Low-density macroarray for rapid detection and identification of Crimean-Congo hemorrhagic fever virus

Roman Wölfel*, Janusz T. Paweska, Nadine Petersen, Antoinette A. Grobbelaar, Patricia A. Leman, Roger Hewson, M. C. Georges-Courbot, Anna Papa, Volker Heiser, Marcus Panning, Stephan Günther, Christian Dro Sten

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

31 Citations (Scopus)

Abstract

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 streptavidinhorseradish 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.

Original languageEnglish
Pages (from-to)1025-1030
Number of pages6
JournalJournal of Clinical Microbiology
Volume47
Issue number4
DOIs
Publication statusPublished - Apr 2009

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