Quantitative analysis of human- and cow-specific 16S rRNA gene markers for assessment of fecal pollution in river waters by real-time PCR

Ju Yong Jeong, Hee Deung Park, Kyong Hee Lee, Jae Hong Hwang, Jong Ok Ka

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The base sequences representing human- and cow-specific 16S rRNA gene markers identified in a T-RFLP analysis were recovered from clone libraries. The human- and cowspecific primers were designed from these sequences and their specificities were analyzed with fecal DNAs from human, cow, and pig. The AllBac primer set showed positive results for all human, cow, and pig samples, whereas the human-specific primer set showed positive result only for the human sample but not for the cow or pig samples. Likewise, the cow-specific primer set showed positive results only for the cow sample but not for the human or pig samples. Real-time PCR assay with these primers was developed for the identification and quantification of fecal pollution in the river water. The human- and cow-specific markers were detected in the order of 9 log10 copies per gram wet feces, which were two orders of magnitude lower than those of total Bacteroidales. For the river water samples, the human-specific marker was detected in 1.7-6.2 log10 copies/100 ml water, which was 2.4-4.9 orders of magnitude lower than those of total Bacteroidales. There was no significant correlation between total Bacteroidales and conventional fecal indicators, but there was a high correlation between Bacteroidales and the human-specific marker. This assay could reliably identify and quantify the fecal pollution sources, enabling effective measures in the watersheds and facilitating water quality management.

Original languageEnglish
Pages (from-to)245-253
Number of pages9
JournalJournal of microbiology and biotechnology
Volume20
Issue number2
DOIs
Publication statusPublished - 2010 Feb 1

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Keywords

  • 16S rRNA gene markers
  • Bacteroidales
  • Fecal pollution
  • Real-time PCR

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

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