Detection of Legionella pneumophila in Cooling Water Systems of Hospitals and Nursing Homes of Kerman City, Iran by Semi- Nested PCR

Abstract

� Abstract—Legionella pneumophila is involved in more than 95% cases of severe atypical pneumonia. Infection is mainly by inhalation the indoor aerosols through the water-coolant systems. Because some Legionella strains may be viable but not culturable, therefore, Taq polymerase, DNA amplification and semi-nested-PCR were carried out to detect Legionella-specific 16S-rDNA sequence. For this purpose, 1.5 litter of water samples from 77 water-coolant system were collected from four different hospitals, two nursing homes and one student hostel in Kerman city of Iran, each in a brand new plastic bottle during summer season of 2006 (from April to August). The samples were filtered in the sterile condition through the Millipore Membrane Filter. DNA was extracted from membrane and used for PCR to detect Legionella spp. The PCR product was then subjected to semi-nested PCR for detection of L. pneumophila. Out of 77 water samples that were tested by PCR, 30 (39%) were positive for most species of Legionella. However, L. pneumophila was detected from 14 (18.2%) water samples by semi-nested PCR. From the above results it can be concluded that water coolant systems of different hospitals and nursing homes in Kerman city of Iran are highly contaminated with L. pneumophila spp. and pose serious concern. So, we recommend avoiding such type of coolant system in the hospitals and nursing homes. Keywords—Legionella pneumophila, water-coolant system,