title: Rapid detection of human pathogenic orthobunyaviruses.
abstract: 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 10(4) to 10(1) 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 10(7) 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.
authors.affiliation: Department of Virology, Institute of Medical Microbiology and Hygiene, University of Freiburg, 79104 Freiburg, Germany. weidmann@ukl.uni-freiburg.de
authors.forename: Manfred
authors.initials: M
authors.lastname: Weidmann
authors.forename: Veronique
authors.initials: V
authors.lastname: Rudaz
authors.forename: Marcio R T
authors.initials: MR
authors.lastname: Nunes
authors.forename: Pedro F C
authors.initials: PF
authors.lastname: Vasconcelos
authors.forename: Frank T
authors.initials: FT
authors.lastname: Hufert
mesh_terms: D000818:Animals; D002043:Bunyaviridae / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002044:Bunyaviridae Infections / Q000175:diagnosis* / Q000821:virology; D002460:Cell Line; D002522:Chlorocebus aethiops; D017931:DNA Primers; D006801:Humans; D012367:RNA, Viral / Q000097:blood; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012840:Simbu virus / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D019914:Taq Polymerase; D013997:Time Factors; D014709:Vero Cells
chemical_list: D017931:DNA Primers; D012367:RNA, Viral; D019914:Taq Polymerase