Visual detection of pandemic influenza A H1N1 Virus 2009 by reverse-transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye

J Virol Methods. 2010 Aug;167(2):214-7. doi: 10.1016/j.jviromet.2010.03.027. Epub 2010 Apr 8.

Abstract

A sensitive reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of pandemic influenza A H1N1 virus infection. The reaction was performed in one step in a single tube at 65 degrees C for 60 min with the addition of hydroxynaphthol blue (HNB) dye prior to amplification. The detection limit of the RT-LAMP assay was approximately 60 copies, and no cross-detection was observed. The assay was evaluated further with 50 clinical specimens diagnosed clinically with seasonal influenza or pandemic influenza A H1N1 virus infection. RT-LAMP with HNB dye was demonstrated to be a sensitive and easy assay for rapid detection of pandemic influenza A H1N1 virus.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Influenza A Virus, H1N1 Subtype / isolation & purification*
  • Influenza, Human / diagnosis*
  • Influenza, Human / virology*
  • Naphthalenesulfonates / metabolism*
  • Nucleic Acid Amplification Techniques / methods*
  • Sensitivity and Specificity
  • Staining and Labeling / methods*
  • Virology / methods*

Substances

  • Naphthalenesulfonates
  • trisodium 3-hydroxy-4-((2Z)-2-(2-oxo-4-sulfonatonaphthalen-1-ylidene)hydrazinyl)naphthalene-2,7-disulfonate