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Massively parallel pathogen identification using high-density microarrays

Authors :
Giulia C. Kennedy
Ana Maria Burguiere
Walter Lee
Hervé Bourhy
Philip Dickinson
Gilberte Coralie
Laurent Dacheux
Nicolas Berthet
Christopher Davies
Stewart T. Cole
Christophe Batéjat
Anita K. Reinhardt
Ingrid Filliol
Tatiana Vallaeys
Iain G. Old
Yaron Turpaz
Beate Heym
Shenglan Zhang
Katherine A. Kong
Jean Claude Manuguerra
Génotypage des Pathogènes et Santé Publique (Plate-forme) (PF8)
Institut Pasteur [Paris] (IP)
Affymetrix
Cellule d'Intervention Biologique d'Urgence - Laboratory for Urgent Response to Biological Threats (CIBU)
Hôpital Ambroise Paré [AP-HP]
Dynamique des Lyssavirus et Adaptation à l'Hôte (DyLAH)
Génétique Moléculaire Bactérienne
This study was supported by Grant No. UC1 AI062613 (Kennedy) from the National Institute of Allergy and Infectious Diseases, National Institutes of Health. We thank A. Baldwin, S. Brisse, E. Carniel, J.-Y. Coppée, J.-M. Fournier, P. Glaser, P. Grimont, M. Mock, M. Popoff, O. Chesneau, P. Courvalin, P. Despres, C. LeBouguenec, B. Regnault, M.-L. Quilici and their staff for helpful suggestions and for providing nucleic acids from various bacteria and viruses. Vesicular stomatitis virus was a gift from Danielle Blondel, Unité Mixte de Virologie Moléculaire et Structurale, UMR 2472, CNRS, 91198 Gif sur Yvette Cedex, France. We thank C. Drosten (Bernhard Nocht Institute, Hamburg, Germany) for kindly making primer sequences and plasmids available for this study. We thank Guoliang Xing, Gangwu Mei, Brant Wong and Michael Mittman for assistance in array design.
Institut Pasteur [Paris]
Source :
Microbial Biotechnology, Microbial Biotechnology, 2008, 1 (1), pp.79-86. ⟨10.1111/j.1751-7915.2007.00012.x⟩, Microbial Biotechnology, Wiley, 2008, 1 (1), pp.79-86. ⟨10.1111/j.1751-7915.2007.00012.x⟩
Publication Year :
2011

Abstract

International audience; Identification of microbial pathogens in clinical specimens is still performed by phenotypic methods that are often slow and cumbersome, despite the availability of more comprehensive genotyping technologies. We present an approach based on whole-genome amplification and resequencing microarrays for unbiased pathogen detection. This 10 h process identifies a broad spectrum of bacterial and viral species and predicts antibiotic resistance and pathogenicity and virulence profiles. We successfully identify a variety of bacteria and viruses, both in isolation and in complex mixtures, and the high specificity of the microarray distinguishes between different pathogens that cause diseases with overlapping symptoms. The resequencing approach also allows identification of organisms whose sequences are not tiled on the array, greatly expanding the repertoire of identifiable organisms and their variants. We identify organisms by hybridization of their DNA in as little as 1-4 h. Using this method, we identified Monkeypox virus and drug-resistant Staphylococcus aureus in a skin lesion taken from a child suspected of an orthopoxvirus infection, despite poor transport conditions of the sample, and a vast excess of human DNA. Our results suggest this technology could be applied in a clinical setting to test for numerous pathogens in a rapid, sensitive and unbiased manner.

Details

ISSN :
17517915
Volume :
1
Issue :
1
Database :
OpenAIRE
Journal :
Microbial biotechnology
Accession number :
edsair.doi.dedup.....6b146890379f91e05bdb71de44bf1e72
Full Text :
https://doi.org/10.1111/j.1751-7915.2007.00012.x⟩