title: Development of H5-RT-LAMP (loop-mediated isothermal amplification) system for rapid diagnosis of H5 avian influenza virus infection.
abstract: We developed a rapid and sensitive diagnosis system for H5N1 highly pathogenic avian influenza (HPAI) virus infection using an unique gene amplification method, reverse transcriptase loop-mediated isothermal amplification (RT-LAMP). The sensitivity of the system was found to be 100-fold higher than that of ordinary one-step RT-PCR. Moreover, by using viral RNAs extracted from influenza viruses of all 15 HA subtypes, the RT-LAMP system was confirmed to amplify only the RNA of H5 subtype virus. In the surveillance of H5N1 virus infection of wild birds, we detected two positive cases from dead crows found near the affected area with H5N1-HPAI by using RT-LAMP system, although one of two positive cases was missed by RT-PCR. These results suggested that our newly developed RT-LAMP system specific for H5 virus would be a beneficial diagnostic tool for surveillance of recent outbreaks caused by H5N1-HPAI viruses.
authors.affiliation: Laboratory of Influenza Viruses, Department of Virology 3, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashi-Murayama, Tokyo 208-0011, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Imai
authors.forename: Ai
authors.initials: A
authors.lastname: Ninomiya
authors.forename: Harumi
authors.initials: H
authors.lastname: Minekawa
authors.forename: Tsugunori
authors.initials: T
authors.lastname: Notomi
authors.forename: Toru
authors.initials: T
authors.lastname: Ishizaki
authors.forename: Masato
authors.initials: M
authors.lastname: Tashiro
authors.forename: Takato
authors.initials: T
authors.lastname: Odagiri
mesh_terms: D000818:Animals; D002645:Chickens; D017931:DNA Primers; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis / Q000821:virology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011200:Poultry; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D012367:RNA, Viral