Jpn. J. Infect. Dis., 59 (1), 46-51, 2006
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Original Article
Development of a Real-Time PCR Assay for Detection and Quantification of Francisella tularensis
Osamu Fujita*, Masashi Tatsumi, Kiyoshi Tanabayashi and Akio Yamada
Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo 162-8640, Japan
(Received October 12, 2005. Accepted January 5, 2006)
*Corresponding author: Mailing address: Department of Veterinary Science, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan. Tel: +81-3-5285-1111 ext. 2623, Fax: +81-3-5285-1179, E-mail: esperanz@nih.go.jp
SUMMARY: The facultative intracellular bacterium, Francisella tularensis, is an etiological agent of tularemia and is also considered to be a potential biological threat agent due to its extreme infectivity. We established a real-time PCR assay using the LightCycler (LC) system to detect a Francisella-specific sequence of the outer membrane protein (fopA) gene. Twenty-five F. tularensis strains including 16 Japanese isolates were subjected to this LC-PCR assay, and were tested positive, whereas Francisella philomiragia and other bacteria species did not show any specific fluorescent signal. A linear response was observed using F. tularensis genomic DNAs of between 20 fg and 2 ng, corresponding to 1.2 to 1.2 x 105 bacteria. The newly established real-time PCR allows the detection of the F. tularensis genome specifically, sensitively, and rapidly. This assay may contribute to the standardization of the laboratory diagnosis of tularemia.