[Rapid and simple detection of Legionella species by LAMP, a mew DNA amplification method]

Abstract

Legionella is a common cause of community-acquired respiratory tract infections and occasionally causes nosocomial pneumonia. Rapid and accurate detection of legionellae is important for diagnosis and treatment of patients. In order to detect legionellae, a new DNA amplification method was designed and evaluated. Loop-mediated isothermal amplification (LAMP) method amplifies DNA with high specificity, sensitivity, and rapidity under isothermal conditions at 65 degrees C. This method employs a DNA polymerase with strand displacement activity and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA. The primers targeting 16S rRNA gene were designed in order to detect a wide range of Legionella species. We could specifically detect Legionella species including Legionella pneumophila, Legionella anisa, Legionella bozemanii, Legionella dumoffii, Legionella erythra, Legionella feeleii, Legionella gormanii, Legionella longbeachae, Legionella micdadei, Legionella oakridgensis, and Legionella sainthelensi. The detection limit of the assay was 6 cfu per test of L. pneumophila strain. Furthermore, all of the positive LAMP results could be obtained within 50 minutes. The LAMP method was able to detect a wide range of Legionella species with high specificity, sensitivity, rapidity, and a simple procedure.