A quantitative real-time PCR assay for the highly sensitive and specific detection of human faecal influence in spring water from a large alpine catchment area

G. H. Reischer, D. C. Kasper, R. Steinborn, A. H. Farnleitner*, R. L. Mach

*Corresponding author for this work

Research output: Journal article (peer-reviewed)Journal article

108 Citations (Scopus)

Abstract

Aims: The aim of the study was the development of a sensitive human-specific quantitative real-time PCR assay for microbial faecal source tracking (MST) in alpine spring water. The assay detects human-specific faecal DNA markers (BacH) from 16S rRNA gene sequences from the phylum Bacteroidetes using TaqMan® minor groove binder probes. Methods and Results: The qualitative and quantitative detection limits of the PCR assay were 6 and 30 marker copies, respectively. Specificity was proved by testing 41 human faeces and waste water samples and excluding cross-amplification from 302 animal faecal samples from Eastern Austria. Marker concentrations in human faecal material were in the range from 6·6 × 109 to 9·1 × 1010 marker equivalents per gram. The method was sensitive enough to detect a few 100 pg of faeces in faecal suspensions. The assay was applied on water samples from an alpine karstic spring catchment area and the results reflected the expected levels of human faecal influence. Conclusions: The method exhibited sufficient sensitivity to allow quantitative source tracking of human faecal impact in the investigated karstic spring water. Significance and Impact of the Study: The developed method constitutes the first quantitative human-specific MST tool sensitive enough for investigations in ground and spring water.

Original languageEnglish
Pages (from-to)351-356
Number of pages6
JournalLetters in Applied Microbiology
Volume44
Issue number4
DOIs
Publication statusPublished - Apr 2007
Externally publishedYes

Keywords

  • Bacteroidetes
  • Genetic marker
  • Karstic spring water
  • Quantitative real-time PCR
  • Source tracking
  • Water quality

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology

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