribute towards the development of novel screening methods to enable rapid identification of carbapenem-resistant organism or presence of resistance genes. There are currently three classes of carbopenem-resistance enzymes known (Class A carbapenemases, Class B Metallo-B-lactamases, and Class D OXA carbapenemases), which are further divided into several subgroups dependant on the microorganism. Techniques such as MALDI-MS, LC-MS, and GC-MS have already shown to be useful in discriminating bacteria and have been widely adopted by microbiology laboratories. In this research project, the student will use high resolution mass spectrometry to target and characterise the molecular phenotype of drug resistance mechanisms from the extracellular production of proteins, lipids, and metabolites (both volatile and non-volatile) with the aim of discovering metabolite biomarkers with diagnostic potential. The student will sample from axenic cultures and co-cultures of the same species to identify core and resistant microbial footprint, and metabolites produced from dynamic population change in acquiring resistance genes when in co-culture with drug sensitive strains. To confirm association with metabolic pathways, the student will also perform functional analysis by isotopic enrichment of culture media. The student will then perform complex univariate and multivariate analyses to develop robust classification algorithms and compare these to commonly used microbiology assays. Prospective metabolite biomarkers will then directly contribute to the formation of rapid mass-spectrometry or pre-cast identification tests which can be used for clinical identification, environment analysis (e.g. agriculture or wastewater), and applications in the food industry, thereby contributing to global surveillance of AMR.

耐碳青霉烯肠杆菌科被世界卫生组织（WHO）列为开发新药和治疗监测战略的关键优先事项。菌种包括大肠杆菌和肺炎克雷伯菌，这两种菌种都已发展出多重耐药菌株。碳青霉烯类被世卫组织称为最后的抗菌素，如果不立即采取行动，到2050年，抗菌素耐药性将成为每年死亡的主要原因，造成1000万人死亡，经济损失超过100万亿美元。该项目将有助于开发新的筛选方法，以快速鉴定碳青霉烯抗性生物或抗性基因的存在。目前已知的碳青霉烯耐药酶有三类（A类碳青霉烯酶、B类金属-B-内酰胺酶和D类OXA碳青霉烯酶），并根据微生物的不同进一步分为几个亚群。MALDI-MS、LC-MS和GC-MS等技术已经被证明在区分细菌方面很有用，并被微生物实验室广泛采用。在这个研究项目中，学生将使用高分辨率质谱法从细胞外蛋白质、脂质和代谢物（挥发性和非挥发性）的产生中定位和表征耐药机制的分子表型，目的是发现具有诊断潜力的代谢物生物标志物。学生将从同一物种的无菌培养和共培养中取样，以确定核心和耐药微生物足迹，以及在与药物敏感菌株共培养时获得耐药基因的动态种群变化产生的代谢物。为了确认与代谢途径的关联，学生还将通过培养基的同位素富集进行功能分析。然后，学生将执行复杂的单变量和多变量分析，以开发强大的分类算法，并将其与常用的微生物分析进行比较。未来代谢物生物标记物将直接有助于形成快速质谱法或预制件鉴定测试，可用于临床鉴定、环境分析（例如农业或废水）和食品工业中的应用，从而有助于AMR的全球监测。