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Sensitivity Comparison of Different 16s rDNA- Specific  Primers for Detection of Legionella Species in Aquatic Samples

Author(s): Farzaneh Baghal Asghari | Mahnaz Nikaeen

Journal: Iranian Journal of Health and Environment
ISSN 2008-2029

Volume: 5;
Issue: 3;
Start page: 263;
Date: 2012;
Original page

Keywords: PCR | Water | Detection | Legionella

Background and Objectives: Legionella are gram-negative bacteria widely dispersed in natural and man-made water sources. Some Legionella species are pathogenic and could cause respiratory infections. Cultivation technique is the conventional method for the detection of Legionella spp. in aquatic samples. However, the method has low sensitivity and require prolonged incubation period. Therefore, Polymerase chain reaction (PCR) as a rapid method with extreme sensitivity is used. The present study was designed to evaluate the feasibility and sensitivity of PCR method for detection of Legionellas pp. in aquatic samples using three sets of primers.Materials and Methods: In this study, 60 water samples were investigated for the presence of Legionella species using Nested- PCR technique. The sensitivity of this technique was evaluated for the detection of Legionella species in aquatic samples using three primer sets, including (LEG225-LEG858), (LEG448-LEG858), and (LEG448-JRP).Results: The nested PCR assay revealed that detection percentage of Legionella in samples was 70 when LEG448-JRP primers were used, whereas this percentage reduced to 50 and 45 when we applied prime sets of LEG225-LEG858 and LEG448 - LEG858, respectively.Conclusion: The results of the study showed that contamination of aquatic samples to the Legionella spp. could be easily and rapidly detected by nested PCR. However, selecting appropriate method for DNA extraction and choosing the primers are important factors in efficiency and sensitivity of detection method.
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