title: Rapid typing of influenza viruses using super high-speed quantitative real-time PCR.
abstract: The development of a rapid and sensitive system for detecting influenza viruses is a high priority for controlling future epidemics and pandemics. Quantitative real-time PCR is often used for detecting various kinds of viruses; however, it requires more than 2h per run. Detection assays were performed with super high-speed RT-PCR (SHRT-PCR) developed according to a newly designed heating system. The new method uses a high-speed reaction (18s/cycle; 40 cycles in less than 20min) for typing influenza viruses. The detection limit of SHRT-PCR was 1 copy/reaction and 10(-1) plaque-forming unit/reaction for viruses in culture supernatants during 20min. Using SHRT-PCR, 86 strains of influenza viruses isolated by the Tokyo Metropolitan Institute of Public Health were tested; the results showed 100% sensitivity and specificity for each influenza A and B virus, and swine-origin influenza virus. Twenty-seven swabs collected from the pharyngeal mucosa of outpatients were also tested, showing positive signs for influenza virus on an immunochromatographic assay; the results between SHRT-PCR and immunochromatography exhibited 100% agreement for both positive and negative results. The rapid reaction time and high sensitivity of SHRT-PCR makes this technique well suited for monitoring epidemics and pre-pandemic influenza outbreaks.
authors: Department of Molecular Medical Research, Tokyo Metropolitan Institute of Medical Science, 2-1-6, Kamikitazawa, Setagaya-ku, Tokyo 156-8506, Japan.
authors: Akira
authors: A
authors: Sakurai
authors: Namiko
authors: N
authors: Nomura
authors: Reiko
authors: R
authors: Nanba
authors: Takayuki
authors: T
authors: Sinkai
authors: Tsunehito
authors: T
authors: Iwaki
authors: Taminori
authors: T
authors: Obayashi
authors: Kazuhiro
authors: K
authors: Hashimoto
authors: Michiya
authors: M
authors: Hasegawa
authors: Yoshihiro
authors: Y
authors: Sakoda
authors: Akihiro
authors: A
authors: Naito
authors: Yoshihito
authors: Y
authors: Morizane
authors: Mitsugu
authors: M
authors: Hosaka
authors: Kunio
authors: K
authors: Tsuboi
authors: Hiroshi
authors: H
authors: Kida
authors: Akemi
authors: A
authors: Kai
authors: Futoshi
authors: F
authors: Shibasaki
mesh_terms: D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D009975:Orthomyxoviridae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D010614:Pharynx / Q000821:virology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014041:Tokyo; D014773:Virology / Q000379:methods*