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The Journal of the Japanese Society for Clinical Microbiology |
Biblioraphy Information
| ArticleTitle |
Rapid and simple detection of Legionella species by LAMP, a new DNA amplification method |
| Language |
J |
| AuthorList |
Toshimitsu Annaka1), Manabu Yoshino1), Takayoshi Momoda1), Jiro Nemoto1), Atsuko Sunada1), Tadashi Kojima1), Masanari Ikedo1), Yoshikazu Ishii2), Keizo Yamaguchi2) |
| Affiliation |
1) Eiken Chemical Co., Ltd.
2) Toho Univ. School of Medicine |
| Publication |
J.J.C.M.: 13 (1), 19-25, 2003 |
| Received |
November 29, 2002 |
| Accepted |
March 14, 2003 |
| 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 ° 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 the clinically important Legionella species including Legionella pneumophila serogroups 1 to 6, Legionella bozemanii, Legionella dumoffii, Legionella gormanii, and Legionella longbeachae. The detection limit of the assay was 60 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. |
| Keywords |
Legionella, DNA amplification, rapid detection, LAMP, loop-mediated isothermal amplification |
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