title: Development of a new method for diagnosis of rubella virus infection by reverse transcription-loop-mediated isothermal amplification.
abstract: We developed a useful method for the detection of rubella virus genome RNA by reverse transcription loop-mediated isothermal amplification (RT-LAMP) and compared the sensitivity of RT-LAMP with that of other virological tests: reverse transcription-PCR (RT-PCR) and virus isolation. The rubella virus genome was amplified by RT-LAMP from clinical isolates obtained between 1987 and 2004 with similar sensitivities to the Takahashi vaccine strain. The detection limit of RT-LAMP was compared with that of RT-PCR using the Takahashi vaccine strain. We detected rubella virus genome material corresponding to 30 PFU/ml in a culture fluid sample by RT-LAMP within 60 min after the extraction of RNA with equal sensitivity to RT-nested PCR. The positive result rates of RT-LAMP, RT-PCR, and virus isolation were also compared using throat swabs obtained from patients who were clinically diagnosed with acute rubella virus infection in 2004 in Tochigi, Japan. Among nine patients with clinical rubella, the positive result rates were three/nine (33.3%) for virus isolation, six/nine (66.7%) for RT-PCR, and seven/nine (77.8%) for RT-LAMP. Consequently, RT-LAMP for rubella virus would be expected to be a reliable rapid diagnostic tool in the clinical setting.
authors.affiliation: Laboratory of Viral Infection I, Kitasato Institute for Life Science, 5-9-1 Shirokane, Tokyo 108-8641, Japan.
authors.forename: Nobuo
authors.initials: N
authors.lastname: Mori
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Motegi
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Shimamura
authors.forename: Takashi
authors.initials: T
authors.lastname: Ezaki
authors.forename: Tomo
authors.initials: T
authors.lastname: Natsumeda
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Yonekawa
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Ota
authors.forename: Tsugunori
authors.initials: T
authors.lastname: Notomi
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nakayama
mesh_terms: D016679:Genome, Viral; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012409:Rubella / Q000175:diagnosis* / Q000821:virology; D012412:Rubella virus / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral