Kiyoko Okamoto1, Yoshio Mori2, Rika Komagome3, Hideki Nagano3, Masahiro Miyoshi3, Motohiko Okano3, Yoko Aoki4, Atsushi Ogura5, Chiemi Hotta5, Tomoko Ogawa5, Miwako Saikusa6, Hiroe Kodama7, Yoshihiro Yasui8, Hiroko Minagawa8, Takako Kurata9, Daiki Kanbayashi9, Tetsuo Kase9, Sachiko Murata10, Komei Shirabe10, Mitsuhiro Hamasaki11, Takashi Kato12, Noriyuki Otsuki1, Masafumi Sakata1, Katsuhiro Komase1, Makoto Takeda1. 1. Department of Virology III, National Institute of Infectious Diseases, Tokyo 208-0011, Japan. 2. Department of Virology III, National Institute of Infectious Diseases, Tokyo 208-0011, Japan. Electronic address: yoshiom@nih.go.jp. 3. Hokkaido Institute of Public Health, Sapporo 060-0819, Japan. 4. Yamagata Prefectural Institute of Public Health, Yamagata 990-0031, Japan. 5. Chiba Prefectural Institute of Public Health, Chiba 260-8715, Japan. 6. Yokohama City Institute of Public Health, Yokohama 236-0051 Japan. 7. Ishikawa Prefectural Institute of Public Health and Environmental Science, Ishikawa 920-1154, Japan. 8. Aichi Prefectural Institute of Public Health, Aichi 462-8576, Japan. 9. Osaka Prefectural Institute of Public Health, Osaka, 537-0025, Japan. 10. Yamaguchi Prefectural Institute of Public Health and Environment, Yamaguchi, 753-0821, Japan. 11. Fukuoka Institute of Health and Environmental Sciences, Fukuoka 818-0135, Japan. 12. Okinawa Prefectural Institute of Health and Environment, Okinawa 901-1202, Japan.
Abstract
BACKGROUND: 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. OBJECTIVES: 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. STUDY DESIGN: 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. RESULTS: 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. CONCLUSIONS: 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.
BACKGROUND: 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. OBJECTIVES: 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. STUDY DESIGN: 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. RESULTS: 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. CONCLUSIONS: 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: H Tumani; H F Petereit; A Gerritzen; C C Gross; A Huss; S Isenmann; S Jesse; M Khalil; P Lewczuk; J Lewerenz; F Leypoldt; N Melzer; S G Meuth; M Otto; K Ruprecht; E Sindern; A Spreer; M Stangel; H Strik; M Uhr; J Vogelgsang; K-P Wandinger; T Weber; M Wick; B Wildemann; J Wiltfang; D Woitalla; I Zerr; T Zimmermann Journal: Neurol Res Pract Date: 2020-03-16