RESUMO
Legionella pneumophila is the primary cause of the legionellosis diseases (90 %) (Yu et al. in J Infect Dis 186:127-128, 2002; Doleans et al. in J Clin Microbiol 42:458-460, 2004; Den Boer et al. in Clin Microbiol Infect 14:459-466, 2008). In this study, methodologies based on molecular biology were developed in order to provide a quick diagnosis of the bacterial presence in water samples of Spain. Multiplex real-time polymerase chain reaction assays were realized to target the 16S rRNA and macrophage infectivity potentiator (mip) genes of, respectively, Legionella spp. and L. pneumophila including in the design of an internal control. The results obtained by the culture and the gene amplification methods agreed in 94.44 % for the 16S rRNA gene, and a concordance of 66.67 % of the cases was obtained for the mip gene.
Assuntos
Monitoramento Ambiental/métodos , Legionella pneumophila/fisiologia , Legionella/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Microbiologia da Água , Proteínas de Bactérias/genética , Humanos , Legionella/genética , Legionella/isolamento & purificação , Legionella pneumophila/genética , Legionella pneumophila/isolamento & purificação , Peptidilprolil Isomerase/genética , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade , EspanhaRESUMO
Rapid and more sensitive methods for the detection and quantification of viable Legionella cells have been developed. In this paper, a comparative analysis of environmental water samples using the ScanVIT-Legionella™ method and the traditional "gold standard" method of culturing is realised indicating the usefulness of the ScanVIT method. The ScanVIT-Legionella™ method was performed on environmental water samples from different locations of Huesca region (Spain). Legionella micro-colonies should appear green colour and Legionella pneumophila micro-colonies appear red. Twenty-one environmental water samples analysed by standard culture plus five control samples (Two sterile water samples with Legionella as positive controls and three sterile water samples as negative controls). All of them were used to apply ScanVIT-Legionella™ method. From of 21 environmental samples eleven were positive, six negative with both methods and four samples were negative for culture method and positive for ScanVIT-Legionella™ method. The positive control samples were positive and the negative were negative for both methods. A comparative analysis of the results obtained with two methods showed a strong positive determination coefficient (R(2) = 0.99753). The results demonstrate the usefulness of the ScanVIT-Legionella™ method as a rapid diagnostic tool in order to provide a diagnosis as quick as possible. ScanVIT-Legionella™ method offers a series of advantages such as quickly diagnosis, higher sensitivity and the possibility to identify Legionella spp. and L. pneumophila simultaneously.
