Microbiome and Resistome of Mycobacterium ulcerans lesions

布鲁里溃疡是一种被忽视的细菌性疾病，主要发生在西非。致病菌溃疡分枝杆菌主要在幼儿中引起严重的溃疡性皮肤病变。治疗的选择是利福平和克拉霉素的抗生素联合治疗支持手术清创。治疗成功取决于几个因素，包括受影响皮肤的大小，细菌伤口菌群的组成，以及治疗依从性。然而，尽管抗生素治疗取得了成功，但很大一部分患者对治疗没有充分的反应，伤口愈合也有很大差异。在相当多的病例中，治疗被延迟或完全失败。除了患者依从性可能较差之外，溃疡分枝杆菌病变的细菌组成可能是影响愈合过程的驱动因素，也是伤口内细菌（包括溃疡分枝杆菌）的抗微生物药物耐药性的驱动因素。迄今为止，在受影响最严重的国家的流行地区，这些因素几乎没有得到重视。因此，本项目旨在通过基于培养的方法和下一代测序分析（NGS）分析布鲁里溃疡病变的伤口微生物组，其中宏基因组分析将补充调查，以揭示细菌多样性的全部范围。布鲁里溃疡患者将在加纳库马西及周边城市的7家医院登记。如果伤口没有愈合，将从患者身上采集样本长达9个月。拟议的研究将允许识别相关细菌参与布鲁里溃疡病变的发展，以及他们的愈合过程。NGS筛选还可以识别无法通过标准方法培养的细菌，并且以前与伤口愈合延迟无关。此外，基于测序的抗生素耐药性测试将得到表型抗性筛选的支持。将布鲁里溃疡伤口的细菌组成进行比较，尽管有充分的抗生素治疗，但观察到病变愈合不良，以及对治疗反应良好的患者。具有长期和不良愈合史的个体有被特殊微生物菌群定植和/或携带抗生素耐药细菌（包括溃疡分枝杆菌菌株）的风险。因此，为耐药细菌的出现做好准备是必要的，以充分应对不断变化的治疗方案。

Buruli ulcer (BU) is a neglected bacterial disease, predominantly occurring in West Africa. The causative agent Mycobacterium ulcerans induces severe ulcerative skin lesions mainly in young children. The treatment of choice is an antibiotic combination therapy of rifampicin and clarithromycin supported by surgical debridement. Treatment success depends on several factors comprising the size of the affected skin, the composition of the bacterial wound flora, and also treatment compliance. However, despite successes of antibiotic therapy, a significant proportion of patients do not respond adequately to treatment, and wound healing varies substantially. In a considerable number of cases, therapy is delayed or has failed completely. Except from possible poor patient compliance, the bacterial composition of BU lesions might be a driving factor affecting the healing process, as well as antimicrobial resistance of bacteria within the wounds including those of M. ulcerans. These factors have so far received little attention in endemic areas in the most affected countries. Therefore, this project aims to analyse the wound microbiome of BU lesions by culture-based methods in addition to next generation sequencing analysis (NGS), where metagenomic analyses will complement the investigations to uncover the full range of bacterial diversity. BU patients will be enrolled in seven hospitals of Kumasi, Ghana, and surrounding cities. Samples will be collected from patients up to 9 months if the wounds do not heal. The proposed study will allow identification of relevant bacteria involved in the development of BU lesion, and their healing processes. NGS screens may also identify bacteria that cannot be cultured by standard methods, and have previously not been associated with delayed wound healing. In addition, sequencing based antibiotic resistance tests will be supported by phenotypic resistance screens. The bacterial composition of BU wounds will be compared amongst patients where despite adequate antibiotic therapy poor healing of the lesions was observed, and those that responded well to therapy. Individuals with a long and poor healing history are at risk to be colonized by a peculiar microbial flora and/or carrying antibiotic resistant bacteria including M. ulcerans strains. Thus, preparedness for the emergence of resistant bacteria is necessary to respond adequately to changing therapy regimes.