TY - JOUR
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
PY - 2010/2
Y1 - 2010/2
N2 - 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.
AB - 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.
KW - Influenza A/H1N1pdm
KW - Influenza pandemic
KW - Molecular diagnosis
KW - RT-PCR
KW - S-OIV
KW - Sequencing
KW - Typing
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U2 - 10.1016/j.jviromet.2009.10.004
DO - 10.1016/j.jviromet.2009.10.004
M3 - Article
C2 - 19836420
AN - SCOPUS:73049107509
VL - 163
SP - 486
EP - 488
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 2
ER -