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Journal of Clinical Microbiology, July 2003, p. 3299-3305, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3299-3305.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Rapid Detection of Human Pathogenic Orthobunyaviruses

Department of Virology, Institute of Medical Microbiology and Hygiene, University of Freiburg, 79104 Freiburg, Germany,¹ Department of Arbovirus, Instituto Evandro Chagas, 66090-000 Belém, PA, Brazil²

Received 30 September 2002/ Returned for modification 22 December 2002/ Accepted 12 April 2003

Modern detection and identification tools can help to provide answers to urgent questions about the incidence, prevalence, and epidemiology of currently emerging diseases. We developed highly sensitive one-step TaqMan reverse transcription-PCR assays with sensitivities ranging from 10⁴ to 10¹ molecules for 11 human pathogens of the orthobunyaviruses. We compared the performances of these assays on three currently available cyclers (ABI-PRISM 7700, LightCycler, and SmartCycler). The assay for Oropouche virus (OROV) was tested using sera collected from days 1 to 5 after onset of OROV disease and was found to be greatly superior to an established nested PCR system. A mean copy number of 1.31 x 10⁷ OROV RNA/ml of serum was detected. Diagnostic RNA detection can be used as early as day 1 after onset of OROV disease. The use of a mobile SmartCycler and a hands-on time of less than 3 h could help to intensify outbreak surveillance and control, especially in field studies.

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