Department of Veterinary Pathobiology, University of Missouri, Columbia, MO, USA.
J Appl Microbiol. 2009 Feb;106(2):634-41. doi: 10.1111/j.1365-2672.2008.04037.x.
The focus of this study was to identify a bacterial 16S rRNA gene sequence, unique to microbiota in the human gut, for use in development of a dependable PCR assay to detect human faecal pollution in water.
Suppression subtractive hybridization (SSH) and bioinformatics were used to identify a genetic marker, within the 16S rRNA gene of Faecalibacterium, for the detection of human faeces. DNA sequencing analysis demonstrated that a majority (16) of 74 clones of the SSH library contained insertion sequences identified as Faecalibacterium 16S rRNA genes. Human faeces-specific sequences were derived and six PCR primer sets designed and tested against faecal DNA samples from human and nonhuman sources. One PCR primer set, HFB-F3 and HFB-R5, was exclusively associated with human faeces. These primers generated a human faeces-specific amplicon of 399 bp from 60.2% of human faecal samples and 100% of sewage samples.
The subject Faecalibacterium marker is specific for sewage.
This study represents the initial report of a Faecalibacterium marker for human faeces, which may prove useful for microbial source tracking.
本研究的重点是鉴定人类肠道微生物群中独特的细菌 16S rRNA 基因序列，用于开发可靠的 PCR 检测方法，以检测水中的人类粪便污染。
采用抑制性消减杂交（SSH）和生物信息学技术，鉴定出粪便短链脂肪酸细菌 16S rRNA 基因中的遗传标记，用于检测人类粪便。DNA 测序分析表明，SSH 文库的 74 个克隆中有 16 个（16）克隆包含插入序列，鉴定为粪便短链脂肪酸细菌 16S rRNA 基因。从粪便 DNA 样本中衍生出人类粪便特异性序列，并设计和测试了 6 对 PCR 引物对非人类来源的粪便 DNA 样本。一对 PCR 引物对 HFB-F3 和 HFB-R5 与人类粪便特异性相关。这些引物从 60.2%的人类粪便样本和 100%的污水样本中产生了 399bp 的人类粪便特异性扩增子。
该粪便短链脂肪酸细菌标记物是特异性的，用于检测污水。
本研究首次报道了一种用于人类粪便的粪便短链脂肪酸细菌标记物，它可能对微生物源追踪有用。
