Design and clinical application of a molecular method for detection and typing of the influenza A/H1N1pdm virus.
J Virol Methods, 2010/2;163(2):486-8.
Lalle E[1], Bordi L, Castilletti C, Meschi S, Selleri M, Carletti F, Lapa D, Travaglini D, Ippolito G, Capobianchi MR, Di Caro A
Affiliations
PMID: 19836420DOI: 10.1016/j.jviromet.2009.10.004
Impact factor: 2.623
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.
MeSH terms
DNA Primers; Humans; Influenza A Virus, H1N1 Subtype; Influenza, Human; Polymerase Chain Reaction; Sensitivity and Specificity; Sequence Analysis, DNA
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