Detection of Legionella pneumophila by PCR-ELISA method in industrial cooling tower water.
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.
Soheili
Majid
M
Department of Biotechnology, Research Institute of Petroleum Industry, Tehran, Iran.
Nejadmoghaddam
Mohammad Reza
MR
Babashamsi
Mohammad
M
Ghasemi
Jamileh
J
Jeddi Tehrani
Mahmood
M
D019921:Biotinylation; D017931:DNA Primers / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D016952:Legionella pneumophila / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000737:chemistry; D014871:Water Microbiology; D018508:Water Purification / Q000379:methods*; D014881:Water Supply
D017931:DNA Primers; D012336:RNA, Ribosomal, 16S