Evaluation of sensitivity of TaqMan RT-PCR for rubella virus detection in clinical specimens
Autor: | Hideki Nagano, Yoshihiro Yasui, Hiroe Kodama, Yoshio Mori, Takashi Kato, Katsuhiro Komase, Sachiko Murata, Hiroko Minagawa, Motohiko Okano, Tomoko Ogawa, Masahiro Miyoshi, Atsushi Ogura, Daiki Kanbayashi, Noriyuki Otsuki, Rika Komagome, Takako Kurata, Komei Shirabe, Masafumi Sakata, Yoko Aoki, Miwako Saikusa, Chiemi Hotta, Makoto Takeda, Mitsuhiro Hamasaki, Kiyoko Okamoto, Tetsuo Kase |
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Rok vydání: | 2016 |
Předmět: |
Male
0301 basic medicine Urine Real-Time Polymerase Chain Reaction medicine.disease_cause Sensitivity and Specificity Measles Rubella 03 medical and health sciences 0302 clinical medicine Japan Virology Genotype medicine TaqMan Humans Viral rna 030212 general & internal medicine Rapid response business.industry Taqman rt pcr Rubella virus medicine.disease 030104 developmental biology Infectious Diseases Pharynx RNA Viral Female business |
Zdroj: | Journal of Clinical Virology. 80:98-101 |
ISSN: | 1386-6532 |
Popis: | 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 5 days 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. |
Databáze: | OpenAIRE |
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