Genetic Homology between Bacteria Isolated from Pulmonary Abscesses or Pyothorax and Bacteria from the Oral Cavity.
Genetic Homology between Bacteria Isolated from Pulmonary Abscesses or Pyothorax and Bacteria from the Oral Cavity.
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DOI：
10.1128/spectrum.00974-21
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发表时间：
2022-02-23
影响因子：
3.7
通讯作者：
Nakahara Y
中科院分区：
文献类型：
--
作者：
Katsuda R;Inubushi J;Tobata H;Eguchi T;Terada K;Kagami R;Kawamura T;Kajiwara Y;Nakahara Y
肺脓肿和脓胸被认为是主要由口腔微生物引起的细菌感染。然而，过去关于此类感染的报道并没有提供遗传证据，也缺乏准确性，因为他们使用的是通过口腔的样本。本研究的目的是确定在没有口腔污染的情况下，经皮从肺脓肿和脓胸获得的口腔微生物群和脓标本中是否存在遗传相同的细菌菌株。首先，通过16S rRNA基因测序对脓液中分离的细菌进行鉴定。然后使用细菌物种特异性引物通过定量PCR确定从配对患者口腔拭子样本悬浮液中提取的DNA含有相同的物种。这表明在31个菌株中有8个菌株的目标物种DNA水平足够用于进一步分析。因此，这8个菌株的全基因组序列随后被确定并与同一物种的开放数据库进行比较。分别为8株菌株合成了5条菌株特异性引物。从相应患者的配对口腔拭子样本悬浮液中提取DNA，使用5种菌株特异性引物进行PCR扩增。结果提供了强有力的证据，证明某些痰源细菌菌株是口腔来源的。此外，这两步鉴定过程提供了一种新的方法，将有助于研究微生物群的某些病原体。我们提出直接的遗传证据，一些细菌在肺脓肿和脓胸是来自口腔菌群。这是首次报道在肺脓肿和脓胸脓液以及口腔拭子样本中均存在遗传同源菌株。我们开发了一种新的方法，结合定量PCR和下一代测序，成功地防止了脓液标本被口腔细菌污染。新的遗传方法对其他杂区系的研究也有一定的指导意义。例如，在菌株水平上检测肠道菌群中的病原体。
Pulmonary abscesses and pyothorax are bacterial infections believed to be caused primarily by oral microbes. However, past reports addressing such infections have not provided genetic evidence and lack accuracy, as they used samples that had passed through the oral cavity. The aim of this study was to determine whether genetically identical bacterial strains exist in both the oral microbiota and pus specimens that were obtained percutaneously from pulmonary abscesses and pyothorax, without oral contamination. First, bacteria isolated from pus were identified by 16S rRNA gene sequencing. It was then determined by quantitative PCR using bacterial-species-specific primers that DNA extracted from paired patient oral swab sample suspensions contained the same species. This demonstrated sufficient levels of bacterial DNA of the targeted species to use for further analysis in 8 of 31 strains. Therefore, the whole-genome sequences of these eight strains were subsequently determined and compared against an open database of the same species. Five strain-specific primers were synthesized for each of the eight strains. DNA extracted from the paired oral swab sample suspensions of the corresponding patients was PCR amplified using five strain-specific primers. The results provided strong evidence that certain pus-derived bacterial strains were of oral origin. Furthermore, this two-step identification process provides a novel method that will contribute to the study of certain pathogens of the microbiota. IMPORTANCE We present direct genetic evidence that some of the bacteria in pulmonary abscesses and pyothorax are derived from the oral flora. This is the first report describing the presence of genetically homologous strains both in pus from pulmonary abscesses and pyothorax and in swab samples from the mouth. We developed a new method incorporating quantitative PCR and next-generation sequencing and successfully prevented contamination of pus specimens with oral bacteria by percutaneous sample collection. The new genetic method would be useful for enabling investigations on other miscellaneous flora; for example, detection of pathogens from the intestinal flora at the strain level.
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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