title: Visual detection of human infection with influenza A (H7N9) virus by subtype-specific reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye.
abstract: A rapid and sensitive H7 and N9 subtype-specific reverse transcription loop-mediated isothermal amplification assay was developed respectively for visual detection of human-infected influenza A (H7N9) virus. The reaction was performed in one step in a single tube at 63°C for 60 min with the addition of hydroxynaphthol blue dye before amplification. The detection limits of both subtype-specific assays were comparable to those of validated H7 and N9 real-time PCR assays respectively and no cross-detection was observed with influenza A pandemic H1N1, H3N2, H5N1, H9N2 or influenza B virus. The assays were evaluated further with H7N9 virus-infected clinical specimens.
authors.affiliation: Key Laboratory for Medical Virology, Ministry of Health, National Institute for Viral Disease Control and Prevention, Chinese Centre for Disease Control and Prevention, Beijing, China.
authors.forename: K
authors.initials: K
authors.lastname: Nie
authors.forename: X
authors.initials: X
authors.lastname: Zhao
authors.forename: X
authors.initials: X
authors.lastname: Ding
authors.forename: X D
authors.initials: XD
authors.lastname: Li
authors.forename: S M
authors.initials: SM
authors.lastname: Zou
authors.forename: J F
authors.initials: JF
authors.lastname: Guo
authors.forename: D Y
authors.initials: DY
authors.lastname: Wang
authors.forename: R B
authors.initials: RB
authors.lastname: Gao
authors.forename: X Y
authors.initials: XY
authors.lastname: Li
authors.forename: W J
authors.initials: WJ
authors.lastname: Huang
authors.forename: Y L
authors.initials: YL
authors.lastname: Shu
authors.forename: X J
authors.initials: XJ
authors.lastname: Ma
mesh_terms: D000818:Animals; D006801:Humans; D064766:Influenza A Virus, H7N9 Subtype / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009282:Naphthalenesulfonates / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*; D013696:Temperature; D014773:Virology / Q000379:methods*
chemical_list: D009282:Naphthalenesulfonates; C540250:trisodium 3-hydroxy-4-((2Z)-2-(2-oxo-4-sulfonatonaphthalen-1-ylidene)hydrazinyl)naphthalene-2,7-disulfonate
keywords: Colorimetric detection; hydroxynaphthol blue dye; influenza A H7N9 virus; reverse transcription loop-mediated isothermal amplification; subtype-specific