Evaluation of reverse transcription loop-mediated isothermal amplification assays for rapid diagnosis of pandemic influenza A/H1N1 2009 virus

J Med Virol. 2011 Jan;83(1):10-5. doi: 10.1002/jmv.21934.

Abstract

Two genetic diagnosis systems using reverse transcription-loop-mediated isothermal amplification (RT-LAMP) technology were evaluated: one for detecting the HA gene of the pandemic influenza A/H1N1 2009 virus (H1pdm RT-LAMP) and the other for detecting the matrix gene of the influenza A virus (TypeA RT-LAMP). The competence of these two RT-LAMP assay kits for the diagnosis of the pandemic influenza A/H1N1 2009 virus was compared using real-time RT-PCR assays developed recently on viruses isolated and clinical specimens collected from patients with suspected infection. TypeA RT-LAMP and H1pdm RT-LAMP showed almost the same sensitivity as real-time RT-PCR for viruses isolated. The sensitivity and specificity of TypeA RT-LAMP and H1pdm RT-LAMP were 96.3% and 88.9%, respectively, for clinical specimens. Considering that the ability of the two RT-LAMP assay kits for detection of the pandemic influenza A/H1N1 2009 virus was comparable to that of the real-time RT-PCR assays, and that the assays were completed within 1 hr and did not require any expensive equipment, these two RT-LAMP assays are promising rapid diagnostic tests for the pandemic influenza A/H1N1 2009 virus at the hospital bedside.

Publication types

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

MeSH terms

  • Hemagglutinins, Viral / genetics
  • Humans
  • Influenza A Virus, H1N1 Subtype / genetics
  • Influenza A Virus, H1N1 Subtype / isolation & purification*
  • Influenza, Human / diagnosis*
  • Influenza, Human / virology*
  • Nucleic Acid Amplification Techniques / methods*
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification*
  • Reverse Transcription
  • Sensitivity and Specificity
  • Temperature
  • Viral Matrix Proteins / genetics
  • Virology / methods*

Substances

  • Hemagglutinins, Viral
  • M1 protein, Influenza A virus
  • RNA, Viral
  • Viral Matrix Proteins
  • hemagglutinin fusogenic peptide, influenza virus