title: Mobile and accurate detection system for infection by the 2009 pandemic influenza A (H1N1) virus with a pocket-warmer reverse-transcriptase loop-mediated isothermal amplification.
abstract: The 2009 pandemic H1N1 influenza A virus spread quickly worldwide in 2009. Since most of the fatal cases were reported in developing countries, rapid and accurate diagnosis methods that are usable in poorly equipped laboratories are necessary. In this study, a mobile detection system for the 2009 H1N1 influenza A virus was developed using a reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) kit with a disposable pocket-warmer as a heating device (designated as pwRT-LAMP). The pwRT-LAMP can detect as few as 100 copies of the virus--which is nearly as sensitive as real-time reverse-transcription polymerase chain reaction (RT-PCR)--and does not cross-react with RNA of seasonal influenza viruses. To evaluate the usefulness of the pwRT-LAMP system, nasal swab samples were collected from 56 patients with flu-like symptoms and were tested. Real-time RT-PCR confirmed that the 2009 H1N1 influenza A virus was present in 27 of the 56 samples. Of these 27 positive samples, QuickVue Influenza A+B immunochromatography detected the virus in only 11 samples (11/27; 40.7%), whereas the pwRT-LAMP system detected the virus in 26 of the 56 samples (26/27 of the positive samples; 96.3%). These findings indicate that the mobile pwRT-LAMP system is an accurate diagnostic system for the 2009 H1N1 influenza A virus, and has great potential utility in diagnosing future influenza pandemics.
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan.
authors.forename: Ben
authors.initials: B
authors.lastname: Hatano
authors.forename: Megumi
authors.initials: M
authors.lastname: Goto
authors.forename: Hitomi
authors.initials: H
authors.lastname: Fukumoto
authors.forename: Takeyuki
authors.initials: T
authors.lastname: Obara
authors.forename: Takayuki
authors.initials: T
authors.lastname: Maki
authors.forename: Go
authors.initials: G
authors.lastname: Suzuki
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Kohsuke
authors.initials: K
authors.lastname: Hagisawa
authors.forename: Yoshitaro
authors.initials: Y
authors.lastname: Matsushita
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Fujii
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Imakiire
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Kikuchi
authors.forename: Ryota
authors.initials: R
authors.lastname: Takahashi
authors.forename: Mie
authors.initials: M
authors.lastname: Kanai
authors.forename: Kaku
authors.initials: K
authors.lastname: Tamura
authors.forename: Tomoko
authors.initials: T
authors.lastname: Izumi
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Yujiro
authors.initials: Y
authors.lastname: Iwamoto
authors.forename: Satoshi
authors.initials: S
authors.lastname: Mimura
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Mukai
authors.forename: Kazue
authors.initials: K
authors.lastname: Takita
authors.forename: Hiroki
authors.initials: H
authors.lastname: Takeo
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Kitamura
authors.forename: Eiichi
authors.initials: E
authors.lastname: Shimizu
authors.forename: Koji
authors.initials: K
authors.lastname: Fukushima
authors.forename: Yukiya
authors.initials: Y
authors.lastname: Hakozaki
authors.forename: Akimi
authors.initials: A
authors.lastname: Uehata
authors.forename: Masao
authors.initials: M
authors.lastname: Sakai
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ohshima
authors.forename: Toshiki
authors.initials: T
authors.lastname: Shirotani
authors.forename: Kunihiro
authors.initials: K
authors.lastname: Oba
authors.forename: Hideki
authors.initials: H
authors.lastname: Hasegawa
authors.forename: Tetsutaro
authors.initials: T
authors.lastname: Sata
authors.forename: Harutaka
authors.initials: H
authors.lastname: Katano
mesh_terms: D000328:Adult; D005260:Female; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology*; D008297:Male; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019095:Point-of-Care Systems; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*