Analysis of the microbiota of sputum samples from patients with lower respiratory tract infections
Analysis of the microbiota of sputum samples from patients with lower respiratory tract infections
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下呼吸道感染患者痰标本微生物群分析
DOI：
10.1093/abbs/gmq081
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发表时间：
2010-10-01
影响因子：
3.7
通讯作者：
Guo, Xiaokui
中科院分区：
文献类型：
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作者：
Zhou, Yuhua;Lin, Ping;Guo, Xiaokui
痰是从下呼吸道感染患者身上采集的最常见的样本，对于这些感染的细菌鉴定至关重要。在这项研究中，我们收集了101例下呼吸道感染患者的101份痰标本。最初，痰中细菌的16S rDNA V3高变区的焦磷酸测序被用作微生物区系分析的一种独立于培养物的方法。为了进行比较，还对同一队列的痰样本进行了临床实验室测试，使用的是依赖培养的自动细菌鉴定系统。通过焦磷酸测序，发现了70,000个DNA片段，经核糖体数据库项目(RDP)过程分析后，将其归入129个细菌属。大多数序列属于几个优势属，如链球菌和葡萄球菌，表明这些属在下呼吸道感染中起重要作用。此外，还发现了一些属于潜在病原体的序列，如支原体、嗜血杆菌和莫拉氏菌，但临床实验室测试没有发现这些序列。对两种方法检测的9个属的敏感性进行了比较，结果表明焦磷酸测序法较敏感，克雷伯氏菌和分枝杆菌除外。值得注意的是，这种方法揭示了更复杂的细菌群落，显示了一种有希望的细菌检测能力。
Sputum is the most common sample collected from patients suffering from lower respiratory tract infections and it is crucial for the bacterial identification of these infections. In this study, we enrolled 101 sputum samples from 101 patients with lower respiratory tract infections. Initially, pyrosequencing of the 16S rDNA V3 hypervariable regions of the bacteria contained in the sputum was utilized as a culture-independent approach for microbiota analysis. For comparison, clinical laboratory tests using a culture-dependent automated bacterial identification system for the same cohort of sputum samples were also done. By pyrosequencing, > 70,000 DNA fragments were found and classified into 129 bacterial genera after being analyzed by the Ribosomal Database Project (RDP) process. Most sequences belonged to several predominant genera, such as Streptococcus and Staphylococcus, indicating that these genera play an important role in lower respiratory tract infections. In addition, some sequences belonging to potential causative agents, such as Mycoplasma, Haemophilus, and Moraxella, were also found, but these sequences were not found by clinical laboratory tests. For the nine genera detected by both methods, the methods' sensitivities were compared and the results showed that pyrosequencing was more sensitive, except for Klebsiella and Mycobacterium. Significantly, this method revealed much more complicated bacterial communities and it showed a promising ability for the detection of bacteria.