title: Evaluation of sensitivity of TaqMan RT-PCR for rubella virus detection in clinical specimens.
abstract: An easy and reliable assay for detection of the rubella virus is required to strengthen rubella surveillance. Although a TaqMan RT-PCR assay for detection of the rubella virus has been established in Japan, its utility for diagnostic purposes has not been tested.
  
  
  
  To allow introduction of the TaqMan RT-PCR into the rubella surveillance system in Japan, the sensitivity of the assay was determined using representative strains for all genotypes and clinical specimens.
  
  
  
  The detection limits of the method for individual genotypes were examined using viral RNA extracted from 13 representative strains. The assay was also tested at 10 prefectural laboratories in Japan, designated as local reference laboratories for measles and rubella, to allow nationwide application of the assay.
  
  
  
  The detection limits and amplification efficiencies of the assay were similar among all the representative strains of the 13 genotypes. The TaqMan RT-PCR could detect approximately 90% of throat swab and urine samples taken up to 5days of illness. These samples were determined positive by a highly sensitive nested RT-PCR.
  
  
  
  The TaqMan RT-PCR could detect at least 10 pfu of rubella virus. Although the sensitivity was somewhat lower than that of the conventional nested RT-PCR, the TaqMan RT-PCR could be more practical to routine tests for rubella laboratory diagnosis and detection in view of the rapid response and reducing risks of contamination.
authors.affiliation: Department of Virology III, National Institute of Infectious Diseases, Tokyo 208-0011, Japan.
authors.forename: Kiyoko
authors.initials: K
authors.lastname: Okamoto
authors.affiliation: Department of Virology III, National Institute of Infectious Diseases, Tokyo 208-0011, Japan. Electronic address: yoshiom@nih.go.jp.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Mori
authors.affiliation: Hokkaido Institute of Public Health, Sapporo 060-0819, Japan.
authors.forename: Rika
authors.initials: R
authors.lastname: Komagome
authors.affiliation: Hokkaido Institute of Public Health, Sapporo 060-0819, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Nagano
authors.affiliation: Hokkaido Institute of Public Health, Sapporo 060-0819, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Miyoshi
authors.affiliation: Hokkaido Institute of Public Health, Sapporo 060-0819, Japan.
authors.forename: Motohiko
authors.initials: M
authors.lastname: Okano
authors.affiliation: Yamagata Prefectural Institute of Public Health, Yamagata 990-0031, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Aoki
authors.affiliation: Chiba Prefectural Institute of Public Health, Chiba 260-8715, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ogura
authors.affiliation: Chiba Prefectural Institute of Public Health, Chiba 260-8715, Japan.
authors.forename: Chiemi
authors.initials: C
authors.lastname: Hotta
authors.affiliation: Chiba Prefectural Institute of Public Health, Chiba 260-8715, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Ogawa
authors.affiliation: Yokohama City Institute of Public Health, Yokohama 236-0051 Japan.
authors.forename: Miwako
authors.initials: M
authors.lastname: Saikusa
authors.affiliation: Ishikawa Prefectural Institute of Public Health and Environmental Science, Ishikawa 920-1154, Japan.
authors.forename: Hiroe
authors.initials: H
authors.lastname: Kodama
authors.affiliation: Aichi Prefectural Institute of Public Health, Aichi 462-8576, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Yasui
authors.affiliation: Aichi Prefectural Institute of Public Health, Aichi 462-8576, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Minagawa
authors.affiliation: Osaka Prefectural Institute of Public Health, Osaka, 537-0025, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Kurata
authors.affiliation: Osaka Prefectural Institute of Public Health, Osaka, 537-0025, Japan.
authors.forename: Daiki
authors.initials: D
authors.lastname: Kanbayashi
authors.affiliation: Osaka Prefectural Institute of Public Health, Osaka, 537-0025, Japan.
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Kase
authors.affiliation: Yamaguchi Prefectural Institute of Public Health and Environment, Yamaguchi, 753-0821, Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Murata
authors.affiliation: Yamaguchi Prefectural Institute of Public Health and Environment, Yamaguchi, 753-0821, Japan.
authors.forename: Komei
authors.initials: K
authors.lastname: Shirabe
authors.affiliation: Fukuoka Institute of Health and Environmental Sciences, Fukuoka 818-0135, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Hamasaki
authors.affiliation: Okinawa Prefectural Institute of Health and Environment, Okinawa 901-1202, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kato
authors.affiliation: Department of Virology III, National Institute of Infectious Diseases, Tokyo 208-0011, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Otsuki
authors.affiliation: Department of Virology III, National Institute of Infectious Diseases, Tokyo 208-0011, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Sakata
authors.affiliation: Department of Virology III, National Institute of Infectious Diseases, Tokyo 208-0011, Japan.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Komase
authors.affiliation: Department of Virology III, National Institute of Infectious Diseases, Tokyo 208-0011, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Takeda
mesh_terms: D005260:Female; D006801:Humans; D007564:Japan; D008297:Male; D010614:Pharynx / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012409:Rubella / Q000175:diagnosis*; D012412:Rubella virus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014556:Urine / Q000821:virology
chemical_list: D012367:RNA, Viral
keywords: Laboratory diagnosis; Rubella; TaqMan real-time RT-PCR