High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in cerebrospinal fluid samples from patients with purulent meningitis.
High-throughput sequencing of 16S rDNA amplicons characterizes bacterial composition in cerebrospinal fluid samples from patients with purulent meningitis.
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16S rDNA 扩增子的高通量测序表征化脓性脑膜炎患者脑脊液样本中的细菌组成
DOI：
10.2147/dddt.s82728
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Wang Z
中科院分区：
文献类型：
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作者：
Liu A;Wang C;Liang Z;Zhou ZW;Wang L;Ma Q;Wang G;Zhou SF;Wang Z
化脓性脑膜炎 (PM) 是一种严重的传染病，发病率和死亡率很高。人们已经认识到细菌感染是PM发病的一个主要因素。然而，由于脑脊液细菌培养阳性率较低，因此缺乏PM中细菌组成的信息。在此，我们的目的是利用高通量测序方法区分和鉴定 PM 患者脑脊液样本中的主要病原体和细菌组成。采集26名PM患者的脑脊液样本，并确定为培养阴性样本。使用454 GS FLX系统对这26个PM样品中16S rDNA基因高变区的聚合酶链反应产物进行测序。结果显示，焦磷酸测序reads为71,440条，其中优势门为变形菌门和厚壁菌门；主要属为链球菌属、不动杆菌属、假单胞菌属和奈瑟菌属。脑脊液中细菌种类复杂，61.5%的样本存在混合病原体。观察到大量属于已知致病潜力的细菌。单个样本的可操作分类单元数量从 6 到 75 个不等，通过 α 和 β 多样性分析计算得出的物种多样性存在可比较的差异。总的来说，数据表明，高通量测序方法有助于表征PM患者脑脊液中的病原体，并确定PM患者脑脊液样本中细菌的丰度和组成，这可能有助于更好地了解PM病原体，并帮助临床医生做出合理有效的治疗决策。
Purulent meningitis (PM) is a severe infectious disease that is associated with high rates of morbidity and mortality. It has been recognized that bacterial infection is a major contributing factor to the pathogenesis of PM. However, there is a lack of information on the bacterial composition in PM, due to the low positive rate of cerebrospinal fluid bacterial culture. Herein, we aimed to discriminate and identify the main pathogens and bacterial composition in cerebrospinal fluid sample from PM patients using high-throughput sequencing approach. The cerebrospinal fluid samples were collected from 26 PM patients, and were determined as culture-negative samples. The polymerase chain reaction products of the hypervariable regions of 16S rDNA gene in these 26 samples of PM were sequenced using the 454 GS FLX system. The results showed that there were 71,440 pyrosequencing reads, of which, the predominant phyla were Proteobacteria and Firmicutes; and the predominant genera were Streptococcus, Acinetobacter, Pseudomonas, and Neisseria. The bacterial species in the cerebrospinal fluid were complex, with 61.5% of the samples presenting with mixed pathogens. A significant number of bacteria belonging to a known pathogenic potential was observed. The number of operational taxonomic units for individual samples ranged from six to 75 and there was a comparable difference in the species diversity that was calculated through alpha and beta diversity analysis. Collectively, the data show that high-throughput sequencing approach facilitates the characterization of the pathogens in cerebrospinal fluid and determine the abundance and the composition of bacteria in the cerebrospinal fluid samples of the PM patients, which may provide a better understanding of pathogens in PM and assist clinicians to make rational and effective therapeutic decisions.