Detection of Legionella pneumophila by PCR-ELISA method in industrial cooling tower water

Pak J Biol Sci. 2007 Nov 15;10(22):4015-21. doi: 10.3923/pjbs.2007.4015.4021.

Abstract

Water supply and Cooling Tower Water (CTW) are among the most common sources of Legionella pneumophila (LP) contamination. A nonradio active method is described to detect LP in industrial CTW samples. DNA was purified and amplified by nested -PCR with amplimers specific for the 16s rRNA gene of LP. The 5' end biotinylated oligomer probe was immobilized on sterptavidin B coated microtiter plates. The nested-PCR product was labeled with digoxigenin and then hybridized with 5'-biotinylated probes. The amplification products were detected by using proxidase-labled anti dioxygenin antibody in a colorimetric reaction. The assay detected LP present in 1 L of 5 CTW samples examined. All of the samples were Legionella positive in both culture and PCR-ELISA methods. The PCR-ELISA assay appears to exhibit high specificity and is a more rapid technique in comparison with bacterial culture method. Thus could prove suitable for use in the routine examination of industrial CTW contamination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotinylation
  • DNA Primers / chemistry
  • Electrophoresis, Agar Gel
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Legionella pneumophila / genetics
  • Legionella pneumophila / isolation & purification*
  • Polymerase Chain Reaction / methods*
  • RNA, Ribosomal, 16S / analysis
  • RNA, Ribosomal, 16S / chemistry
  • Water Microbiology
  • Water Purification / methods*
  • Water Supply

Substances

  • DNA Primers
  • RNA, Ribosomal, 16S