Development and clinical testing of a simple, low-density gel element array for influenza identification, subtyping, and H275Y detection
Identifieur interne : 000A66 ( Ncbi/Checkpoint ); précédent : 000A65; suivant : 000A67Development and clinical testing of a simple, low-density gel element array for influenza identification, subtyping, and H275Y detection
Auteurs : Darrell P. Chandler [États-Unis] ; Sara B. Griesemer [États-Unis] ; Christopher Knickerbocker [États-Unis] ; Julia B. Golova [États-Unis] ; Amine Lambarqui [États-Unis] ; Alexander N. Perov [États-Unis] ; Cynthia Zimmerman [États-Unis] ; Cory Wiles [États-Unis] ; George B. Rudy [États-Unis] ; Kirsten St. George [États-Unis]Source :
- Journal of virological methods [ 0166-0934 ] ; 2014.
Abstract
The objectives of this study were to develop a user-friendly, gel element microarray test for influenza virus detection, subtyping, and neuraminidase inhibitor resistance detection, assess the performance characteristics of the assay, and perform a clinical evaluation on retrospective nasopharyngeal swab specimens. A streamlined microarray workflow enabled a single user to run up to 24 tests in an 8 hour shift. The most sensitive components of the test were the primers and probes targeting the A/H1pdm09 HA gene with an analytical limit of detection (LoD) <100 gene copies (gc) per reaction. LoDs for all targets in nasopharyngeal swab samples were ≤ 1000 gc, with the exception of one target in the seasonal A/H1N1 subtype. Seasonal H275Y variants were detectable in a mixed population when present at > 5% with wild type virus, while the 2009 pandemic H1N1 H275Y variant was detectable at ≤1% in a mixture with pandemic wild type virus. Influenza typing and sub typing results concurred with those obtained with real-time RT-PCR assays on more than 97% of the samples tested. The results demonstrate that a large panel of single-plex, real-time RT-PCR tests can be translated to an easy-to-use, sensitive, and specific microarray test for potential diagnostic use.
Url:
DOI: 10.1016/j.jviromet.2014.07.019
PubMed: 25066276
PubMed Central: 4175443
Affiliations:
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Rudy</name><affiliation wicri:level="2"><nlm:aff id="A1">Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300, Frederick MD 21701</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Maryland</region></placeName><wicri:cityArea>Akonni Biosystems, Inc., 400 Sagner Avenue, Suite 300</wicri:cityArea></affiliation></author><author><name sortKey="St George, Kirsten" sort="St George, Kirsten" uniqKey="St George K" first="Kirsten" last="St. George">Kirsten St. George</name><affiliation wicri:level="2"><nlm:aff id="A2">Laboratory of Viral Diseases, Wadsworth Center, New York State Dept of Health, 120 New Scotland Avenue, Albany NY 12208</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Laboratory of Viral Diseases, Wadsworth Center, New York State Dept of Health, 120 New Scotland Avenue</wicri:cityArea></affiliation></author></analytic><series><title level="j">Journal of virological methods</title><idno type="ISSN">0166-0934</idno><idno type="eISSN">1879-0984</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="P2">The objectives of this study were to develop a user-friendly, gel element microarray test for influenza virus detection, subtyping, and neuraminidase inhibitor resistance detection, assess the performance characteristics of the assay, and perform a clinical evaluation on retrospective nasopharyngeal swab specimens. A streamlined microarray workflow enabled a single user to run up to 24 tests in an 8 hour shift. The most sensitive components of the test were the primers and probes targeting the A/H1pdm09 HA gene with an analytical limit of detection (LoD) <100 gene copies (gc) per reaction. LoDs for all targets in nasopharyngeal swab samples were ≤ 1000 gc, with the exception of one target in the seasonal A/H1N1 subtype. Seasonal H275Y variants were detectable in a mixed population when present at > 5% with wild type virus, while the 2009 pandemic H1N1 H275Y variant was detectable at ≤1% in a mixture with pandemic wild type virus. Influenza typing and sub typing results concurred with those obtained with real-time RT-PCR assays on more than 97% of the samples tested. The results demonstrate that a large panel of single-plex, real-time RT-PCR tests can be translated to an easy-to-use, sensitive, and specific microarray test for potential diagnostic use.</p></div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Maryland</li><li>État de New York</li></region></list><tree><country name="États-Unis"><region name="Maryland"><name sortKey="Chandler, Darrell P" sort="Chandler, Darrell P" uniqKey="Chandler D" first="Darrell P." last="Chandler">Darrell P. Chandler</name></region><name sortKey="Golova, Julia B" sort="Golova, Julia B" uniqKey="Golova J" first="Julia B." last="Golova">Julia B. Golova</name><name sortKey="Griesemer, Sara B" sort="Griesemer, Sara B" uniqKey="Griesemer S" first="Sara B." last="Griesemer">Sara B. Griesemer</name><name sortKey="Knickerbocker, Christopher" sort="Knickerbocker, Christopher" uniqKey="Knickerbocker C" first="Christopher" last="Knickerbocker">Christopher Knickerbocker</name><name sortKey="Lambarqui, Amine" sort="Lambarqui, Amine" uniqKey="Lambarqui A" first="Amine" last="Lambarqui">Amine Lambarqui</name><name sortKey="Perov, Alexander N" sort="Perov, Alexander N" uniqKey="Perov A" first="Alexander N." last="Perov">Alexander N. Perov</name><name sortKey="Rudy, George B" sort="Rudy, George B" uniqKey="Rudy G" first="George B." last="Rudy">George B. Rudy</name><name sortKey="St George, Kirsten" sort="St George, Kirsten" uniqKey="St George K" first="Kirsten" last="St. George">Kirsten St. George</name><name sortKey="Wiles, Cory" sort="Wiles, Cory" uniqKey="Wiles C" first="Cory" last="Wiles">Cory Wiles</name><name sortKey="Zimmerman, Cynthia" sort="Zimmerman, Cynthia" uniqKey="Zimmerman C" first="Cynthia" last="Zimmerman">Cynthia Zimmerman</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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