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 language | English |
|---|---|
| Pages (from-to) | 486-488 |
| Number of pages | 3 |
| Journal | Journal of Virological Methods |
| Volume | 163 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Feb 2010 |
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Keywords
- Influenza A/H1N1pdm
- Influenza pandemic
- Molecular diagnosis
- RT-PCR
- S-OIV
- Sequencing
- Typing
ASJC Scopus subject areas
- Virology
Cite this
Design and clinical application of a molecular method for detection and typing of the influenza A/H1N1pdm virus. / Lalle, Eleonora; Bordi, Licia; Castilletti, Concetta; Meschi, Silvia; Selleri, Marina; Carletti, Fabrizio; Lapa, Daniele; Travaglini, Damiano; Ippolito, Giuseppe; Capobianchi, Maria Rosaria; Di Caro, Antonino.
In: Journal of Virological Methods, Vol. 163, No. 2, 02.2010, p. 486-488.Research output: Contribution to journal › Article
}
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
UR - http://www.scopus.com/inward/record.url?scp=73049107509&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=73049107509&partnerID=8YFLogxK
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 -