Design and clinical application of a molecular method for detection and typing of the influenza A/H1N1pdm virus

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In March/April 2009, Mexico experienced an outbreak of respiratory illness, due to a new influenza of swine origin virus, which spread rapidly via human-to-human transmission, and became pandemic (A/H1N1pdm). Because of its unique genome composition, which includes gene segments of swine, avian and human origin, and to the considerable differences to the human influenza A viruses that have circulated so far, the currently used molecular methods proved inadequate. Based on published sequences, a primer set targeting the nucleoprotein gene was designed, which provided enhanced sensitivity for the new strain and proved suitable for sequence-based strain identification. The novel nucleoprotein reverse-transcription-PCR showed higher sensitivity for A/H1N1pdm than a commercial test for influenza A, and was comparable to the real-time-based method developed by the Centers for Disease Control and Prevention. It was used to screen 177 clinical samples referred to the laboratory for suspected A/H1N1pdm infection, detecting 17 (9.6%) infections that were confirmed by sequence analysis (100% sensitivity as compared to the real-time kit). The novel method is suitable for the diagnosis of A/H1N1pdm, and is also suitable, at least in the screening phase, for laboratories not equipped with the real-time PCR technology.

Original languageEnglish
Pages (from-to)486-488
Number of pages3
JournalJournal of Virological Methods
Issue number2
Publication statusPublished - Feb 2010


  • Influenza A/H1N1pdm
  • Influenza pandemic
  • Molecular diagnosis
  • RT-PCR
  • S-OIV
  • Sequencing
  • Typing

ASJC Scopus subject areas

  • Virology


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