title: Rapid detection of European orthobunyaviruses by reverse transcription loop-mediated isothermal amplification assays.
abstract: The development of reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assays are described herein for the detection of two orthobunyaviruses (Bunyaviridae), which represent the two main serogroups found in mosquitoes in Central Europe. The RT-LAMP assays were optimized for the detection of Ťahyňa virus (a California encephalitis group virus found in Aedes sp or Ochlerotatus sp mosquitoes) and Batai virus (also called Čalovo virus, a Bunyamwera group virus found in Anopheles maculipennis s.l. mosquitoes) nucleic acid using endemic European virus isolates. The sensitivity of the RT-LAMP assays was determined to be comparable to that of conventional tests, with a limit of detection<0.1 pfu per reaction. The assays can be performed in 60min under isothermal conditions using very simple equipment. Furthermore, it was possible to proceed with the assays without nucleic acid extraction, albeit at a 100-fold loss of sensitivity. The RT-LAMP assays are a sensitive, cost-efficient method for both arbovirus surveillance as well as diagnostic laboratories to detect the presence of these endemic orthobunyaviruses.
authors.affiliation: Institute for Virology, University of Veterinary Medicine, Vienna, Austria. Electronic address: Jeremy.Camp@vetmeduni.ac.at.
authors.forename: Jeremy V
authors.initials: JV
authors.lastname: Camp
authors.affiliation: Institute for Virology, University of Veterinary Medicine, Vienna, Austria; Department of Basic Medical Sciences, College of Medicine, Mohammed Bin Rashid University of Medicine and Health Sciences, Dubai, United Arab Emirates.
authors.forename: Norbert
authors.initials: N
authors.lastname: Nowotny
mesh_terms: D000818:Animals; D003362:Cost-Benefit Analysis; D005060:Europe; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016851:Orthobunyavirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D012367:RNA, Viral
keywords: Arbovirus; Bunyaviridae; Loop-mediated isothermal amplification; Mosquito; Orthobunyavirus; RT-LAMP