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Journal of Clinical Microbiology, April 2009, p. 1025-1030, Vol. 47, No. 4
0095-1137/09/$08.00+0     doi:10.1128/JCM.01920-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Low-Density Macroarray for Rapid Detection and Identification of Crimean-Congo Hemorrhagic Fever Virus{triangledown}

Roman Wölfel,1* Janusz T. Paweska,2 Nadine Petersen,3 Antoinette A. Grobbelaar,2 Patricia A. Leman,2 Roger Hewson,4 Marie-Claude Georges-Courbot,5 Anna Papa,6 Volker Heiser,7 Marcus Panning,3 Stephan Günther,3 and Christian Drosten3

Bundeswehr Institute of Microbiology, Munich, Germany,1 National Institute for Communicable Diseases, Sandringham, South Africa,2 Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany,3 Health Protection Agency, Porton Down, Salisbury, United Kingdom,4 Unit of Biology of Viral Emerging Infections, Institute Pasteur, Lyon, France,5 Microbiology Department, Medical School, Aristotle University of Thessaloniki, Thessaloniki, Greece,6 Chipron GmbH, Berlin, Germany7

Received 4 October 2008/ Returned for modification 26 November 2008/ Accepted 23 January 2009

Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral zoonosis which occurs throughout Africa, Eastern Europe, and Asia and results in an approximately 30% fatality rate. A reverse transcription-PCR assay including a competitive internal control was developed on the basis of the most up-to-date genome information. Biotinylated amplification products were hybridized to DNA macroarrays on the surfaces of polymer supports, and hybridization events were visualized by incubation with a streptavidin-horseradish peroxidase conjugate and the formation of a visible substrate precipitate. Optimal assay conditions for the detection of as few as 6.3 genome copies per reaction were established. Eighteen geographically and historically diverse CCHF virus strains representing all clinically relevant isolates were detected. The feasibility of the assay for clinical diagnosis was validated with acute-phase patient samples from South Africa, Iran, and Pakistan. The assay provides a specific, sensitive, and rapid method for CCHF virus detection without requiring sophisticated equipment. It has usefulness for the clinical diagnosis and surveillance of CCHF infections under limited laboratory conditions in developing countries or in field situations.

* Corresponding author. Mailing address: Bundeswehr Institute of Microbiology, Dept. for Medical Biological Reconnaissance & Verification, Neuherbergstrasse 11, 80937 Munich, Germany. Phone: 49-89-3168-3894. Fax: 49-89-3168-3292. E-mail: romanwoelfel{at}bundeswehr.org

{triangledown} Published ahead of print on 18 February 2009.

Journal of Clinical Microbiology, April 2009, p. 1025-1030, Vol. 47, No. 4
0095-1137/09/$08.00+0     doi:10.1128/JCM.01920-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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