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

*Korrespondierende:r Autor:in für diese Arbeit

Publikation: Beitrag in Fachzeitschrift (peer-reviewed)Artikel in Fachzeitschrift

112 Zitate (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.

OriginalspracheEnglisch
Seiten (von - bis)351-356
Seitenumfang6
FachzeitschriftLetters in Applied Microbiology
Jahrgang44
Ausgabenummer4
DOIs
PublikationsstatusVeröffentlicht - Apr. 2007
Extern publiziertJa

ASJC Scopus Sachgebiete

  • Angewandte Mikrobiologie und Biotechnologie

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