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Journal of Clinical Microbiology, March 2009, p. 666-673, Vol. 47, No. 3
0095-1137/09/$08.00+0     doi:10.1128/JCM.01625-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Practical Considerations for High-Throughput Influenza A Virus Surveillance Studies of Wild Birds by Use of Molecular Diagnostic Tests {triangledown}

Vincent J. Munster,* Chantal Baas, Pascal Lexmond, Theo M. Bestebroer, Judith Guldemeester, Walter E. P. Beyer, Emmie de Wit, Martin Schutten, Guus F. Rimmelzwaan, Albert D. M. E. Osterhaus, and Ron A. M. Fouchier

National Influenza Center and Department of Virology, Erasmus Medical Center, Rotterdam, The Netherlands

Received 21 August 2008/ Returned for modification 11 November 2008/ Accepted 15 December 2008

Influenza A virus surveillance studies of wild bird populations are essential to improving our understanding of the role of wild birds in the ecology of low-pathogenic avian influenza viruses and their potential contribution to the spread of H5N1 highly pathogenic avian influenza viruses. Whereas the primary results of such surveillance programs have been communicated extensively, practical considerations and technical implementation options generally receive little attention. In the present study, the data obtained from 39,490 samples were used to compare the impacts of variables such as the sampling procedure, storage and transport conditions, and the choice of molecular and classical diagnostic tests on the outcome of the results. Molecular diagnostic tests allowed estimation of the virus load in samples, which has implications for the ability to isolate virus. Virus isolation in embryonated eggs was more sensitive than virus isolation in cell cultures. Storage and transport conditions had less of an impact on diagnostics by the use of molecular tests than by the use of classical approaches. These findings indicate that molecular diagnostic tests are more sensitive and more reliable than classical tests. In addition, molecular diagnostic tests facilitated analyses in real time and allowed the discrimination of H5 influenza viruses with low and high pathogenicities without the need for virus isolation. Critical assessment of the methods used in large surveillance studies like this will facilitate comparison of the results between studies. Moreover, the lessons learned from current large-scale influenza A virus surveillance activities could be valuable for other pathogen surveillance programs in the future.

* Corresponding author. Mailing address: Department of Virology, Erasmus Medical Center Rotterdam, P.O. Box 2040, Rotterdam 3000 CA, The Netherlands. Phone: 31-10-7044150. Fax: 31-10-7044760. E-mail: v.munster{at}erasmusmc.nl

{triangledown} Published ahead of print on 24 December 2008.

Journal of Clinical Microbiology, March 2009, p. 666-673, Vol. 47, No. 3
0095-1137/09/$08.00+0     doi:10.1128/JCM.01625-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Gall, A., Hoffmann, B., Harder, T., Grund, C., Ehricht, R., Beer, M. (2009). Rapid and Highly Sensitive Neuraminidase Subtyping of Avian Influenza Viruses by Use of a Diagnostic DNA Microarray. J. Clin. Microbiol. 47: 2985-2988 [Abstract] [Full Text]  


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