Differentiation of Legionella pneumophila and Non-Legionella pneumophila Species by a Multiplex PCR Method

Abstract

Molecular techniques can be utilized as a rapid method to detect Legionella species in water samples with high accuracy. This study aimed to investigate the presence of Legionella species, including Legionella pneumophila, in environmental water samples using both culturing and molecular techniques. A total of 23 water samples were cultured on buffered charcoal yeast extract (BCYE) agar, with all samples (100%) showing bacterial growth. The samples were then analyzed by multiplex PCR targeting the 16S rRNA gene and the L. pneumophila-specific Lpmip gene. Results revealed that 10 samples (43.5%) tested positive for the 16S rRNA gene, indicating the presence of Legionella species, while 6 samples (26.1%) tested positive for the L. pneumophila Lpmip gene. These findings highlight the prevalence of Legionella species in the samples, but also suggest a discrepancy between culture-based detection and molecular identification, with a lower proportion of samples testing positive for L. pneumophila. The study emphasizes the importance of using both culture and PCR methods for comprehensive detection and identification of Legionella species, as culture alone may not fully reflect the diversity of Legionella in environmental water sources