title: Development of Faecalibacterium 16S rRNA gene marker for identification of human faeces.
abstract: OBJECTIVE
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  The subject Faecalibacterium marker is specific for sewage.
  
  
  CONCLUSIONS
  This study represents the initial report of a Faecalibacterium marker for human faeces, which may prove useful for microbial source tracking.
authors.affiliation: Department of Veterinary Pathobiology, University of Missouri, Columbia, MO, USA.
authors.forename: G
authors.initials: G
authors.lastname: Zheng
authors.forename: H
authors.initials: H
authors.lastname: Yampara-Iquise
authors.forename: J E
authors.initials: JE
authors.lastname: Jones
authors.forename: C
authors.initials: C
authors.lastname: Andrew Carson
mesh_terms: D000818:Animals; D004784:Environmental Monitoring / Q000379:methods*; D005243:Feces / Q000382:microbiology*; D005798:Genes, Bacterial; D020459:Genes, rRNA; D005819:Genetic Markers; D006801:Humans; D057230:Limit of Detection; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000032:analysis; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D017422:Sequence Analysis, DNA; D012722:Sewage / Q000382:microbiology*; D013045:Species Specificity; D014871:Water Microbiology; D014873:Water Pollutants / Q000032:analysis*
chemical_list: D005819:Genetic Markers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012722:Sewage; D014873:Water Pollutants