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

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Abstract

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
Volume163
Issue number2
DOIs
Publication statusPublished - Feb 2010

Fingerprint

Influenza A virus
Human Influenza
Nucleoproteins
Swine
Gene Targeting
Pandemics
Centers for Disease Control and Prevention (U.S.)
Mexico
Infection
Reverse Transcription
Disease Outbreaks
Sequence Analysis
Real-Time Polymerase Chain Reaction
Genome
Viruses
Technology
Polymerase Chain Reaction
Genes

Keywords

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

ASJC Scopus subject areas

  • Virology

Cite this

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title = "Design and clinical application of a molecular method for detection and typing of the influenza A/H1N1pdm virus",
abstract = "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.",
keywords = "Influenza A/H1N1pdm, Influenza pandemic, Molecular diagnosis, RT-PCR, S-OIV, Sequencing, Typing",
author = "Eleonora Lalle and Licia Bordi and Concetta Castilletti and Silvia Meschi and Marina Selleri and Fabrizio Carletti and Daniele Lapa and Damiano Travaglini and Giuseppe Ippolito and Capobianchi, {Maria Rosaria} and {Di Caro}, Antonino",
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T1 - Design and clinical application of a molecular method for detection and typing of the influenza A/H1N1pdm virus

AU - Lalle, Eleonora

AU - Bordi, Licia

AU - Castilletti, Concetta

AU - Meschi, Silvia

AU - Selleri, Marina

AU - Carletti, Fabrizio

AU - Lapa, Daniele

AU - Travaglini, Damiano

AU - Ippolito, Giuseppe

AU - Capobianchi, Maria Rosaria

AU - Di Caro, Antonino

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KW - Sequencing

KW - Typing

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