Rapid and sensitive quantification of Vibrio cholerae and vibrio mimicus cells in water samples by use of catalyzed reporter deposition fluorescence in situ hybridization combined with solid-phase cytometry

Sonja Schauer, Regina Sommer, Andreas H. Farnleitner, Alexander K.T. Kirschner*

*Corresponding author for this work

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

30 Citations (Scopus)

Abstract

A new protocol for rapid, specific, and sensitive cell-based quantification of Vibrio cholerae/Vibrio mimicus in water samples was developed. The protocol is based on catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) in combination with solid-phase cytometry. For pure cultures, we were able to quantify down to 6 V. cholerae cells on one membrane with a relative precision of 39% and down to 12 cells with a relative precision of 17% after hybridization with the horseradish peroxidase (HRP)-labeled probe Vchomim1276 (specific for V. cholerae and V. mimicus) and signal amplification. The corresponding position of the probe on the 16S rRNA is highly accessible even when labeled with HRP. For the first time, we were also able to successfully quantify V. cholerae/V. mimicus via solid-phase cytometry in extremely turbid environmental water samples collected in Austria. Cell numbers ranged from 4.5×101 cells ml-1 in the large saline lake Neusiedler See to 5.6×104 cells ml-1 in an extremely turbid shallow soda lake situated nearby. We therefore suggest CARD-FISH in combination with solid-phase cytometry as a powerful tool to quantify V. cholerae/V. mimicus in ecological studies as well as for risk assessment and monitoring programs.

Original languageEnglish
Pages (from-to)7369-7375
Number of pages7
JournalApplied and Environmental Microbiology
Volume78
Issue number20
DOIs
Publication statusPublished - Oct 2012
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Ecology
  • Applied Microbiology and Biotechnology

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