Simultaneous detection of Legionella species and Legionella pneumophila by duplex PCR (dPCR) assay in cooling tower water samples from Jakarta, Indonesia

• Main Authors: Andi Yasmon, Yusmaniar Yusmaniar, Anis Anis, Budiman Bela
• Format: Article
• Language: English
• Published: Faculty of Medicine Universitas Indonesia 2010-11-01
• Series: Medical Journal of Indonesia
• Online Access: http://mji.ui.ac.id/journal/index.php/mji/article/view/408

<p><strong>Aim:</strong> Since culture method is time-consuming and has low  sensitivity, we developed a duplex PCR (dPCR) assay for the detection of Legionella sp. and L. pneumophila in cooling tower samples. We used culture method as a gold standard.</p><p><strong>Methods:</strong> Optimization of dPCR method was performed to obtain an assay with high sensitivity and specifi city. The optimized method was used to detect Legionella sp. dan L. pneumophila in 9 samples obtained from 9 buildings in Jakarta. For culture method, the bacteria were grown or isolated on selective growth factor supplemented-buffered charcoal yeast extract (BCYE) media.</p><p><strong>Results:</strong> Of 9 samples tested by dPCR assay, 6 were positive for Legionella species,1 was positive for L. pneumophila, and 2 showed negative results. For the same samples, no Legionella sp. was detected by the culture method.</p><p><strong>Conclusion</strong>: dPCR assay was much more sensitive than the culture method and was potentially used as a rapid, specifi c and sensitive test for routine detection of Legionella sp. dan for L. pneumophila in water samples. <em><strong>(Med J Indones 2010; 19:223-7)</strong></em></p><p><strong>Keywords:</strong><em> BCYE media, mip gene, 16S-rRNA gene</em></p>