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Journal of Clinical Microbiology, May 2008, p. 1747-1751, Vol. 46, No. 5
0095-1137/08/$08.00+0     doi:10.1128/JCM.02292-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Simultaneous Genotyping of All Hemagglutinin and Neuraminidase Subtypes of Avian Influenza Viruses by Use of Padlock Probes {triangledown} ,{dagger}

Péter Gyarmati,1* Tim Conze,2 Siamak Zohari,1 Neil LeBlanc,1 Mats Nilsson,2 Ulf Landegren,2 Johan Banér,3 and Sándor Belák1

Joint Research and Development Division, Department of Virology, National Veterinary Institute and Swedish University of Agricultural Sciences, Ulls väg 2B, SE-751 89 Uppsala, Sweden,1 Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala University, SE-751 85 Uppsala, Sweden,2 Stanford Genome Technology Center, Stanford University, Palo Alto, California 943043

Received 28 November 2007/ Returned for modification 23 January 2008/ Accepted 3 March 2008

A subtyping assay for both the hemagglutinin (HA) and neuraminidase (NA) surface antigens of the avian influenza virus (AIV) has been developed. The method uses padlock probe chemistry combined with a microarray output for detection. The outstanding feature of this assay is its capability to designate both the HA and the NA of an AIV sample from a single reaction mixture. A panel of 77 influenza virus strains was tested representing the entire assortment of the two antigens. One hundred percent (77/77) of the samples tested were identified as AIV, and 97% (75/77) were subtyped correctly in accordance with previous examinations performed by classical diagnostic methods. Testing of heterologous pathogens verified the specificity of the assay. This assay is a convenient and practical tool for the study of AIVs, providing important HA and NA data more rapidly than conventional methods.

* Corresponding author. Mailing address: Department of Virology, Swedish National Veterinary Institute, Travvägen 20-22, 75189 Uppsala, Sweden. Phone: 46 18 674384. Fax: 46 18 674669. E-mail: peter.gyarmati{at}sva.se

{triangledown} Published ahead of print on 19 March 2008.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, May 2008, p. 1747-1751, Vol. 46, No. 5
0095-1137/08/$08.00+0     doi:10.1128/JCM.02292-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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Antimicrob. Agents Chemother. Clin. Microbiol. Rev.
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