title: Uracil-DNA glycosylase-treated reverse transcription loop-mediated isothermal amplification for rapid detection of avian influenza virus preventing carry-over contamination.
abstract: Here, we describe a uracil-DNA glycosylase (UNG)-treated reverse transcription loop-mediated isothermal amplification (uRT-LAMP) for the visual detection of all subtypes of avian influenza A virus (AIV). The uRT-LAMP assay can prevent unwanted amplification by carryover contamination of the previously amplified DNA, although the detection limit of the uRT-LAMP assay is 10-fold lower than that of the RT-LAMP without a UNG treatment. To the best of our knowledge, this is the first successful application of deoxyuridine triphosphate/UNG strategy in RT-LAMP for AIV detection, and the assay can be applied for the rapid, and reliable diagnosis of AIVs, even in contaminated samples.
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Korea.|RAD Inc., Daegu 39852, Korea.
authors.forename: Eun-Mi
authors.initials: EM
authors.lastname: Kim
authors.affiliation: M Monitor Inc., Daegu 41914, Korea.
authors.forename: Hyo-Sung
authors.initials: HS
authors.lastname: Jeon
authors.affiliation: M Monitor Inc., Daegu 41914, Korea.
authors.forename: Ji-Jung
authors.initials: JJ
authors.lastname: Kim
authors.affiliation: Virology Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.
authors.forename: Yeun-Kyung
authors.initials: YK
authors.lastname: Shin
authors.affiliation: Avian Disease Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea.
authors.forename: Youn-Jeong
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Korea.
authors.forename: Sang-Geon
authors.initials: SG
authors.lastname: Yeo
authors.affiliation: College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Korea.|RAD Inc., Daegu 39852, Korea.
authors.forename: Choi-Kyu
authors.initials: CK
authors.lastname: Park
mesh_terms: D000818:Animals; D001717:Birds; D059018:DNA Contamination; D009980:Influenza A virus / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D051981:Uracil-DNA Glycosidase / Q000737:chemistry*
chemical_list: D051981:Uracil-DNA Glycosidase
keywords: avian influenza virus; contamination; loop-mediated isothermal amplification; uracil-DNA glycosylase