牛皮关节炎中的皮肤微生物组:方法的发展和试验数据
Madhura Castelino1, Stephen Eyre2, John Moat3
1NIHR Manchester Musculoskeletal Biomedical Research Unit, Manchester Academic Health Science Centre, Manchester, UK.
Lancet (London, England)
|August 28, 2015
概括
在MiSeq平台上测序的V3-V4 16SrRNA超变区为皮肤微生物分析提供了与Roche454平台相匹配的结果. 这种方法为研究皮肤微生物组提供了可靠的方法,
科学领域:
- 微生物学
- 基因组学
- 生物信息学
背景情况:
- 皮肤微生物群在诸如牛皮关节炎等炎症病的发展中起着至关重要的作用.
- 了解牛皮斑中的皮肤微生物组合对于推动这一领域的研究至关重要.
研究的目的:
- 使用MiSeq平台确定皮肤微生物组研究的最佳研究设计.
- 使用模拟细菌群体,比较两对原料和两种测序平台 (MiSeq和Roche454) 产生的数据.
主要方法:
- 从模拟社区提取DNA和健康志愿者的皮肤抽样.
- 使用特定的原始对进行16S rRNA基因超变区 (V1-V3和V3-V4) 的放大.
- 在MiSeq和Roche454平台上进行测序,然后使用QIIME软件进行数据处理.
主要成果:
- 与V1-V3区域相比,V3-V4超变区显示出更高的细菌多样性捕获.
- 在每个平台中观察到高一致性 (98.9-99.8%),人类DNA污染的影响最小.
- 在使用V3-V4区域时,MiSeq和Roche454平台在家庭层面显示出高一致性 (87%).
结论:
- 在MiSeq平台上对V3-V4 16SrRNA高变区进行测序,可以获得与Roche454平台相匹配的结果.
- MiSeq平台和选择的原料提供了一种可靠和可比的皮肤微生物群分析方法.
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