Objective: To analyze the microbiome of diabetic foot osteomyelitis (DFO) by means of metagenome sequencing and provide evidence for identification of pathogenic bacteria in DFO. Methods: A total of 5 patients (3 males and 2 females) with DFO hospitalized at the Department of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University were enrolled and infected bone specimens were obtained between September 2016 and April 2017. The mean age was (55.8±9.5) years. Metagenome sequencing was performed to explore the characteristics of microbiome, and compared with the results of 16S rRNA sequencing. Results: The results of metagenome sequencing showed that DFO contained diverse microorganism. Totally, 22 dominant species were obtained, Klebsiella pneumoniae (69.66%) was the most abundant, followed by Veillonella parvula (36.93%) and Prevotella intermedia (34.19%). Compared with the 16S rRNA sequencing, metagenome sequencing could obtain more species information on the basis of fewer samples. At the genus level, both sequencing techniques suggested the most dominant pathogen in DFO was anaerobe. All bone specimens had polymicrobial communities. Conclusions: More microecological diversity and abundance of DFO can be found by using metagenome sequencing. At the species level, more bacteria, even bacterial strains can be identified by metagenome sequencing. At the genus level, the most abundant bacteria is anaerobe, however, at the species level, it is facultative anaerobe.
目的： 分析糖尿病足骨髓炎（DFO）的微生物群落，为DFO病原菌的鉴定提供依据。 方法： 选取2016年9月至2017年4月于南方医科大学南方医院内分泌科住院治疗的5例确诊为DFO患者的感染骨组织，其中男3例，女2例，年龄（55.8±9.5）岁。利用宏基因组测序技术分析其微生物特点，并与16S rRNA测序结果进行比较。 结果： 宏基因组测序显示DFO微生物多样性明显，种水平共获得优势细菌22种，丰度最高的菌种是肺炎克雷伯菌（69.66%），其次是小韦永球菌（36.93%）、中间普雷沃菌（34.19%）。与16S rRNA测序相比，宏基因组测序能在更少样本的基础上得到更多物种信息。在属水平，两种测序方法均显示DFO以厌氧菌为主。所有样本均为多细菌感染。 结论： 宏基因组测序技术能发现更多DFO菌群微生态的多样性及丰度特点。在种水平，能鉴定出更多16S rRNA测序技术无法分辨的细菌，甚至可以鉴定到菌株。属水平丰度最高的细菌是厌氧菌，种水平丰度最高的细菌是兼性厌氧菌。.
Keywords: Diabetic foot; Metagenome; Microbiome; Osteomyelitis; Sequence analysis.